Ⅰ型胶原/聚己内酯/凹凸棒石复合支架材料体外诱导成骨的研究  

The Study of Osteogenic Induction of TypeⅠCollagen/Poly( caprolactone)/Attapulgite Composite Scaffold Materials in Vitro

作　　者：张晓敏[1,2] 王世勇[2] 李根[2] 赵红斌[1,2] 

机构地区：[1]甘肃农业大学生命科学技术学院,兰州730070 [2]兰州军区兰州总医院骨科研究所,兰州730050

出　　处：《中国生物工程杂志》2016年第5期27-33,共7页China Biotechnology

基　　金：甘肃省重大科技专项资助项目(1203FKDA036)

摘　　要：目的:研究Ⅰ型胶原(ColⅠ)/聚己内酯(PCL)/凹凸棒石(ATP)复合支架材料的生物相容性及体外骨诱导性。方法:采用溶液浇铸-粒子滤沥法制备三种不同ATP含量(0%wt、10%wt、30%wt)的ColⅠ/PCL/ATP复合支架材料;将D1细胞与三种支架材料共培养,扫描电镜、鬼笔环肽和H&E染色、CCK-8法评价支架材料的生物相容性;D1细胞复合三种支架材料培养7天、14天、21天后RT-q PCR检测其成骨相关基因(Runx-2、Osterix、ALP、Col I、OPN、OC)的相对表达量,分别评价比较三种支架材料的成骨诱导效应。结果:SEM、鬼笔环肽和H&E染色显示D1细胞在三种支架材料表面均呈现良好的黏附;CCK-8结果显示,细胞在ATP含量30%wt的支架材料上增殖率显著高于其他两组,RT-q PCR检测结果显示,与0%wt、10%wt ATP相比,30%wt ATP组的Runx-2相对表达量在7天时显著升高,14天、21天降低;ALP相对表达量在14天时显著升高,21天时显著降低;Osterix、Col I、OPN、OC的相对表达量随时间和ATP剂量的增加显著上调(P<0.05)。结论:ColⅠ/PCL/ATP复合支架材料具有良好的生物相容性及骨诱导性,有望成为一种理想的骨组织工程支架材料。Objective： To study the biocompatibility and osteoinductivity of type Ⅰ collagen / poly（ caprolactone） /attapulgite composite scaffolds in vitro. Methods： The Col Ⅰ/PCL/ATP composite scaffolds materials of three different content（ 0%,10%,and 30% wt） ATP were fabricated by the solution perfusion-salt-leaching process. D1 cells were co-cultured with the materials for 7d or 1 - 6d,the biocompatibility of scaffold was evaluated by scanning electron microscopy,Phalloidin and HE staining and CCK-8,respectively.D1 cells were co-cultured with the materials for 7d,14 d,and 21d; the relative expression of bone-related genes including Runx-2,Osterix,ALP,Col I,OPN,and OC were measured by RT-q PCR,and evaluated osteogenic effect comparison of three scaffolds. Results： Three scaffolds had suitable cell adhesion and appreciation properties,especially30% wt ATP materials. The results of RT-q PCR show that Runx-2 expression of 30% wt ATP was significant increased in7 d,but decreased in 14 d,21d; and ALP expression was significant increased in 14 d,but significant decreased in 21d;and there were significant increased in the relative mRNA levels of Osterix,Col I,OPN,OC,compared with the 0% and10% wt ATP materials（ P〈0. 05）,which had time and dose dependent manner. Conclusion： ColⅠ / PCL / ATP composite scaffolds may be used as an ideal scaffold for bone tissue engineering.

关 键 词：凹凸棒石 复合支架材料 生物相容性 骨诱导性 

分 类 号：R318.08[医药卫生—生物医学工程]