Osteogenesis enhanced by chitosan (poly-N-acetyl glucosaminoglycan) in vitro

Journal of Periodontology
P R KlokkevoldG W Bernard

Abstract

Chitosan, with a chemical structure similar to hyaluronic acid, has been implicated as a wound healing agent. The purpose of this research was to evaluate the effect of chitosan on osteoblast differentiation and bone formation in vitro. Mesenchymal stem cells were harvested from fetal Swiss Webster mice calvariae prior to osteoblast differentiation and calcification (12 to 13 days in utero). Stem cells were seeded into 6-well culture plates at a density of 350,000 cells per well. Using this model, it was possible to quantify the influence of chitosan on osteoprogenitor differentiation and osteogenesis. Experimental wells were pretreated with 200 microliters chitosan (2 mg/ml in 0.2% acetic acid vehicle). Control wells were pretreated with 200 microliters vehicle (0.2% acetic acid) or remained untreated. Cells were allowed to grow under optimal conditions for 14 days. Cell cultures were fixed with glutaraldehyde and stained with Von Kossa stain to identify bone forming colonies. Positive staining colonies were identified and counted under light microscopy. Histologic cross-sections of representative positively stained colonies identified osteoblasts and confirmed bone formation. Examination of control wells revealed 3.6 +/- 0.6 ...Continue Reading

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