自体骨移植联合生龙接骨方剂修复兔骨缺损实验研究

Experimental study on autologous bone graft combined with Shenglong Jiegu Formula in repairing rabbit bone defects

目的观察自体骨移植联合生龙接骨方剂对新西兰兔骨缺损模型修复效果的影响。方法选取54只SPF级6月龄雄性新西兰兔作为研究对象,随机分为单纯自体骨移植组(A组)、单纯生龙接骨方剂组(B组)、生龙接骨方剂联合自体骨移植组(C组),每组18只。各组均采用单侧桡骨中段临界尺寸法构建骨缺损模型,A、C组均予以自体骨移植,B、C组均灌胃生龙接骨方剂20 mg/(kg·d),A组灌胃等量生理盐水,持续干预1个月。于术后第1、2、3个月后分别分批处死实验兔,每组每次6只,收集桡骨组织,观察其损伤情况,采用苏木素-伊红(HE)染色、Masson染色检测桡骨愈合情况,经Micro-CT扫描测定各组实验兔骨组织体积(BV)、骨小梁数(Tb.N)、骨小梁厚度(Tb.Th)、骨小梁间距(Th.Sp)和骨密度(BMD)。结果术后各组大体观察到缺损断端骨痂形成至骨皮质连接的骨愈合过程,HE染色显示1~3个月各组炎性细胞浸润减少,C组较其他两组有较好的成骨作用。Masson染色结果显示,C组颜色对比鲜明,结构清晰,可以清晰地区分成熟骨质与新生骨质。Micro-CT检测结果显示,三组术后第2、3个月的BV、Tb.N、Tb.Th、BMD均高于本组术后1个月,B、C组术后3个月的BV、Tb.N、Tb.Th、BMD均高于本组术后2个月,差异有统计学意义(P<0.05)。术后第1、2、3个月,B、C组BV、Tb.N高于A组,C组BMD高于A组,BV、Tb.N、BMD高于B组,差异有统计学意义(P<0.05);术后第1、2、3个月,三组Tb.Th、Tb.Sp比较,差异无统计学意义(P>0.05)。结论自体骨移植联合生龙接骨方剂修复骨缺损效果明显,可提高新西兰兔成骨能力。

Objective To observe the effect of autologous bone graft combined with Shenglong Jiegu Formula in repairing rabbit bone defects.Methods A total of 546-month-old male New Zealand rabbits with SPF grade were selected as the study subjects and they were divided into simple autogenous bone grafting group(group A),simple Shenglong Jiegu Formula group(group B)and Shenglong Jiegu Formula combined with autogenous bone grafting group(group C),with 18 rabbits in each group.The bone defects model was constructed by the critical size method in the middle of the radius in each group.Group A and C were given autologous bone graft,group B and C were given 20 mg/(kg·d)of Shenglong Jiegu Formula,group A was given the same amount of normal saline,and the intervention lasted for 1 month.The experimental rabbits were killed in batches after 1,2 and 3 months,with 6 rabbits in each group.The radius tissue was collected and the damage was observed.Hematoxylin-eosin staining and Masson staining were used to detect the healing of the radius.Bone volume(BV),trabecular number(Tb.N),trabecular thickness(Tb.Th),trabecular space(Th.Sp)and bone mineral density(BMD)were measured by Micro-CT scan.Results The process of bone healing from callus formation to cortical junction was observed in all groups after surgery.Hematoxylin-eosin staining showed that inflammatory cell infiltration decreased in each group from 1 to 3 months,and group C had better osteogenic effect than other two groups.Masson staining results showed that the color of group C was in sharp contrast,the structure was clear,and the mature bone and new bone could be clearly distinguished.Micro-CT detection results showed that the BV,Tb.N,Tb.Th and BMD of the three groups after surgery at 2,3 months after surgery were higher than those of 1 month after surgery,and the BV,Tb.N,Tb.Th and BMD of the group B and C at 3 months after surgery were higher than those of 2 months after surgery,with statistical significances(P<0.05).At 1,2 and 3 months after surgery,BV and Tb.N in group B and C were higher than those in group A,BMD in group C was higher than that in group A,and BV,Tb.N and BMD in group C were higher than those in group B,the differences were statistically significant(P<0.05).There were no significant differences in Tb.Th and Tb.Sp among the three groups at 1,2 and 3 months after operation(P>0.05).Conclusion Autologous bone graft combined with Shenglong Jiegu Formula has obvious effect on repairing bone defects and can improve the osteogenic ability of New Zealand rabbits.