Ⅰ型胶原修饰微球对骨髓间充质干细胞粘附增殖及成骨行为的影响  

Effects of microspheres modified with collagen Ⅰ on the adhesion,proliferation and osteogenesis of BMSCs

作　　者：竺鑫晨 刘俊[1] 严佳 胡克 陈刚[1] 刘梅[1] 章非敏[1] ZHU Xinchen;LIU Jun;YAN Jia;HU Ke;CHEN Gang;LIU Mei;ZHANG Feimin(Jiangsu Key Laboratory of Oral Diseases,Department of Prosthodontics,the Affiliated Stomatological Hospital of Nanjing Medical University,Nanjing 210029,China)

机构地区：[1]南京医科大学口腔疾病研究江苏省重点实验室,南京医科大学附属口腔医院修复科,江苏南京210029 [2]南京医科大学生物医学工程与信息学院,临床医学工程校重点实验室,江苏南京211166

出　　处：《口腔医学》2020年第4期299-303,共5页Stomatology

基　　金：国家自然科学基金面上项目(81870807);江苏省高校优势学科建设工程资助项目(2018-87)。

摘　　要：目的研究不同浓度Ⅰ型胶原(COLⅠ)修饰的聚己内酯(Polycaprolactone,PCL)微球的结构性能及细胞学行为的差异,以指导制备出更利于骨组织再生的支架材料。方法以PCL制备微球,作为对照组;并用不同浓度(0.1%、0.5%、2.0%)COLⅠ对其进行表面修饰,作为实验组。通过场发射扫描电镜观察微球表面形貌;干湿质量法检测材料的吸水率;CCK-8法检测大鼠骨髓间充质干细胞(BMSCs)增殖情况;激光共聚焦显微镜评价细胞在材料表面的粘附情况;检测第3、7天细胞的碱性磷酸酶(ALP)活性。结果扫描电镜观察到胶原修饰组胶原交叉排列呈网状覆盖于微球表面;胶原修饰组的吸水率高于对照组(P<0.05);CCK-8结果显示四组微球上的BMSCs细胞在第1、3、5、7天均持续增殖,0.1%COLⅠ胶原修饰组较其他组细胞增殖能力高(P<0.05);激光共聚焦显微镜观察到0.1%COLⅠ和0.5%COLⅠ组细胞在粘附6 h后的铺展形态较另外两组好;ALP活性检测显示0.1%COLⅠ胶原修饰组的ALP活性高于其他组(P<0.05)。结论本实验制备的0.1%COLⅠ修饰的微球能促进BMSCs细胞在体外的粘附增殖及早期成骨分化。Objective To study the differences of structural properties and cytological behaviors of these microspheres modified with different concentrations of type Ⅰ collagen(COLⅠ),to guide the preparation of scaffolds conducive to bone tissue regeneration.Methods PCL microspheres were prepared(as the control group),then modified with COLⅠ of different concentrations(as experimental groups).The surface morphology was observed by field emission scanning electron microscope(FESEM).Water absorption rate of materials was measured by dry-wet mass method.The proliferation of BMSCs was tested by Cell counting Kit-8(CCK-8)assay,while the adhesion of cells was estimated by confocal laser scanning microscopy(CLSM)and Alkaline phosphatase(ALP)activity of cells was measured on the 3rd and 7th days.Results FESEM pictures showed that collagen fibers were crisscross arranged to wrap the surface of microspheres in collagen modified groups.The water absorption rate of collagen modified group was higher than that of control group(P<0.05).CCK-8 results showed that BMSCs in all groups were proliferating continuously within 7 days,and the proliferation rate of the 0.1% COLⅠ group was higher than the other groups(P<0.05).Cells in the 0.1% and 0.5% COLⅠgroups spread better than the other two groups at 6 h by CLSM.ALP activity of BMSCs cells in 0.1% COLⅠ group was higher than that of the other groups(P<0.05).Conclusion The 0.1% COLⅠ modified microspheres prepared in this study can promote adhesion,proliferation and early osteogenic differentiation of BMSCs in vitro.

关 键 词：Ⅰ型胶原 微球 表面修饰 骨髓间充质干细胞 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]