钛种植体表面纳米羟基磷灰石改性对骨结合及RANKL、OPG表达的影响

发布时间：2018-01-30 16:43

本文关键词： 种植体纳米羟基磷灰石骨结合 RANKL OPG　出处：《山西医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:观察钛种植体表面纳米羟基磷灰石改性(nHA)对骨结合及RANKL、OPG表达的影响,探讨钛种植体表面nHA改性在促进骨结合过程中的作用,进而为临床种植体表面设计提供相关研究基础。方法:取健康的新西兰大白兔15只(均为6月龄左右),随机分成3组,每组5只,每只兔子采用双侧自体对照,即在右侧后肢股骨髁下外侧植入1枚实验组(nHA)种植体,在左侧同样的部位植入1枚对照组(Ti组)种植体。术后2w、4w、8w分别处死一组动物,获取约15mm3大小含种植体的标本。将标本进行大体观察及X线片观察,之后制作脱钙切片进行HE染色以及用免疫组化方法检测种植体周围骨组织中RANKL、OPG的蛋白表达水平。实验所测得的数据均运用SPSS17.0进行分析,P0.05认为差异有统计学意义。结果:大体观察:所有种植体与周围骨结合良好,均无松动脱落现象,种植体周围也无感染、化脓及异常生长的软组织。X线片观察:随着种植体植入体内时间的增加,两组种植体周围的低密度影像范围均逐渐减小甚至消失,但在2w和4w时,实验组种植体周围的低密度影像范围均比对照组小,而8w时,两组种植体周围的骨密度与正常的骨组织密度均无明显差别。HE染色观察:随着种植体植入体内时间的增加,两组种植体周围的新生骨及骨小梁均呈增加的趋势,但在2w和4w时,实验组种植体周围骨小梁面积百分比均比对照组大,差异有统计学意义,说明nHA可以促进早期骨结合;8w时,两组种植体周围均可见连续红染的新生骨,两组的骨小梁面积百分比均很高,且两组之间无统计学差异,说明两组种植体周围骨组织生长在8w时达到稳定水平。免疫组化检测RANKL、OPG蛋白表达:所有种植体周围均可检测到RANKL、OPG蛋白表达,2w、4w、8w时,实验组和对照组RANKL、OPG的MOD值分别进行比较,差异均有统计学差异。在2w和4w时,两组动物RANKL的MOD值均大于OPG,而8w时结果相反,反映早期破骨活跃程度大于晚期阶段。2w时,实验组RANKL/OPG的比值大于对照组,差异有统计学意义,说明早期阶段实验组的破骨活跃程度更高;4w时,实验组RANKL/OPG的比值小于对照组,差异有统计学意义,说明实验组的破骨活跃程度下降,而对照组的破骨活跃程度仍处于升高状态;8w时,实验组RANKL/OPG的比值略小于对照组,且均小于1,差异无统计学意义,说明晚期阶段两组之间破骨活跃程度无明显差异,且已转化为成骨阶段。结论:nHA具有良好的生物相容性,且可以促进早期骨结合。在种植体与周围骨组织结合的过程中RANKL与OPG蛋白均有表达,且RANKL/OPG比值的变化规律与骨结合过程一致,提示钛种植体nHA改性后促进早期骨结合可能与其促进骨结合中破骨细胞的形成与分化有关。
[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.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R783.6

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