同种异体骨共价枝接精氨酸-甘氨酸-天冬氨酸多肽对成骨细胞黏附和增殖的影响

目的观察同种异体骨共价枝接精氨酸-甘氨酸-天冬氨酸（Arg-Gly-Asp，RGD）多肽对大鼠成骨细胞黏附和增殖的影响。方法以RGD多肽在同种异体骨片表面进行共价枝接反应，用X线光电子能谱分析仪（XPS）进行表面分析，以大鼠成骨细胞在骨片表面进行组织培养，MTY法及细胞计数法评价成骨细胞的黏附效力及增殖活性，通过相差显微镜、扫描电镜观察细胞形态及增殖情况。结果以碳二亚胺（EDC）为交联剂能够将RGD多肽有效固定于同种异体骨片表面，成骨细胞在同种异体骨片表面培养4h、加h后，实验组细胞黏附效力（0．322±0．014、0．447±0．030）均明显强于对照组（0．240±0．012、0．337±0．048），培养4d、6d、8d后，实验组成骨细胞增殖情况（0．510±0．054、0．612±0．075、0．629±0．098）亦强于对照组（0．405±0．054、0．340±0．030、0．339±0．018）。结论RGD多肽共价修饰同种异体骨能够有效促进成骨细胞的黏附和增殖。

Covalently modification with Arg-Gly-Asp polypeptedes containing peptides of allogenic bone for osteoblastic adhesion and proliferation

Objective To observe the effect of RGD polypeptides covalently grafted on the allogenie bone for the adhesion and proliferation of the osteoblast. Methods The RGD-containing polypeptides were immobilized on the surface of allogenic bone covalently, followed by the surface analysis though XPS. The murine osteoblasts were seeded on the RGD - modified allogenic and the blank bone. The ostoeblastic adhesive efficiency and proliferative activity were estimated by MTT and cytometry after culture. The cellular morphous and proliferative condition was observed under the phase contrast microscope and scanning electron microscope. Results Using the EDC as cross-linking agent, the RGD polypeptides were immobilized on the allogenic surface effectively. After 4 hours and 20 hours of culture, the efficiencies of cell adhesion on the modified bone （0. 322±0. 014 and 0. 447±0. 030） were higher than the blank （0. 240±0. 012, 0. 337±0. 048） ; also, cellular proliferation on the RGD - modified bone （0. 510±0. 054, 0. 612±0. 075, 0. 629±0. 098） was better than in unmodified group （0. 405±0. 054, 0. 340±0. 030, 0. 339±0. 018）. Conclusion The modification with RGD - containing peptides on allogenie bone can promote the adhesion and proliferation of osteoblasts.