不同种属来源骨髓间充质干细胞修复喉软骨缺损的实验研究

发布时间：2018-05-18 20:48

  本文选题：骨髓间充质干细胞 + 异种　； 参考：《辽宁医学院》2011年硕士论文

【摘要】：目的 探讨异种来源的骨髓间充质干细胞(BMSCs)复合聚乳酸/羟基乙酸共聚物(PLGA)生物支架,加入转化生长因子-β1(TGF-β1)和软骨形态发生蛋白(CDMP1)联合诱导后修复甲状软骨缺损的能力,评价修复效果,为应用软骨组织工程技术修复软骨缺损提供新的思路和方法。 方法 分别取20周人胚胎及刚出生乳兔的四肢骨骨髓,以密度梯度离心法分离出BMSCs,体外培养扩增至第三代。倒置显微镜观察BMSCs生长状态,MTT检测细胞活率。分别取第三代的BMSCs,制成细胞悬液,以5×108/ml的密度接种到PLGA生物支架上,将细胞支架复合物加入CDMP1和TGF-β1联合诱导21天。取24只3月龄新西兰白兔,体重3-4公斤,雌雄不限,随机分为三组。全麻下制作甲状软骨缺损的模型,异种实验组植入人胚胎BMSCs支架复合物,同种实验组植入兔BMSCs支架复合物,空白对照组植入单纯支架。术后4周,8周进行大体观察,HE染色,阿利新蓝染色,免疫组化观察缺损修复的效果。 结果 两种BMSCs在生长速度和形态上无明显差异。细胞支架复合物植入缺损区后均有软骨细胞生成,缺损处表面光滑平整,阿利新蓝染色和免疫组化见两组的新生细胞分泌软骨细胞基质糖氨聚糖(GAG)和Ⅱ型胶原量无明显差异。单纯支架组缺损处只有少量纤维组织修复,表面凹陷粗糙,阿利新蓝染色和免疫组化检测几乎无软骨细胞基质GAG和Ⅱ型胶原分泌。统计学显示两种BMSCs支架复合物修复喉软骨缺损的效果差异无统计学意义(P 0.05),且均好于单纯支架组,差异有统计学意义(P 0.05)。 结论 异种来源的BMSCs复合PLGA在TGF-β1和CDMP1联合诱导下所得的组织工程化软骨与同种来源的BMSCs所获得的组织工程化软骨修复喉软骨缺损具有可比性。
[Abstract]:Purpose To explore the ability of bone marrow mesenchymal stem cells (BMSCs) combined with poly (lactic acid) / glycolic acid (PLGA) scaffold to repair thyroid cartilage defect induced by TGF- 尾 1 (transforming growth factor- 尾 1) and CDMP1 (cartilage morphogenetic protein). To evaluate the effect of cartilage repair and provide new ideas and methods for repairing cartilage defects by using cartilage tissue engineering technology. Method Bone marrow of limbs of 20 week human embryo and newborn rabbit were isolated by density gradient centrifugation. BMSCs were cultured and amplified to the third generation in vitro. The growth state of BMSCs was observed by inverted microscope. Cell suspensions were prepared from the third generation BMSCs and inoculated into PLGA scaffolds at the density of 5 脳 108/ml. The scaffold complexes were added to CDMP1 and TGF- 尾 1 for 21 days. 24 3-month old New Zealand white rabbits weighing 3-4 kg were randomly divided into three groups. The model of thyroid cartilage defect was made under general anesthesia. Human embryonic BMSCs scaffold complex was implanted in xenogeneic experimental group, rabbit BMSCs scaffold complex was implanted in allogenic experimental group, and pure scaffold was implanted in blank control group. At 4 weeks and 8 weeks after operation, HE staining, Alixin blue staining and immunohistochemistry were used to observe the effect of repairing defects. Result There was no significant difference in growth rate and morphology between the two kinds of BMSCs. Chondrocytes were formed after the scaffold complex was implanted into the defect area, and the surface of the defect was smooth and smooth. There was no significant difference between the two groups in the secretion of matrix glycosaminoglycan (GAG) and type 鈪，

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