本文選題：A-PRF + 骨缺損�。� 參考：《西南醫(yī)科大學(xué)》2016年碩士論文

【摘要】：目的:通過建立兔顱骨的改良型富血小板纖維蛋白(AdvancedPlatelet-rich fibrin,A-PRF)骨誘導(dǎo)模型,觀察A-PRF成骨效果,探討A-PRF誘導(dǎo)骨再生過程中誘導(dǎo)新生骨的微觀結(jié)構(gòu)和組織學(xué)特點(diǎn),評(píng)價(jià)A-PRF修復(fù)骨缺損的特點(diǎn),為A-PRF在誘導(dǎo)骨再生及頜骨重建的臨床應(yīng)用提供實(shí)驗(yàn)參考。方法:標(biāo)準(zhǔn)實(shí)驗(yàn)動(dòng)物日本大耳兔30只隨機(jī)分為A-PRF組和空白組各15只,在兔顱骨骨中縫兩側(cè)均制造一個(gè)6.0 mm的全層洞穿性骨缺損,A-PRF組植入A-PRF膜,空白組不填入任何材料,術(shù)后當(dāng)日、2周、4周、8周、12周術(shù)區(qū)行大體觀察后分別處死實(shí)驗(yàn)動(dòng)物,取下術(shù)區(qū)骨標(biāo)本行X線檢查、Micro-CT分析、HE染色、Masson染色、免疫組織化學(xué)分析。結(jié)果:30只實(shí)驗(yàn)動(dòng)物手術(shù)順利完成,愈合良好,成功建立兔顱骨的A-PRF骨誘導(dǎo)模型,術(shù)區(qū)觀察見A-PRF組A-PRF膜隨著新生骨的增加而逐漸減少,A-PRF膜術(shù)后8周后消失,新生骨術(shù)后4周后形成明顯,術(shù)后8周后逐漸成熟與周圍骨同。空白組骨缺損區(qū)邊界清楚,隨著時(shí)間的延長(zhǎng),軟組織逐漸長(zhǎng)入骨缺損區(qū),至術(shù)后12周,未見明顯新骨形成。X線觀察可見:術(shù)后當(dāng)日骨缺損明顯,隨著時(shí)間的延長(zhǎng),A-PRF組的骨缺損邊緣逐漸模糊,呈現(xiàn)白色高密度影,且呈向心性生長(zhǎng)態(tài)勢(shì)�？瞻捉M至術(shù)后8周骨缺損邊緣霧化,術(shù)后12周骨缺損邊緣見少量白色高密度影。Micro-CT測(cè)量分析:A-PRF組術(shù)后當(dāng)日骨缺損內(nèi)未見骨小梁生成,術(shù)后2周邊緣見少量新骨形成,隨著時(shí)間的延長(zhǎng),骨小梁逐漸清晰可見且排列規(guī)律,術(shù)后12周時(shí)與周圍正常骨小梁接近。空白組術(shù)后12周時(shí)仍然清晰可見骨缺損區(qū),呈低密度影像,骨小梁極少。統(tǒng)計(jì)分析BV/TV、Tb.N、Tb.Th、Tb.Sp參數(shù)可得出同一時(shí)間節(jié)點(diǎn)實(shí)驗(yàn)組與空白組的差異有統(tǒng)計(jì)學(xué)差異(P0.05)。HE染色:A-PRF組術(shù)后當(dāng)日骨缺損區(qū)見大量紅細(xì)胞,白細(xì)胞,血小板等結(jié)構(gòu),術(shù)后2周,骨缺損區(qū)見一種接近骨組織的物質(zhì)形成,術(shù)后4周骨缺損區(qū)見新生的骨小梁,術(shù)后8周、12周骨缺損區(qū)見少量新生骨組織中形成骨髓腔和骨髓�？瞻捉M:術(shù)后當(dāng)日骨缺損未見任何物質(zhì)。術(shù)后2周見少量纖維結(jié)蹄組織長(zhǎng)入,術(shù)后4周纖維結(jié)蹄組織增多,仍未見新骨形成,術(shù)后8周見少量骨細(xì)胞游離期間,未見新骨形成,術(shù)后12周見少量新骨形成,骨小梁細(xì)小,未見骨髓腔和骨髓。Masson染色:A-PRF組和空白組均成紅-藍(lán)相間,A-PRF組在術(shù)后各時(shí)間節(jié)點(diǎn)較空白組紅染區(qū)域更多,更廣。免疫組織化學(xué)分析:RANKL、OPG免疫組織化學(xué)平均光密度分析結(jié)果顯示:A-PRF組RANKL表達(dá)術(shù)后當(dāng)日至術(shù)后2周釋放較平穩(wěn),術(shù)后2周后逐漸增多,至術(shù)后12周時(shí)仍成上升趨勢(shì),同一時(shí)間節(jié)點(diǎn)與空白組比較有統(tǒng)計(jì)學(xué)差異(P0.05),OPG表達(dá)在術(shù)后當(dāng)日釋放逐漸增多,8周達(dá)到高峰后逐漸下降,同一時(shí)間節(jié)點(diǎn)與空白組比較有統(tǒng)計(jì)學(xué)差異(P0.05)�？瞻捉MRANKL、OPG表達(dá)均在術(shù)后8周后表達(dá)增多,至觀察期結(jié)束,表達(dá)仍成上升趨勢(shì)。結(jié)論:1.本實(shí)驗(yàn)通過制備兔顱骨的臨界性骨缺損,成功建立兔顱骨的A-PRF骨誘導(dǎo)模型2.A-PRF可誘導(dǎo)新生骨形成。3.A-PRF在修復(fù)骨缺損的過程中,大體觀察、X線影像和Micro-CT測(cè)量分析均表現(xiàn)為向心性成骨態(tài)勢(shì)。4.A-PRF在體內(nèi)的完全降解時(shí)間約為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.
【學(xué)位授予單位】：西南醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R683

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