本文關(guān)鍵詞： 種植體 納米羥基磷灰石 骨結(jié)合 RANKL OPG　出處：《山西醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:觀察鈦種植體表面納米羥基磷灰石改性(nHA)對(duì)骨結(jié)合及RANKL、OPG表達(dá)的影響,探討鈦種植體表面nHA改性在促進(jìn)骨結(jié)合過程中的作用,進(jìn)而為臨床種植體表面設(shè)計(jì)提供相關(guān)研究基礎(chǔ)。方法:取健康的新西蘭大白兔15只(均為6月齡左右),隨機(jī)分成3組,每組5只,每只兔子采用雙側(cè)自體對(duì)照,即在右側(cè)后肢股骨髁下外側(cè)植入1枚實(shí)驗(yàn)組(nHA)種植體,在左側(cè)同樣的部位植入1枚對(duì)照組(Ti組)種植體。術(shù)后2w、4w、8w分別處死一組動(dòng)物,獲取約15mm3大小含種植體的標(biāo)本。將標(biāo)本進(jìn)行大體觀察及X線片觀察,之后制作脫鈣切片進(jìn)行HE染色以及用免疫組化方法檢測(cè)種植體周圍骨組織中RANKL、OPG的蛋白表達(dá)水平。實(shí)驗(yàn)所測(cè)得的數(shù)據(jù)均運(yùn)用SPSS17.0進(jìn)行分析,P0.05認(rèn)為差異有統(tǒng)計(jì)學(xué)意義。結(jié)果:大體觀察:所有種植體與周圍骨結(jié)合良好,均無松動(dòng)脫落現(xiàn)象,種植體周圍也無感染、化膿及異常生長的軟組織。X線片觀察:隨著種植體植入體內(nèi)時(shí)間的增加,兩組種植體周圍的低密度影像范圍均逐漸減小甚至消失,但在2w和4w時(shí),實(shí)驗(yàn)組種植體周圍的低密度影像范圍均比對(duì)照組小,而8w時(shí),兩組種植體周圍的骨密度與正常的骨組織密度均無明顯差別。HE染色觀察:隨著種植體植入體內(nèi)時(shí)間的增加,兩組種植體周圍的新生骨及骨小梁均呈增加的趨勢(shì),但在2w和4w時(shí),實(shí)驗(yàn)組種植體周圍骨小梁面積百分比均比對(duì)照組大,差異有統(tǒng)計(jì)學(xué)意義,說明nHA可以促進(jìn)早期骨結(jié)合;8w時(shí),兩組種植體周圍均可見連續(xù)紅染的新生骨,兩組的骨小梁面積百分比均很高,且兩組之間無統(tǒng)計(jì)學(xué)差異,說明兩組種植體周圍骨組織生長在8w時(shí)達(dá)到穩(wěn)定水平。免疫組化檢測(cè)RANKL、OPG蛋白表達(dá):所有種植體周圍均可檢測(cè)到RANKL、OPG蛋白表達(dá),2w、4w、8w時(shí),實(shí)驗(yàn)組和對(duì)照組RANKL、OPG的MOD值分別進(jìn)行比較,差異均有統(tǒng)計(jì)學(xué)差異。在2w和4w時(shí),兩組動(dòng)物RANKL的MOD值均大于OPG,而8w時(shí)結(jié)果相反,反映早期破骨活躍程度大于晚期階段。2w時(shí),實(shí)驗(yàn)組RANKL/OPG的比值大于對(duì)照組,差異有統(tǒng)計(jì)學(xué)意義,說明早期階段實(shí)驗(yàn)組的破骨活躍程度更高;4w時(shí),實(shí)驗(yàn)組RANKL/OPG的比值小于對(duì)照組,差異有統(tǒng)計(jì)學(xué)意義,說明實(shí)驗(yàn)組的破骨活躍程度下降,而對(duì)照組的破骨活躍程度仍處于升高狀態(tài);8w時(shí),實(shí)驗(yàn)組RANKL/OPG的比值略小于對(duì)照組,且均小于1,差異無統(tǒng)計(jì)學(xué)意義,說明晚期階段兩組之間破骨活躍程度無明顯差異,且已轉(zhuǎn)化為成骨階段。結(jié)論:nHA具有良好的生物相容性,且可以促進(jìn)早期骨結(jié)合。在種植體與周圍骨組織結(jié)合的過程中RANKL與OPG蛋白均有表達(dá),且RANKL/OPG比值的變化規(guī)律與骨結(jié)合過程一致,提示鈦種植體nHA改性后促進(jìn)早期骨結(jié)合可能與其促進(jìn)骨結(jié)合中破骨細(xì)胞的形成與分化有關(guān)。
[Abstract]:Objective: to observe the effect of nano-hydroxyapatite modified nHAs on bone binding and expression of RANKL OPG on the surface of titanium implants. To investigate the effect of nHA modification on the surface of titanium implants in the process of promoting bone bonding. Methods: 15 healthy New Zealand white rabbits (all about 6 months old) were randomly divided into 3 groups with 5 rabbits in each group. Each rabbit was implanted with bilateral autogenous control, that is, a subcondylar subcondylar implant was implanted on the right hind limb, and a control group was implanted at the same position on the left. A group of animals were killed for 8 weeks, and about 15 mm3 of implant specimens were obtained. The specimens were observed in general and X-ray. Then decalcified sections were made for HE staining and immunohistochemical method was used to detect RANKL in bone tissue around implants. The protein expression level of OPG was analyzed by SPSS17.0. Results: gross observation: all implants and surrounding bone well combined, no loosening and falling off phenomenon, no infection around the implant. X ray observation of abscess and abnormal growth of soft tissue: with the increase of implant implantation time, the low density imaging range around implants gradually decreased or even disappeared in both groups, but at 2 and 4 weeks. The range of low-density images around the implants in the experimental group was smaller than that in the control group, but at 8w. There was no significant difference between the bone density around the implants and the normal bone tissue density in the two groups. Observation by HE staining: with the increase of implant implantation time in vivo. The new bone and trabeculae around implants in both groups showed an increasing trend, but at 2w and 4w, the percentage of trabecular area around implants in the experimental group was larger than that in the control group, and the difference was statistically significant. These results suggest that nHA can promote early bone bonding. At 8 weeks, continuous red-stained new bone was found around the implants in both groups, and the trabecular area percentage was very high in both groups, and there was no statistical difference between the two groups. The results showed that the bone tissue around the implants reached a stable level at 8 weeks. The expression of RANKL- OPG protein was detected by immunohistochemistry: RANKL could be detected around all implants. The MOD values of RANKL OPG in the experimental group and the control group were significantly different at 2 weeks and 4 weeks after the expression of OPG protein, and at 2 weeks and 4 weeks, the MOD values of the experimental group and the control group were significantly higher than those of the control group. The MOD value of RANKL in the two groups was higher than that in the latter group, but the result was opposite at 8w, which showed that the activity of early osteoclast was higher than that of late stage. The ratio of RANKL/OPG in the experimental group was higher than that in the control group, and the difference was statistically significant, which indicated that the osteoclast activity was higher in the experimental group at the early stage. At 4 weeks, the ratio of RANKL/OPG in the experimental group was lower than that in the control group, the difference was statistically significant, which indicated that the osteoclast activity of the experimental group was decreased, while the osteoclast activity of the control group was still in an elevated state. At 8 weeks, the ratio of RANKL/OPG in the experimental group was slightly smaller than that in the control group, and all of them were less than 1, the difference was not statistically significant, indicating that there was no significant difference in the osteoclast activity between the two groups at the late stage. It has been transformed into osteogenic stage. Conclusion the RANKL and OPG proteins are expressed in the process of osseous binding between implants and surrounding bone tissues, which has good biocompatibility and can promote early bone binding. The change of RANKL/OPG ratio is consistent with the process of bone binding, which suggests that the early bone binding after nHA modification of titanium implants may be related to the formation and differentiation of osteoclasts in the process of bone binding.
【學(xué)位授予單位】：山西醫(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)】：R783.6

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