改良型富血小板纤维蛋白促进兔颅骨骨再生的实验研究

发布时间：2018-07-01 19:14

本文选题：A-PRF + 骨缺损　；参考：《西南医科大学》2016年硕士论文

【摘要】：目的:通过建立兔颅骨的改良型富血小板纤维蛋白(AdvancedPlatelet-rich fibrin,A-PRF)骨诱导模型,观察A-PRF成骨效果,探讨A-PRF诱导骨再生过程中诱导新生骨的微观结构和组织学特点,评价A-PRF修复骨缺损的特点,为A-PRF在诱导骨再生及颌骨重建的临床应用提供实验参考。方法:标准实验动物日本大耳兔30只随机分为A-PRF组和空白组各15只,在兔颅骨骨中缝两侧均制造一个6.0 mm的全层洞穿性骨缺损,A-PRF组植入A-PRF膜,空白组不填入任何材料,术后当日、2周、4周、8周、12周术区行大体观察后分别处死实验动物,取下术区骨标本行X线检查、Micro-CT分析、HE染色、Masson染色、免疫组织化学分析。结果:30只实验动物手术顺利完成,愈合良好,成功建立兔颅骨的A-PRF骨诱导模型,术区观察见A-PRF组A-PRF膜随着新生骨的增加而逐渐减少,A-PRF膜术后8周后消失,新生骨术后4周后形成明显,术后8周后逐渐成熟与周围骨同。空白组骨缺损区边界清楚,随着时间的延长,软组织逐渐长入骨缺损区,至术后12周,未见明显新骨形成。X线观察可见:术后当日骨缺损明显,随着时间的延长,A-PRF组的骨缺损边缘逐渐模糊,呈现白色高密度影,且呈向心性生长态势。空白组至术后8周骨缺损边缘雾化,术后12周骨缺损边缘见少量白色高密度影。Micro-CT测量分析:A-PRF组术后当日骨缺损内未见骨小梁生成,术后2周边缘见少量新骨形成,随着时间的延长,骨小梁逐渐清晰可见且排列规律,术后12周时与周围正常骨小梁接近。空白组术后12周时仍然清晰可见骨缺损区,呈低密度影像,骨小梁极少。统计分析BV/TV、Tb.N、Tb.Th、Tb.Sp参数可得出同一时间节点实验组与空白组的差异有统计学差异(P0.05)。HE染色:A-PRF组术后当日骨缺损区见大量红细胞,白细胞,血小板等结构,术后2周,骨缺损区见一种接近骨组织的物质形成,术后4周骨缺损区见新生的骨小梁,术后8周、12周骨缺损区见少量新生骨组织中形成骨髓腔和骨髓。空白组:术后当日骨缺损未见任何物质。术后2周见少量纤维结蹄组织长入,术后4周纤维结蹄组织增多,仍未见新骨形成,术后8周见少量骨细胞游离期间,未见新骨形成,术后12周见少量新骨形成,骨小梁细小,未见骨髓腔和骨髓。Masson染色:A-PRF组和空白组均成红-蓝相间,A-PRF组在术后各时间节点较空白组红染区域更多,更广。免疫组织化学分析:RANKL、OPG免疫组织化学平均光密度分析结果显示:A-PRF组RANKL表达术后当日至术后2周释放较平稳,术后2周后逐渐增多,至术后12周时仍成上升趋势,同一时间节点与空白组比较有统计学差异(P0.05),OPG表达在术后当日释放逐渐增多,8周达到高峰后逐渐下降,同一时间节点与空白组比较有统计学差异(P0.05)。空白组RANKL、OPG表达均在术后8周后表达增多,至观察期结束,表达仍成上升趋势。结论:1.本实验通过制备兔颅骨的临界性骨缺损,成功建立兔颅骨的A-PRF骨诱导模型2.A-PRF可诱导新生骨形成。3.A-PRF在修复骨缺损的过程中,大体观察、X线影像和Micro-CT测量分析均表现为向心性成骨态势。4.A-PRF在体内的完全降解时间约为4周至8周。
[Abstract]:Objective: To observe the effect of A-PRF osteogenesis by establishing a modified AdvancedPlatelet-rich fibrin (A-PRF) bone induction model of rabbit skull, and to explore the microstructure and histological characteristics of the induced bone in the process of A-PRF induced bone regeneration, evaluate the characteristics of A-PRF repair of bone defect, and to induce the bone regeneration and the weight of the jaws for A-PRF in the induction of bone regeneration. The clinical application provides experimental reference. Methods: 30 Japanese big ear rabbits were randomly divided into A-PRF group and 15 blank group. A 6 mm full layer penetrating bone defect was made on both sides of the rabbit skull bone. Group A-PRF was implanted with A-PRF membrane, and the blank group was not filled with any material. After the operation, 2 weeks, 4 weeks, 8 weeks, and 12 weeks were large. After body observation, the experimental animals were sacrificed respectively. The X-ray examination of bone mark, Micro-CT analysis, HE staining, Masson staining and immunohistochemical analysis were taken. Results: 30 experimental animals were successfully completed and healed well. The A-PRF bone induction model of the rabbit skull was successfully established. The operation area observed that the A-PRF membrane in the group A-PRF was gradually increased with the increase of the new bone. Reduction, 8 weeks after the operation of A-PRF membrane disappeared, the formation of new bone after 4 weeks was obvious, 8 weeks after the operation gradually mature with the surrounding bone. The blank group bone defect area is clear, as the time prolongs, the soft tissue gradually grows to the bone defect area, to 12 weeks after the operation, no obvious new bone formation is found. X - ray observation can be seen that the bone defect is obvious and with time after the operation. The edge of bone defect in group A-PRF was gradually blurred, showing a white high density shadow and a centripetal growth situation. The blank group was nebulized at the edge of the bone defect 8 weeks after the operation. A small amount of white high density shadow was observed at the edge of the bone defect at the 12 week after the operation. There was no bone Liang Shengcheng in the bone defect on the day after operation in group A-PRF, and the margin of the bone defect was less in the 2 weeks after the operation. The bone trabecula was clearly visible and arranged with time, and the bone trabecula was close to the normal bone trabecula around 12 weeks after the operation. The bone defect area was clearly visible at 12 weeks after the operation. The low density image and the bone trabecula were very few at the 12 week after the operation. The statistical analysis of BV/TV, Tb.N, Tb.Th, and Tb.Sp parameters can be used to obtain the same time node experimental group and space. The differences in the white group were statistically different (P0.05).HE staining: a large number of red cells, leukocytes and platelets were found in the bone defect area of group A-PRF on the day after operation, and 2 weeks after the operation, a kind of material formation near the bone tissue was found in the bone defect area, and the new bone trabecula was seen in the bone defect area at the 4 week after the operation. A small amount of new bone tissue was found in the bone defect area at 8 weeks after the operation. Bone marrow cavity and bone marrow. Blank group: no material was found on the bone defect on the day after operation. A small amount of fibrous hoofed tissue was seen 2 weeks after operation, and fibrous hoofed tissue increased at 4 weeks after operation. No new bone formation was found. No new bone formation was found during the 8 weeks after the operation. A small amount of new bone was found at the end of the operation. The bone trabecula was small and no bone marrow cavity was found at 12 weeks after the operation. Bone marrow.Masson staining: both group A-PRF and blank group were red and blue, and group A-PRF was more and more extensive in each time node after operation than that in blank group. Immunohistochemical analysis: RANKL, OPG immunohistochemical mean light density analysis showed that the release of RANKL expression in A-PRF group was more stable after 2 weeks after operation, and gradually increased after 2 weeks after operation. At the same time, there was a statistical difference between the nodes and the blank group (P0.05) at the same time (P0.05). The expression of OPG increased gradually on the day after the operation, and gradually decreased after the peak of the 8 week. There was a statistical difference between the node and the blank group at the same time (P0.05). The expression of the blank group was RANKL, and the expression of OPG increased after 8 weeks after the operation. The expression still became a rising trend at the end of the observation period. Conclusion: 1. the critical bone defect of rabbit skull was prepared by the 1. experiment. The successful establishment of the A-PRF bone induction model 2.A-PRF of the rabbit skull can induce the formation of.3.A-PRF in the process of repairing bone defect. The gross observation, X-ray image and Micro-CT analysis all show the tendency of centripetal osteogenesis.4.A. The complete degradation time of -PRF in vivo is about 4 weeks to 8 weeks.
【学位授予单位】：西南医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R683

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