自体BMSCs复合改建脱细胞真皮基质体内构建组织工程软骨的实验研究

发布时间：2018-07-02 10:47

本文选题：脱细胞真皮基质 + BMSCs　；参考：《中国矫形外科杂志》2014年12期

【摘要】：[目的]目的:探讨BMSCs复合脱细胞真皮基质(ADM)三维多孔支架体内构建组织工程软骨的可行性。[方法]首先对支架材料进行大体及扫描电镜观察,分别测定支架的孔隙直径、孔隙率、降解率;其次,分离培养兔骨髓基质细胞(BMSCs),采用扫描电镜及MTT法检测BMSCs在支架材料上的生长、增殖情况;最后取3个月龄清洁级新西兰大白兔27只,雌性,体质量(2.4±0.2)kg,随机分为3组,即A组:空白对照组(仅髁部造缺损),B组:单纯ADM覆盖,C组:ADM+BMSCs,细胞支架复合物体外培养1周后植入兔股骨髁软骨缺损处,分别在4、8、12周后对标本进行组织学、免疫化学检测及ICRS评分。[结果]测量示支架孔隙直径为(101.2±9.3)um,孔隙率为88.6%±2.7%,降解率2周为(13.83±7.12)%,4周为(25.66±8.19)%。MTT法显示细胞在支架上生长良好,与对照组相比,吸光度值差异无统计学意义(P0.05),提示支架相容性良好,无细胞毒性。经过最长12周培养后取材进行组织学、免疫化学检测,结果显示ADM+BMSCs组软骨缺损获得了更为满意的修复效果:关节表面平坦光滑,与周围组织未见明显界限;组织学染色证实软骨基质分泌旺盛并有典型软骨细胞形成,组织结构类似正常关节软骨,ADM+BMSCs组术后第4、8、12周ICRS评分均高于其他两组,差异具有统计学意义(P0.05)。[结论]ADM孔隙均匀,组织相容性好,适于细胞粘附及长期生长,是一种适用于软骨组织工程的良好载体。BMSCs-ADM支架复合体在体内成功构建组织工程软骨,可进一步用于软骨组织工程研究。
[Abstract]:Objective: to investigate the feasibility of constructing tissue engineered cartilage with BMSCs combined with acellular dermal matrix (ADM) in vivo. [methods] the pore diameter, porosity and degradation rate of the scaffold were measured by scanning electron microscope (SEM), and the pore diameter of the scaffold was measured by scanning electron microscope (SEM). Rabbit bone marrow stromal cells (BMSCs) were isolated and cultured. The growth and proliferation of BMSCs on scaffold materials were detected by scanning electron microscope and MTT method. At last, 27 New Zealand white rabbits of 3 months old, female, with body weight of (2.4 卤0.2) kg, were randomly divided into 3 groups. Group A: blank control group (condylar defect only) group B: ADM overlay C group: group C: ADM BMSCsThe scaffold complex was cultured in vitro for 1 week, then implanted into rabbit femoral condylar cartilage defect, and the specimens were histologically carried out after 812 weeks. Immunohistochemical examination and ICRS score. [results] the pore diameter of scaffold was (101.2 卤9.3) umum, porosity was 88.6% 卤2.7, degradation rate was (13.83 卤7.12) weeks and (25.66 卤8.19) 路MTT assay showed that the cells grew well on the scaffold. There was no significant difference in absorbance between the two groups (P0.05), suggesting that the scaffold had good compatibility. No cytotoxicity. The results showed that the cartilage defect of ADM BMSCs group was more satisfactory: the articular surface was flat and smooth, and there was no obvious boundary between the cartilage defect of ADM BMSCs group and surrounding tissue. Histological staining confirmed that cartilage matrix secretion exuberant and typical chondrocyte formation, tissue structure similar to the normal articular cartilage ADM BMSCs group were higher than the other two groups at 12 weeks after operation, the difference was statistically significant (P0.05). [conclusion] ADM has uniform pore, good histocompatibility, suitable for cell adhesion and long-term growth. It is a good carrier for cartilage tissue engineering. BMSCs-ADM scaffold complex can successfully construct tissue engineered cartilage in vivo. It can be further used in cartilage tissue engineering.
【作者单位】：天津医科大学一中心临床学院;天津医科大学一中心临床学院卫生部危重病急救医学重点实验室;天津市第一中心医院骨科;
【基金】：天津市卫生局科技基金重点攻关课题(编号:11KG109)
【分类号】：R318.08

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