珊瑚转化羟基磷灰石应用于牙周组织工程的细胞相容性和细胞毒性

背景:经处理后的珊瑚转化羟基磷灰石具有良好的生物相容性和可降解性,其三维相通的孔隙结构不仅可为种子细胞的黏附、增殖提供足够的内部空间和表面积,而且有利于营养成分的渗透和血管化形成。目的:对珊瑚转化羟基磷灰石进行细胞相容性和细胞毒性的研究,以初步探讨其应用于牙周组织工程的可行性。方法:将体外培养的人牙周膜细胞接种到珊瑚转化羟基磷灰石支架上体外三维复合培养,通过细胞计数方法和扫描电镜观察人牙周膜细胞在珊瑚转化羟基磷灰石支架上的附着、生长情况,并设立阴性对照组和阳性对照组,采用MTT测试法和碱性磷酸酶活性检测法评估稀释浓度分别为100%,50%,10%,1%的珊瑚转化羟基磷灰石浸提液对人牙周膜细胞增殖及功能表达的影响。结果与结论:珊瑚转化羟基磷灰石具有良好的多孔网状结构,人牙周膜细胞在珊瑚转化羟基磷灰石支架上生长旺盛,伸展充分,而不同浓度的材料浸提液对人牙周膜细胞的生长、增殖及碱性磷酸酶活性的影响差异均无显著性意义。提示珊瑚转化羟基磷灰石具有良好的三维空间结构和细胞相容性,且无细胞毒性,有望用作牙周组织工程的支架材料。

Cytocompatibility and cytotoxicity of coral hydroxyapatite in the periodontal tissue engineering

BACKGROUND: With excellent biocompatibility and degradability, porous coral hydroxyapatite can provide the enough three-dimensional space for the adherence and proliferation of seed cells, which is beneficial for the vascularization and the infiltration of nutrients. OBJECTIVE: To investigate the cytocompatibility and cytotoxicity of coral hydroxyapatite so as to evaluate the feasibility of coral hydroxyapatite as a scaffold of periodontal tissue engineering. METHODS: Human periodontal ligament cells (HPDLCs) cultured in vitro were collected and seeded on the three-dimensional framework of coral hydroxyapatite. The HPDLCs growth on coral hydroxyapatite scaffolds was observed by cell counting and scanning electronic microscope. Compared with the negative and positive controls, the influences of coral hydroxyapatite extracts at dilution concentrations of 100%, 50%, 10%, 1% on cell proliferation and differentiation were observed by 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay and alkaline phosphatase activity assay. RESULTS AND CONCLUSION: The cell counting results showed that human periodontal ligament cells adhered and proliferated on the three-dimensional framework of coral hydroxyapatite, and the flourishing growth of cells in the porous coral hydroxyapatite scaffolds was observed by scanning electronic microscope. The cytotoxicity detection showed that the different concentrations of coral hydroxyapatite extracts had no influences on the cell proliferation and differentiation. Then the porous coral hydroxyapatite was proved to be feasible as the periodontal tissue engineering scaffold with satisfactory cytocompatibility and with no cytotoxicity.