壳聚糖-Ⅰ型胶原复合膜的制备及其生物相容性实验研究  

Physico-chemical properties of chitosan-collagen typeⅠcomposite membrane and its biocompatibility with MG63 osteoblast-like cells

作　　者：周敏[1] 徐淑兰[1] 钟星华[1] 杨烁[1] 姚中雄[1] 

机构地区：[1]南方医科大学附属口腔医院·广东省口腔医院,广东广州510280

出　　处：《广东牙病防治》2012年第4期193-198,共6页Journal of Dental Prevention and Treatment

基　　金：海珠区科技计划项目(2010-Y-03)

摘　　要：目的了解制备的壳聚糖-Ⅰ型胶原复合膜的理化性能及其与MG63成骨样细胞的生物相容性,以初步探讨其作为引导骨再生屏障膜的可行性。方法采用提纯的牛肌腱Ⅰ型胶原和壳聚糖,经冷冻干燥,使用热交联、化学交联技术制成壳聚糖-Ⅰ型胶原复合膜(实验组)和Ⅰ型胶原膜(对照组)。采用氨基酸含量检测、差示量热扫描法分析牛肌腱Ⅰ型胶原性质和表征,红外光谱、扫描电镜分析2种膜的理化性能。将MG63成骨样细胞接种于2种膜上,MTS法检测细胞1 d、3 d、5 d、7 d的增殖情况。结果牛肌腱Ⅰ型胶原的胶原3股螺旋结构保持较完整,其热变性温度为99.04℃,提热吸收峰为53.54℃,与标准品指标接近,其成分比例及性质表征符合Ⅰ型胶原特征。红外光谱结果表明实验组具备胶原和壳聚糖特征峰表现,并且2种材料之间形成大量氢键,分子间结合良好。扫描电镜显示实验组和对照组膜表面和横切面为多孔结构,实验组膜孔径较大,为10～100μm。MG63成骨样细胞在2组膜上呈明显的增殖趋势,具有较好的细胞相容性。结论制备的壳聚糖-Ⅰ型胶原复合膜具有优良的结构及生物学性能,可应用于下一步引导骨再生膜材料的研究。Objective To prepare Chitosan-collagen typeⅠcomposite membrane and investigate its physico-chemical properties and biocompatibility with MG63 osteoblast-like cells,in order to evaluate the feasibility of Chitosan-collagen typeⅠcomposite membrane as the barrier membrane of guided bone regeneration.Methods To extract collagen from bovine tendons and to prepare Chitosan-collagen typeⅠcomposite membrane,through lyophilization,thermal cross-linking and chemical crosslinking with EDC-NHS.Chitosan-collagen typeⅠcomposite membrane was used as experimental group and collagen typeⅠmembrane as control group.Properties of collagen typeⅠfrom bovine tendons was examined through amino acid content analysis and DSC.To analyse the physical and chemical properties of group and control group with Fourier transform infrared spectroscopy（FTIR） and scanning electron microscopy（SEM）.The biocompatibility with MG63 osteoblast-like cells was checked by MTS on day 1,3,5,7.Results The amino acid composition analysis and DSC results showed that the three helical structure of collagen typeⅠfrom bovine tendons had been retained completely.Its component,proportion and the properties accord with the characteristic of collagen typeⅠ.FTIR results showed that the experimental group had characteristic peak of the collagen and chitosan;hydrogen bond formed between this two materials;indicating the binding force between molecules was strong.The SEM results of experimental group and control group showed that surface and cross section of both two membranes had porous structure;the pore diameter of experimental group was bigger,for 10-100 μm.The MTS results showed that the optic density（OD） value of experimental groupand control group both increased with times,signifying the growth trend was obvious.Conclusion Chitosan-collagen type Ⅰ composite membrane exhibits good biocompatibility with MG63 osteoblast-like cells and has favorable structure and biological performance,showing wide applications prospects as GBR membrane.

关 键 词：壳聚糖 Ⅰ型胶原 复合膜 MG63成骨样细胞 生物相容性 

分 类 号：R783.1[医药卫生—口腔医学]