联合应用bFGF、PDGF对大鼠胫骨骨折愈合的影响

发布时间：2018-06-20 08:23

本文选题：碱性成纤维细胞生长因子 + 血小板衍生生长因子　；参考：《河北医科大学》2017年硕士论文

【摘要】：目的:研究局部联合应用外源性碱性成纤维细胞生长因子(basic fibroblast growth factor,b FGF)、血小板衍生生长因子(platelet derived growth factor,PDGF)对大鼠胫骨骨折愈合过程的影响作用,为临床早期治疗骨折延迟愈合或不愈合提供实验参考。方法:清洁级成熟雄性SD大鼠48只,随机分成4组:A组(b FGF)、B组(PDGF)、C组(b FGF+PDGF)、D组(空白对照组)。每组各12只。各组实验动物按无菌操作要求建立左胫骨中段骨折模型并应用1mm克氏针行髓内内固定治疗。A组、B组及C组大鼠均于术后第3天,在骨折断端周围经皮注射药物,1次/d,连续7天,各组给药方式如下:A组(100ng b FGF+0.3ml生理盐水);B组(100ng PDGF+0.3ml生理盐水);C组(100ng b FGF+100ng PDGF+0.3ml生理盐水)。D组则于骨折断端周围经皮注射等量生理盐水,给药时间及次数均同于其他组。各组分别于术后2周及4周随机抽选6只大鼠行股动脉取血并处死,X线照片观察骨折愈合情况,取骨折局部大体标本部分骨痂进行研磨、离心处理,留取上清液,并采用ELISA法测定骨痂标本中ALP的含量。另取部分骨痂脱钙后进行HE染色组织学观察,并测定骨痂中骨小梁成骨指数IOB及平均骨小梁宽度。采用蛋白质芯片技术测定2周及4周时大鼠血清中VEGF、BMP-2及IGF-1的含量。结果:1术后2周及4周,大体标本表明:A组、B组及C组在新生质硬组织大小、硬度方面及骨痂成熟程度、稳定性方面大于D组,C组则明显优于其他各组。2术后2周X线示:C组大鼠骨折处局部骨折线模糊,骨折间隙较其他组小,断端可见片状模糊高密度影;A组和B组大鼠骨折处局部骨折线可见,较D组稍模糊,其断端可见条索状低密度影。术后4周X线示:C组大鼠骨折线消失,可见梭行外生骨痂影,形成骨桥并桥接骨折两端;A组和B组大鼠骨折线基本消失,骨折断端变钝,可见高密度外骨痂形成并连接骨折两端;D组大鼠骨折线仍清楚可见,骨折断端间隙较2周时缩小。3组织学观察发现:术后2周时,C组切片有软骨骨痂和纤维骨痂形成,其中软骨痂较明显,软骨细胞增生活跃,新生骨小梁及血管较其他组明显;A组和B组也见纤维骨痂和软骨骨痂形成,其纤维骨痂相对明显;D组则以纤维骨痂为主。术后4周时,C组大鼠骨小梁基本成熟,可见较多成熟板层骨,髓腔内出现大量骨髓细胞;A组和B组大鼠可见明显的条索状软骨钙化,骨小梁增多,可见部分成熟板层骨形成,髓腔内可见部分骨髓细胞;D组纤维骨痂较2周时减少,软骨骨痂增多,成骨细胞增生活跃。骨痂分析:术后2周及4周时,C组骨痂内骨小梁成骨指数明显高于其他各组且有统计学意义(P0.05),A组及B组高于对照组D组,但小于C组,差异显著存在且有统计学意义(P0.05)。4 ELISA法测定骨痂中ALP的含量示:C组大鼠患肢骨痂内ALP的含量于术后2周及4周时在各组中均为最高,且比较差异具有统计学意义(P0.05);术后2周时A组及B组高于D组并小于C组,且差异存在统计学意义(P0.05)。术后4周时A组、B组及D组各组间无统计学差异(P0.05)。5血清生化指标结果示:术后2周时,A、B、C三组大鼠血清中VEGF的含量均高于对照组D组,且差异具有统计学意义(P0.05);C组最高,其次为BAD组。术后4周时:C组血清中VEGF的含量仍为四组中最高(P0.05),余三组间无明显统计学差异(P0.05)。术后2周时C组大鼠血清BMP-2的含量高于其他组(P0.05),B组大鼠血清BMP-2的含量高于A组和D组(P0.05)。术后4周时各组间无统计学差异。术后2周时,大鼠血清中IGF-1的含量为C组B组A组D组,(P0.05)。术后4周时A、B及D组小于C组(P0.05),但A、B、D三组间无明显统计差异。结论:1局部单独注射外源性碱性成纤维细胞生长因子或血小板衍生生长因子可以促进大鼠胫骨骨折愈合,且联合注射优于单独注射。2局部联合注射外源性碱性成纤维细胞生长因子和血小板衍生生长因子可以增加血清中VEGF、BMP-2、IGF-1及骨痂中ALP的表达,这可能是其促进大鼠胫骨骨折愈合的一种途径。3外源性碱性成纤维细胞生长因子和血小板衍生生长因子对增强大鼠血清中VEGF、IGF-1的表达有协同作用。
[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.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R683

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