HCF-PRP在兔股骨头缺血坏死修复中促进血管再生的研究

发布时间：2018-04-11 06:55

  本文选题：富血小板血浆 + 肝素结合纤维蛋白　； 参考：《昆明医科大学》2017年硕士论文

【摘要】：[目的]富血小板血浆(platelet-rich plasma,PRP)被发现富含多种生长因子且具有促进组织修复等多方面作用,目前已被广泛应用于临床多学科和领域。研究发现,PRP在促进血管生成方面显示良好的潜能,且与生物载体结合后其有效作用时间更长,效果更显著。所以,本项目旨在通过研究肝素结合纤维蛋白负载富含血小板血浆(heparin-conjugated fibrin loaded platelet-rich plasma,HCF-PRP)在激素联合脂多糖诱发的兔股骨头缺血坏死的修复中促进血管再生及组织修复的作用,以期为临床治疗肢体慢性缺血性疾病开拓新的思路。[方法]取6-8周龄健康的新西兰大耳兔32只,予以注射脂多糖和地塞米松,以构建股骨头缺血坏死的动物模型,并于药物注射6周后,通过MRI检查判断股骨头缺血坏死情况,筛选出建模成功兔。将建模成功实验动物(n=27)随机分成5组,A组(n=3)为空白对照组,不做任何处理,B组(n=6)单纯行右(R)后肢股骨头穿刺术,左(L)后肢不做任何处理,C组(n=6)行右后肢股骨头穿刺术+肝素结合纤维蛋白(heparin-conjugated fibrin,HCF)注射,左后肢不做任何处理,D组(n=6)行右后肢股骨头穿刺术+PRP注射,左后肢不做任何处理,E组(n=6)行右后肢股骨头穿刺术+HCF-PRP注射,左后肢不做任何处理,进行对照研究。取治疗后第8周作为研究的时间点,行MRI影像学检查,判断各组治疗效果。然后取兔股骨头标本分别做苏木精-伊红(hematoxylin and eosin,HE)染色及血管内皮生长因子(vascular endothelial growth factor,VEGF)免疫组织化学染色,然后进行病理学观察、微血管计数(microvascularcount,MVC)及VEGF表达半定量分析,来观察组织内血管增生情况。利用SPSS17.0统计学软件对上述资料进行统计分析和比较,P0.05为差异有统计学意义,P0.01为差异有显著统计学意义。[结果](1)实验大白兔行激素联合内毒素注射建立股骨头缺血坏死动物模型,经MRI评估,显示成功率为84.36%(27/32)。(2)实验动物于术后第8周再次行MRI检查,显示各组实验动物股骨头缺血坏死的恢复情况不同,E组实验兔R侧股骨头疗效最好,D组R侧次之,A组双侧及B、C、D、E组L侧股骨头疗效最差,B、C组R侧疗效位于A、D组之间。(3)股骨头病理组织切片显示,其坏死表现为E组实验兔R侧股骨头最轻,D组R侧次之,A组双侧及B、C、D、E组L侧股骨头最重,B、C组R侧坏死程度介于A、D组之间。(4)组间R侧MVC两两比较,E 组 MVC 最多(P0.01),D 组次之(P0.01),A 组最少(P0.05),B、C组介于A、D组之间(P0.05);组间L侧MVC两两比较,无差异(P0.05);组内R侧与L侧MVC相比,A组无差异(P0.05),B、D、E组差异显著(P0.01),C组有差异(P0.05)。(5)VEGF表达量比较,E组R侧最高(P0.05),D组R侧次之(P0.05),A组双侧及B、C、D、E组L侧最少(DvsAE,P0.05,其余P0.05),B、C组R侧介于之间(P0.05)。(6)兔股骨头组织内微血管数与VEGF表达量之间呈显著相关,相关系数0.956,P0.01。[结论]1、激素联合内毒素注射可以成功建立股骨头缺血坏死动物模型,且成功率较高;2、在激素联合内毒素诱发的兔股骨头缺血坏死模型中,通过直接穿刺注射HCF-PRP,能促进组织中血管再生,加速侧枝循环建立,促进缺血坏死股骨头组织修复。这为治疗肢体慢性缺血性疾病提供了一种新的思路和方法。
[Abstract]:[Objective] the platelet rich plasma (platelet-rich plasma, PRP) was found to be rich in a variety of growth factors which can promote tissue repair and so on, it has been widely used in many clinical disciplines and fields. The study found that PRP in promoting angiogenesis show good potential, and the effect of combined biological carrier a longer period of time, the effect is more significant. So, this project aims to study the heparin binding protein fiber load of platelet rich plasma (heparin-conjugated fibrin loaded platelet-rich plasma, HCF-PRP) to promote angiogenesis and tissue repair in the repair effect induced by hormone combined with lipid polysaccharide in rabbit ischemic necrosis of the femoral head, in order for the clinical treatment of chronic ischemic limb the disease develop new methods. Thinking] 32 6-8 week old healthy New Zealand rabbits, treated with lipopolysaccharide and dexamethasone, to build stocks The animal model of avascular necrosis, and drug 6 weeks after injection, the judgment of avascular necrosis of the femoral head by MRI examination, screening out the successful modeling. The success of modeling experimental animal rabbits (n=27) were randomly divided into 5 groups, group A (n=3) as control group, without any treatment, B group (n=6) only the right hind limb (R) femoral head puncture, left hindlimb (L) without any treatment, C group (n=6) underwent right femoral bone puncture + heparin combined with fibrin (heparin-conjugated fibrin HCF) injection, the left hind without any treatment, D group (n=6) underwent right femoral head puncture of the left hind limb +PRP injection, without any treatment, E group (n=6) underwent right femoral bone puncture +HCF-PRP injection, the left hind without any treatment, were enrolled in the study. The study time as eighth weeks after treatment, MRI imaging examination, determine the treatment. Then take the rabbit the femoral head specimens respectively. Hematoxylin eosin (hematoxylin and, eosin, HE) staining and vascular endothelial growth factor (vascular endothelial, growth factor, VEGF) immunohistochemical staining, and pathological observation, microvessel count (microvascularcount, MVC) and the expression of VEGF semi quantitative analysis, to observe the proliferation of vascular tissue. The statistical analysis and comparison of the data with statistics software of SPSS17.0 P0.05 is statistically significant, P0.01 had significant difference. Results: (1) experimental rabbits underwent hormone combined with endotoxin injection to establish animal model of avascular necrosis of the femoral head, evaluated by MRI, showed that the success rate is 84.36% (27/32) (2) animal experiments. Again MRI examination eighth weeks after operation, the recovery of each animal experiment showed avascular necrosis of the femoral head, the best E group rabbits R lateral femoral head effect, group D side R times, A group and C D, bilateral B, E缁凩渚ц偂楠ㄥご鐤楁晥鏈，

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