MSCs附和PLGA电纺支架修复兔跟腱损伤的实验研究

发布时间：2018-04-19 17:30

本文选题：骨髓间充质干细胞 + 静电纺丝　；参考：《中国人民解放军军医进修学院》2012年博士论文

【摘要】：目的：研究兔骨髓间充质干细胞附和PLGA电纺支架修复跟腱损伤的作用。方法：利用静电纺丝技术制备PLGA电纺丝膜状支架,对PLGA支架结构进行观察和测量,并使用高温高压、低温等离子、钻6。照射、紫外线照射、酒精浸泡等五种不同方式消毒,进行细菌培养检测消毒后效果并观察支架结构的破坏情况；采用骨髓穿刺法获取兔骨髓,并用密度梯度离心法分离并体外培养骨髓间充质干细胞(MSCs);将第3代骨髓间充质干细胞结合PLGA支架复合培养；将45只成年新西兰大白兔,随机分为3组,每组15只,将左后肢跟腱中央束切断,改良Kessler法缝合,MSCs组使用MSCs附和PLGA电纺丝膜进行包裹,PLGA支架组使用单纯PLGA电纺丝膜进行包裹,对照组只做单纯缝合。分别在2、4、6周时间点各组处死5只兔取材,组织学和生物力学分析评估跟腱的愈合情况。结果：PLGA电纺丝膜状支架纤维直径均匀,结构规整,孔隙率82.6±3.45%；五种消毒方式均可达到灭菌目的；其中钻6。照射和紫外线照射对支架结构的破坏最小最适合PLGA电纺丝膜状支架的消毒；使用梯度离心法分离获得的MSCs纯度高,生物学特性明显,体外增殖能力较强,可以与PLGA电纺丝膜状支架较好结合;MSCs组和PLGA电纺支架组组织学显示肌腱愈合好,与周围组织粘连较轻,生物力学优于对照组。结论：静电纺丝技术是获得纳米级纤维的一种简便方法,使用梯度离心法分离获得的MSCs纯度较高,生物学特性明显,体外增殖能力较强,可以与PLGA电纺丝膜状支架结合;MSCs附和PLGA电纺支架能够促进兔跟腱损伤的愈合,减少外源性疤痕组织长入,减少肌腱与周围组织的粘连。
[Abstract]:Aim: to study the effect of rabbit bone marrow mesenchymal stem cells (BMSCs) and PLGA electrospun scaffold on the repair of Achilles tendon injury.Methods: the PLGA electrospun membrane scaffolds were prepared by electrospinning technique. The structure of PLGA scaffolds was observed and measured. High temperature and high pressure, low temperature plasma, drilling 6.Five different disinfection methods, such as irradiation, ultraviolet radiation and alcohol immersion, were carried out to detect the disinfection effect and observe the destruction of the scaffold structure, and the rabbit bone marrow was obtained by bone marrow puncture.Bone marrow mesenchymal stem cells (MSCs) were isolated by density gradient centrifugation and cultured in vitro. Bone marrow mesenchymal stem cells (BMSCs) of the third generation were co-cultured with PLGA scaffolds. 45 adult New Zealand white rabbits were randomly divided into 3 groups, 15 in each group.The left hind limb was cut off from the central tendon of the Achilles tendon. The modified Kessler method was used to sew the MSCs and PLGA electrospinning membrane. The PLGA electrospun membrane was used to wrap the scaffold group, while the control group was sutured only.Five rabbits were sacrificed at the time of 2 weeks and 4 weeks, respectively. The healing of Achilles tendon was evaluated by histological and biomechanical analysis.Results the diameter of the fiber was uniform, the structure was regular, the porosity was 82.6 卤3.45, and all the five disinfection methods could be used to sterilize the fibers.The minimal damage to the scaffold structure caused by irradiation and ultraviolet irradiation is the most suitable for disinfection of PLGA electrospinning membrane scaffold. The MSCs isolated by gradient centrifugation has high purity, obvious biological characteristics and strong proliferative ability in vitro.It could be combined with PLGA electrospinning membrane scaffold and PLGA electrospinning scaffold to show that tendon healed well, adhesion to surrounding tissue was lighter, biomechanics was superior to control group.Conclusion: electrostatic spinning is a simple and convenient method to obtain nanofibers. MSCs obtained by gradient centrifugation has high purity, obvious biological characteristics and strong proliferative ability in vitro.PLGA and PLGA can promote the healing of Achilles tendon injury, reduce the growth of exogenous scar tissue and reduce the adhesion between tendon and surrounding tissues.
【学位授予单位】：中国人民解放军军医进修学院
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R318.08

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