纳米多孔β-磷酸三钙复合自体骨折血肿的异位成骨潜能的初步实验研究

A preliminary experimental study on the ectopic osteogenesis potential of nano-porousβ-tricalcium phosphate composite autologous fracture hematoma

目的探究纳米多孔β-磷酸三钙(β-tricalcium phosphate,β-TCP)复合自体骨折血肿的异位成骨潜能。方法将78只新西兰大白兔随机分为A、B、C、D和E共5组,髂骨翼上截取约5 mm×10 mm×10 mm的骨块,造成人工骨折并形成血肿,然后分别将血肿/β-TCP、β-TCP、血肿/髂骨、髂骨种植于背阔肌下,于模型建立后第1、4和8周取下背阔肌下标本,采用病理染色技术和免疫组织化学技术评估新骨生成、支架材料降解情况及进行生物力学测试。E组用于观察血肿在β-TCP支架内部的吸附观察,E组动物健侧髂骨设为正常对照组(F组),仅行生物力学测试。结果髂骨骨折后第4天大量的细胞和细胞外基质充填在支架内,此时的血肿/β-TCP机械性能稍优于单纯β-TCP支架。A、C两组标本有周围有较多的新生骨组织及血管组织,而B、D两组标本却只能见到极少量散在的类骨质形成,且D组标本吸收明显;在残余支架体积方面,A、B两组之间差异无统计学意义(P>0.05)。A、C两组新生骨量随着时间的延长而增加(P均<0.01),而B、D两组新生骨量随时间的变化差异无统计学意义(P>0.05)。结论β-TCP与骨折血肿内活性成分具有良好的相容性,β-TCP复合骨折血肿后具有确切的成骨潜能,有望作为一种新型的陈旧性骨折及骨缺损的治疗策略。

Objective To investigate the ectopic osteogenesis potential of nano-porousβ-tricalcium phosphate complex autogenous fracture hematoma.Methods Seventy-eight New Zealand white rabbits were randomly divided into 5 groups:Group A,B,C,D and E.Bone fragments of approximately 5 mm×10 mm×10 mm were extracted from the upper part of the iliac bone to create an artificial fracture and form hematoma,then hematoma/β-TCP,β-TCP,hematoma/iliac crest and iliac crest were implanted under latissimus dorsal muscle,and sublatissimus dorsal muscle specimens were removed at 1,4 and 8 week after the establishment of the model.New bone formation,degradation of scaffold materials and biomechanical tests were evaluated by pathological staining and immunohistochemistry.Group E was used to observe the hematoma while observing the adsorption inside theβ-TCP stent.The healthy iliac bone of group E animals was set as a normal control(group F),and only biomechanical tests were performed.Results A large number of cells and extracellular matrix were filled in the scaffold on day 4 after iliac fracture,and the mechanical properties of hematoma/β-TCP were slightly better than that ofβ-TCP scaffold alone.There were more new bone tissue and vascular tissue around the specimens of group A and group C,while only a small amount of scattered osteoid formation could be seen in the specimens of group B and group D,and the absorption of specimens of group D was obvious.There was no significant difference in the residual scaffold volume between group A and group B(P>0.05).The new bone mass of group A and group C increased with the extension of time(P<0.01),but the change of new bone mass of group B and group D had no statistical significance with the extension of time(P>0.05).Conclusionβ-TCP has good compatibility with the active ingredients in fracture hematoma,andβ-TCP has definite osteogenic potential after fracture hematoma,which is expected to be a new treatment strategy for old fractures and bone defects.