Application_of_chitosan_and_chitosan_derivatives_as_biomaterials1


    Review
    Application
    of
    chitosan
    and
    chitosan
    derivatives
    as
    biomaterials
    Changyong
    Choi
    JoungPyo
    Nam
    JaeWoon
    Nah *
    Department
    of
    Polymer
    Science
    and
    Engineering
    Sunchon
    National
    University
    255
    Jungangro
    Suncheon
    Jeollanamdo
    Republic
    of
    Korea
    Contents
    Introduction







































































































    1
    Genetic
    materials
    delivery
    by
    chitosan

















































































    2
    ChitosanDNA
    polyplexes























































































    3
    ChitosanRNA
    polyplexes























































































    3
    Genetic
    materials
    delivery
    by
    chitosan
    derivatives








































































    3
    Hydrophilic
    material
    modified
    chitosan











































































    4
    Hydrophobic
    material
    modified
    chitosan










































































    4
    Cationic
    material
    modified
    chitosan














































































    5
    Targeting
    ligand
    modified
    chitosan















































































    6
    Thiol
    group
    modified
    chitosan



















































































    7
    Amino
    acid
    and
    peptide
    modified
    chitosan









































































    7
    Genetic
    materials
    delivery
    by
    anionic
    materialchitosan
    complexes


























































    7
    Anion
    polymer
    and
    chitosan
    complexes











































































    8
    Anionic
    biopolymer
    and
    chitosan
    complexes







































































    8
    Polypeptide
    and
    chitosan
    complexes














































































    8
    Conclusion








































































































    9
    Acknowledgements

































































































    9
    References








































































































    9
    Introduction
    Gene
    therapy
    uses
    genetic
    materials
    (eg
    deoxyribonucleic
    acids
    (DNA)
    or
    ribonucleic
    acid
    (RNA))
    as
    a
    pharmaceutical
    agent
    to
    treat
    various
    diseases
    Gene
    therapy
    has
    the
    following
    three
    main
    mechanisms
    (1)
    delivering
    missing
    genes
    (2)
    replacing
    defective
    genes
    and
    (3)
    gene
    silencing
    undesired
    gene
    expression
    [1]
    Through
    these
    mechanisms
    gene
    therapy
    treats
    a
    wide
    range
    of
    diseases
    Consequently
    the
    interest
    in
    gene
    therapy
    is
    increasing
    Despite
    these
    advantages
    use
    of
    genetic
    materials
    in
    gene
    therapy
    is
    limited
    due
    to
    rapid
    degradation
    by
    nuclease
    large
    size
    poor
    cellular
    uptake
    high
    anionic
    charge
    density
    and
    Journal
    of
    Industrial
    and
    Engineering
    Chemistry
    33
    (2016)
    1–10
    A
    R
    T
    I
    C
    L
    E
    I
    N
    F
    O
    Article
    history
    Received
    24
    August
    2015
    Received
    in
    revised
    form
    13
    October
    2015
    Accepted
    20
    October
    2015
    Available
    online
    24
    October
    2015
    Keywords
    Chitosan
    Chitosan
    derivatives
    Biomaterials
    Delivery
    system
    A
    B
    S
    T
    R
    A
    C
    T
    Chitosan
    is
    a
    linear
    polysaccharide
    composed
    of
    randomly
    distributed
    b(14)linked
    Dglucosamine
    and
    NacetylDglucosamine
    It
    is
    one
    of
    the
    major
    cationic
    polymers
    and
    the
    second
    most
    abundant
    polysaccharides
    in
    nature
    It
    is
    extensively
    used
    to
    the
    biomedical
    and
    the
    industrial
    fields
    Specially
    chitosan
    has
    been
    studied
    much
    in
    the
    field
    of
    gene
    therapy
    during
    the
    last
    decade
    for
    its
    biocompatibility
    and
    noncytotoxicity
    However
    it
    has
    several
    problems
    such
    as
    solubility
    low
    transfection
    efficiency
    and
    low
    specialty
    on
    targeted
    disease
    To
    solve
    these
    problems
    various
    strategies
    have
    been
    reported
    to
    enhance
    them
    This
    review
    briefly
    introduces
    various
    strategies
    of
    chitosan
    carrier
    ß
    2015
    The
    Korean
    Society
    of
    Industrial
    and
    Engineering
    Chemistry
    Published
    by
    Elsevier
    BV
    All
    rights
    reserved
    * Corresponding
    author
    Tel
    +82
    61
    750
    3566
    fax
    +82
    61
    750
    5423
    Email
    address
    jwnah@sunchonackr
    (JW
    Nah)
    Contents
    lists
    available
    at
    ScienceDirect
    Journal
    of
    Industrial
    and
    Engineering
    Chemistry
    jou
    r
    n
    al
    h
    o
    mep
    ag
    e
    w
    ww
    elsevier
    co
    m
    loc
    atejiec
    httpdxdoiorg101016jjiec201510028
    1226086Xß
    2015
    The
    Korean
    Society
    of
    Industrial
    and
    Engineering
    Chemistry
    Published
    by
    Elsevier
    BV
    All
    rights
    reservednonspecificity
    [1–4]
    To
    overcome
    these
    problems
    vectors
    are
    used
    for
    safe
    delivery
    of
    genetic
    materials
    in
    gene
    therapy
    Generally
    vectors
    can
    be
    classified
    into
    two
    types
    One
    of
    them
    is
    a
    viral
    vector
    which
    is
    commonly
    used
    to
    deliver
    genetic
    material
    into
    the
    cells
    The
    viral
    vectors
    such
    as
    retroviruses
    lentiviruses
    adenoviruses
    and
    adenoassociated
    viruses
    are
    very
    effective
    in
    achieving
    high
    transfection
    efficiency
    however
    their
    availability
    for
    therapeutic
    use
    in
    the
    human
    body
    is
    limited
    because
    of
    immune
    responses
    safety
    problems
    high
    cost
    and
    low
    transgenic
    size
    [5–
    9]
    The
    other
    type
    is
    nonviral
    vectors
    which
    are
    preferred
    as
    safer
    alternatives
    to
    viral
    vectors
    for
    gene
    therapy
    The
    viral
    vectors
    such
    as
    liposome
    protein
    and
    cationic
    polymer
    have
    many
    advantages
    including
    stability
    safety
    low
    immune
    response
    and
    cell
    targeting
    properties
    [510]
    Thus
    in
    recent
    years
    the
    interest
    in
    nonviral
    vector
    is
    increasing
    and
    active
    research
    has
    been
    reported
    Cationic
    polymers
    are
    widely
    used
    as
    carriers
    for
    nonviral
    genetic
    materials
    delivery
    [11–13]
    They
    can
    condense
    with
    genetic
    materials
    through
    electrostatic
    interaction
    to
    form
    polyplexes
    and
    facilitate
    the
    cellular
    uptake
    by
    cells
    [1213]
    In
    addition
    the
    amine
    group
    of
    polyplexes
    is
    quick
    on
    the
    uptake
    of
    the
    cell
    absorbing
    protons
    facilitating
    the
    escape
    of
    the
    polyplexes
    from
    endosome
    or
    lysosome
    through
    a
    triggered
    osmotic
    swelling
    effect
    [121415]
    Chitosan
    is
    a
    linear
    polysaccharide
    composed
    of
    randomly
    distributed
    b(14)linked
    Dglucosamine
    (deacetylated
    unit)
    and
    NacetylDglucosamine
    (acetylate
    unit)

    a
    structure
    very
    similar
    to
    that
    of
    cellulose
    As
    such
    chitosan
    is
    one
    of
    the
    major
    cationic
    polymers
    [1617]
    It
    is
    obtained
    by
    the
    alkaline
    deacetylation
    of
    chitin
    which
    is
    the
    second
    most
    abundant
    polysaccharides
    in
    nature
    after
    cellulose
    Chitosan
    forms
    inter
    and
    intramolecular
    hydrogen
    bonding
    owing
    to
    amine
    and
    hydroxyl
    groups
    therefore
    it
    has
    a
    rigid
    crystalline
    structure
    [18]
    Chitosan
    has
    a
    various
    bioactivities
    due
    to
    the
    abundant
    primary
    amino
    groups
    in
    the
    chitosan
    main
    chain
    For
    this
    reasons
    the
    chitosan
    is
    extensively
    used
    to
    the
    biomedical
    fields
    such
    as
    drug
    andor
    gene
    delivery
    and
    the
    industrial
    fields
    such
    as
    water
    treatment
    (eg
    harmful
    algae
    control)
    heavy
    metal
    flocculants
    and
    functional
    foods
    [19]
    Chitosan
    is
    soluble
    in
    an
    acid
    solution
    but
    insoluble
    at
    natural
    and
    alkaline
    pH
    values
    because
    of
    the
    pKa
    value
    of
    chitosan
    of
    about
    65
    [20]
    The
    solubility
    of
    chitosan
    is
    significantly
    dependent
    on
    the
    degree
    of
    deacetylation
    (DDA)
    When
    DDA
    of
    chitosan
    is
    40
    chitosan
    is
    soluble
    up
    to
    a
    pH
    of
    9
    Whereas
    DDA
    of
    chitosan
    is
    80
    it
    is
    soluble
    only
    up
    to
    a
    pH
    of
    65
    [18]
    Moreover
    the
    molecular
    weight
    (MW)
    of
    chitosan
    and
    the
    ionic
    strength
    of
    the
    solution
    influence
    the
    solubility
    of
    chitosan
    Reporting
    on
    the
    chemical
    properties
    of
    chitosan
    including
    cationic
    properties
    Sanford
    pointed
    out
    that
    the
    high
    charge
    density
    at
    pH
    <
    65
    forms
    gels
    with
    polyanions
    adheres
    to
    negatively
    charged
    surfaces
    chelates
    certain
    transitional
    metals
    and
    is
    readily
    susceptible
    to
    chemical
    modification
    [21]
    During
    the
    last
    decade
    chitosan
    has
    been
    extensively
    used
    as
    a
    gene
    carrier
    for
    gene
    therapy
    by
    applying
    the
    chemical
    properties
    described
    above
    It
    has
    also
    been
    extensively
    studied
    as
    nonviral
    derived
    cationic
    natural
    polymers
    for
    a
    number
    of
    pharmaceutical
    and
    biomedical
    applications
    due
    to
    its
    biocompatibility
    biodegradability
    to
    normal
    body
    constituents
    nontoxic
    hemostatic
    bacteriostatic
    fungistatic
    spermicidal
    anticancerogen
    anticholesteremic
    properties
    easily
    susceptible
    to
    chemical
    modification
    [21–24]
    In
    addition
    chitosan
    is
    tightly
    condensed
    with
    negatively
    charged
    genetic
    materials
    protecting
    genetic
    materials
    against
    nuclease
    degradation
    due
    to
    cationic
    property
    as
    a
    positive
    charge
    [2526]
    ChitosanDNA
    polyplexes
    have
    been
    reported
    to
    transfect
    into
    various
    cell
    types
    (eg
    human
    embryonic
    kidney
    cells
    (HEK293)
    [13]
    cervical
    cancer
    cells
    (HeLa
    cell)
    [13]
    primary
    chondrocytes
    [27]
    Chinese
    hamster
    ovary
    cells
    (CHOK1)
    [28]
    fibroblast
    cells
    (NIH
    3T3)
    [29]
    and
    epithelioma
    papulosum
    cyprinid
    cells
    (EPC)
    [30])
    This
    review
    briefly
    introduces
    the
    strategies
    of
    chitosan
    and
    chitosan
    derivatives
    that
    have
    been
    reported
    as
    genetic
    materials
    delivery
    carrier
    in
    various
    gene
    therapies
    Genetic
    materials
    delivery
    by
    chitosan
    The
    amine
    groups
    of
    chitosan
    are
    positively
    charged
    in
    acidic
    mediums
    and
    provide
    a
    strong
    electrostatic
    interaction
    with
    negatively
    charged
    mucosal
    surfaces
    or
    other
    macromolecules
    such
    as
    genetic
    materials
    [18]
    Therefore
    chitosan
    has
    been
    used
    as
    a
    delivery
    carrier
    for
    gene
    delivery
    in
    order
    to
    enhance
    transfection
    efficiency
    and
    protect
    genetic
    materials
    against
    nuclease
    Howev
    er
    to
    effectively
    transfer
    genetic
    material
    by
    using
    chitosan
    the
    MW
    and
    DDA
    of
    chitosan
    must
    be
    considered
    Over
    the
    years
    many
    studies
    have
    reported
    on
    the
    effect
    of
    various
    MW
    and
    DDA
    of
    chitosan
    to
    transfer
    genetic
    material
    into
    the
    cells
    effectively
    The
    MW
    and
    DDA
    of
    chitosan
    have
    an
    influence
    on
    its
    biological
    and
    physicochemical
    properties
    The
    DDA
    affects
    biodegradability
    and
    results
    in
    very
    low
    rates
    of
    enzymatic
    degradation
    of
    acetyl
    groups
    [3132]
    The
    transfection
    efficiency
    and
    binding
    affinity
    of
    chitosanDNA
    system
    were
    found
    to
    increase
    with
    the
    increase
    of
    MW
    and
    DDA
    [3133]
    Kiang
    et
    al
    have
    demonstrated
    that
    destabilization
    of
    particles
    cause
    the
    decreased
    DDA
    in
    a
    decrease
    in
    luciferase
    expression
    levels
    in
    HEK293
    HeLa
    and
    cervical
    carcinoma
    (SW756)
    cells
    in
    vitro
    [13]
    The
    high
    MW
    (100
    kDa)
    of
    chitosan
    was
    found
    to
    have
    several
    advantages
    in
    genetic
    materials
    delivery
    forming
    extremely
    stable
    polyplexes
    with
    genetic
    materials
    delaying
    the
    release
    of
    the
    genetic
    materials
    and
    forming
    the
    physical
    shape
    of
    the
    polyplexes
    [2534]
    However
    high
    MW
    of
    chitosan
    has
    pharmaceutical
    drawbacks
    such
    as
    low
    solubility
    at
    physiological
    pH
    slow
    dissociation
    and
    release
    of
    genetic
    materials
    and
    high
    viscosity
    at
    concentrations
    used
    for
    in
    vivo
    delivery
    [183435]
    These
    factors
    of
    chitosan
    with
    high
    MW
    led
    to
    slow
    onset
    of
    action
    [34]
    To
    overcome
    these
    problems
    of
    chitosan
    with
    high
    Mw
    numerous
    studies
    have
    been
    conducted
    by
    using
    low
    MW
    chitosan
    Jang
    et
    al
    have
    reported
    that
    low
    molecular
    weight
    watersoluble
    chitosan
    (LMWSC)
    is
    prepared
    using
    a
    novel
    salt
    removedmethod
    to
    enhance
    the
    solubility
    of
    chitosan
    under
    physiological
    pH
    [36]
    They
    characterized
    the
    structure
    of
    LMWSC
    having
    free
    amine
    group
    by
    using
    fourier
    transform
    infrared
    (FTIR)
    and
    nuclear
    magnetic
    resonance
    (NMR)
    and
    investigated
    DDA
    and
    MW
    of
    LMWSC
    from
    which
    salt
    is
    removed
    by
    using
    NMR
    and
    viscometer
    The
    results
    demonstrated
    that
    the
    LMWSC
    is
    successfully
    prepared
    and
    it
    can
    be
    used
    in
    pharmaceutical
    and
    food
    industry
    [36]
    Bloomfield
    [37]
    and
    Strand
    et
    al
    [38]
    demonstrated
    that
    the
    low
    MW
    chitosan
    provides
    stable
    polyplexes
    without
    aggregation
    soluble
    at
    neutral
    pH
    reduces
    viscosity
    and
    is
    more
    easily
    dissociated
    polyplexes
    than
    using
    high
    MW
    chitosan
    in
    gene
    therapy
    The
    pDNA
    loaded
    chitosan
    microspheres
    are
    prepared
    by
    using
    a
    precipitation
    technique
    for
    in
    vitro
    tranfection
    in
    rat
    prostate
    adenocarcinoma
    cell
    line
    (MATLyLu)
    [39]
    The
    MW
    of
    chitosan
    and
    the
    amount
    of
    plasmid
    influenced
    the
    in
    vitro
    transfection
    in
    the
    cells
    The
    pDNA
    loaded
    chitosan
    microspheres
    prepared
    with
    low
    MW
    chitosan
    expressed
    slightly
    higher
    transfection
    than
    the
    high
    molecular
    weight
    chitosan
    [39]
    Ko¨pingHo¨gga˚ rd
    et
    al
    reported
    on
    the
    properties
    and
    efficiency
    of
    low
    MW
    (<5
    kDa)
    and
    they
    found
    that
    24mer
    (approximately
    3
    kDa–4
    kDa)
    of
    chitosan
    forms
    stable
    complexes
    and
    gives
    a
    high
    level
    of
    gene
    expression
    comparable
    to
    the
    high
    MW
    of
    chitosan
    [40]
    Furthermore
    it
    was
    found
    that
    the
    stability
    and
    transfection
    efficiency
    of
    chitosanused
    polyplexes
    has
    been
    dependent
    not
    only
    on
    the
    MW
    and
    DDA
    of
    chitosan
    but
    also
    on
    the
    pH
    of
    transfection
    medium
    [30313440–43]
    serum
    concentration
    [1343–45]
    and
    concentrations
    of
    genetic
    materials
    [30313846]
    The
    pH
    of
    transfection
    medium
    was
    affected
    to
    C
    Choi
    et
    al

    Journal
    of
    Industrial
    and
    Engineering
    Chemistry
    33
    (2016)
    1–102form
    the
    shape
    in
    coils
    (physiological
    pH)
    or
    globules
    in
    acid
    pH
    condition
    The
    amount
    of
    globules
    of
    polyplexes
    formed
    with
    chitosan
    increased
    as
    the
    pH
    decreased
    ranging
    from
    65
    to
    35
    and
    led
    to
    high
    level
    transfection
    efficiency
    In
    addition
    amine
    group
    of
    chitosan
    of
    pKa
    value
    (about
    65)
    was
    activated
    in
    low
    pH
    environment
    and
    formed
    with
    globules
    by
    charge
    inversion
    [40]
    Guliyeva
    et
    al
    reported
    that
    pDNA
    is
    released
    from
    chitosan
    microparticles
    with
    pH
    21
    mediums
    in
    shorter
    periods
    than
    pH
    647
    mediums
    and
    the
    released
    amounts
    are
    higher
    than
    other
    pH
    conditions
    in
    pH
    45
    and
    pH
    647
    because
    of
    different
    solubilities
    of
    chitosan
    in
    acidic
    and
    basic
    pHs
    of
    mediums
    [42]
    Lavertu
    et
    al
    [31]
    Nydert
    et
    al
    [41]
    and
    Nimesh
    et
    al
    [43]
    demonstrated
    that
    transfection
    efficiency
    is
    more
    effective
    at
    acidic
    pH
    than
    natural
    pH
    in
    HEK293
    cell
    Through
    the
    results
    of
    transfection
    efficiency
    in
    vitro
    Lavertu
    et
    al
    demonstrated
    that
    transfection
    efficiency
    is
    highly
    sensitive
    to
    DDA
    MW
    concentrations
    of
    genetic
    materials
    and
    medium
    pH
    [31]
    For
    efficient
    gene
    delivery
    system
    that
    uses
    chitosan
    the
    optimal
    conditions
    have
    yet
    to
    be
    established
    including
    MW
    DDA
    medium
    pH
    concentrations
    of
    genetic
    materials
    and
    serum
    stability
    ChitosanDNA
    polyplexes
    Chellat
    et
    al
    experimented
    on
    the
    metalloproteinase
    and
    cytokine
    production
    by
    the
    human
    monocytic
    leukemia
    cell
    line
    (THP1)
    macrophages
    using
    chitosanDNA
    (ChDNA)
    nanoparticle
    and
    their
    results
    indicated
    that
    the
    ChDNA
    nanoparticles
    do
    not
    induce
    the
    release
    of
    proinflammatory
    cytokines
    whereas
    MMP9
    was
    significantly
    increased
    [47]
    ChitosanDNA
    nanoparticles
    were
    prepared
    for
    nonviral
    gene
    transfer
    in
    animal
    models
    of
    fetal
    gene
    therapy
    to
    characterize
    the
    chitosanDNA
    nanoparticle
    stability
    within
    amniotic
    fluid
    in
    vitro
    [48]
    Chitosan
    protected
    pDNA
    from
    enzymatic
    degradation
    despite
    the
    chitosan
    aggregate
    in
    amniotic
    fluid
    The
    transfection
    efficiency
    of
    chitosanDNA
    nanoparticles
    shows
    high
    levels
    in
    vivo
    than
    in
    vitro
    transfection
    due
    to
    bioavailable
    property
    of
    chitosan
    in
    vivo
    and
    the
    long
    exposure
    time
    of
    chitosanDNA
    nanoparticles
    in
    vivo
    [48]
    Niu
    et
    al
    investigated
    human
    insulin
    band
    by
    using
    the
    gel
    electrophoresis
    system
    in
    harvested
    tissues
    such
    as
    stomachs
    intestines
    and
    rectums
    after
    human
    insulin
    gene
    transfection
    using
    gene
    wrapped
    chitosan
    nanoparticles
    and
    they
    detected
    that
    the
    human
    insulin
    bands
    are
    found
    only
    in
    the
    harvested
    intestines
    of
    diabetic
    rats
    [29]
    Yang
    et
    al
    evaluated
    the
    potential
    of
    chitosan
    in
    DNA
    vaccine
    delivery
    via
    mucosa
    and
    their
    results
    show
    that
    low
    MW
    of
    chitosan
    has
    lower
    binding
    affinity
    to
    DNA
    but
    higher
    transfection
    efficiency
    than
    the
    high
    MW
    of
    chitosan
    and
    the
    intranasal
    vaccination
    of
    chitosan
    (low
    MW)DNA
    polyplexes
    elicits
    signifi
    cant
    systemic
    immune
    responses
    [49]
    Roy
    et
    al
    investigated
    the
    effect
    of
    chitosanDNA
    nanoparticles
    on
    food
    allergy
    therapy
    and
    demonstrated
    that
    chitosanDNA
    nanoparticles
    are
    effective
    in
    controlling
    murine
    anaphylactic
    responses
    [50]
    This
    result
    indicates
    that
    the
    chitosan
    has
    potential
    utility
    in
    treating
    food
    allergy
    ChitosanDNA
    nanospheres
    were
    prepared
    to
    investigate
    the
    potential
    of
    chitosanDNA
    nanospheres
    against
    acute
    respira
    tory
    syncytial
    virus
    (RSV)
    infection
    and
    they
    were
    found
    to
    have
    a
    greater
    potential
    against
    acute
    RSV
    infection
    than
    the
    controls
    owing
    to
    the
    results
    of
    induction
    of
    RSVspecific
    immunoglobulin
    G
    (IgG)
    antibodies
    nasal
    immunoglobulin
    A
    (IgA)
    antibodies
    interferong
    and
    cytotoxic
    T
    lymphocytes
    production
    in
    lung
    and
    splenocytes
    [51]
    ChitosanRNA
    polyplexes
    Recently
    small
    interfering
    ribonucleic
    acid
    (siRNA)
    has
    been
    studied
    as
    a
    new
    therapeutic
    tool
    for
    gene
    expressionimplicated
    disease
    [5253]
    To
    silence
    target
    genes
    siRNA
    offers
    a
    potentially
    new
    therapeutic
    strategy
    with
    high
    specificity
    by
    reducing
    undesirable
    gene
    expression
    [54–56]
    However
    using
    siRNA
    should
    consider
    the
    limitations
    such
    as
    rapid
    degradation
    short
    halflife
    and
    low
    internalization
    in
    the
    therapeutic
    [5457]
    Therefore
    in
    gene
    therapy
    that
    uses
    siRNA
    a
    carrier
    system
    is
    required
    for
    the
    protection
    against
    nuclease
    degrada
    tion
    and
    delivery
    of
    siRNA
    into
    target
    cell
    Kenneth
    et
    al
    reported
    that
    the
    chitosansiRNA
    nanoparticles
    enhance
    green
    fluorescent
    protein
    (EGFP)
    gene
    knockdown
    in
    both
    human
    lung
    carcinoma
    cells
    (H1299)
    and
    murine
    peritoneal
    macrophages
    also
    showing
    similar
    results
    in
    transgenic
    EGFP
    mice
    after
    nasal
    administration
    [58]
    In
    another
    research
    EGFP
    gene
    knockdown
    was
    enhanced
    by
    increasing
    nitrogenphosphorus
    (NP)
    ratios
    ranging
    from
    50
    to
    150
    and
    high
    MW
    of
    chitosan
    [59]
    Malmo
    et
    al
    reported
    that
    the
    results
    of
    flow
    cytometry
    and
    the
    knockdown
    efficiency
    assay
    in
    H1299
    cells
    indicated
    the
    most
    efficient
    gene
    silencing
    is
    achieved
    by
    using
    the
    fully
    deNacetylated
    chitosan
    with
    intermediate
    chin
    lengths
    (degrees
    of
    polymerization
    (DPn)
    100–300)
    [60]
    Aerosolised
    chitosansiRNA
    nanoparticles
    were
    prepared
    to
    detect
    pulmonary
    gene
    silencing
    in
    transgenic
    EGFP
    mice
    [61]
    This
    research
    demonstrated
    that
    in
    vivo
    study
    results
    showed
    significant
    EGFP
    gene
    silencing
    above
    the
    68
    reduction
    of
    fluorescence
    ratio
    compared
    to
    the
    mismatch
    group
    in
    transgenic
    EGFP
    mice
    dosed
    with
    the
    aerosolized
    chitosansiRNA
    nanoparticle
    and
    the
    exact
    siRNA
    dosage
    improved
    the
    effect
    of
    gene
    silencing
    more
    than
    the
    intranasal
    administration
    did
    [61]
    Ji
    et
    al
    showed
    that
    chitosansiRNA
    nanoparticle
    which
    is
    four
    and
    a
    half
    LIM
    domains
    protein
    2
    (FHL2)
    siRNA
    formulated
    with
    chitosan
    could
    knock
    down
    about
    696
    FHL2
    gene
    expression
    This
    result
    is
    very
    similar
    to
    the
    reduced
    FHL2
    gene
    expression
    transfected
    by
    the
    commercial
    transfection
    agent
    called
    Lipo
    fectamine
    [52]
    Alameh
    et
    al
    prepared
    chitosansiRNA
    nanocom
    plexes
    to
    enhance
    gene
    silencing
    with
    dipeptidyl
    peptidase
    IV
    (DPPIV)
    siRNA
    This
    research
    revealed
    an
    80
    silencing
    of
    the
    DPPIV
    gene
    compared
    to
    nontransfected
    cells
    [62]
    To
    induce
    mucosal
    secretory
    IgA
    (SIgA)
    secretion
    the
    VPIencoded
    chitosan
    DNA
    (pcDNA3VP1)
    which
    is
    the
    major
    structural
    protein
    of
    coxsackievirus
    B3
    (CVB3)
    was
    prepared
    by
    vortexing
    DNA
    with
    chitosan
    [63]
    ChitosanDNA
    (pcDNA3VP1)
    successfully
    induced
    mucosal
    SIgA
    secretion
    and
    significantly
    reduced
    the
    viral
    load
    after
    acute
    CVB3
    infection
    in
    mice
    Thus
    this
    research
    indicates
    that
    chitosan
    may
    be
    a
    promising
    vaccine
    candidate
    for
    protection
    against
    infection
    Genetic
    materials
    delivery
    by
    chitosan
    derivatives
    Despite
    the
    excellent
    properties
    of
    biodegradability
    biocom
    patibility
    and
    nontoxicity
    chitosan
    has
    been
    mainly
    limited
    in
    biomedical
    field
    because
    of
    several
    disadvantages
    One
    of
    them
    is
    low
    solubility
    at
    natural
    pH
    and
    alkaline
    pH
    [2064]
    Another
    disadvantage
    is
    the
    low
    transfection
    efficiency
    due
    to
    the
    relatively
    low
    cationic
    density
    in
    chitosan
    that
    causes
    less
    compact
    of
    chitosangenetic
    material
    complexes
    [64–66]
    Other
    disadvan
    tages
    are
    lack
    of
    cell
    specificity
    and
    low
    escape
    property
    from
    endosomes
    into
    the
    cytoplasm
    [6467]
    To
    overcome
    these
    disadvantages
    many
    research
    efforts
    have
    been
    reported
    in
    recent
    years
    to
    enhance
    solubility
    transfection
    efficiency
    release
    and
    cell
    specificity
    of
    chitosan
    derivatives
    The
    following
    sections
    discuss
    various
    derivatives
    including
    hydrophilic
    moiety
    hydrophobic
    moiety
    cationic
    moiety
    anion
    moiety
    and
    target
    moiety
    modified
    chitosan
    which
    aim
    at
    enhancing
    the
    property
    of
    chitosan
    for
    effective
    genetic
    materials
    delivery
    Chitosan
    derivatives
    have
    been
    prepared
    using
    a
    variety
    of
    methods
    and
    materials
    to
    enhance
    application
    of
    chitosan
    The
    classification
    of
    various
    strategies
    to
    prepare
    chitosan
    derivatives
    is
    shown
    in
    Table
    1
    C
    Choi
    et
    al

    Journal
    of
    Industrial
    and
    Engineering
    Chemistry
    33
    (2016)
    1–10
    3Hydrophilic
    material
    modified
    chitosan
    Hydrophilic
    modification
    to
    chitosan
    backbone
    was
    introduced
    to
    improve
    transfection
    efficiency
    for
    increased
    water
    solubility
    at
    physiological
    pH
    and
    improved
    intracellular
    DNA
    release
    [6869]
    Polyethylene
    glycolation
    (PEGylation)
    and
    trimethylation
    are
    the
    strategies
    used
    most
    to
    improve
    the
    solubility
    of
    chitosan
    PEG
    has
    a
    high
    solubility
    in
    water
    low
    cytotoxicity
    and
    high
    cell
    permeability
    properties
    therefore
    it
    is
    a
    good
    candidate
    to
    induce
    hydrophilic
    part
    to
    chitosan
    backbone
    In
    addition
    PEG
    can
    increase
    plasma
    halflives
    and
    shield
    them
    from
    inactivation
    by
    the
    immune
    system
    [7071]
    Trimethylation
    of
    chitosan
    backbone
    enhances
    water
    solubility
    of
    chitosan
    at
    broader
    pH
    range
    which
    leads
    to
    compact
    interaction
    with
    pDNA
    [72–75]
    Trimethylation
    chitosan
    (TMC)
    is
    capable
    of
    opening
    the
    tight
    junctions
    of
    cells
    at
    physiological
    pH
    which
    led
    to
    enhance
    paracellular
    permeability
    [727476]
    Zhang
    et
    al
    prepared
    PEGconjugated
    chitosanDNA
    (CSDNA
    PEG)
    nanoparticles
    and
    demonstrated
    that
    the
    transfection
    efficiency
    of
    the
    PEGylated
    nanopartecles
    improved
    more
    than
    PEG
    unconjugated
    chitosanDNA
    nanoparticles
    both
    in
    vitro
    and
    in
    vivo
    experiment
    [70]
    Kean
    et
    al
    investigated
    the
    transfection
    efficiency
    of
    the
    trimethylated
    oligomeric
    chitosan
    (TMO)
    and
    trimethylated
    polymeric
    chitosan
    (TMC)
    prepared
    with
    low
    molecular
    weight
    [75]
    The
    transfection
    efficiency
    of
    TMO
    and
    TMC
    with
    various
    degrees
    of
    quaternization
    showed
    a
    greater
    efficiency
    than
    high
    molecular
    weight
    (25
    kDa)
    polyethyleneimine
    (PEI)
    in
    epithelial
    breast
    cancer
    (MCF7)
    cells
    Moreover
    TMO
    and
    TMC
    also
    showed
    appreciable
    transfection
    in
    monkey
    kidney
    fibroblasts
    (COS7)
    cells
    [75]
    Unfortunately
    chitosan
    which
    has
    at
    higher
    degrees
    of
    quaternization
    led
    to
    increased
    cytotoxicity
    [75–
    78]
    PEGylated
    TMC
    was
    conjugated
    with
    different
    grafting
    ratios
    and
    PEG
    chain
    lengths
    to
    increase
    solubility
    and
    decrease
    cytotoxicity
    [77]
    The
    half
    maximal
    inhibitory
    concentration
    (IC50)
    value
    of
    PEGylated
    TMC
    was
    421
    mgmL
    while
    PEG
    unmodified
    TMC
    showed
    higher
    toxicity
    with
    an
    IC50 value
    of
    96
    mgmL
    in
    NIH
    3T3
    cells
    Hydrophobic
    material
    modified
    chitosan
    Hydrophobic
    moiety
    modified
    chitosan
    polyplexes
    formed
    with
    DNA
    have
    several
    advantages
    such
    as
    alleviation
    of
    serum
    inhibition
    facilitated
    intracellular
    DNA
    dissociation
    efficient
    protection
    from
    enzymatic
    degradation
    and
    improved
    cell
    membrane
    permeation
    over
    the
    unmodified
    chitosan
    polyplexes
    [79–81]
    Hydrophobic
    moiety
    modified
    chitosan
    forms
    amphi
    philic
    polymer
    by
    chemically
    attaching
    hydrophobic
    functional
    moieties
    to
    chitosan
    The
    modified
    hydrophobic
    group
    can
    be
    a
    charge
    neutralization
    or
    even
    a
    charge
    inversion
    of
    chitosanDNA
    polyplexes
    [82]
    The
    strategy
    of
    introducing
    hydrophobic
    group
    to
    chitosan
    has
    been
    tried
    with
    various
    hydrophobic
    moiety
    including
    alkyl
    group
    (as
    steric
    acid
    alkyl
    bromide
    and
    caproic
    acid)
    [64657983–85]
    bile
    acid
    (as
    a
    deoxycholic
    acid)
    [8687]
    and
    5b
    cholanic
    acid
    [66]
    Stearic
    acid
    grafted
    chitosan
    oligosaccharide
    (CSOSA)
    is
    synthesized
    by
    coupling
    reaction
    with
    a
    1ethyl3(3dimethyla
    minopropyl)
    carbodiimde
    (EDC)
    [65]
    CSOSA
    formed
    micelle
    by
    selfaggregation
    in
    aqueous
    solution
    The
    critical
    micelle
    concen
    tration
    (CMC)
    of
    CSOSA
    is
    about
    0035
    mgmL
    with
    154
    amino
    substituted
    degree
    of
    CSO
    The
    transfection
    efficiency
    of
    CSOSA
    micelles
    with
    pDNA
    (pEGFPC1)
    is
    about
    15
    higher
    than
    that
    of
    chitosan
    oligosaccharide
    (CSO)pDNA
    particles
    (about
    2)
    In
    addition
    CSOSApDNA
    micelles
    are
    not
    interfered
    in
    the
    presence
    of
    10
    serum
    Zhu
    et
    al
    prepared
    dodecylated
    chitosanpDNA
    nanoparticles
    (DCDNPs)
    with
    a
    mean
    diameter
    of
    approximately
    90–180
    nm
    for
    local
    gene
    delivery
    via
    endovascular
    stents
    coated
    [85]
    The
    DCDNPs
    containing
    pDNA
    (EGFPC1)coated
    stents
    showed
    high
    level
    of
    green
    fluorescent
    protein
    (GFP)
    expression
    in
    cells
    which
    grew
    on
    the
    stent
    surface
    and
    along
    the
    adjacent
    area
    Table
    1
    The
    classification
    of
    various
    strategies
    to
    provide
    the
    advantage
    in
    chitosan
    Modification
    Materials
    Advantage
    Ref
    Hydrophilic
    material
    modification
    Trimethyl
    group
    High
    solubility
    low
    cytotoxicity
    high
    transfection
    efficiency
    [71–77]
    PEG
    [697076]
    Hydrophobic
    material
    modification
    Alkyl
    group
    High
    transfection
    efficiency
    alleviate
    serum
    inhibition
    enhanced
    protection
    efficiency
    improved
    cell
    membrane
    permeation
    [63647882–84]
    Bile
    acid
    [8586]
    5bcholanic
    acid
    [65]
    Cationic
    material
    modification
    PEI
    Improve
    cationic
    density
    enhance
    condensation
    capability
    high
    transfection
    efficiency
    effectively
    escape
    from
    endosome
    [87–92]
    Urocanic
    acid
    [6693]
    Imidazole
    [9495]
    Diethyleneamine
    [96]
    Spermine
    [4]
    Targeting
    ligand
    modification
    Saccharide
    Hepatocyte
    targeting
    (Galactose)
    To
    achieve
    cellspecificity
    High
    cellular
    uptake
    High
    transfection
    efficiency
    [101–106]
    Macrophages
    targeting
    Dendritic
    cell
    targeting
    (Mannose)
    [107–110]
    Hepatocyte
    targeting
    (Lactose)
    [111]
    Folic
    acid
    Folate
    receptor
    on
    present
    various
    tumor
    (ovarian
    lung
    breast
    colon
    etc)
    [112–114]
    Transferrin
    Transferrin
    receptor
    on
    present
    on
    malignant
    cells
    [115]
    RGD
    peptide
    Integrin
    anb3
    targeting
    [116]
    Thiol
    group
    modification
    Thioglycolic
    acid
    Increase
    extracellular
    stability
    high
    cellular
    uptake
    improved
    intracellular
    release
    high
    transfection
    efficiency
    [138–140]
    Cystamine
    [132]
    2Iminothiolane
    [141]
    Amino
    acid
    and
    peptide
    modification
    TAT
    peptide
    Enhance
    cell
    penetrating
    High
    cellular
    uptake
    improved
    intracellular
    release
    high
    transfection
    efficiency
    [144145]
    Cystein
    Induced
    thiol
    group
    [134147]
    Arginine
    Induced
    hydrophilicity
    [146]
    Nonpolar
    amino
    acids
    (alanine
    valine
    leucine
    isoleucine)
    Induced
    hydrophobicity
    [1]
    C
    Choi
    et
    al

    Journal
    of
    Industrial
    and
    Engineering
    Chemistry
    33
    (2016)
    1–104Caproic
    acid
    grafted
    chitosan
    (CGC)
    was
    prepared
    to
    investigate
    the
    effect
    of
    the
    degree
    of
    substitutions
    of
    hydrophobic
    group
    on
    binding
    affinity
    with
    pDNA
    cellular
    uptake
    transfection
    efficiency
    and
    biocompatibility
    [79]
    The
    transfection
    efficiency
    of
    CGCpDNA
    (gWizGFP
    or
    gWizbGal)
    nanoparticles
    exhibited
    a
    higher
    gene
    expression
    compared
    to
    chitosanpDNA
    nanoparticles
    but
    did
    not
    depend
    on
    the
    degree
    of
    substitution
    of
    caproic
    acid
    group
    ranging
    from
    5
    to
    25
    The
    optimal
    formulation
    is
    CGC15
    because
    substitution
    of
    a
    large
    percentage
    of
    hydrophobic
    groups
    induced
    unstable
    polyplexes
    and
    led
    to
    low
    levels
    of
    transfection
    efficiency
    [79]
    Liu
    et
    al
    investigated
    the
    gene
    transfection
    of
    Nalkylated
    chitosan
    prepared
    with
    alkyl
    bromide
    (various
    alkyl
    chain
    lengths
    as
    butyl
    octyl
    dodecyl
    and
    hexadecyl
    bromide)
    [83]
    Various
    N
    alkylated
    chitosanplasmid
    encoding
    chloramphenicol
    acetyl
    transferase
    (pcDNA
    31CAT)
    complexes
    exhibited
    a
    greater
    transfection
    efficiency
    than
    naked
    DNA
    and
    unmodified
    chito
    sanpcDNA
    31CAT
    complexes
    and
    increased
    the
    transfection
    efficiency
    by
    increasing
    alkyl
    chin
    ranging
    from
    4
    to
    16
    in
    mouse
    skeletal
    muscle
    cell
    lines
    (C2C12)
    Jang
    et
    al
    had
    reported
    previously
    that
    deoxycholic
    acid
    conjugated
    chitosan
    oligosaccharide
    nanoparticles
    (COSDs)
    pre
    pared
    by
    coupling
    reaction
    between
    primary
    amine
    groups
    in
    COS
    and
    Nhydroxysuccinimide
    (NHS)activated
    deoxycholic
    acid
    (DOCA)
    [86]
    COSDspDNA
    nanoparticles
    whose
    optimal
    formula
    tion
    is
    COS3D25
    (with
    1–3
    kDa
    and
    5
    degree
    of
    substitution
    of
    DOCA)
    showed
    a
    great
    potential
    for
    gene
    carrier
    with
    high
    level
    of
    gene
    transfection
    efficiencies
    even
    in
    the
    presence
    of
    serum
    compared
    to
    the
    deoxycholic
    acid
    unmodified
    chitosanpDNA
    nanoparticles
    and
    polyLlysine
    (PLL)pDNA
    nanoparticles
    Yoo
    et
    al
    reported
    that
    hydrophobically
    modified
    glycol
    chitosan
    (HGC)
    prepared
    by
    coupling
    reaction
    with
    hydrophobic
    moiety
    (5bcholanic
    acid)
    spontaneously
    formed
    nanoparticles
    by
    a
    hydrophobic
    interaction
    between
    HGC
    and
    hydrophobized
    DNA
    showing
    a
    higher
    transfection
    efficiency
    than
    naked
    DNA
    and
    a
    commercialized
    transfection
    agent
    such
    as
    superfect
    in
    vivo
    [66]
    Fig
    1
    shows
    the
    structures
    of
    chitosan
    derivatives
    which
    are
    modified
    with
    hydrophilic
    group
    or
    hydrophobic
    group
    Cationic
    material
    modified
    chitosan
    Since
    the
    major
    disadvantage
    of
    chitosan
    to
    delivery
    of
    genetic
    materials
    is
    the
    low
    transfection
    efficiency
    due
    to
    the
    relatively
    low
    cationic
    density
    in
    chitosan
    [64–66]
    many
    studies
    reported
    the
    cationic
    densityenhanced
    chitosan
    through
    various
    cationic
    groups
    including
    PEI
    [88–93]
    Urocanic
    acid
    [6794]
    imidazole
    [9596]
    diethylethylamine
    [97]
    and
    spermine
    [4]
    modification
    The
    enhanced
    cationic
    density
    of
    chitosan
    can
    play
    a
    crucial
    role
    in
    endosomal
    rupture
    through
    proton
    sponge
    mechanism
    which
    can
    lead
    to
    high
    transfection
    efficiency
    Although
    PEI
    is
    the
    most
    effective
    nonviral
    vector
    on
    cationic
    polymers
    because
    of
    its
    high
    pH
    buffering
    capacity
    [98]
    it
    can
    be
    highly
    toxic
    depending
    on
    the
    dose
    and
    molecular
    weight
    and
    its
    nonbiodegradable
    property
    is
    also
    the
    main
    concern
    when
    PEI
    is
    used
    for
    a
    longterm
    [92]
    Therefore
    to
    enhance
    the
    transfection
    efficiency
    and
    decrease
    toxicity
    the
    method
    of
    inducing
    PEI
    to
    chitosan
    backbone
    has
    been
    studied
    Wong
    et
    al
    demonstrated
    that
    PEIgchitosan
    synthesized
    by
    performing
    cationic
    polymeri
    zation
    of
    aziridine
    to
    watersoluble
    oligochitosan
    (MW
    34
    kDa)
    showed
    good
    DNA
    condensation
    capability
    low
    cytotoxicity
    and
    high
    gene
    transfection
    efficiency
    both
    in
    vitro
    and
    in
    vivo
    [88]
    Lu
    et
    al
    prepared
    LM
    PEI
    grafted
    chitosan
    (Nmaleated
    form)
    for
    gene
    delivery
    [89]
    The
    DNA
    condensation
    capability
    of
    PEI
    grafted
    chitosan
    (NMCgPEI)
    was
    increased
    by
    increasing
    chitosan
    molecular
    weight
    ranging
    from
    5
    to
    50
    kDa
    under
    the
    same
    condition
    as
    condensation
    ratios
    (ww)
    NMCgPEI
    improved
    more
    buffering
    capacity
    than
    ungrafted
    chitosan
    but
    it
    is
    still
    lower
    than
    that
    of
    PEI
    In
    addition
    the
    buffering
    capacity
    of
    NMCgPEI
    was
    decreased
    by
    increasing
    the
    molecular
    weight
    of
    chitosan
    Therefore
    the
    transfection
    efficiency
    of
    NMC5kgPEI
    and
    NMC10k
    gPEI
    copolymers
    has
    a
    relatively
    higher
    gene
    transfection
    ability
    Fig
    1
    Chitosan
    derivatives
    with
    hydrophilic
    group
    (A)
    or
    hydrophobic
    group
    (B)
    A
    (a)
    trimethylated
    chitosan
    [74]
    (b)
    pegylated
    chitosan
    [69]
    B
    (a)
    deoxycolic
    acid
    modified
    chitosan
    [85]
    (b)
    5bcholanic
    acid
    modified
    chitosan
    [65]
    C
    Choi
    et
    al

    Journal
    of
    Industrial
    and
    Engineering
    Chemistry
    33
    (2016)
    1–10
    5than
    NMC50kgPEI
    in
    293
    T
    cell
    and
    HeLa
    cell
    Lu
    et
    al
    prepared
    N
    succinyl
    chitosan
    (NSC)
    to
    graft
    with
    LM
    PEI
    (800
    Da)
    [91]
    NSC
    graftedPEI
    (NSCgPEI)
    was
    synthesized
    by
    coupling
    reaction
    using
    different
    amounts
    (02
    and
    04mmol)
    of
    coupling
    agent
    as
    EDC
    The
    grafted
    degree
    (GD)
    of
    PEI
    molecules
    grafted
    to
    the
    saccharide
    unit
    of
    NSCgPEI
    was
    measured
    to
    be
    0167
    (NSCg
    PEI1
    used
    02mmol
    of
    EDC)
    and
    0266
    (NSCgPEI1
    used
    04
    mmol
    of
    EDC)
    respectively
    The
    transfection
    efficiency
    of
    NSCg
    PEIDNA
    complexes
    was
    higher
    than
    PEI
    (25
    kDa)DNA
    complexes
    and
    it
    was
    increased
    by
    increasing
    GD
    in
    HeLa
    and
    Chinese
    hamster
    ovary
    (CHO)
    cells
    Wang
    et
    al
    demonstrated
    that
    urocanic
    acid
    (UAC)modified
    chitosanmediated
    efficient
    p53
    gene
    transfer
    which
    is
    hepato
    cellular
    carcinoma
    (HCC)
    growth
    inhibiting
    genes
    [99–101]
    could
    induce
    apoptosis
    significantly
    inhibiting
    the
    growth
    of
    human
    hepatoblastoma
    (HepG2)
    cells
    in
    vitro
    [67]
    Kim
    et
    al
    also
    demonstrated
    the
    transfection
    efficiency
    of
    urocanic
    acidmodified
    chitosan
    (UAC)
    in
    293
    T
    cells
    [94]
    UACDNA
    complex
    showed
    good
    DNA
    binding
    ability
    high
    protection
    of
    DNA
    from
    nuclease
    attack
    cytotoxicity
    lower
    than
    PEI
    and
    high
    transfection
    efficiency
    with
    increased
    UA
    contents
    in
    the
    UAC
    ranging
    from
    20
    to
    70
    The
    imidazole
    ring
    of
    urocanic
    acid
    may
    be
    playing
    a
    role
    in
    endosomal
    rupture
    through
    proton
    sponge
    mechanism
    [94]
    In
    addition
    Moreira
    et
    al
    [95]
    and
    Ghosn
    et
    al
    [96]
    used
    imidazole
    moiety
    to
    enhance
    transfection
    efficiency
    through
    proton
    sponge
    mechanism
    of
    imidazole
    Moreira
    et
    al
    investigated
    the
    transfection
    efficiency
    of
    imidazole
    moiety
    modified
    chitosan
    (CHimi)
    in
    the
    presence
    of
    bafiomycin
    A1
    the
    vacuolar
    type
    H+ATPase
    inhibitor
    [96]
    To
    improve
    cationic
    density
    Jiang
    et
    al
    prepared
    spermine
    induced
    chitosan
    (CHIgSPE)
    through
    imine
    reaction
    between
    periodateoxidized
    chitosan
    and
    spermine
    as
    a
    gene
    carrier
    in
    vitro
    and
    in
    vivo
    [4]
    CHIgSPEDNA
    complexes
    showed
    a
    good
    transfection
    efficiency
    while
    CHIgSPEGFP
    showed
    a
    GFP
    expression
    higher
    than
    that
    of
    the
    chitosanGFP
    complexes
    without
    toxicity
    Fig
    2
    shows
    the
    cationic
    groupmodified
    chitosan
    structure
    Targeting
    ligand
    modified
    chitosan
    Targeting
    ligand
    provides
    cellspecificity
    to
    chitosan
    whereby
    the
    target
    ligand
    modified
    chitosan
    has
    high
    cellular
    uptake
    and
    high
    transfection
    efficiency
    Various
    targeting
    ligands
    such
    as
    galactose
    [102–107]
    mannose
    [108–111]
    lactose
    [112]
    folate
    [113–115]
    transferrin
    [116]
    arginineglycineaspartic
    acid
    (RGD)
    [117]
    have
    been
    reported
    to
    provide
    cellspecificity
    and
    high
    transfection
    efficiency
    to
    chitosan
    Hepatocyte
    possesses
    asialoglycoprotein
    receptor
    (ASGR)
    that
    is
    known
    to
    be
    present
    in
    hepatocytes
    at
    a
    high
    density
    of
    500000
    receptors
    per
    cell
    and
    internalize
    galactoseterminal
    (asialo)
    glycoproteins
    also
    retained
    on
    several
    human
    hepatoma
    cell
    lines
    [118–120]
    Kim
    et
    al
    demonstrated
    that
    galactosylated
    water
    soluble
    chitosan
    (GC)
    coupled
    between
    lactobionic
    acid
    and
    chitosan
    showed
    a
    very
    high
    luciferase
    activity
    with
    DNA
    (pCI
    Luc)
    in
    HepG2
    cells
    wellknown
    model
    cells
    of
    parenchymal
    cells
    in
    the
    liver
    which
    have
    rich
    ASGPR
    receptors
    on
    the
    surface
    of
    cells
    [103]
    Furthermore
    the
    result
    of
    a
    competition
    assay
    clearly
    demonstrated
    that
    GC
    transferred
    the
    gene
    through
    receptor
    mediated
    transfection
    system
    Gao
    et
    al
    also
    prepared
    galactosy
    lated
    low
    molecular
    weight
    chitosan
    (galLMWC)
    through
    a
    method
    similar
    to
    Kim
    et
    al
    [103]
    and
    demonstrated
    the
    transfection
    efficiency
    of
    galLMWCDNA
    complexes
    in
    various
    cell
    lines
    [106]
    The
    bGalactosidase
    activity
    of
    galLMWCDNA
    complexes
    has
    shown
    a
    high
    transfection
    efficiency
    in
    ASGPR
    over
    expressed
    cell
    as
    a
    HepG2
    while
    the
    bGalactosidase
    activity
    of
    galLMWCDNA
    complexes
    showed
    a
    very
    low
    transfection
    efficiency
    on
    HeLa
    cells
    because
    HeLa
    cells
    have
    no
    ASGRR
    To
    enhance
    the
    transfection
    efficiency
    Lu
    et
    al
    [104]
    and
    Kim
    et
    al
    [105]
    prepared
    the
    PEI
    and
    galactose
    induced
    to
    chitosan
    The
    cellular
    uptake
    of
    Nsuccinylchitosangraftpolyethylenimine
    lactobionic
    acid
    (NSCgPEILA)YoYo1
    labeled
    DNA
    (pGL3)
    complexes
    was
    found
    to
    be
    located
    mainly
    in
    the
    cytoplasm
    of
    most
    cells
    (HepG2)
    and
    a
    few
    green
    dots
    were
    found
    in
    the
    nuclei
    using
    a
    confocal
    microscope
    [104]
    In
    addition
    the
    cellular
    uptake
    of
    (NSCgPEILA)YoYo1
    labeled
    DNA
    complexes
    increased
    the
    degree
    of
    LA
    substitution
    Macrophages
    express
    a
    mannose
    receptor
    that
    is
    used
    for
    mannosemediated
    endocytosis
    or
    phagocytosis
    [121]
    of
    various
    antigens
    and
    drug
    delivery
    systems
    [122]
    to
    target
    macrophages
    Hashimoto
    et
    al
    reported
    that
    DNAmannosylated
    chitosan
    (DNA
    manchitosan)
    complexes
    were
    found
    through
    mannose
    receptor
    mediated
    gene
    transfer
    whereby
    the
    transfection
    efficiency
    is
    enhanced
    [109]
    pDNAmanchitosan
    or
    pDNAchitosan
    complexes
    were
    observed
    in
    different
    intracellular
    transport
    in
    macrophages
    using
    the
    confocal
    laser
    microscope
    pDNAmanchitosan
    com
    plexes
    are
    delivered
    inside
    the
    cells
    while
    they
    are
    localized
    in
    early
    phogosomes
    near
    the
    plasma
    membrane
    In
    addition
    the
    transfection
    efficiency
    of
    pDNAmanchitosan
    (5
    substitution
    degree
    of
    mannose)
    was
    observed
    to
    be
    higher
    than
    chitosan
    and
    pDNAmanchitosan
    (10)
    Therefore
    the
    low
    substitution
    degree
    of
    mannose
    at
    5
    in
    chitosan
    is
    sufficient
    for
    gene
    delivery
    to
    macrophages
    Jiang
    et
    al
    prepared
    the
    mannose
    modified
    chitosangPEI
    (ManChigPEI)
    as
    a
    gene
    carrier
    for
    murine
    Fig
    2
    Cationic
    group
    modification
    chitosan
    A
    PEI
    modified
    chitosan
    [90]
    B
    urocanic
    acid
    modified
    chitosan
    [93]
    C
    imidazole
    modified
    chitosan
    [94]
    D
    diethyleneamine
    modified
    chitosan
    [96]
    and
    E
    spermine
    modified
    chitosan
    [4]
    C
    Choi
    et
    al

    Journal
    of
    Industrial
    and
    Engineering
    Chemistry
    33
    (2016)
    1–106macrophage
    cells
    (Raw
    2647)
    [110]
    ManChigPEI
    was
    observed
    to
    have
    high
    transfection
    efficiency
    in
    Raw
    2647
    cell
    and
    the
    mannose
    receptor
    expressed
    cell
    (antigen
    presenting
    cells
    (APCs))
    in
    contrast
    to
    ManChigPEI
    that
    was
    observed
    to
    have
    a
    very
    low
    transfection
    efficiency
    in
    HeLa
    cell
    the
    untargeted
    cell
    This
    result
    of
    transfection
    efficiency
    with
    ManChigPEI
    through
    mannose
    receptormediate
    gene
    transfer
    is
    similar
    to
    the
    results
    of
    the
    competition
    assay
    in
    the
    presence
    50
    mM
    of
    mannose
    Kim
    et
    al
    prepared
    the
    mannosylated
    chitosan
    (MC)
    which
    exhibited
    much
    enhanced
    interleukin12
    (IL12)
    gene
    transfer
    efficiency
    to
    dendritic
    cells
    that
    reside
    within
    the
    tumor
    through
    mannose
    receptormediated
    endocytosis
    between
    MC
    and
    mannose
    recep
    tor
    of
    dendritic
    by
    directly
    injection
    into
    tumor
    [111]
    The
    delivered
    pmIL12
    genes
    induced
    high
    expression
    levels
    of
    cytokines
    such
    as
    IL12
    p70
    and
    interferong
    (IFNg)
    As
    a
    result
    the
    growth
    and
    angiogenesis
    of
    the
    tumor
    were
    clearly
    suppressed
    Folate
    receptors
    (FR)
    are
    known
    to
    be
    overexpressed
    in
    various
    cancer
    cells
    surface
    such
    as
    ovarian
    lung
    breast
    colon
    and
    kidney
    cells
    and
    rarely
    found
    on
    normal
    cell
    surface
    [115123]
    The
    folate
    conjugated
    polymer
    including
    cationic
    liposomes
    [124]
    PLL
    [125]
    and
    PEI
    [126]
    have
    been
    reported
    to
    be
    cellular
    uptake
    into
    the
    FR
    present
    on
    tumor
    by
    receptormediated
    endocytosis
    Mansouri
    et
    al
    [113]
    and
    Chan
    et
    al
    [114]
    reported
    that
    folatechitosanDNA
    nanoparticles
    and
    chitosangPEGfolateDNA
    complex
    are
    char
    acterized
    for
    gene
    therapy
    Viola
    et
    al
    reported
    on
    the
    enhanced
    transfection
    efficiency
    of
    histidinetrimethylated
    chitosanfolate
    PEG
    (HTFP)
    polymers
    as
    a
    gene
    delivery
    vector
    [115]
    HTFPDNA
    (pGL3luc)
    polyplexes
    showed
    more
    enhanced
    transfection
    efficiency
    than
    histidinetrimethylated
    chitosan
    (HTMC)
    and
    HTFP
    with
    excess
    folate
    Transferrin
    (TF)
    has
    been
    widely
    applied
    with
    many
    advantages
    as
    a
    targeting
    ligand
    to
    achieve
    targeting
    of
    anticancer
    agent
    proteins
    and
    therapeutic
    genes
    to
    primary
    proliferating
    malignant
    cells
    that
    overexpressed
    transferrin
    receptors
    (TfR)
    [127–
    129]
    Kadiyala
    et
    al
    prepared
    transferrinconjugated
    chitosan
    DNA
    nanoparticle
    (NP
    ratio
    of
    3)
    to
    clarify
    the
    influence
    of
    the
    intrinsic
    properties
    including
    polymer
    chain
    length
    charge
    ratio
    nanoparticle
    size
    surface
    charge
    and
    ligand
    conjugation
    [116]
    The
    presence
    of
    TF
    in
    chitosanDNA
    nanoparticle
    enhances
    the
    transport
    of
    nanoparticles
    Han
    et
    al
    prepared
    RGDlabeled
    chitosan
    (RGDCH)
    to
    enhance
    targeted
    gene
    silencing
    [117117]
    RGD
    peptide
    binds
    with
    anb3
    integrin
    which
    is
    overexpressed
    in
    a
    wide
    range
    of
    tumors
    and
    rarely
    found
    on
    normal
    cell
    [130–132]
    siRNARGDCH
    nanopar
    ticles
    showed
    the
    targeted
    silencing
    of
    multiple
    growhpromoting
    genes
    (eg
    POSTN
    FAK
    and
    PLXDC1)
    in
    the
    human
    epithelial
    ovarian
    cancer
    models
    such
    as
    SKOV3ip1
    HeyA8
    and
    A2780
    In
    addition
    the
    PLXDC1
    siRNARGDCH
    nanoparticles
    delivered
    into
    the
    anb3
    integrinpositive
    tumor
    endothelial
    cells
    in
    the
    A2780
    tumorbearing
    mice
    were
    observed
    to
    have
    a
    higher
    gene
    silencing
    efficiency
    than
    control
    (siRNARGDCH
    nanoparticles)
    and
    PLXDC1
    siRNACH
    nanoparticles
    [117]
    Thiol
    group
    modified
    chitosan
    The
    thiol
    group
    introduced
    into
    several
    carrier
    systems
    increases
    the
    extracellular
    stability
    and
    improves
    the
    intracellular
    release
    properties
    owing
    to
    its
    enhanced
    properties
    such
    as
    the
    formation
    of
    reducible
    disulfide
    bonds
    between
    introduced
    thiol
    groups
    in
    carrier
    [133–136]
    In
    addition
    the
    thiol
    group
    introduced
    into
    chitosan
    as
    a
    carrier
    can
    promote
    its
    mucoadhesive
    potential
    because
    of
    the
    formation
    of
    disulfide
    bonds
    between
    the
    thiol
    group
    of
    polymer
    and
    mucin
    glycoproteins
    on
    the
    cell
    membrane
    whereby
    thiol
    groupmodified
    polymer
    enhances
    the
    cellular
    uptake
    [137138]
    Various
    materials
    such
    as
    cystamine
    [133]
    thiglycolic
    acid
    (TGA)
    [139–141]
    and
    2iminothiolane
    [142]
    have
    been
    used
    to
    introduce
    thiol
    group
    into
    chitosan
    for
    efficient
    gene
    transfer
    Varkouhi
    et
    al
    reported
    that
    the
    introduction
    of
    thiol
    groups
    to
    trimethylated
    chitosan
    (TMCSH)
    by
    cystamine
    modification
    enhances
    the
    extracellular
    stability
    of
    the
    complexes
    (siRNA
    TMCSH)
    and
    promotes
    the
    intracellular
    release
    of
    siRNA
    [133]
    siRNATMCSH
    polyplexes
    were
    observed
    to
    have
    a
    high
    gene
    silencing
    efficiency
    and
    good
    stability
    against
    competing
    anionic
    macromolecules
    as
    a
    hyaluronic
    acid
    while
    siRNATMC
    polyplexes
    hardly
    show
    any
    silencing
    activity
    in
    the
    same
    condition
    Thiolated
    chitosan
    (CSH)
    nanocomplexes
    TGAmodified
    to
    chitosan
    and
    DNA
    are
    prepared
    to
    enhance
    transfection
    efficiency
    [139]
    CSH360
    (thiol
    groups
    360
    mmol
    of
    chitosan)DNA
    nano
    complexes
    induced
    a
    significantly
    higher
    GFP
    expression
    than
    the
    thiol
    group
    unmodified
    chitosanDNA
    nanocomplexes
    in
    various
    cell
    lines
    including
    HEK293
    madindarby
    canine
    kidney
    (MDCK)
    and
    human
    larynx
    carcinoma
    cell
    (Hep2)
    In
    addition
    disulphide
    crosslinked
    CSH360DNA
    nanocomplexes
    are
    observed
    to
    show
    a
    sustained
    DNA
    release
    and
    continuous
    expression
    in
    cultured
    cells
    for
    over
    60
    h
    after
    transfection
    Martien
    et
    al
    reported
    that
    thiolated
    chitosan
    when
    synthesized
    by
    introducing
    TGA
    to
    chitosan
    via
    amide
    bond
    formation
    mediated
    by
    a
    carbodiimide
    showed
    a
    higher
    transfection
    efficiency
    than
    the
    thiol
    group
    unmodified
    chitosan
    in
    human
    colorectal
    carcinoma
    cell
    lines
    (Caco2)
    [140]
    and
    Caco2
    differentiated
    cell
    culture
    system
    [141]
    Amino
    acid
    and
    peptide
    modified
    chitosan
    Recently
    chitosanpeptide
    derivatives
    have
    received
    increasing
    interest
    in
    the
    drug
    and
    gene
    delivery
    system
    due
    to
    their
    beneficial
    property
    such
    as
    enhanced
    cell
    adsorption
    and
    excellent
    safety
    profile
    [1143144]
    Malhotra
    et
    al
    [145]
    and
    Katas
    et
    al
    [146]
    demonstrated
    that
    TAT
    peptide
    (cellpenetrating
    peptide
    (RKKRRQRRR))
    chitosan
    can
    be
    used
    to
    enhance
    gene
    therapeutic
    as
    a
    gene
    carrier
    To
    introduce
    the
    function
    of
    cellpenetrating
    hydrophilicity
    hydrophobicity
    and
    thiol
    group
    to
    chitosan
    amino
    acids
    are
    used
    in
    several
    studies
    [1136147148]
    Gao
    et
    al
    prepared
    arginine
    chitosan
    (ArgCs)DNA
    selfassemble
    nanoparticles
    (ACSNs)
    to
    overcome
    its
    poor
    water
    solubility
    and
    low
    transfection
    efficiency
    [147]
    Arginine
    of
    ArgCs
    can
    increase
    the
    pKa
    value
    of
    chitosan
    which
    leads
    to
    high
    solubility
    The
    transfection
    efficiency
    of
    ACSNs
    is
    found
    not
    only
    to
    be
    much
    higher
    than
    that
    of
    chitosanDNA
    nanoparticles
    (CSNs)
    but
    also
    similar
    level
    as
    Lipofectamine
    Loretz
    et
    al
    [136]
    and
    Zhao
    et
    al
    [148]
    reported
    that
    thiolated
    chitosan
    improves
    the
    transfection
    efficiency
    through
    cysteine
    modification
    because
    of
    the
    sulfide
    group
    introduced
    to
    chitosan
    Layek
    et
    al
    reported
    on
    the
    hydrophobic
    amino
    acid
    grafted
    chitosan
    (AGC)
    with
    Lalanine
    Lvaline
    Lleucine
    and
    Lisoleucine
    to
    enhance
    membrane
    permeability
    [1]
    The
    cellular
    uptakes
    of
    AGCA
    AGCV
    AGCL
    and
    AGCI
    polylexes
    with
    pDNA
    are
    624
    846
    978
    and
    983
    respectively
    The
    GFP
    expression
    efficiency
    of
    AGCamino
    sampDNA
    polyplexes
    is
    similar
    to
    the
    results
    of
    cellular
    uptake
    at
    AGCI
    
    AGCL
    >
    AGCV
    >
    AGCA
    Fig
    3
    shows
    the
    structure
    of
    chitosan
    derivatives
    which
    are
    modified
    with
    targeting
    ligand
    thiol
    group
    or
    amino
    acid
    Genetic
    materials
    delivery
    by
    anionic
    materialchitosan
    complexes
    Several
    anionic
    materials
    have
    been
    used
    to
    improve
    the
    properties
    (eg
    low
    transfection
    efficiency
    low
    solubility
    and
    serum
    stability)
    of
    chitosan
    as
    a
    genetic
    materials
    delivery
    vector
    Anionic
    materials
    formed
    the
    complexes
    with
    chitosan
    through
    gelation
    or
    electrostatic
    interaction
    C
    Choi
    et
    al

    Journal
    of
    Industrial
    and
    Engineering
    Chemistry
    33
    (2016)
    1–10
    7Anion
    polymer
    and
    chitosan
    complexes
    Tripolyphosphate
    (TPP)
    a
    poly
    anion
    can
    form
    nanoparticles
    with
    chitosan
    by
    using
    the
    ionotropic
    gelation
    between
    positively
    charged
    chitosan
    and
    negatively
    charged
    TPP
    [149150]
    In
    addition
    TPP
    was
    used
    as
    crosslinker
    like
    a
    polyanionic
    linker
    because
    of
    its
    unique
    properties
    of
    nontoxicity
    and
    instant
    gelling
    ability
    [151]
    Gaspar
    et
    al
    reported
    that
    the
    particle
    size
    of
    chitosanTPPpDNA
    nanocapsules
    influences
    the
    parameters
    like
    the
    chitosan
    degree
    of
    deacetylation
    and
    formulation
    ratio
    (chitosan
    to
    TPP)
    [152]
    The
    particle
    size
    of
    chitosanTPPpDNA
    nanocapsules
    was
    increased
    when
    the
    chitosan
    degree
    of
    deacetylation
    increased
    in
    the
    presence
    of
    pDNA
    Katas
    et
    al
    reported
    on
    the
    chitosanTPP
    nanoparticles
    prepared
    through
    ionic
    gelation
    with
    two
    different
    types
    of
    chitosan
    (hydrochloride
    form
    and
    glutamate
    form)
    and
    each
    type
    with
    two
    different
    MW
    [153]
    siRNA
    (targeting
    against
    pGL3
    luciferase
    gene)
    associated
    with
    chitosanTPP
    nanoparticles
    that
    used
    hydrochloride
    form
    of
    chitosan
    showed
    a
    high
    gene
    silencing
    efficiency
    In
    addition
    the
    gene
    silencing
    efficiency
    of
    siRNA
    associated
    with
    chitosanTPP
    nanoparticles
    increased
    with
    the
    increasing
    MW
    (270–470
    kDa)
    Wang
    et
    al
    demonstrated
    the
    potential
    of
    chitosanTPP
    nanopar
    ticle
    to
    deliver
    short
    hairpin
    RNA
    (shRNA)
    [154]
    ChitosanTPP
    mediator
    successfully
    delivered
    GFP
    expression
    vector
    pSUPER
    into
    the
    human
    rhabdomyosarcoma
    cell
    (RD)
    and
    showed
    a
    higher
    GFP
    expression
    than
    the
    chitosan
    mediator
    In
    addition
    transform
    ing
    growth
    factor
    (TGFB1)specific
    shRNA
    was
    effectively
    delivered
    using
    chitosanTPP
    mediator
    into
    the
    RD
    cells
    that
    reduce
    the
    TGFB1
    level
    When
    delivered
    into
    the
    mice
    through
    chitosanTPP
    mediator
    with
    four
    type
    shRNA

    shRNAun
    (negative
    control)
    shRNAa
    shRNAb
    and
    shRNAc

    after
    RD
    cell
    inoculation
    in
    nude
    mice
    for
    2
    weeks
    the
    tumor
    volume
    in
    the
    four
    groups
    was
    decreased
    to
    939
    767
    455
    and
    625
    of
    the
    control
    tumor
    size
    respectively
    Anionic
    biopolymer
    and
    chitosan
    complexes
    Hyaluronic
    acid
    (HA)
    is
    another
    anionic
    biopolymer
    widely
    used
    in
    biomedical
    application
    for
    its
    excellent
    cytocompatibility
    cell
    adhesion
    morphogenesis
    inflammation
    regulation
    and
    biocompatibility
    [155–158]
    In
    addition
    HA
    can
    also
    interact
    with
    the
    CD44
    receptor
    which
    is
    expressed
    in
    the
    human
    cornea
    and
    conjunctiva
    [159160]
    that
    led
    to
    regeneration
    of
    corneal
    and
    conjunctiva
    epithelial
    cells
    [161]
    Fuente
    et
    al
    demonstrated
    that
    optimal
    formulation
    of
    HAchitosan
    nanoparticles
    (HACS
    NP)
    was
    found
    to
    be
    12
    (HAchitosan
    oligomer
    (CS)
    of
    various
    HACS
    NP
    formulations
    including
    HACS
    (11
    12
    and
    21)
    hyaluronic
    acid
    oligomer
    (HAO)CS
    (11
    12
    and
    21)
    HACSO
    (12)
    and
    HAOCSO
    (11
    12))
    [160]
    Ravin˜a
    et
    al
    reported
    that
    the
    cell
    toxicity
    of
    HA
    CSgrafted
    PEG
    (HACSgPEG
    21
    of
    formulation
    ratio)
    showed
    a
    decrease
    with
    high
    amount
    of
    nanoparticles
    (HACSgPEGDNA)
    at
    10181
    mgcm2 due
    to
    HA’s
    high
    biocompatibility
    compared
    to
    CSgPEG
    or
    HACSgPEG
    12
    or
    HACSgPEG
    12
    formulations
    [162]
    In
    addition
    HACSgPEGDNA
    (pEGFP)
    was
    found
    to
    be
    effective
    gene
    transfection
    efficiency
    in
    all
    formulations
    (11
    12
    and
    21)
    and
    HACSgPEGsiRNA
    (against
    the
    EGFP)
    significantly
    silenced
    the
    EGFP
    expression
    compared
    to
    nontreated
    cells
    Lin
    et
    al
    demonstrated
    that
    the
    prepared
    HAchitosanDNA
    complex
    multilayer
    not
    only
    has
    good
    cytocompatibility
    but
    also
    possesses
    the
    in
    vitro
    gene
    transfection
    ability
    [163]
    Lu
    et
    al
    investigated
    the
    pDNA
    delivered
    by
    HACS
    vectors
    to
    enhance
    transfection
    optimal
    environment
    [164]
    Various
    factors
    including
    transfection
    medium
    pH
    NP
    ratio
    pDNA
    concentrations
    and
    MW
    of
    chitosan
    were
    investigated
    The
    transfection
    efficiency
    of
    HACSpDNA
    nanopar
    ticles
    was
    significantly
    higher
    than
    that
    of
    CSDNA
    nanoparticles
    at
    the
    following
    condition
    medium
    pH
    68
    5
    of
    NP
    ratio
    4
    mgmL
    of
    pDNA
    concentration
    and
    50
    kDa
    of
    chitosan
    MW
    respectively
    Polypeptide
    and
    chitosan
    complexes
    Poly(gglutamic
    acid)
    (PGA)
    a
    naturally
    occurring
    peptide
    is
    biodegradable
    watersoluble
    and
    nontoxic
    [54165]
    Therefore
    PGA
    and
    its
    derivatives
    have
    been
    employed
    as
    a
    carrier
    in
    biomedical
    field
    such
    as
    oral
    delivery
    of
    insulin
    [166167]
    and
    protein
    vaccines
    delivery
    [168]
    Liao
    et
    al
    reported
    that
    chitosan
    siRNAPGA
    complexes
    significantly
    expedite
    the
    onset
    of
    gene
    knockdown
    and
    also
    enhance
    their
    inhibition
    efficiency
    and
    prolong
    the
    duration
    of
    gene
    silencing
    [54]
    In
    addition
    the
    cellular
    uptake
    of
    chitosansiRNAPGA
    is
    increased
    when
    the
    PGA
    ratio
    increases
    from
    0
    to
    50
    Generally
    anionic
    complexes
    are
    not
    taken
    up
    well
    by
    cells
    owing
    to
    the
    electrostatic
    repulsion
    between
    anionic
    complexes
    and
    negatively
    charged
    cell
    membranes
    [54]
    However
    PGAcoated
    cationic
    polymers
    significantly
    enhance
    their
    cellular
    uptake
    suggesting
    that
    there
    might
    be
    a
    PGAspecific
    receptormediated
    pathway
    [54165169]
    In
    addi
    tion
    Peng
    groups
    reported
    on
    the
    cellular
    uptake
    and
    transfection
    Fig
    3
    Chitosan
    derivatives
    with
    targeting
    ligand
    (A)
    thiol
    group
    (B)
    or
    amino
    acid
    (C)
    A
    (a)
    galactosylated
    chitosan
    [106]
    (b)
    mannosylated
    chitosan
    [109]
    and
    (c)
    lactosylated
    chitosan
    [111]
    B
    (a)
    thioglycolic
    acid
    conjugated
    chitosan
    [140]
    and
    (b)
    cystamine
    modified
    trimethylated
    chitosan
    [132]
    C
    amino
    acid
    modified
    chitosan
    [1134146]
    C
    Choi
    et
    al

    Journal
    of
    Industrial
    and
    Engineering
    Chemistry
    33
    (2016)
    1–108efficiency
    [165]
    and
    the
    mechanisms
    of
    cellular
    uptake
    and
    intracellular
    trafficking
    [170]
    of
    chitosanDNAPGA
    (CSDNAPGA)
    complexes
    as
    a
    gene
    delivery
    vector
    Transfection
    and
    intracellular
    uptake
    of
    CSDNAPGA
    which
    is
    prepared
    at
    different
    chitosan
    DNAPGA
    (NPC)
    ratios
    increased
    via
    a
    specific
    proteinmediated
    endocytosis
    when
    NPC
    ratios
    increased
    ranging
    from
    1010
    to
    1014
    [165]
    The
    mechanisms
    of
    cellular
    uptake
    and
    intracellular
    trafficking
    of
    CSDNAPGA
    were
    investigated
    with
    various
    inhibi
    tors
    on
    the
    uptake
    such
    as
    chlorpromazine
    (clathrinmediated
    uptake
    inhibitor)
    wortamannin
    (phosphatidyl
    inositol3phos
    phate
    inhibitor)
    cytochalasin
    D
    (actin
    polymerization
    and
    membrane
    ruffling
    or
    macropinocytosis
    inhibitor)
    filipin
    (caveo
    laemediated
    endocytosis
    inhibitor)
    or
    genistein
    (caveolaemedi
    ated
    endocytosis
    inhibitor)
    respectively
    [170]
    As
    a
    result
    of
    the
    uptake
    test
    with
    endocytosis
    inhibitor
    the
    CSDNAPGA
    complexes
    are
    internalized
    via
    macropinocytosis
    and
    caveolaemediated
    pathway
    Conclusion
    Chitosan
    chitosan
    derivatives
    and
    chitosananionic
    materials
    complexes
    can
    be
    designed
    by
    enhancing
    the
    physicochemical
    properties
    for
    genetic
    materials
    delivery
    for
    gene
    therapy
    The
    MW
    DDA
    genetic
    materials
    concentration
    and
    serum
    stability
    of
    chitosan
    and
    the
    various
    modifications
    (hydrophilic
    group
    hydrophobic
    group
    cationic
    group
    targeting
    ligand
    thiol
    group
    and
    amino
    acid)
    are
    very
    important
    factors
    in
    preparing
    chitosan
    derivatives
    to
    enhance
    the
    efficiency
    of
    gene
    therapy
    In
    addition
    anionic
    materials
    such
    as
    TPP
    HA
    and
    PGA
    also
    enhance
    the
    cellular
    uptake
    and
    transfection
    efficiency
    of
    chitosangenetic
    materials
    This
    review
    focused
    on
    the
    chitosan
    derivatives
    chitosananionic
    materials
    preparing
    techniques
    and
    sitespecific
    targeting
    to
    improve
    the
    chitosan
    properties
    for
    genetic
    materials
    delivery
    These
    factors
    modifications
    and
    complexation
    with
    anionic
    materials
    effectively
    improved
    the
    chitosan
    properties
    including
    solubility
    in
    aqueous
    solution
    toxicity
    in
    HM
    chitosan
    buffering
    capacity
    escapes
    in
    endosome
    cellular
    uptake
    genetic
    material
    release
    from
    chitosan
    based
    polyplexes
    transfection
    efficiency
    and
    silencing
    efficiency
    However
    most
    of
    these
    results
    were
    obtained
    from
    experiments
    in
    vitro
    Therefore
    further
    research
    is
    needed
    on
    chitosan
    for
    gene
    therapy
    in
    vivo
    Research
    on
    chitosan
    chitosan
    derivatives
    and
    chitosananionic
    materials
    complexes
    still
    needs
    to
    understand
    the
    effects
    of
    the
    character
    istics
    of
    the
    gene
    carriers
    on
    cellular
    entry
    and
    intracellular
    trafficking
    processes
    Moreover
    most
    of
    the
    researches
    used
    HM
    chitosan
    Research
    on
    in
    vivo
    and
    LM
    chitosan
    is
    called
    for
    the
    development
    of
    genetic
    materials
    delivery
    Acknowledgements
    This
    work
    was
    supported
    by
    National
    Research
    Foundation
    of
    Korea
    (NRF)
    grant
    funded
    by
    the
    Ministry
    of
    Science
    ICT
    &
    Future
    Planning
    (No
    NRF2014R1A2A1A10053027)
    References
    [1]
    B
    Layek
    J
    Singh
    Biomacromolecules
    14
    (2013)
    485
    [2]
    C
    Louise
    Methods
    Mol
    Biol
    333
    (2006)
    201
    [3]
    A
    Rolland
    Adv
    Drug
    Deliv
    Rev
    57
    (2005)
    669
    [4]
    HL
    Jiang
    HT
    Lim
    YK
    Kim
    R
    Arote
    JY
    Shin
    JT
    Kwon
    JE
    Kim
    JH
    Kim
    D
    Kim
    C
    Chae
    JW
    Nah
    YJ
    Choi
    CS
    Cho
    MH
    Cho
    Eur
    J
    Pharm
    Biopharm
    77
    (2011)
    36
    [5]
    WF
    Anderson
    Science
    256
    (1992)
    808
    [6]
    S
    Nayak
    RW
    Herzog
    Gene
    Ther
    17
    (2010)
    295
    [7]
    HS
    Zhou
    DP
    Liu
    CC
    Liang
    Med
    Res
    Rev
    24
    (2004)
    748
    [8]
    AE
    Smith
    Annu
    Rev
    Microbiol
    49
    (1995)
    807
    [9]
    M
    Morille
    C
    Passirani
    A
    Vonarbourg
    A
    Clavreul
    JP
    Benoit
    Biomaterials
    29
    (2008)
    3477
    [10]
    S
    Li
    L
    Huang
    Gene
    Ther
    7
    (2000)
    31
    [11]
    Y
    Liang
    Z
    Liu
    X
    Shuai
    W
    Wang
    J
    Liu
    W
    Bi
    C
    Wang
    X
    Jing
    Y
    Liu
    E
    Tao
    Biochem
    Biophys
    Res
    Commun
    421
    (2012)
    690
    [12]
    X
    Gao
    R
    Kuruba
    K
    Damodaran
    BW
    Day
    D
    Liu
    S
    Li
    J
    Control
    Release
    137
    (2009)
    38
    [13]
    T
    Kiang
    J
    Wen
    HW
    Lim
    KW
    Leong
    Biomaterials
    25
    (2004)
    5293
    [14]
    O
    Boussif
    F
    Lezoualc’h
    MA
    Zanta
    MD
    Mergny
    D
    Scherman
    B
    Demeneix
    JP
    Behr
    Proc
    Natl
    Acad
    Sci
    U
    S
    A
    92
    (1995)
    7297
    [15]
    J
    Suh
    HJ
    Paik
    BK
    Hwang
    Bioorg
    Chem
    22
    (1994)
    318
    [16]
    F
    HoppeSeiler
    Ber
    Dtsch
    Chem
    Ges
    27
    (1994)
    3329
    [17]
    A
    Domard
    N
    Cartier
    Int
    J
    Biol
    Macromol
    14
    (1992)
    100
    [18]
    R
    Hejazi
    M
    Amiji
    J
    Control
    Release
    89
    (2003)
    151
    [19]
    C
    Dong
    W
    Chen
    C
    Liu
    Bioresour
    Technol
    170
    (2014)
    239
    [20]
    W
    Liu
    S
    Sun
    Z
    Cao
    X
    Zhang
    K
    Yao
    WW
    Lu
    KD
    Luk
    Biomaterials
    26
    (2005)
    2705
    [21]
    PA
    Sandford
    Am
    Chem
    Soc
    Div
    Polym
    Chem
    31
    (1990)
    628
    [22]
    S
    Mansouri
    P
    Lavigne
    K
    Corsi
    M
    Benderdour
    E
    Beaumont
    JC
    Fernandes
    Eur
    J
    Pharm
    Biopharm
    57
    (2004)
    1
    [23]
    H
    Struszczyk
    D
    Wawro
    A
    Niekraszewicz
    in
    CJ
    Brine
    PA
    Sandford
    JP
    Zikakis
    (Eds)
    Advances
    in
    Chitin
    and
    Chitosan
    Elsevier
    Applied
    Science
    London
    1991
    p
    580
    [24]
    T
    Chandy
    CP
    Sharma
    Biomater
    Artif
    Cells
    Artif
    Organs
    18
    (1990)
    1
    [25]
    FC
    MacLaughlin
    RJ
    Mumper
    J
    Wang
    JM
    Tagliaferri
    I
    Gill
    M
    Hinchcliffe
    AP
    Rolland
    J
    Control
    Release
    56
    (1998)
    259
    [26]
    Z
    Cui
    RJ
    Mumper
    J
    Control
    Release
    75
    (2001)
    409
    [27]
    X
    Zhao
    SB
    Yu
    FL
    Wu
    ZB
    Mao
    CL
    Yu
    J
    Control
    Release
    112
    (2006)
    223
    [28]
    W
    Weecharangsan
    P
    Opanasopit
    T
    Ngawhirunpat
    A
    Apirakaramwong
    T
    Rojanarata
    U
    Ruktanonchai
    RJ
    Lee
    Int
    J
    Pharm
    348
    (2008)
    161
    [29]
    L
    Niu
    YC
    Xu
    HY
    Xie
    Z
    Dai
    HQ
    Tang
    Acta
    Pharmacol
    Sin
    29
    (2008)
    1342
    [30]
    K
    Romøren
    S
    Pedersen
    G
    Smistad
    Ø
    Evensen
    BJ
    Thu
    Int
    J
    Pharm
    261
    (2003)
    115
    [31]
    M
    Lavertu
    S
    Methot
    N
    TranKhanh
    MD
    Buschmann
    Biomaterials
    27
    (2006)
    4815
    [32]
    Si
    Aiba
    Int
    J
    Biol
    Macromol
    14
    (1992)
    225
    [33]
    PL
    Ma
    M
    Lavertu
    FoM
    Winnik
    MD
    Buschmann
    Biomacromolecules
    10
    (2009)
    1490
    [34]
    M
    KopingHoggard
    KM
    Varum
    M
    Issa
    S
    Danielsen
    BE
    Christensen
    BT
    Stokke
    P
    Artursson
    Gene
    Ther
    11
    (2004)
    1441
    [35]
    M
    KopingHoggard
    I
    Tubulekas
    H
    Guan
    K
    Edwards
    M
    Nilsson
    KM
    Varum
    P
    Artursson
    Gene
    Ther
    8
    (2001)
    1108
    [36]
    MK
    Jang
    YI
    Jeong
    CS
    Cho
    SH
    Yang
    YE
    Kang
    JW
    Nah
    Bull
    Korean
    Chem
    Soc
    23
    (2002)
    914
    [37]
    VA
    Bloomfield
    Biopolymers
    44
    (1997)
    269
    [38]
    SP
    Strand
    S
    Lelu
    NK
    Reitan
    C
    de
    Lange
    Davies
    P
    Artursson
    KM
    Varum
    Biomaterials
    31
    (2010)
    975
    [39]
    J
    Akbuga
    S
    OzbasTuran
    N
    Erdogan
    Eur
    J
    Pharm
    Biopharm
    58
    (2004)
    501
    [40]
    M
    KopingHoggard
    YS
    Mel’nikova
    KM
    Varum
    B
    Lindman
    P
    Artursson
    J
    Gene
    Med
    5
    (2003)
    130
    [41]
    P
    Nydert
    A
    Dragomir
    L
    Hjelte
    Biotechnol
    Appl
    Biochem
    51
    (2008)
    153
    [42]
    U
    Guliyeva
    F
    Oner
    S
    Ozsoy
    R
    Haziroglu
    Eur
    J
    Pharm
    Biopharm
    62
    (2006)
    17
    [43]
    S
    Nimesh
    MM
    Thibault
    M
    Lavertu
    MD
    Buschmann
    Mol
    Biotechnol
    46
    (2010)
    182
    [44]
    MN
    Centelles
    C
    Qian
    MA
    Campanero
    JM
    Irache
    Int
    J
    Nanomed
    3
    (2008)
    451
    [45]
    Y
    Yuan
    J
    Tan
    Y
    Wang
    C
    Qian
    M
    Zhang
    Acta
    Biochim
    Biophys
    Sin
    (Shanghai)
    41
    (2009)
    515
    [46]
    XW
    Li
    DKL
    Lee
    ASC
    Chan
    HO
    Alpar
    BBA

    Gene
    Struct
    Expr
    1630
    (2003)
    7
    [47]
    F
    Chellat
    A
    GrandjeanLaquerriere
    R
    Le
    Naour
    J
    Fernandes
    L
    Yahia
    M
    Guenounou
    D
    LaurentMaquin
    Biomaterials
    26
    (2005)
    961
    [48]
    PT
    Yang
    L
    Hoang
    WW
    Jia
    ED
    Skarsgard
    J
    Surg
    Res
    171
    (2011)
    691
    [49]
    X
    Yang
    X
    Yuan
    D
    Cai
    S
    Wang
    L
    Zong
    Int
    J
    Pharm
    375
    (2009)
    123
    [50]
    K
    Roy
    HQ
    Mao
    SK
    Huang
    KW
    Leong
    Nat
    Med
    5
    (1999)
    387
    [51]
    M
    Kumar
    AK
    Behera
    RF
    Lockey
    J
    Zhang
    G
    Bhullar
    CP
    De
    La
    Cruz
    LC
    Chen
    KW
    Leong
    SK
    Huang
    SS
    Mohapatra
    Hum
    Gene
    Ther
    13
    (2002)
    1415
    [52]
    SM
    Elbashir
    J
    Harborth
    W
    Lendeckel
    A
    Yalcin
    K
    Weber
    T
    Tuschl
    Nature
    411
    (2001)
    494
    [53]
    AM
    Ji
    D
    Su
    O
    Che
    WS
    Li
    L
    Sun
    ZY
    Zhang
    B
    Yang
    F
    Xu
    Nanotechnology
    20
    (2009)
    405103
    [54]
    ZX
    Liao
    YC
    Ho
    HL
    Chen
    SF
    Peng
    CW
    Hsiao
    HW
    Sung
    Biomaterials
    31
    (2010)
    8780
    [55]
    E
    Song
    SK
    Lee
    DM
    Dykxhoorn
    C
    Novina
    D
    Zhang
    K
    Crawford
    J
    Cemy
    PA
    Sharp
    J
    Lieberman
    N
    Manjunath
    P
    Shankar
    J
    Virol
    77
    (2003)
    7174
    [56]
    MA
    Behlke
    Mol
    Ther
    13
    (2006)
    644
    [57]
    M
    Sioud
    Expert
    Opin
    Drug
    Deliv
    2
    (2005)
    639
    [58]
    KA
    Howard
    UL
    Rahbek
    X
    Liu
    CK
    Damgaard
    SZ
    Glud
    MO
    Andersen
    MB
    Hovgaard
    A
    Schmitz
    JR
    Nyengaard
    F
    Besenbacher
    J
    Kjems
    Mol
    Ther
    14
    (2006)
    476
    [59]
    X
    Liu
    KA
    Howard
    M
    Dong
    MO
    Andersen
    UL
    Rahbek
    MG
    Johnsen
    OC
    Hansen
    F
    Besenbacher
    J
    Kjems
    Biomaterials
    28
    (2007)
    1280
    [60]
    J
    Malmo
    H
    Sorgard
    KM
    Varum
    SP
    Strand
    J
    Control
    Release
    158
    (2012)
    261
    [61]
    EJ
    Nielsen
    JM
    Nielsen
    D
    Becker
    A
    Karlas
    H
    Prakash
    SZ
    Glud
    J
    Merrison
    F
    Besenbacher
    TF
    Meyer
    J
    Kjems
    KA
    Howard
    Pharm
    Res
    27
    (2010)
    2520
    [62]
    M
    Alameh
    M
    Jean
    D
    Dejesus
    MD
    Buschmann
    A
    Merzouki
    Int
    J
    Nanomed
    5
    (2010)
    473
    [63]
    W
    Xu
    Y
    Shen
    Z
    Jiang
    Y
    Wang
    Y
    Chu
    S
    Xiong
    Vaccine
    22
    (2004)
    3603
    [64]
    YZ
    Du
    P
    Lu
    JP
    Zhou
    H
    Yuan
    FQ
    Hu
    Int
    J
    Pharm
    391
    (2010)
    260
    [65]
    FQ
    Hu
    MD
    Zhao
    H
    Yuan
    J
    You
    YZ
    Du
    S
    Zeng
    Int
    J
    Pharm
    315
    (2006)
    158
    C
    Choi
    et
    al

    Journal
    of
    Industrial
    and
    Engineering
    Chemistry
    33
    (2016)
    1–10
    9[66]
    HS
    Yoo
    JE
    Lee
    H
    Chung
    IC
    Kwon
    SY
    Jeong
    J
    Control
    Release
    103
    (2005)
    235
    [67]
    W
    Wang
    J
    Yao
    JP
    Zhou
    Y
    Lu
    Y
    Wang
    L
    Tao
    YP
    Li
    Biochem
    Biophys
    Res
    Commun
    377
    (2008)
    567
    [68]
    B
    Wang
    C
    He
    C
    Tang
    C
    Yin
    Biomaterials
    32
    (2011)
    4630
    [69]
    S
    Mao
    W
    Sun
    T
    Kissel
    Adv
    Drug
    Deliv
    Rev
    62
    (2010)
    12
    [70]
    Y
    Zhang
    J
    Chen
    Y
    Zhang
    Y
    Pan
    J
    Zhao
    L
    Ren
    M
    Liao
    Z
    Hu
    L
    Kong
    J
    Wang
    Biotechnol
    Appl
    Biochem
    46
    (2007)
    197
    [71]
    F
    Kreppel
    S
    Kochanek
    Mol
    Ther
    16
    (2008)
    16
    [72]
    Z
    Mao
    L
    Ma
    Y
    Jiang
    M
    Yan
    C
    Gao
    J
    Shen
    Macromol
    Biosci
    7
    (2007)
    855
    [73]
    A
    Kotze´
    H
    Lueben
    B
    de
    Leeuw
    B
    de
    Boer
    JC
    Verhoef
    H
    Junginger
    Pharm
    Res
    14
    (1997)
    1197
    [74]
    SM
    van
    der
    Merwe
    JC
    Verhoef
    JHM
    Verheijden
    AF
    Kotze´
    HE
    Junginger
    Eur
    J
    Pharm
    Biopharm
    58
    (2004)
    225
    [75]
    T
    Kean
    S
    Roth
    M
    Thanou
    J
    Control
    Release
    103
    (2005)
    643
    [76]
    S
    Mao
    X
    Shuai
    F
    Unger
    M
    Wittmar
    X
    Xie
    T
    Kissel
    Biomaterials
    26
    (2005)
    6343
    [77]
    O
    Germershaus
    S
    Mao
    J
    Sitterberg
    U
    Bakowsky
    T
    Kissel
    J
    Control
    Release
    125
    (2008)
    145
    [78]
    RJ
    Verheul
    M
    Amidi
    S
    van
    der
    Wal
    E
    van
    Riet
    W
    Jiskoot
    WE
    Hennink
    Biomaterials
    29
    (2008)
    3642
    [79]
    B
    Layek
    J
    Singh
    Int
    J
    Pharm
    447
    (2013)
    182
    [80]
    Z
    Liu
    Z
    Zhang
    C
    Zhou
    Y
    Jiao
    Prog
    Polym
    Sci
    35
    (2010)
    1144
    [81]
    M
    Piest
    JF
    Engbersen
    J
    Control
    Release
    148
    (2010)
    83
    [82]
    PS
    Kuhn
    Y
    Levin
    MC
    Barbosa
    Physica
    A
    274
    (1999)
    8
    [83]
    WG
    Liu
    X
    Zhang
    SJ
    Sun
    GJ
    Sun
    KD
    Yao
    DC
    Liang
    G
    Guo
    JY
    Zhang
    Bioconjugate
    Chem
    14
    (2003)
    782
    [84]
    J
    Yan
    YZ
    Du
    FY
    Chen
    J
    You
    H
    Yuan
    FQ
    Hu
    Mol
    Pharm
    (2013)
    [85]
    D
    Zhu
    X
    Jin
    X
    Leng
    H
    Wang
    J
    Bao
    W
    Liu
    K
    Yao
    C
    Song
    Int
    J
    Nanomed
    5
    (2010)
    1095
    [86]
    SY
    Chae
    S
    Son
    M
    Lee
    MK
    Jang
    JW
    Nah
    J
    Control
    Release
    109
    (2005)
    330
    [87]
    JY
    Lee
    SH
    Lee
    MH
    Oh
    JS
    Kim
    TG
    Park
    YS
    Nam
    J
    Control
    Release
    162
    (2012)
    407
    [88]
    K
    Wong
    G
    Sun
    X
    Zhang
    H
    Dai
    Y
    Liu
    C
    He
    KW
    Leong
    Bioconjugate
    Chem
    17
    (2006)
    152
    [89]
    B
    Lu
    XD
    Xu
    XZ
    Zhang
    SX
    Cheng
    RX
    Zhuo
    Biomacromolecules
    9
    (2008)
    2594
    [90]
    QQ
    Zhao
    JL
    Chen
    M
    Han
    WQ
    Liang
    Y
    Tabata
    JQ
    Gao
    J
    Biosci
    Bioeng
    105
    (2008)
    65
    [91]
    B
    Lu
    YX
    Sun
    YQ
    Li
    XZ
    Zhang
    RX
    Zhuo
    Mol
    Biosyst
    5
    (2009)
    629
    [92]
    JQ
    Gao
    QQ
    Zhao
    TF
    Lv
    WP
    Shuai
    J
    Zhou
    GP
    Tang
    WQ
    Liang
    Y
    Tanata
    YL
    Hu
    Int
    J
    Pharm
    387
    (2010)
    286
    [93]
    B
    Lu
    CF
    Wang
    DQ
    Wu
    C
    Li
    XZ
    Zhang
    RX
    Zhuo
    J
    Control
    Release
    137
    (2009)
    54
    [94]
    TH
    Kim
    JE
    Ihm
    YJ
    Choi
    JW
    Nah
    CS
    Cho
    J
    Control
    Release
    93
    (2003)
    389
    [95]
    C
    Moreira
    H
    Oliveira
    LR
    Pires
    S
    Simoes
    MA
    Barbosa
    AP
    Pego
    Acta
    Biomater
    5
    (2009)
    2995
    [96]
    B
    Ghosn
    A
    Singh
    M
    Li
    AV
    Vlassov
    C
    Burnett
    N
    Puri
    L
    Roy
    Oligonucleotides
    20
    (2010)
    163
    [97]
    F
    de
    Paula
    Pansani
    Oliveira
    IP
    Dalla
    Picola
    Q
    Shi
    HF
    Barbosa
    VA
    Tiera
    JC
    Fernandes
    MJ
    Tiera
    Nanotechnology
    24
    (2013)
    055101
    [98]
    S
    Zhang
    Y
    Xu
    B
    Wang
    W
    Qiao
    D
    Liu
    Z
    Li
    J
    Control
    Release
    100
    (2004)
    165
    [99]
    KF
    Pirollo
    L
    Xu
    EH
    Chang
    Curr
    Opin
    Mol
    Ther
    2
    (2000)
    168
    [100]
    RR
    Mitry
    CE
    Sarraf
    R
    Havlik
    NA
    Habib
    Hepatology
    31
    (2000)
    885
    [101]
    GW
    Xu
    ZT
    Sun
    K
    Forrester
    XW
    Wang
    J
    Coursen
    CC
    Harris
    Hepatology
    24
    (1996)
    1264
    [102]
    S
    Gao
    J
    Chen
    L
    Dong
    Z
    Ding
    YH
    Yang
    J
    Zhang
    Eur
    J
    Pharm
    Biopharm
    60
    (2005)
    327
    [103]
    TH
    Kim
    IK
    Park
    JW
    Nah
    YJ
    Choi
    CS
    Cho
    Biomaterials
    25
    (2004)
    3783
    [104]
    B
    Lu
    DQ
    Wu
    H
    Zheng
    CY
    Quan
    XZ
    Zhang
    RX
    Zhuo
    Mol
    Biosyst
    6
    (2010)
    2529
    [105]
    TH
    Kim
    SI
    Kim
    T
    Akaike
    CS
    Cho
    J
    Control
    Release
    105
    (2005)
    354
    [106]
    S
    Gao
    J
    Chen
    X
    Xu
    Z
    Ding
    YH
    Yang
    Z
    Hua
    J
    Zhang
    Int
    J
    Pharm
    255
    (2003)
    57
    [107]
    B
    Song
    W
    Zhang
    R
    Peng
    J
    Huang
    T
    Nie
    Y
    Li
    Q
    Jiang
    R
    Gao
    Colloids
    Surf
    B
    Biointerfaces
    70
    (2009)
    181
    [108]
    MM
    Issa
    M
    KopingHoggard
    K
    Tommeraas
    KM
    Varum
    BE
    Christensen
    SP
    Strand
    P
    Artursson
    J
    Control
    Release
    115
    (2006)
    103
    [109]
    M
    Hashimoto
    M
    Morimoto
    H
    Saimoto
    Y
    Shigemasa
    H
    Yanagie
    M
    Eriguchi
    T
    Sato
    Biotechnol
    Lett
    28
    (2006)
    815
    [110]
    HL
    Jiang
    YK
    Kim
    R
    Arote
    D
    Jere
    JS
    Quan
    JH
    Yu
    YJ
    Choi
    JW
    Nah
    MH
    Cho
    CS
    Cho
    Int
    J
    Pharm
    375
    (2009)
    133
    [111]
    TH
    Kim
    H
    Jin
    HW
    Kim
    MH
    Cho
    CS
    Cho
    Mol
    Cancer
    Ther
    5
    (2006)
    1723
    [112]
    M
    Hashimoto
    M
    Morimoto
    H
    Saimoto
    Y
    Shigemasa
    T
    Sato
    Bioconjugate
    Chem
    17
    (2006)
    309
    [113]
    S
    Mansouri
    Y
    Cuie
    F
    Winnik
    Q
    Shi
    P
    Lavigne
    M
    Benderdour
    E
    Beaumont
    JC
    Femandes
    Biomaterials
    27
    (2006)
    2060
    [114]
    P
    Chan
    M
    Kurisawa
    JE
    Chung
    YY
    Yang
    Biomaterials
    28
    (2007)
    540
    [115]
    VB
    Morris
    CP
    Sharma
    Int
    J
    Pharm
    389
    (2010)
    176
    [116]
    I
    Kadiyala
    Y
    Loo
    K
    Roy
    J
    Rice
    KW
    Leong
    Eur
    J
    Pharm
    Sci
    39
    (2010)
    103
    [117]
    HD
    Han
    LS
    Mangala
    JW
    Lee
    MM
    Shahzad
    HS
    Kim
    D
    Shen
    EJ
    Nam
    EM
    Mora
    RL
    Stone
    C
    Lu
    SJ
    Lee
    JW
    Roh
    AM
    Nick
    G
    LopezBerestein
    AK
    Sood
    Clin
    Cancer
    Res
    16
    (2010)
    3910
    [118]
    DA
    Wall
    G
    Wilson
    AL
    Hubbard
    Cell
    21
    (1980)
    79
    [119]
    G
    Ashwell
    J
    Harford
    Annu
    Rev
    Biochem
    51
    (1982)
    531
    [120]
    RJ
    Fallon
    AL
    Schwartz
    J
    Biol
    Chem
    263
    (1988)
    13159
    [121]
    P
    Stahl
    PH
    Schlesinger
    E
    Sigardson
    JS
    Rodman
    YC
    Lee
    Cell
    19
    (1980)
    207
    [122]
    T
    Sato
    J
    Sunamoto
    Prog
    Lipid
    Res
    31
    (1992)
    345
    [123]
    Y
    Lu
    PS
    Low
    J
    Control
    Release
    91
    (2003)
    17
    [124]
    N
    Kamaly
    T
    Kalber
    M
    Thanou
    JD
    Bell
    AD
    Miller
    Bioconjugate
    Chem
    20
    (2009)
    648
    [125]
    S
    Hwa
    Kim
    J
    Hoon
    Jeong
    K
    Chul
    Cho
    S
    Wan
    Kim
    T
    Gwan
    Park
    J
    Control
    Release
    104
    (2005)
    223
    [126]
    B
    Liang
    ML
    He
    ZP
    Xiao
    Y
    Li
    CY
    Chan
    HF
    Kung
    XT
    Shuai
    Y
    Peng
    Biochem
    Biophys
    Res
    Commun
    367
    (2008)
    874
    [127]
    PM
    Friden
    Neurosurgery
    35
    (1994)
    294
    [128]
    G
    Quick
    JV
    Zyl
    A
    Haqtrey
    M
    Ariatti
    Drug
    Deliv
    7
    (2000)
    231
    [129]
    M
    Singh
    Curr
    Pharm
    Des
    5
    (1999)
    443
    [130]
    JH
    Park
    S
    Kwon
    JO
    Nam
    RW
    Park
    H
    Chung
    SB
    Seo
    IS
    Kim
    IC
    Kwon
    SY
    Jeing
    J
    Control
    Release
    95
    (2004)
    579
    [131]
    JH
    Kim
    YS
    Kim
    K
    Park
    E
    Kang
    S
    Lee
    HY
    Nam
    K
    Kim
    JH
    Park
    DY
    Chi
    RW
    Park
    IS
    Kim
    K
    Choi
    I
    Chan
    Kwon
    Biomaterials
    29
    (2008)
    1920
    [132]
    J
    Xie
    Z
    Shen
    KC
    Li
    N
    Danthi
    Int
    J
    Nanomed
    2
    (2007)
    479
    [133]
    AK
    Varkouhi
    RJ
    Verheul
    RM
    Schiffelers
    T
    Lammers
    G
    Storm
    WE
    Hennink
    Bioconjugate
    Chem
    21
    (2010)
    2339
    [134]
    F
    Meng
    WE
    Hennink
    Z
    Zhong
    Biomaterials
    30
    (2009)
    2180
    [135]
    M
    Breunig
    C
    Hozsa
    U
    Lungwitz
    K
    Watanabe
    I
    Umeda
    H
    Kato
    A
    Goepferich
    J
    Control
    Release
    130
    (2008)
    57
    [136]
    B
    Loretz
    M
    Thaler
    A
    BernkopSchnurch
    Bioconjugate
    Chem
    18
    (2007)
    1028
    [137]
    N
    Langoth
    H
    Kahlbacher
    G
    Schoffmann
    I
    Schmerold
    M
    Schuh
    S
    Franz
    P
    Kurka
    A
    BernkopSchnurch
    Pharm
    Res
    23
    (2006)
    573
    [138]
    L
    Yin
    J
    Ding
    C
    He
    L
    Cui
    C
    Tang
    C
    Yin
    Biomaterials
    30
    (2009)
    5691
    [139]
    D
    Lee
    W
    Zhang
    SA
    Shirley
    X
    Kong
    GR
    Hellermann
    RF
    Lockey
    SS
    Mohapatra
    Pharm
    Res
    24
    (2007)
    157
    [140]
    R
    Martien
    B
    Loretz
    M
    Thaler
    S
    Majzoob
    A
    BernkopSchnurch
    J
    Biomed
    Mater
    Res
    A
    82
    (2007)
    1
    [141]
    R
    Martien
    B
    Loretz
    AM
    Sandbichler
    AB
    Schnurch
    Nanotechnology
    19
    (2008)
    045101
    [142]
    H
    Oliveira
    LR
    Pires
    R
    Fernandez
    MC
    Martins
    S
    Simoes
    AP
    Pego
    J
    Biomed
    Mater
    Res
    A
    95
    (2010)
    801
    [143]
    MA
    Kay
    JC
    Glorioso
    L
    Naldini
    Nat
    Med
    7
    (2001)
    33
    [144]
    N
    CarrilloCarrasco
    RJ
    Chandler
    S
    Chandrasekaran
    CP
    Venditti
    Hum
    Gene
    Ther
    21
    (2010)
    1147
    [145]
    M
    Malhotra
    C
    TomaroDuchesneau
    S
    Prakash
    Biomaterials
    34
    (2013)
    1270
    [146]
    H
    Katas
    NNS
    Nik
    Dzulkefli
    SJ
    Sahudin
    Nanomaterials
    2012
    (2012)
    1
    [147]
    Y
    Gao
    Z
    Xu
    S
    Chen
    W
    Gu
    L
    Chen
    Y
    Li
    Int
    J
    Pharm
    359
    (2008)
    241
    [148]
    X
    Zhao
    L
    Yin
    J
    Ding
    C
    Tang
    S
    Gu
    C
    Yin
    Y
    Mao
    J
    Control
    Release
    144
    (2010)
    46
    [149]
    N
    Csaba
    M
    KopingHoggard
    E
    FernandezMegia
    R
    NovoaCarballal
    R
    Riguera
    MJ
    Alonso
    J
    Biomed
    Nanotechnol
    5
    (2009)
    162
    [150]
    AH
    Krauland
    MJ
    Alonso
    Int
    J
    Pharm
    340
    (2007)
    134
    [151]
    MR
    de
    Moura
    FA
    Aouada
    RJ
    AvenaBustillos
    TH
    McHugh
    JM
    Krochta
    LHC
    Mattoso
    J
    Food
    Eng
    92
    (2009)
    448
    [152]
    VM
    Gaspar
    F
    Sousa
    JA
    Queiroz
    IJ
    Correia
    Nanotechnology
    22
    (2011)
    015101
    [153]
    H
    Katas
    HO
    Alpar
    J
    Control
    Release
    115
    (2006)
    216
    [154]
    SL
    Wang
    HH
    Yao
    LL
    Guo
    L
    Dong
    SG
    Li
    YP
    Gu
    Cancer
    Genet
    Cytogenet
    190
    (2009)
    8
    [155]
    A
    Almond
    Cell
    Mol
    Life
    Sci
    64
    (2007)
    1591
    [156]
    EJ
    Menzel
    C
    Farr
    Cancer
    Lett
    131
    (1998)
    3
    [157]
    C
    Picart
    Curr
    Med
    Chem
    15
    (2008)
    685
    [158]
    A
    Dierich
    E
    Le
    Guen
    N
    Messaddeq
    JF
    Stoltz
    P
    Netter
    P
    Schaaf
    JC
    Voegel
    N
    BenkiraneJessel
    Adv
    Mater
    19
    (2007)
    693
    [159]
    LE
    Lerner
    DM
    Schwartz
    DG
    Hwang
    EL
    Howes
    R
    Stern
    Exp
    Eye
    Res
    67
    (1998)
    481
    [160]
    SN
    Zhu
    B
    Nolle
    G
    Duncker
    Br
    J
    Ophthalmol
    81
    (1997)
    80
    [161]
    M
    de
    la
    Fuente
    B
    Seijo
    MJ
    Alonso
    Invest
    Ophthalmol
    Vis
    Sci
    49
    (2008)
    2016
    [162]
    M
    Ravina
    E
    Cubillo
    D
    Olmeda
    R
    NovoaCarballal
    E
    FernandezMegia
    R
    Riguera
    A
    Sanchez
    A
    Cano
    MJ
    Alonso
    Pharm
    Res
    27
    (2010)
    2544
    [163]
    QK
    Lin
    KF
    Ren
    J
    Ji
    Colloids
    Surf
    B
    Biointerfaces
    74
    (2009)
    298
    [164]
    HD
    Lu
    HQ
    Zhao
    K
    Wang
    LL
    Lv
    Int
    J
    Pharm
    420
    (2011)
    358
    [165]
    SF
    Peng
    MJ
    Yang
    CJ
    Su
    HL
    Chen
    PW
    Lee
    MC
    Wei
    HW
    Sung
    Biomaterials
    30
    (2009)
    1797
    [166]
    YH
    Lin
    K
    Sonaje
    KM
    Lin
    JH
    Juang
    FL
    Mi
    HW
    Yang
    HW
    Sung
    J
    Control
    Release
    132
    (2008)
    141
    [167]
    YH
    Lin
    CT
    Chen
    HF
    Liang
    AR
    Kulkarni
    PW
    Lee
    CH
    Chen
    HW
    Sung
    Nanotechnology
    18
    (2007)
    105102
    [168]
    X
    Wang
    T
    Uto
    T
    Akagi
    M
    Akashi
    M
    Baba
    J
    Med
    Virol
    80
    (2008)
    11
    [169]
    T
    Kurosaki
    T
    Kitahara
    S
    Fumoto
    K
    Nishida
    J
    Nakamura
    T
    Niidome
    Y
    Kodama
    H
    Nakagawa
    H
    To
    H
    Sasaki
    Biomaterials
    30
    (2009)
    2846
    [170]
    SF
    Peng
    MT
    Tseng
    YC
    Ho
    MC
    Wei
    ZX
    Liao
    HW
    Sung
    Biomaterials
    32
    (2011)
    239
    C
    Choi
    et
    al

    Journal
    of
    Industrial
    and
    Engineering
    Chemistry
    33
    (2016)
    1–1010

    《香当网》用户分享的内容,不代表《香当网》观点或立场,请自行判断内容的真实性和可靠性!
    该内容是文档的文本内容,更好的格式请下载文档

    下载pdf到电脑,查找使用更方便

    pdf的实际排版效果,会与网站的显示效果略有不同!!

    需要 2 香币 [ 分享pdf获得香币 ]

    下载pdf

    相关文档

    下载需要 2 香币 [香币充值 ]
    亲,您也可以通过 分享原创pdf 来获得香币奖励!

    相关pdf

    相关ppt