本文選題：富血小板血漿 + 肝素結(jié)合纖維蛋白��； 參考：《昆明醫(yī)科大學(xué)》2017年碩士論文

【摘要】：[目的]富血小板血漿(platelet-rich plasma,PRP)被發(fā)現(xiàn)富含多種生長(zhǎng)因子且具有促進(jìn)組織修復(fù)等多方面作用,目前已被廣泛應(yīng)用于臨床多學(xué)科和領(lǐng)域。研究發(fā)現(xiàn),PRP在促進(jìn)血管生成方面顯示良好的潛能,且與生物載體結(jié)合后其有效作用時(shí)間更長(zhǎng),效果更顯著。所以,本項(xiàng)目旨在通過(guò)研究肝素結(jié)合纖維蛋白負(fù)載富含血小板血漿(heparin-conjugated fibrin loaded platelet-rich plasma,HCF-PRP)在激素聯(lián)合脂多糖誘發(fā)的兔股骨頭缺血壞死的修復(fù)中促進(jìn)血管再生及組織修復(fù)的作用,以期為臨床治療肢體慢性缺血性疾病開(kāi)拓新的思路。[方法]取6-8周齡健康的新西蘭大耳兔32只,予以注射脂多糖和地塞米松,以構(gòu)建股骨頭缺血壞死的動(dòng)物模型,并于藥物注射6周后,通過(guò)MRI檢查判斷股骨頭缺血壞死情況,篩選出建模成功兔。將建模成功實(shí)驗(yàn)動(dòng)物(n=27)隨機(jī)分成5組,A組(n=3)為空白對(duì)照組,不做任何處理,B組(n=6)單純行右(R)后肢股骨頭穿刺術(shù),左(L)后肢不做任何處理,C組(n=6)行右后肢股骨頭穿刺術(shù)+肝素結(jié)合纖維蛋白(heparin-conjugated fibrin,HCF)注射,左后肢不做任何處理,D組(n=6)行右后肢股骨頭穿刺術(shù)+PRP注射,左后肢不做任何處理,E組(n=6)行右后肢股骨頭穿刺術(shù)+HCF-PRP注射,左后肢不做任何處理,進(jìn)行對(duì)照研究。取治療后第8周作為研究的時(shí)間點(diǎn),行MRI影像學(xué)檢查,判斷各組治療效果。然后取兔股骨頭標(biāo)本分別做蘇木精-伊紅(hematoxylin and eosin,HE)染色及血管內(nèi)皮生長(zhǎng)因子(vascular endothelial growth factor,VEGF)免疫組織化學(xué)染色,然后進(jìn)行病理學(xué)觀察、微血管計(jì)數(shù)(microvascularcount,MVC)及VEGF表達(dá)半定量分析,來(lái)觀察組織內(nèi)血管增生情況。利用SPSS17.0統(tǒng)計(jì)學(xué)軟件對(duì)上述資料進(jìn)行統(tǒng)計(jì)分析和比較,P0.05為差異有統(tǒng)計(jì)學(xué)意義,P0.01為差異有顯著統(tǒng)計(jì)學(xué)意義。[結(jié)果](1)實(shí)驗(yàn)大白兔行激素聯(lián)合內(nèi)毒素注射建立股骨頭缺血壞死動(dòng)物模型,經(jīng)MRI評(píng)估,顯示成功率為84.36%(27/32)。(2)實(shí)驗(yàn)動(dòng)物于術(shù)后第8周再次行MRI檢查,顯示各組實(shí)驗(yàn)動(dòng)物股骨頭缺血壞死的恢復(fù)情況不同,E組實(shí)驗(yàn)兔R側(cè)股骨頭療效最好,D組R側(cè)次之,A組雙側(cè)及B、C、D、E組L側(cè)股骨頭療效最差,B、C組R側(cè)療效位于A、D組之間。(3)股骨頭病理組織切片顯示,其壞死表現(xiàn)為E組實(shí)驗(yàn)兔R側(cè)股骨頭最輕,D組R側(cè)次之,A組雙側(cè)及B、C、D、E組L側(cè)股骨頭最重,B、C組R側(cè)壞死程度介于A、D組之間。(4)組間R側(cè)MVC兩兩比較,E 組 MVC 最多(P0.01),D 組次之(P0.01),A 組最少(P0.05),B、C組介于A、D組之間(P0.05);組間L側(cè)MVC兩兩比較,無(wú)差異(P0.05);組內(nèi)R側(cè)與L側(cè)MVC相比,A組無(wú)差異(P0.05),B、D、E組差異顯著(P0.01),C組有差異(P0.05)。(5)VEGF表達(dá)量比較,E組R側(cè)最高(P0.05),D組R側(cè)次之(P0.05),A組雙側(cè)及B、C、D、E組L側(cè)最少(DvsAE,P0.05,其余P0.05),B、C組R側(cè)介于之間(P0.05)。(6)兔股骨頭組織內(nèi)微血管數(shù)與VEGF表達(dá)量之間呈顯著相關(guān),相關(guān)系數(shù)0.956,P0.01。[結(jié)論]1、激素聯(lián)合內(nèi)毒素注射可以成功建立股骨頭缺血壞死動(dòng)物模型,且成功率較高;2、在激素聯(lián)合內(nèi)毒素誘發(fā)的兔股骨頭缺血壞死模型中,通過(guò)直接穿刺注射HCF-PRP,能促進(jìn)組織中血管再生,加速側(cè)枝循環(huán)建立,促進(jìn)缺血壞死股骨頭組織修復(fù)。這為治療肢體慢性缺血性疾病提供了一種新的思路和方法。
[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緇凩渚ц偂楠ㄥご鐤楁晥鏈，

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