키토산이 배양중인 치주인대세포와 치은섬유아세포에 미치는 영향
Effects of Chitosan on Human Periodontal Ligament Cells and Gingival Fibroblasts in Vitro
구강과학- 전남대학교 2001년 13권 1호 p.16 ~ 34
김옥수, 김지영,
소속 상세정보
김옥수 ( ) - 전남대학교 치과대학 치주과학교실
김지영 ( ) - 전남대학교 치과대학 치주과학교실
KMID : 0988420010130010016
Abstract
The aim of this study was to evaluate the effects of chitosan coating on the
attachment, proliferation, functional and morphological change of periodontal ligament cells and gingival fibroblasts.
Primary culture of human periodontal ligament cells and gingival fibroblasts were grown in Dulbecco´s modified Eagle´s medium with 10% fetal bovine serum and 1% antibiotics. In experimental group, cells were inoculated in the multiwell plates coated with chitosan in concentration of 0.02, 0.2 and 2㎎/㎖. Cell counting and MTT assay were done after 0.5, 1.5, 3, 6 and 24 hours of incubation to evaluate the cell attachment, and then after 2 and 7 days of culture th evalute the cell proliferation.
The alkaline phosphatase activity was measured after 4 and 7 days of culture and the ability to produce mineralized nodules was evaluated after 21 days of culture.
The results were as follows:
1. The morphology of both cells on the chitosan coating was round or spheric. Round cells were aggregated arger 6 hours of culture. Aggregated cells on the chitosan coated surface showed nodule like appearance after 24 hours of culture and not achieved confluency at 7 days.
2. During early period of culture, the attachment of periodontal ligament cells and gingical fibroblasts were inhibited by chitosan coating Inhibition of cell attachment tended to increase with the concentration of chitosan.
3. At the chitosan concentration of 0.02 and 0.2㎎/㎖, periodontal ligament cells were more rapidly proliferated at 7 days, compared to the control group. At the concentration of 2㎎/㎖, the proliferation of gingival fibroblasts and periodontal ligament cells was inhibited(p<0.01).
4. Alkaline phosphatase activity of periodontal ligament cells was increased in
chitosan coated group, especially at the concentration of 0.02㎎/㎖ after 4 days of culture. Alkaline phosphatase activity of gingival fibroblasts was inhibited by chitosan coating and decreased in course of time.
5. Periodontal ligament cells produced mineralized nodules on chitosancoated wells without the addition of mineralized nodule forming materials (ascorbic acid, β glycerophosphate, dexamethasone). With the addition of mineralized nodule, forming materials, periodontal ligament cells produced more mineralized nodules at the concentration of 0.02㎎/㎖, compared to the control. Gingival fibroblasts also produced the mineralized nodule at the concentration of 2㎎/㎖ of chitosan.
In summary, the attachment, proliferation, cell activity, and alkaline phosphatase activity of periodontal ligament cells depended on the concentration of coated chitosan and those of gingival fibroblasts were inhibited by chitosan. Chitosan stimulated mineralized nodule formation by periodontal ligament cells. At the appropriate concentration (0.02㎎/㎖), chitosam could increase alkaline phosphatase activity, stimulate the formation of mineralized nodule by periodontal ligament cells have antifibroblastic activity on gingival fibroblasts. These results suggest that chitosan can be used as an adjunct of regeneration, bone graft and the matrix of tissue engineering for periodontal regeneration, especially bone regeneration.
키워드
원문 및 링크아웃 정보
등재저널 정보