富血小板血浆诱导骨髓基质干细胞联合软骨支架治疗兔激素性股骨头坏死

发布时间：2018-03-08 20:26

本文选题：富血小板血浆　切入点：骨髓基质干细胞　出处：《成都医学院》2017年硕士论文　论文类型：学位论文

【摘要】：股骨头缺血性坏死是常见的骨科疾病,常常会降低患者生活质量,给家庭经济带来负担,其常见的病因有创伤、酒精性、激素等,该病的发病率呈现一个上升趋势[1]。近年来由于激素的广泛应用,激素性股骨头坏死的发生率较高,而且有实验研究表明,激素原因造成的股骨头坏死中股骨头内的骨髓基质干细胞(Bone marrow stromal cells,BMSCs)的数量减少,而且也已经证实激素会影响骨髓基质干细胞的活性,使其分化能力降低,所以针对激素性股骨头坏死的骨髓基质干细胞的研究不断深入,甚至认为激素性股骨头坏死是与骨髓基质干细胞相关的一类疾病。目的观察自体富血小板血浆诱导骨髓基质干细胞联合同种骨软骨支架治疗早期兔激素性股骨头坏死的疗效,探索早期股骨头坏死治疗的新方法。方法采用含有抗凝剂的真空采血管从实验兔子的耳中央动脉采取动脉血5 ml,再通过离心方法获取富血小板血浆(platelet rich plasma,PRP),用16号骨髓穿刺针从实验兔自体股骨髓腔抽取骨髓5 ml,把抽取的骨髓进行密度梯度离心,即可获取兔骨髓基质干细胞,并进行细胞的原代培养和成骨诱导。对已经获取富血小板血浆和骨髓基质干细胞的实验兔进行激素性股骨头坏死造模,取同种股骨髁部骨软骨制作软骨支架,最后以制备的软骨支架为载体,将经过成骨诱导的骨髓基质干细胞移植到软骨支架上,形成细胞-支架复合物,用16号骨髓穿刺针对坏死的股骨头进行髓芯减压术,术中将细胞-支架复合物植入坏死的股骨头下。对实验动物进行分组,一共4组,不同组别采取不同的干预措施。A组空白对照;B组仅仅进行髓芯减压;C组髓芯减压后植入经过成骨诱导培养的骨髓基质干细胞;D组髓芯减压后植入细胞-支架复合物,每组20只,试验后在第2、4、8 W各组分别处死6、6、8只,每只实验兔子的股骨头行大体标本及组织学检查。结果通过各组间在不同时间段的治疗效果比较,D组治疗效果最显著,通过空骨陷窝计数发现D组与其他各组的差异明显,有统计学意义(p0.05)。结论骨髓基质干细胞通过富血小板血浆的诱导向成骨细胞分化;软骨支架材料为骨髓基质干细胞向成骨细胞分化提供良好的空间结构,有利于其向成骨细胞分化;富血小板血浆诱导培养的骨髓基质干细胞和软骨支架材料联合应用可以促进早期兔激素性股骨头坏死的修复。
[Abstract]:Avascular necrosis of the femoral head is a common orthopedic disease, which often reduces the quality of life of the patients and brings a burden to the family economy. Its common causes include trauma, alcohol, hormones, etc. The incidence of the disease is on the rise [1]. In recent years, due to the widespread use of hormones, the incidence of steroid-induced necrosis of the femoral head is relatively high, and some experimental studies have shown that, The number of bone marrow stromal cells in the femoral head of osteonecrosis caused by hormones has been reduced, and it has also been shown that hormones affect the activity of bone marrow stromal cells and reduce their differentiation ability. So the research on bone marrow stromal cells caused by steroid-induced femoral head necrosis is getting deeper and deeper. It is even considered that steroid-induced osteonecrosis of femoral head is a kind of disease associated with bone marrow stromal cells. Objective to observe the effect of autologous platelet-rich plasma induced bone marrow stromal cells combined with osteochondral stents in the treatment of early steroid-induced femoral head necrosis in rabbits. A new method for the treatment of early femoral head necrosis was explored. Methods A vacuum vessel containing anticoagulant was used to take 5 ml of arterial blood from the central ear artery of experimental rabbits, and then centrifugation was used to obtain platelet rich plasma-protopril. Bone marrow was extracted from rabbit autologous femoral medullary cavity by bone marrow puncture needle, and the bone marrow was centrifuged by density gradient centrifugation. Rabbit bone marrow stromal stem cells (BMSCs) were obtained, primary culture and osteogenesis induction were performed. The experimental rabbits which had obtained platelet-rich plasma and bone marrow stromal stem cells (BMSCs) were treated with steroid-induced osteonecrosis of femoral head. The cartilage scaffold was made from the osteochondral cartilage of the femoral condyle. Finally, the bone marrow stromal stem cells (BMSCs) induced by osteogenesis were transplanted onto the cartilage scaffold to form the cell-scaffold complex. The necrotic femoral head was decompressed with 16 bone marrow puncture. The cell-stent composite was implanted under the necrotic femoral head during the operation. The experimental animals were divided into 4 groups. Different intervention measures were taken in different groups. Group A, blank control group B was treated with core decompression and group C, after decompression, bone marrow stromal stem cells cultured in group D were implanted with cell-stent complex after decompression, 20 rats in each group. After the experiment, 8 rats were killed in each group at the 2nd hour and 4th week respectively. The femoral head of each rabbit was examined by gross specimen and histology. Results the therapeutic effect of group D was the most significant compared with that of group D at different time intervals. The difference between group D and other groups was significant (P 0.05). Conclusion Bone marrow stromal cells can differentiate into osteoblasts by platelet-rich plasma. Cartilage scaffolds provide a good spatial structure for the differentiation of bone marrow stromal cells into osteoblasts, which is beneficial to the differentiation of bone marrow stromal cells into osteoblasts. Combined use of platelet-rich plasma induced bone marrow stromal stem cells and cartilage scaffolds can promote the repair of early steroid-induced osteonecrosis of femoral head in rabbits.
【学位授予单位】：成都医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R681.8

【相似文献】