载EPO仿生Gel/HA纳米纤维促骨再生研究  

Effects of EPO loaded biomimetic Gel/HA nanofibers on bone regeneration

作　　者：李道伟 史册 张雪[1,3,2] 胡月 赵欢[1,3,2] 倪世磊 孙宏晨[1,3,2] 

机构地区：[1]吉林省口腔科学与技术纳米工程重点实验室,吉林长春130021 [2]吉林省牙发育及颌骨重塑与再生重点实验室,吉林长春130021 [3]吉林大学口腔医院病理科,吉林长春130021

出　　处：《口腔生物医学》2017年第3期117-121,共5页Oral Biomedicine

基　　金：国家自然科学基金项目(81320108011;81271111;81600823;81600843);国家重点研发项目(2016YFC1102804);吉林省科技计划项目(20170520009JH;20170520016JH);中国博士后科学基金(2017M611332);吉林省省级产业创新专项资金项目(2016c044-3)

摘　　要：目的:通过组织工程策略构建载促红细胞生成素(EPO)的纳米纤维,并研究其促骨形成作用。方法:制备仿生明胶/羟基磷灰石(Gel/HA)纳米纤维支架,通过冻干的方法将EPO直接与Gel/HA纤维支架复合;采用ELISA方法检测EPO的缓释效果;应用实时定量RT-PCR检测纳米纤维对大鼠骨髓间充质干细胞(BMSCs)成骨相关基因表达的影响;体内建立大鼠颅骨极量骨缺损模型,通过锥形束CT和组织学评价其骨缺损修复效果。结果:扫描电镜显示交联前后纳米纤维呈现交织的网状结构;EPO释放呈现明显的突释现象,仅有少量EPO存留于支架内缓慢释放;定量RT-PCR结果显示EPO能够促进大鼠BMSCs中Col1和Osterix的表达,而对Runx2和Alp的表达无明显影响;体内结果显示在6周时和12周时Gel/HA/EPO组有大量的新骨形成,明显高于空白对照组和Gel/HA组,Gel/HA组的新形成骨量也显著高于空白对照组。结论:载EPO仿生Gelatin/HA的纤维支架材料能够促进大鼠BMSCs的成骨分化,能够很好的修复骨缺损。Objective： Construction of erythropoietin（ EPO） contained nanofibers by tissue engineering strategies and subsequent study of its pro-osteogenic effects. Methods： Biomimetic gelatin/hydroxyapatite（ Gel/HA） nanofibrous scaffold was fabricated by electrostatic spinning method. Erythropoietin（ EPO） loaded Gel/HA scaffold was prepared by freeze-dried method after crosslinking of Gel/HA scaffold. The release profile of EPO was detected by ELISA. The osteogenic effects of Gel/HA/EPO were detected by q-PCR.A 5mm diameter calvarial bone defect model was created in rat and scaffolds were implanted. Cone-Beam CT and histological technique were used to evaluate the bone repair effects. Results： SEM results showed that all the scaffolds were reticular and intertwined structure. There was some deposition of EPO protein on the surface of Gel/HA/EPO scaffold. The release profile of EPO showed an obvious burst release phenomenon,and only a small amount of EPO remnants kept releasing slowly. q-PCR results showed that EPO can promote osteoblast differentiation by promoting the expression of Col1 and Osterix. In vivo study showed that the Gel/HA/EPO group had a large amount of new bone formation both in 6-week and 12-week post implantation,which obviously was higher than those of the blank control group and Gel/HA group; Besides,the new bone formation in the Gel/HA group was also significantly higher than that of the blank control group. Conclusions： EPO loaded biomimetic Gelatin/HA nanofibers is an excellent scaffold material for bone tissue engineering.

关 键 词：促红细胞生成素 明胶/羟基磷灰石 仿生纳米纤维支架 骨组织工程 

分 类 号：R782.2[医药卫生—口腔医学]