摘要
目的:观察组织工程骨即重组人骨形态发生蛋白-2(recombinant human bone morphogenetic protein-2,rhBMP-2)/异体骨复合骨不同时间点融合骨组织中相关生长因子基因水平的变化。方法:新西兰大白兔60只,随机分为3组。在L5,L6横突间行后路植骨融合术,分别植入复合骨条,自体骨条及异体骨条,于术后第1、2、3、4、5周取融合标本,用原位杂交及实时荧光定量逆转录聚合酶链反应(Real time RT-PCR)分析内源性BMP-2、BMP-7和血管内皮生长因子(VEGF)的表达。结果:原位杂交染色后BMP-7除在1周复合骨最高外(P>0.01),从第2周起自体骨组高于复合骨组,均优于异体骨组,第4周均达峰值复合骨组(61.12±2.42)和自体骨组(64.64±2.00)及异体骨组(50.20±2.15);而VEGF第3周均达到峰值,复合骨组(65.43±2.67)和自体骨组(63.23±4.63)及异体骨组(45.69±2.61),第3、4、5周复合骨组和自体骨组差异无统计学意义(P>0.05),均高于异体骨组。RT-PCR示:术后第3周,复合骨组BMP-2为(5.3519±1.0384),VEGF为(0.9257±0.2534),均达到峰值且高于异体骨组和自体骨组(P<0.05),之后则缓慢下降。第4周后,内源性BMP-2表达仍保持较高的水平,但VEGF的水平与自体骨组和异体骨组差异无统计学意义(P>0.05)。BMP-7变化趋势和原位杂交变化一致,第4周复合骨组(1.2115±0.4007)和自体骨组(1.7859±0.1033)及异体骨组(1.1322±0.1233),5周三组间均降低,但无统计学差异(P>0.05)。结论:rhBMP-2/异体骨复合骨能缓慢,持续,稳定的释放rhBMP-2,有效地诱导了内源性生长因子的表达,极大地促进了成骨效应和血管化。
Objective: To investigate the endogenous correlative gene expression of the masses taken from the rabbits undergone posterolateral lumbar spinal fusion with rhBMP-2/ allograft composite.Methods: Sixty adult male New Zealand white rabbits were randomly divided into three groups:rhBMP-2/allograft composite group, autograft group and allograft group, and each group included 20 rabbits. The rabbits were undergone L5, L6 intertransverse process fusion using rhBMP-2/allograft composite, autogenous iliac crest bone graft and allograft. Every four rabbits from each group sacrificed at the 1st, 2nd, 3th, 4th, 5th week after operation, the fusion masses were harvested then, then determine BMP -2?BMP-7 and VEGF expression at the transcriptional level by real time RT-PCR and situ hybridization.Results: Situ hybridization results: at the 1st week, the BMP-7 expression of composite was higher than other two groups, while from the soncend week the autograft group was higher than the composite and the BMP-7 expression of these two groups was higher than the aiiograft. At the 4th week, the composite group (61.12±2.42)、autograft group (64.64±2.00) and allograft group (50.20±2.15) reached the highest level. At the 3th week, the VEGF expression of the composite (65.43±2.67), autograft groups (63.23±4.63) and allograft groups (45.69±2.61) reached the highest, at the 3th,4th ,5th week, there was not statistically significant between the composite and autograft. while the VEGF expression of these two groups was higher than the allograft. RT-PCR results: At the 3th week, the endogenous BMP-2 (5.3519±1.0384) and VEGF (0.9257±0.2534) gene transcription of the composite group reached the highest level which was higher in comparison to autograft and allograft groups (P<0.05), and then declined. After 4 weeks, endogenous BMP-2 of the composite still keep high expression, but VEGF level was not statistically significant compared to the other two groups (P>0.05). At the 3th, 4th, 5th week after operation, there was not statistically significant among the composite group and autograft group, But both were superior to the allograft group. The trend of BMP-7 expression change between Real-time PCR and situ hybridization was the same.At the 4th, there was statistically significant among the composite group (1.2115±0.4007), autograft group (1.7859±0.1033) and allograft group (1.1322±0.1233). At the 5th, BMP-7 expression of the three groups declined, while there was not statistically significant (P>0.05).Conclusion: The rhBMP-2/allograft composite could slowly、constantly and steadily release endogenous rhBMP-2 ,boost endogenous BMP-2 ? BMP-7and VEGF gene expression effectively and improve the bone formation and vascularization.
引文
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