本文選題：堿性成纖維細(xì)胞生長(zhǎng)因子 + 血小板衍生生長(zhǎng)因子��； 參考：《河北醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:研究局部聯(lián)合應(yīng)用外源性堿性成纖維細(xì)胞生長(zhǎng)因子(basic fibroblast growth factor,b FGF)、血小板衍生生長(zhǎng)因子(platelet derived growth factor,PDGF)對(duì)大鼠脛骨骨折愈合過程的影響作用,為臨床早期治療骨折延遲愈合或不愈合提供實(shí)驗(yàn)參考。方法:清潔級(jí)成熟雄性SD大鼠48只,隨機(jī)分成4組:A組(b FGF)、B組(PDGF)、C組(b FGF+PDGF)、D組(空白對(duì)照組)。每組各12只。各組實(shí)驗(yàn)動(dòng)物按無菌操作要求建立左脛骨中段骨折模型并應(yīng)用1mm克氏針行髓內(nèi)內(nèi)固定治療。A組、B組及C組大鼠均于術(shù)后第3天,在骨折斷端周圍經(jīng)皮注射藥物,1次/d,連續(xù)7天,各組給藥方式如下:A組(100ng b FGF+0.3ml生理鹽水);B組(100ng PDGF+0.3ml生理鹽水);C組(100ng b FGF+100ng PDGF+0.3ml生理鹽水)。D組則于骨折斷端周圍經(jīng)皮注射等量生理鹽水,給藥時(shí)間及次數(shù)均同于其他組。各組分別于術(shù)后2周及4周隨機(jī)抽選6只大鼠行股動(dòng)脈取血并處死,X線照片觀察骨折愈合情況,取骨折局部大體標(biāo)本部分骨痂進(jìn)行研磨、離心處理,留取上清液,并采用ELISA法測(cè)定骨痂標(biāo)本中ALP的含量。另取部分骨痂脫鈣后進(jìn)行HE染色組織學(xué)觀察,并測(cè)定骨痂中骨小梁成骨指數(shù)IOB及平均骨小梁寬度。采用蛋白質(zhì)芯片技術(shù)測(cè)定2周及4周時(shí)大鼠血清中VEGF、BMP-2及IGF-1的含量。結(jié)果:1術(shù)后2周及4周,大體標(biāo)本表明:A組、B組及C組在新生質(zhì)硬組織大小、硬度方面及骨痂成熟程度、穩(wěn)定性方面大于D組,C組則明顯優(yōu)于其他各組。2術(shù)后2周X線示:C組大鼠骨折處局部骨折線模糊,骨折間隙較其他組小,斷端可見片狀模糊高密度影;A組和B組大鼠骨折處局部骨折線可見,較D組稍模糊,其斷端可見條索狀低密度影。術(shù)后4周X線示:C組大鼠骨折線消失,可見梭行外生骨痂影,形成骨橋并橋接骨折兩端;A組和B組大鼠骨折線基本消失,骨折斷端變鈍,可見高密度外骨痂形成并連接骨折兩端;D組大鼠骨折線仍清楚可見,骨折斷端間隙較2周時(shí)縮小。3組織學(xué)觀察發(fā)現(xiàn):術(shù)后2周時(shí),C組切片有軟骨骨痂和纖維骨痂形成,其中軟骨痂較明顯,軟骨細(xì)胞增生活躍,新生骨小梁及血管較其他組明顯;A組和B組也見纖維骨痂和軟骨骨痂形成,其纖維骨痂相對(duì)明顯;D組則以纖維骨痂為主。術(shù)后4周時(shí),C組大鼠骨小梁基本成熟,可見較多成熟板層骨,髓腔內(nèi)出現(xiàn)大量骨髓細(xì)胞;A組和B組大鼠可見明顯的條索狀軟骨鈣化,骨小梁增多,可見部分成熟板層骨形成,髓腔內(nèi)可見部分骨髓細(xì)胞;D組纖維骨痂較2周時(shí)減少,軟骨骨痂增多,成骨細(xì)胞增生活躍。骨痂分析:術(shù)后2周及4周時(shí),C組骨痂內(nèi)骨小梁成骨指數(shù)明顯高于其他各組且有統(tǒng)計(jì)學(xué)意義(P0.05),A組及B組高于對(duì)照組D組,但小于C組,差異顯著存在且有統(tǒng)計(jì)學(xué)意義(P0.05)。4 ELISA法測(cè)定骨痂中ALP的含量示:C組大鼠患肢骨痂內(nèi)ALP的含量于術(shù)后2周及4周時(shí)在各組中均為最高,且比較差異具有統(tǒng)計(jì)學(xué)意義(P0.05);術(shù)后2周時(shí)A組及B組高于D組并小于C組,且差異存在統(tǒng)計(jì)學(xué)意義(P0.05)。術(shù)后4周時(shí)A組、B組及D組各組間無統(tǒng)計(jì)學(xué)差異(P0.05)。5血清生化指標(biāo)結(jié)果示:術(shù)后2周時(shí),A、B、C三組大鼠血清中VEGF的含量均高于對(duì)照組D組,且差異具有統(tǒng)計(jì)學(xué)意義(P0.05);C組最高,其次為BAD組。術(shù)后4周時(shí):C組血清中VEGF的含量仍為四組中最高(P0.05),余三組間無明顯統(tǒng)計(jì)學(xué)差異(P0.05)。術(shù)后2周時(shí)C組大鼠血清BMP-2的含量高于其他組(P0.05),B組大鼠血清BMP-2的含量高于A組和D組(P0.05)。術(shù)后4周時(shí)各組間無統(tǒng)計(jì)學(xué)差異。術(shù)后2周時(shí),大鼠血清中IGF-1的含量為C組B組A組D組,(P0.05)。術(shù)后4周時(shí)A、B及D組小于C組(P0.05),但A、B、D三組間無明顯統(tǒng)計(jì)差異。結(jié)論:1局部單獨(dú)注射外源性堿性成纖維細(xì)胞生長(zhǎng)因子或血小板衍生生長(zhǎng)因子可以促進(jìn)大鼠脛骨骨折愈合,且聯(lián)合注射優(yōu)于單獨(dú)注射。2局部聯(lián)合注射外源性堿性成纖維細(xì)胞生長(zhǎng)因子和血小板衍生生長(zhǎng)因子可以增加血清中VEGF、BMP-2、IGF-1及骨痂中ALP的表達(dá),這可能是其促進(jìn)大鼠脛骨骨折愈合的一種途徑。3外源性堿性成纖維細(xì)胞生長(zhǎng)因子和血小板衍生生長(zhǎng)因子對(duì)增強(qiáng)大鼠血清中VEGF、IGF-1的表達(dá)有協(xié)同作用。
[Abstract]:Objective: To study the effect of local combined application of exogenous basic fibroblast growth factor (basic fibroblast growth factor, B FGF), platelet derived growth factor (platelet derived growth factor, PDGF) on the healing process of tibial fracture in rats, and to provide experimental reference for the treatment of delayed union or nonunion of fracture in the early clinical period. Methods: 48 mature male SD rats were randomly divided into 4 groups: group A (B FGF), group B (PDGF), C group (B FGF+PDGF), D group (blank control group). Each group had 12 rats in each group. The drugs were injected around the end of the broken end for 7 days for 1 consecutive days. The methods of administration of each group were as follows: group A (100ng B FGF+0.3ml physiological saline), group B (100ng PDGF+0.3ml physiological saline), C group (100ng B FGF+100ng), the group was injected with equal amount of saline around the fracture end, and the time and times of administration were the same in the other groups. The group of 6 rats were randomly selected from the femoral artery to take blood and die at 2 and 4 weeks after the operation. The fracture healing was observed by X-ray photographs. The partial callus of the partial gross specimens of the fracture were lapping, centrifugation and supernatant, and the content of ALP in the callus specimens was measured by ELISA method. The histological observation of HE staining was taken after the decalcification of some callus. Determination of the bone trabecular osteogenesis index IOB and the average bone trabecular width in the callus. The content of VEGF, BMP-2 and IGF-1 in the serum of rats at 2 and 4 weeks was measured by protein chip technique. Results: 2 weeks and 4 weeks after 1, the gross specimens of group A, B and C were found in the size of hard tissue, hardness and the degree of callus maturity and stability. More than group D, group C was obviously superior to other groups in 2 weeks after.2 operation. The local fracture line in group C was blurred, fracture space was smaller than that of other groups, and flaky high density shadow was visible at the broken end; local fracture line in group A and B group was visible, slightly blurred compared with D group, and its broken end showed low density shadow. X ray showed C group rats after operation. Fracture line disappeared, visible fusiform ectic callus shadow, formed bone bridge and bridging both ends of fracture; A group and B group fracture line basically disappeared, fracture end became blunt, high density outer callus formed and connected to both ends of fracture; D group fracture line still clearly visible, fracture end gap between 2 weeks narrowing.3 histological observation found: 2 weeks after operation, C group. Group C after operation, group C group after operation, group C group after 2 weeks of operation, group C group found after operation, group C group, after 2 weeks, C group after operation, group C after operation, group after operation, C group, after 2 weeks, C group The cartilaginous callus and fibrous callus were formed in the section, in which the cartilaginous callus was more obvious, the chondrocytes proliferated active, the new bone trabeculae and blood vessels were more obvious than those in the other groups. The A and B groups also found the fibrous callus and the cartilaginous callus, and the fibrous callus was relatively obvious; the D group was mainly with the fibrous callus. At 4 weeks after the operation, the bone trabecula in group C was basically mature and visible. A large number of bone marrow cells appeared in the intramedullary bone of the multi matured lamellar bone. The A group and the B group showed obvious calcification of the stripe cartilage, the small bone Liang Zengduo, the partial mature lamellar bone formation, and the partial marrow cells in the medullary cavity; the D group was less than 2 weeks, the cartilaginous callus increased and the osteoblasts proliferated active. The bone callus analysis: 2 and 4 weeks after the operation. The osteogenesis index of bone trabecular bone in C group was significantly higher than that of other groups (P0.05), A group and B group were higher than that of control group D, but less than C group, there were significant differences and statistically significant (P0.05).4 ELISA method to determine the content of ALP in the callus: C group rats in the limb callus within 2 weeks and 4 weeks after the operation were all in each group. The difference was statistically significant (P0.05). At 2 weeks after operation, group A and B were higher than group D and less than group C, and the difference was statistically significant (P0.05). There was no statistical difference between group A, B and D group (P0.05).5 serum at 4 weeks after operation (P0.05).5 serum biochemical indicators: 2 weeks after operation, the levels of A, three groups of rats were higher than that of the same In group D, the difference was statistically significant (P0.05); group C was the highest, followed by group BAD. At 4 weeks after operation, the content of VEGF in the serum of C group was still the highest (P0.05), and there was no significant difference between the remaining groups (P0.05). The content of BMP-2 in the serum of group C rats was higher than that of the other groups (P0.05) at 2 weeks after the operation. Group (P0.05). There was no statistical difference between groups at 4 weeks after operation. At 2 weeks after operation, the content of IGF-1 in rat serum was in group C B A group D group, (P0.05). A, B and D groups were less than C group (P0.05) at 4 weeks after operation, but there was no significant statistical difference between groups. Conclusion: the 1 local injection of exogenous basic fibroblast growth factor or platelet derived growth factor by 1 local injection alone The combined injection of.2 locally combined injection of exogenous basic fibroblast growth factor and platelet derived growth factor can increase the expression of ALP in the serum, VEGF, BMP-2, IGF-1 and callus in the serum, which may be a way of promoting the healing of the tibial fracture in rats, which may be a way to promote the healing of the tibial fracture of rats,.3 exogenous. Basic fibroblast growth factor and platelet-derived growth factor play a synergistic role in enhancing the expression of VEGF and IGF-1 in serum of rats.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R683

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