Effects of hyperbaric oxygen combined with percutaneous allogeneic bone marrow mesenchymal stem cells grafting on the repair of rabbit radius open bone defect following seawater immersion

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Objective 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. Key words: Seawater immersion; Open bone defect; Bone marrow mesenchymal stem cell; Hyperbaric oxygen