摘要
目的研究具有纳米结构二嵌段共聚物左旋聚乳酸-聚己内酯（PLLA-b-PCL）与犬骨髓基质干细胞（BMSCs）的体外生物相容性，探讨其作为软骨组织工程支架的可行性。方法开环聚合制备PLLA-b—PCL，液-液相分离制备PLLA-b-PCL纳米支架，扫描电镜观察材料结构。分离培养犬BMSCs，取第3代BMSCs接种于PLLA-b—PCL膜进行复合二维培养，MTT法检测细胞毒性；通过倒置显微镜、Hoechst33342荧光法观察细胞的形态与黏附情况。另取第3代BMSCs与纳米PLLA—b-PCL支架材料（实验组）、PLLA—b—PCL支架材料（对照组）进行三维培养3周，扫描电镜观察BMSCs的形态、黏附、生长情况，Hoechst33258荧光法检测复合物中细胞DNA含量，BCA法测定蛋白质含量。结果PLLA-b-PCL无细胞毒性，BMSCs在纳米PLLA—b—PCL支架上黏附、增殖良好。随时间延长，BMSCs在支架材料上的DNA和蛋白质含量逐渐增加，DNA和蛋白质含量均明显高于对照组，差异有统计学意义（P〈0．05）。结论PLLA-b—PCL纳米支架能为BMSCs的生长分化提供较好的环境，具有良好的生物相容性，有望成为一种较好的软骨组织工程支架材料。
Objective Toevaluate thecellular biocompatibilityofthe nano-poly （L-lactic acid）-block-poly （ε-caprolactone） （Nano-PLLA-b-PCL） with canine bone marrow stromal cells（BMSCs） and its feasibility as a scaffold for the cartilage tissue engineering. Methods Nano-PLLA-b-PCL was made by liquid-liquid phase separation. Canine BMSCs were isolated and multiplied in vitro. The passage 3 cells were seeded onto the PLLA-b-PCL films and cultured in the 2-dimensional environment. The cytotoxicity was measured with MTT assay. Cellular morphological changes were observed by phase-contrast microscopy and Hoechst33342 fluorometry. Other passage 3 cells were seeded onto the Nano-PLLA-b-PCL scaffolds （experiment group） and PLLA-b-PCL scaffolds （control group） and cultured in the 3-dimensional environment for 3 weeks. The ratio of cell adhesion was detected by celi counting method. The morphological changes of ceils were observed by scanning electron microscopy. The protein content in seeded cells was determined by bicin-choninic acid assay （BCA）. The content of DNA was quantified using Hoechst33258 assay. Results MTT assay showed the PLLA-b-PCL had no cytotoxicity. The seeded cells adhered to and proliferated well in the Nano-PLLA-b-PCL scaffolds, and maintained good ceil phenotype. After 21-day cell culture within the Nano-PLLA-b-PCL scaffolds, the BMSCs DNA and protein contents increased with time. Moreover, the contents of DNA and protein were both higher in the experiment group than in the control group （ P 〈 0.05 ）. Conclusions The Nano-PLLA-b-PCL scaffold has a good cellular compatibility. It can provide a suitable environment for BMSCs, and will be an ideal scaffold for the cartilage tissue engineering.
出处
《中华创伤骨科杂志》
CAS
CSCD
2007年第7期665-669,共5页
Chinese Journal of Orthopaedic Trauma
基金
广东省科技计划资助项目（2003A3020102）
关键词
纳米技术
乳酸
骨髓细胞
材料试验
组织工程
Nanotechnology
Lactic acid
Bone marrow cells
Materials testing
Tissue engineering
作者简介
通讯作者：蔡道章，E-mail：daozhang@medmail．com．cn
