带蒂筋膜瓣包裹自体红骨髓接种的组织工程骨修复骨缺损的实验研究

EXPERIMENTAL STUDY OF REPAIRING BONE DEFECT WITH TISSUE ENGINEERED BONE SEEDED WITH AUTOLOGOUS RED BONE MARROW AND WRAPPED BY PEDICLED FASCIAL FLAP

目的研究带蒂筋膜瓣作膜诱导技术,将自体红骨髓(autologous red bone marrow,ARBM)接种的组织工程骨血管化,作为膜内充填物修复骨缺损的效果,为临床应用提供实验依据。方法4～5月龄新西兰大白兔32只,雌雄不限,体重2.0～2.5kg,制备双侧桡骨2cm长骨-骨膜完全缺损模型。将ARBM接种于含BMP的骨诱导活性材料制备组织工程骨。将模型左侧设为对照组(A组),仅植入自体组织工程骨;右侧为实验组(B组),利用显微外科技术在骨缺损邻近制备一5cm×3cm的带无名血管蒂的毛细血管网筋膜瓣,包裹自体组织工程骨充填骨缺损。术后4、8、12、16周行X线检查和吸光度(A)值比测量、大体形态和组织学观察、修复区内骨形态计量分析和交界区血管图像分析。结果X线片、大体形态和组织学观察显示,植入物内部血管的长入、骨小梁及软骨组织形成的数量和速度、成熟骨结构的形成、骨干结构的重塑、骨髓腔的再通、植入物的吸收降解,B组均明显优于A组。A值比测量显示,术后8、12、16周两组间比较及同组内各时间点间比较差异均有统计学意义(P<0.05)。术后4、8、12、16周新生骨小梁面积占修复总面积比值两组间比较,以及B组内不同时间点比较差异均有统计学意义(P<0.05);各时间点骨修复交界区单位面积内血管再生面积B组明显多于A组(P<0.05)。结论带蒂筋膜瓣包裹ARBM接种的组织工程骨,具有构建血管化和膜诱导骨组织再生双重作用,对骨缺损有较好的修复作用。

Objective To investigate the effect of repairing bone defect with tissue engineered bone seeded with the autologous red bone marrow （ARBM） and wrapped by the pedicled fascial flap and provide experimental foundation for clinical application. Methods Thirty-two New Zealand white rabbits （male and/or female） aged 4-5 months old and weighing 2.0-2.5 kg were used to make the experimental model of bilateral 2 cm defect of the long bone and the periosteum in the radius. The tissue engineered bone was prepared by seeding the ARBM obtained from the rabbits on the osteoinductive absorbing material containing BMP. The left side of the experimental model underwent the implantation of autologous tissue engineered bone serving as the control group （group A）. While the right side was designed as the experimental group （group B）,one 5 cm × 3 cm fascial flap pedicled on the nameless blood vessel along with its capillary network adjacent to the bone defect was prepared using microsurgical technology,and the autologous tissue engineered bone wrapped by the fascial flap was used to fill the bone defect. At 4,8,12,and 16 weeks after operation,X-ray exam,absorbance （A） value test,gross morphology and histology observation,morphology quantitative analysis of bone in the reparative area,vascular image analysis on the boundary area were conducted. Results X-ray films,gross morphology observation,and histology observation：group B was superior to group A in terms of the growth of blood vessel into the implant,the quantity and the speed of the bone trabecula and the cartilage tissue formation,the development of mature bone structure,the remolding of shaft structure,the reopen of marrow cavity,and the absorbance and degradation of the implant. A value：there was significant difference between two groups 8,12,and 16 weeks after operation （P 〈 0.05）,and there were significant differences among those three time points in groups A and B （P 〈 0.05）. For the ratio of neonatal trabecula area to the total reparative area,there were significant differences between two groups 4,8,12,and 16 weeks after operation （P 〈 0.05）,and there were signif icant differences among those four time points in group B （P 〈 0.05）. For the vascular regenerative area in per unit area of the junctional zone,group B was superior to group A 4,8,12,and 16 weeks after operation （P 〈 0.05）. Conclusion Tissue engineered bone,seeded with the ARBM and wrapped by the pedicled fascial flap,has a sound reparative effect on bone defect due to its dual role of constructing vascularization and inducing membrane guided tissue regeneration.