髓芯减压联合骨髓基质干细胞移植治疗不同类型血液循环障碍导致的犬早期股骨头坏死的实验研究

Core decompression is widely used in the treatment of early femoral head necrosis(ONFH).We found that some patients recovery well through core decompression, but some patients' conditions have progressed very quickly. ONFH can be caused by two different types of blood circulation disorders, arterial ischemia and venous congestion. We find that the rapeutic effect of core decompression is good for venous congestion ONFH, but not very good for arterial ischemic ONFH through digital subtraction angiography (DSA) of two hundred of early ONFH. In this study, we use lipopolysaccharide (LPS) combined with methylprednisolone (MPS) to imitate the clinical etiology and establish dogs early ONFH model. After that, DSA and anatomy further divide ONFH models into arterial ischemic ONFH and venous stasis ONFH. Two different operations are put for two types of blood circulation disorder models. One is core decompression combined with bone marrow stromal stem cells, the other one is autologous bone grafting with blood vessel pedicle. According to different time points after operations, we observe dynamic revascularization by magnetic resonance imaging (MRI) and X-ray. The formation of new bone was detected by histology. Moreover, DSA and anatomy display the femoral head revascularization. Molecular biology is used to detect vascular endothelial growth factor (VEGF) and bone morphogenetic protein-2 (BMP-2) gene expression in vivo. We explore the repair disparity for the canine ONFH with different surgical operations due to arterial ischemia and venous congestion. This research will put forward a new theory in order to improve the success rate of the early ONFH treatment.

髓芯减压广泛应用于治疗早期股骨头坏死(ONFH)。临床发现该法对有的患者疗效明显，有的效果较差。ONFH是由动脉缺血和静脉回流障碍两种不同的血液循环障碍所引起。我们通过对两百例早期ONFH患者进行数字减影动脉造影术(DSA)检查，发现髓芯减压对静脉回流障碍的患者疗效明显，而对动脉缺血性的患者效果并不十分理想。本研究通过脂多糖(LPS)联合甲基强地松龙(MPS)，建立犬早期ONFH模型，行DSA将模型分为动脉缺血和静脉回流障碍两种类型。分别实施髓芯减压联合骨髓基质干细胞(BMSC)移植术及带血管蒂骨瓣移植。观察股骨头内血运重建及新骨生成情况，分子生物学检测血管内皮因子及骨形成蛋白-2基因在股骨头内的表达。探讨髓芯减压联合BMSC移植对动脉缺血和静脉回流障碍所致犬ONFH的修复作用的差别，为提高保头治疗成功率提出新的理论依据。

项目背景：国内外学者通过对股骨头缺血性坏死(ONFH)的病因和发病机制研究得出，ONFH与多种原因引起的血液循环障碍有关，进而导致股骨头小动脉缺血和小静脉回流障碍。目的：探讨髓芯减压联合骨髓基质干细胞移植对动脉缺血和静脉回流障碍所致犬股骨头坏死的修复作用的差别。研究内容：建立激素性股骨头坏死动物模型，对坏死情况进行X片和MRI等影像学评估，通过DSA造影分为动脉缺血以及静脉回流障碍两种类型。对股骨头标本进行血管灌注和组织学检查，证实血供情况和组织病理学变化。对两种模型分别实施髓芯减压联合骨髓基质干细胞移植和带血管蒂自体骨瓣移植。术后第6周、第12周行大体观察，对模型股骨头再次进行影像学评估。术后12周，行股骨头DSA造影术观察股骨头血供的动态变化。取股骨头标本，行动、静脉血管灌注证实血供情况变化，组织学染色观察组织病理学改变，免疫组化染色及荧光实时定量逆转录聚合酶链式反应PCR检测体内VEGF、BMP-2蛋白以及mRNA的表达变化。..重要结果：激素联合内毒素可以成功制作早期的股骨头坏死模型，其坏死率80%，通过DSA造影将坏死模型分为动脉缺血以及静脉回流障碍两种类型，6周以静脉回流障碍为主，12周时动脉缺血为主；灌注显示骨内血流分别减少33%，55%；分别经带血管蒂骨瓣移植和髓芯减压联合骨髓基质干细胞移植治疗后经病理学证实对于动脉缺血类坏死模型，带血管蒂自体骨瓣移植效果优于髓芯减压联合骨髓基质干细胞移植。同时，对于静脉回流障碍类坏死模型的治疗，髓芯减压联合骨髓基质干细胞移植具有优势。灌注显示术后股骨头内血运较模型对照组有不同程度的增加，其中静脉淤滞期行髓芯减压血运恢复最好。免疫组化显示术后VEGF、BMP-2蛋白以及PCR检测mRNA表达增加。.科学意义：通过实验证实，对于动脉缺血类坏死模型的治疗，带血管蒂自体骨瓣移植效果优于髓芯减压联合骨髓基质干细胞移植。同时，对于静脉回流障碍类坏死模型的治疗，髓芯减压联合骨髓基质干细胞移植具有优势。为提高保头治疗的成功率提出一种新的理论依据。