有骨髓间充质干细胞附着的聚乳酸-羟基乙酸共聚物-Ⅰ型胶原蛋白支架补片在大鼠腹股沟疝模型中的应用

Application of bone marrow mesenchymal stem cells attached poly(lactic-co-glycolic acid)-collagen Ⅰ scaffold patch in rat inguinal hernia model

目的研究骨髓间充质干细胞(BMSCs)覆膜聚乳酸-羟基乙酸共聚物-Ⅰ型胶原蛋白支架(PLGA-collagenⅠ)补片对腹股沟疝的修补效果。方法首先,制备PLGA-collagenⅠ补片;然后提取大鼠骨髓间充质干细胞,并制作腹壁缺损大鼠模型。将覆膜有骨髓间充质干细胞的组织工程补片(实验组)和无细胞组织工程补片(对照组)分别植入腹股沟。ELISA检测IL-6和IL-10。免疫组化检测巨噬细胞表面抗原,分析炎性反应。HE染色观察补片中自身组织的再生情况和材料降解状态。用扫描电子显微镜观察腹膜的完整性。免疫组化检测新生血管状态。Masson三色染色检测胶原蛋白纤维的形成。结果1周后,实验组和对照组的巨噬细胞在补片上的浸润均显著增加(P<0.05)。1个月后,炎性反应逐渐消退,巨噬细胞数量减少(P<0.05)。长期实验表明,BMSCs补片能促进胶原纤维的合成,促进血管再生,修补腹壁损伤。实验组和对照组的大鼠腹壁无肿胀,大鼠健康。术后1、2和6个月进行组织形态学观察和SEM电子显微镜检查。带有BMSCs的PLGA-collagenⅠ补片可促进细胞浸润,血管生成和组织再生,且降解速度更快。6个月后,种植有BMSCs的补片几乎完全降解,没有种植细胞的PLGA-collagenⅠ补片被降解成碎片,并作为异物留在体内。结论植入BMSCs的PLGA-collagenⅠ组织工程材料能更快速可靠的修补疝缺损,达到治疗腹股沟疝的目的。

Objective To study the repairing effect of poly(lactic-co-glycolic acid)-collegenⅠ(PLGA-collagenⅠ)patch on inguinal hernia.Methods The PLGA-collagenⅠpatch was prepared.Then the rat bone marrow mesenchymal stem cells(BMSCs)were extracted,and the rat model of abdominal wall defect was developed.Bone marrow mesenchymal stem cells bearing tissue engineering patches(experimental group)and cell-free tissue engineering patches(control group)were implanted into the groin respectively.The changes of pro-inflammatory factor IL-6(interleukin-6)and IL-10(interleukin-10)were detected by ELISA kit after operation.Immunohistochemistry was used to detect the change of macrophage surface antigen to analyze the inflammatory reaction.HE staining was used to observe the regeneration of tissue in the patch and the degradation state of the material.SEM was used to observe the integrity of peritoneum.The angiogenesis state was detected by immunohistochemistry.The formation of collagen fiber was detected by trichrome staining of Masson.Results After a week,the macrophages invasion in the patch of both the experimental group and the control group was significantly increased.For one month,the inflammatory reaction gradually subsided and the number of macrophages decreased.Long term tests showed that the BMSCs patch promoted the synthesis of collagen fibers,regeneration of blood vessels,and repair of abdominal wall injury.One month after surgery,there was no swelling in the abdominal wall of the experimental group and the control group.In 1,2 and 6 months after the operation,the tissue morphology and SEM electron microscopy were performed.The PLGA-collagenⅠpatch of BMSCs can promote cell infiltration,angiogenesis and tissue regeneration.The degradation rate of PLGA material was faster.In sixth months,the cell-based patch was almost completely degraded.However,PLGA-collagenⅠpatches in which cells were not planted degraded.It degraded into pieces and remained in the body as a foreign body.Conclusions In this study,PLGA-collagenⅠtissue engineering materials planted with BMSCs can repair the defects more quickly and reliably,so as to achieve the purpose of treatment of hernia.