鹿茸多肽纳米复合材料诱导兔骨髓间质细胞向成骨细胞的分化效应

J4 ›› 2011, Vol. 37 ›› Issue (6): 1019-1023.

鹿茸多肽纳米复合材料诱导兔骨髓间质细胞向成骨细胞的分化效应

张郑瑶^1,2,刘晓峰^1,2,3,王珊¹,周秋丽^1,2,田秀丽¹,张瑾¹,史祺云¹,胡进平¹,邓旭明¹,宋宇^1|2

1.吉林大学畜牧兽医学院生物技术系|吉林长春 130062；2.吉林大学药学院微生物与生化药学教研室|吉林长春 130021；3.吉林省肿瘤医院检验科|吉林长春 130012

收稿日期:2011-06-29 出版日期:2011-11-28 发布日期:2011-11-28
通讯作者: 张郑瑶（1984-），女，辽宁省大连市人，在读医学博士，主要从事生物化学与分子生物学研究。 E-mail:applesy@163.com
作者简介:张郑瑶（1984-）|女|辽宁省大连市人|在读医学博士|主要从事生物化学与分子生物学研究。
基金资助:
国家高技术研究发展计划（863计划）项目资助课题（2006AA03Z328）；吉林省长春市科技支撑计划项目资助课题［长科技合（2008217）号］

Differentiation of rabbit mesenchymal stem cells into osteoblasts induced by velvet antler polypeptide/nano TCP/ gelatin material

ZHANG Zheng-yao^1,2|LIU Xiao-feng^1,2,3|WANG Shan¹,ZHOU Qiu-li^1,2,TIAN Xiu-li¹,ZHANG Jin¹,SHI Qi-yun¹,HU Jin-ping¹,DENG Xu-ming¹, SONG Yu^1|2

1. Department of Biotechnology,College of Animal Science and Veterinary Medicine,Jilin University, Changchun 130062,China;2. Department of Microbiology and Biochemical Pharmacy,School of Pharmacy,Jilin University,Changchun 130021,China|3.Department of Laboratory, Tumor Hospital of Jilin Province Changchun 130012,China

Received:2011-06-29 Online:2011-11-28 Published:2011-11-28

摘要/Abstract

摘要：

目的：探讨鹿茸多肽纳米复合材料诱导兔骨髓间质细胞（MSCs）向成骨细胞分化的效应，阐明复合材料作用机制。方法：将含5 mg/g鹿茸多肽、20%纳米β-磷酸三钙（β-TCP）和明胶的复合材料（以下简称复合材料）与第3代MSCs共培养 48 h，用MTT法和流式细胞术检测复合材料对MSCs增殖及细胞周期的影响；AO/EB荧光染色观察MSCs凋亡形态的变化；将MSCs与复合材料共培养，扫描电镜下观察细胞黏附、生长情况；复合材料连续作用于MSCs 21 d，全自动生化分析仪检测细胞中碱性磷酸酶（ALP）活性，茜素红染色法观察复合材料对MSCs矿化结节形成的作用。结果：复合材料作用于MSCs 48 h，可明显促进MSCs增殖，相对增殖率为126.45%,此时的MSCs主要处于S期，占细胞总数45.84%（对照组为7.96%），凋亡率为2.08%（对照组为13.68%）；MSCs在复合材料上黏附、生长良好，呈多角或梭形。复合材料连续作用MSCs 21 d，显著增加了矿化结节形成；随着作用时间的延长，细胞的ALP活性也逐渐增强，21 d 复合材料组与对照组比较明显升高（P<0.001）。结论：复合材料在体外可促进MSCs增殖，并可诱导MSCs向成骨细胞分化，具有良好的成骨效能。

关键词: 纳米结构；骨髓间质细胞；细胞分化

Abstract:

Abstract:Objective To explore the effects of velvet antler polypeptide/ nano β-tricalcium phosphate（β-TCP）/ gelatin material on the osteogenetic differentiation in rabbit mesenchymal stem cells (MSCs),and clarify the mechanism.Methods Cell cycle and proliferation were examined by flow cytometry and MTT assay in the third generation MSCs treated with 5 mg/g velvet antler polypeptide/20 % nano TCP/gelatin material for 48 h.The growth of MSCs cocultured with the material was observed by scanning electron microscope.The morphology of apoptotic MSCs was observed by AO∕EB fluorescence staining assay.The activity of alkaline phosphatase (ALP) was detected with automatic biochemistry analyzer,and the mineralized nodules in MSCs were counted after 21 d treatment.Results The material could significantly promote the proliferation of MSCs,the relative growth rate was 126.45 % after treated with the material for 48 h.The percentage of MSCs in the S phase was 45.84 %（7.96% in control）,and the apoptotic rate was 2.08 %（13.68% in control group）.The calcified nodules and the activity of ALP were increased after treatment with material for 21 d. MSCs could adhere to the surface of the material and go well with it.Conclusion The material has the effects on stimulating the proliferation,differentiation and mineralization of rabbit MSCs.

Key words: nanostructures;mesenchymal stem cells;cell differentiation

中图分类号:

R318.08

张郑瑶|刘晓峰|王珊|周秋丽|田秀丽|张瑾|史祺云|胡进平|邓旭明|宋宇. 鹿茸多肽纳米复合材料诱导兔骨髓间质细胞向成骨细胞的分化效应[J]. J4, 2011, 37(6): 1019-1023.