摘要目的 探讨异体骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)经皮移植联合高压氧修复海水浸泡后兔桡骨开放性骨缺损的效果.方法 48只新西兰大白兔,采用数字表法随机分为4组:空白对照组(A组)、BMSCs移植组(B组)、高压氧治疗组(C组)、BMSCs移植联合高压氧治疗组(D组),每组12只.于兔双侧桡骨中段制作15 mm骨缺损,伤口浸泡于人工海水3h,彻底清创后闭合伤口.高压氧治疗于清创术后立即进行,BMSCs移植于清创术后72 h实施.于伤后第4、8、12周分批处死后取材(每组每批4只兔,共8肢).通过影像学分析、组织学及免疫组化观察,比较各组缺损区修复情况.结果 (1)HE染色观察,兔伤后第4、8周时,D组成骨质量最佳,C组优于B组,A组骨缺损区未见骨性修复,由疤痕组织充填;兔伤后第12周时,D组骨缺损区成骨质量最佳,基本是成熟骨组织.(2)免疫组化观察,兔VEGF及TGF-β1主要表达在成骨细胞、软骨细胞及骨细胞胞浆内,在不同时相点有明显的变化.伤后第4周,两者的表达水平均最高;至第8周,其表达较第4周均明显减弱;至第12周,其表达均进一步回落至低水平.伤后第4、8周,两者的表达水平为D组＞C组＞B组＞A组(各组间两两比较,均为P＜0.05).而兔伤后第12周时,A、B、C、D4组间比较差异无统计学意义(P＞0.05).(3)骨痂灰度值测量,兔伤后第12周时,D组骨痂灰度值为211.84 ±4.31,C组为200.31 ±3.08,B组为152.79±4.43,A组为80.88±3.31,组间差异具有统计学意义(P＜0.05).结论 单一异体BMSCs经皮移植或高压氧治疗修复海水浸泡开放性骨缺损效果并不理想.但是,两者联合应用时具有协同作用,成骨作用明显,是修复海水浸泡开放性骨缺损的可行方法.

abstractsObjective To explore the effects of HBO combined with percutaneous allogeneic bone marrow mesenchymal stem cells (BMSCs) grafting on the repair of rabbit radius open bone defect following seawater immersion.Methods Forty-eight New Zealand rabbits were randomly divided into 4 groups:the blank control group (group A),the(BMSCs) grafting group (group B),the HBO group (group C),and the HBO combined with BMSCs group (group D).A fifteen-millimeter bone defect was made in the mid-section of radius and then was immersed in the artificial seawater for 3 hours,then the wound was closed with debridement.The animals received HBO therapy the moment after completion of debridement.BMSCs grafting was administered 72 hours after debridement.The animals were sacrificed at week 4,8 and 12 to collect the samples (there were 4 rabbits from each batch of the group,and there were altogether 8 limbs).Repairs of the defect areas were compared between groups by means of radiographic scanning,histologic and immunohistochemical analyses.Results HE staining revealed that osteogenesis in the animals of group D at week 4 and 8 was the best,and group C was superior to group B.In the animals of group A,bone repair was not detected in the defect area,which was filled with scar tissue.At week 12 after injury,osteogenesis in the defect areas for the animals of group D was the best,which was mostly mature bone tissue.(2)Immunohistochemical detection indicated that VEGF and TGF-β1 were mostly expressed in the osteoblastic cells,cartilage cells and bone cell cytoplasm,and significant changes could be noted at different time points.At week 4 after injury,the expressions of VEGF and TGF-β1 all reached peak levels.At week 8,their expressions all decreased significantly,as compared with those at week 4 and 12,their expressions all decreased further to a low level.At week 4 and 8 after injury,the expression levels of VEGF and TGF-β1 for the 4 groups were in the following order:group D ＞ group C ＞ group B ＞ group A.Statistical significance could be seen,when comparisons were made between the groups (P ＜0.05).However,at week 12,no significant differences could detected when comparisons were made between the 4 groups (P ＞ 0.05).(3) Osteotylus measurement revealed that osteotylus grayness for the animals of group D,group C,group B and group A at week 12 after injury was 211.84 ± 4.31,200.31 ± 3.08,152.79 ± 4.43 and 80.88 ± 3.31 respectively,and statistical significance could be noted,when comparisons were made between the groups (P ＜ 0.05).Conclusions The effects of simple percutaneous allogeneic bone marrow mesenchymal stem cells grafting or simple HBO therapy on the repair of open bone defect following seawater immersion were not ideal.However,combined applications of these 2 treatment methods could produce synergistic effects,which could promote osteogenetic process of the affected bone and was an applicable method for the repair of open bone defect following seawater immersion.