| 摘要: |
| 目的 制备一种可长时间缓释骨形态发生蛋2（BMP-2）的聚己内酯（PCL）复合支架，并通过检测其对人源骨髓间充质干细胞（BMSC）成骨分化的影响探讨其在骨组织工程中的应用。方法 将磷脂（PL）和BMP-2混合形成的BMP-2/PL混合物（B/P）分散在二氯甲烷中，与PCL混合后，采用相分离法制备负载BMP-2的三维PCL-B/P复合支架和PCL-B传统支架，通过酶联免疫吸附试验（ELISA）检测两种支架的BMP-2缓释效果。将BMSC种植在PCL-B传统支架和PCL-B/P复合支架中，分别采用CCK-8法和实时定量PCR（qPCR）检测两种支架上BMSC的增殖和成骨分化能力。结果 与PCL-B传统支架对比，PCL-B/P复合支架对BMP-2缓释效果更佳，缓释时间更长，可达22 d。在BMSC培养的第7、14和21天，PCL-B/P复合支架上BMSC的增殖能力均优于PCL-B传统支架（P<0.05），且PCL-B/P复合支架上BMSC中碱性磷酸酶和Ⅰ型胶原蛋白、骨钙、骨桥蛋白3种成骨基因mRNA的表达均高于PCL-B传统支架（P<0.05，P<0.01）。结论 成功地制备出一种可长时间缓释BMP-2的高分子三维PCL-B/P复合支架，其较PCL-B传统支架能更好地诱导BMSC的增殖和成骨分化。 |
| 关键词: 骨和骨组织 组织工程 骨形态发生蛋白2 聚己内酯 复合支架 人骨髓 间质干细胞 成骨分化 |
| DOI：10.16781/j.0258-879x.2017.03.0289 |
| 投稿时间：2016-08-29修订日期：2017-01-07 |
| 基金项目:国家自然科学基金（81272026，8167090274）. |
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| Preparation and application of polycaprolactone composite scaffold with long-term slow-release of bone morphogenetic protein 2 |
| LU Chun-wenΔ,WANG ChaoΔ,MAO Ning-fang,DING Mu-chen,YUAN Jia-bin,SHI Zhi-cai* |
(Department of Orthopaedics, Changhai Hospital, Second Military Medical University, Shanghai 200433, China
△Co-first authors.
* Corresponding author) |
| Abstract: |
| Objective To prepare a polycaprolactone (PCL) composite scaffold which can slowly release bone morphogenetic protein 2 (BMP-2) for a long time, and to explore the application value of PCL composite scaffold in bone tissue engineering through studying osteogenic differentiation level of human bone mesenchymal stem cells (BMSCs). Methods The BMP-2/PL compounds (B/P), formed by the BMP-2 and soybean phospholipid (PL), were dispersed in methylene dichloride, and then we mixed it with PCL and prepared the PCL-B/P composite scaffold and PCL-B traditional scaffold loaded with BMP-2 by phase separation method. The slow-release of BMP-2 from two kinds of scaffolds was observed with ELISA assay. The human BMSCs were cultured in the media containing PCL-B/P composite scaffold or PCL-B traditional scaffold, and the proliferation abilities and osteogenic differentiation levels of BMSCs on two kinds of scaffolds were detected and analyzed by CCK-8 assay and qPCR, respectively. Results Compared with the PCL-B traditional scaffold, PCL-B/P composite scaffold in this study showed a better slow-release effect of BMP-2 and a longer slow-release time of 22 days. The proliferation abilities of BMSCs on PCL-B/P composite scaffold were better than that on PCL-B traditional scaffold on the 7th day, 14th day and 21st day of cell culture (P<0.05). The alkaline phosphatase content and mRNA expressions of collagen type Ⅰ, osteocalcin and osteopontin of BMSCs in the PCL-B/P composite scaffolds were significantly higher than those in the PCL-B traditional scaffold (P<0.05, P<0.01). Conclusion We have successfully prepared a PCL-B/P composite scaffold loaded with BMP-2. The PCL-B/P composite scaffold can slowly release BMP-2 for a long time and induce the proliferation and osteogenic differentiation of human BMSCs. |
| Key words: bone and bones tissue engineering bone morphogenetic protein 2 polycaprolactone composite scaffold bone marrow mesenchymal stem cells osteogenic differentiation |
