[Inhibitive effects of chitosan on proliferation of fibroblasts in vitro].
OBJECTIVE
To investigate the effects of chitosan on the cell cycle of the human fibroblasts and on the Ki-67 antigen expression in vitro and to investigate the mechanism of chitosan preventing the postoperative tissue adhesion.


METHODS
The cultured fibroblasts were treated for 48 hours with 0, 0.01, 0.1, 1.0, 10.0 mg/ml of chitosan, respectively; then, the cell cycle of the fibroblasts was measured by the flow cytometry. The cultured fibroblasts were treated for 24 hours with the chitiosan at the above concentrations; then, the Ki-67 antigen in the cell nucleus was detected with the immunohistochemical staining to observe its expression.


RESULTS
The growth of the fibroblasts was obviously suppressed by chitosan, especially in the cell morphology. When the concentrations of chitosan were 1.0 mg/ ml and 10.0 mg/ml, the percentages of the fibroblasts in the proliferation stage were 32.3%+/- 5.2% and 14.7% +/- 2.9%, respectively, which were significantly smaller than the percentage of the fibroblasts when the concentration of chitosan was 0 mg/ml (the control group) (41.9% +/-5.8%, P<005). When the concentrations were 0. 01 mg/ml and 0. 1 mg/ml, the percentages of the fibroblasts in the proliferation stage were 39.0%+/-6.0% and 35.5%+/- 3.4%, respectively, which were smaller than that of the control, but not significantly different from that of the control (P> 0.05). When the concentrations of chitosan were 0.1 mg/ml, 1.0 mg/ml and 10.0 mg/ml, the percentages of the fibroblasts that had the positive Ki-67 antigen were 37.3% +/- 3.4%, 30.5% +/- 6.2% and 17.8% +/- 3.0%, respectively, which were significantly smaller than that of the control (57.6% +/-8.9%, P<0.05). When the concentration was 0. 01 mg/ml, the percentage of the fibroblasts that had the positive Ki-67 antigen was 54.1% +/- 8.0%, which was smaller than that of the control, but not significantly different from that of the control (P>0.05).


CONCLUSIONS
Chitosan can inhibit the proliferation of the fibroblasts and increase the percentage of the fibroblasts in the quiescent stage, which can be considered as one of the mechanisms that chitosan can prevent the postoperative tissue adhesion.
Xia
Pingguang
P
Department of Orthopedic, Changzheng Hospital, Second Military Medical University, Shanghai, China. xiapg@sina.com
Hou
Chunlin
C
Wang
Wanhong
W
D001672:Biocompatible Materials; D002453:Cell Cycle / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D048271:Chitosan / Q000008:administration & dosage / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D019394:Ki-67 Antigen / Q000096:biosynthesis; D008297:Male; D000267:Tissue Adhesions / Q000517:prevention & control*
D001672:Biocompatible Materials; D019394:Ki-67 Antigen; D048271:Chitosan