牙周膜细胞与壳聚糖—聚磷酸三钙复合材料生物相容性研究

Biocompatible study on human periodontal ligament cells and chitosan/tricalcium phosphate

目的研究壳聚糖—聚磷酸三钙复合材料对离体培养的人牙周膜细胞粘附及增殖的影响,探讨二者的生物相容性,为骨组织工程中生物材料的选择提供实验依据。方法采用冻干法制备壳聚糖—聚磷酸三钙复合材料,培养人牙周膜细胞,传代扩增后接种到材料表面,体外继续培养,用倒置光学显微镜、扫描电镜观察细胞的粘附和生长情况,用MTT方法检测种植后2、4、68、d细胞的增殖情况。结果种植2 d后细胞呈梭形纤维细胞样,平均每100倍视野下,有生物材料的实验组与无材料的对照组胞数分别为(380±16)个和(80±20)个,二者比较具有差异性显著(P<0.01)。MTT法检测对照和实验组细胞增殖情况,两组细胞均保持持续增殖。且实验组增殖最快,接种后2、4、6、8 d光吸收值与对照组相比,均有差异性均显著(P<0.01)。扫描电镜下可见材料呈多孔网状结构,人牙周膜细胞紧密贴附在材料表面,细胞可沿材料的孔隙活跃生长。结论壳聚糖-聚磷酸三钙复合材料能促进人牙周膜细胞的增殖,人牙周膜细胞与复合材料具有良好的生物相容性,可作为人牙周膜细胞的载体应用于组织工程。

Objective To study the effect of chitosan/tricalcium phosphate（ CTCP）on the attachment and proliferation of human periodontal ligament cells（PDLCs）eultivated and explore their biocornpatibilities. Methods The CTCP was prepared by freeze-dry- ing. PDLCs were cultivated in vitro, collected and seeded onto the surface of CTCP. Cell attachment and proliferation were observed using inverted light microscope and scanning electronic microscopy（SEM）. The cell proliferation was tracked by MTF method 2,4,6,8 d after seeding. Results PDLCs was fihmblast-like two days after seeding.The mean number of cells was （380 ± 16） and （80 ± 20） respectively in the experimental and control group per 100 eyesight field, with significant difference（ P 〈 0.01 ）. MTF showed that the cells kept on proliferating in both groups, in favor of the experimental group. There was significant difference of the absorption value between two groups 2 d,4 d,6 d,8 d after seeding respectively（ P 〈 0.01）.SEM showed the porous structure of the CTCP and the seeded PDLCs were well attached to the CTCP with cytoplasmic extensions and Lamellipodia. Conclusion CTCP could enhance the proliferation of PDLCs in vitro. The PDLCs have biocompatibility with CTCP, and CTCP could be used as vector for PDLCs in tissue engineering.