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. 2013 Jan 15;18(1):1015–1035. doi: 10.3390/molecules18011015

Table 1.

Insoluble chitosan microparticles: Published examples of in vivo biomedical use, and in vitro investigation of cell responses.

Method of insoluble chitosan microparticle preparation Chitosans analyzed Application
Neutral precipitation of acid-soluble chitosan in vivo or in vitro
Mix acid-soluble chitosan solution or chitosan-glycerol phosphate solution pH 4–6.8 with whole blood and/or directly apply to bleeding tissues 80% DDA

75%–82% DDA, 150–250 kDa
Hemostatic in vivo [17]

Bone and articular cartilage repair in vivo [20]
Inject mixtures of chitosan-DNA, with a molar excess of chitosan, in intramuscular, subcutaneous sites 92% DDA, 10 kDa
80% DDA, 10 kDa 80% DDA, 80 kDa
Gene delivery in vivo [24]
Combine soluble chitosan at 5- to 10-fold molar excess with DNA, pipette into cell culture medium DMEM+10% serum pH 7.6 80% DDA, 15 kDa
92% DDA, >100 kDa
94% DDA, 52 kDa
In vitro DNA delivery: A549, Hela, B16 cells, HEK293 cells [25,26]
Pipette acid-soluble chitosan
pH 5.0 into basal media pH 7.6 (DMEM, αMEM, RPMI±10% to 16% fetal bovine serum)
80% DDA, 179 kDa

81% DDA, 35 kDa
80% DDA, 179 kDa
95% DDA, 168 kDa
92% DDA, 10 kDa
In vitro bone marrow stromal cell osteogenesis [27]
In vitro macrophage activation [28]
In vitro neutrophil chemotaxis, degranulation, chitosan uptake [14]
In vitro chitosan-HEK293 cell adsorption/uptake [26]
Pre-formed chitosan microparticles
Pre-formed microparticles (glutaraldehyde and Tween surfactant) injected into the mouse foot pad or added to DMEM+heat-inactivated 10% fetal bovine serum 75%–85% DDA, 164 kDa Vaccine, lymph node trafficking in vivo [29]
In vitro HEK293, A549, RAW264 cell uptake [29]
1 µm or 3.5 µm pre-formed chitosan microparticles added to RPMI+10% serum, pH 7.2 ≥80% DDA Wound-repair applications: in vitro neutrophil chemotaxis [30] in vitro macrophage activation [31]