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BACKGROUND: The bone marrow stem cell (MSC) transplant treatment have the obvious superiority to tradition graft treatment for bone nonunion, but how to obtain the concentrated and highly effective bone marrow mesenchymal stem cell, as well as the dose-effect relations to fracture healing need further discussions. OBJECTIVE: To observe the curative effect of bone nonunion by using autologous MSC transplant treatment, and to compare with autologous iliac bone graft.DESIGN, TIME AND SETTING: Randomized controlled analysis was performed from January 1999 to June 2005 in the Affiliated Second Hospital of Hebei Northern College.PARTICIPANTS: The admitting 140 patients with humerus and tibia fracture were divided into 2 groups at random, autologous iliac bone graft group and autologous MSC transplant group, with 70 patients in each group. METHODS: Under aseptic condition, autologous MSC transplant group received puncture through posterior superior iliac spine, extracting bone marrow 10-20 mL from different spots, separating MSC using the density gradient centrifugation method, and counting as 4×10~9 nucleated cells/mL under the microscope for later use. In the autologous iliac bone graft group, bone fracture end was implanted with the suitable amount of iliac bone, while autologous MSC transplant group with the mixture of decalcified bone matrix and MSC, followed by suture. After the transplantation, external fixation may assist for 4-6 weeks according to the fixed degree of internal fixation.MAIN OUTCOME MEASURES: ① Bone callus formation and pain conditions in 2 groups at different time points after transplantation. ② Comparison of bone healing time between 2 groups. ③ Adverse events and side effects.RESULTS: According to intention-treatment analysis, experimental adopted 140 patients of humerus and tibia fractures, who all entered the final analysis. ① Bone callus formation and pain at different time points post-surgery: At 1 month after transplantation, bone callus formation in the fracture end was not obvious in autogenous iliac bone graft group, and could be seen in autologous MSC transplant group, both groups of fractures exhibited tenderness. At 2 months after transplantation, bone callus formation was observed in autogenous iliac bone graft group, fracture tenderness was relieved compared with the previous condition; in autologous MSC transplant group, a large number of bone callus formed, fracture tenderness was not obvious. At 3 months after transplantation, there were a large number of bone callus formations in autogenous iliac bone graft group, with slight fracture tenderness; in the autologous MSC transplant group, continuous bone callus formation appeared, without fracture tenderness. ② Bone healing time: The average healing time of autologous MSC transplant group was significantly shorter than autogenous iliac bone graft group [(5.5±1.5), (8.0±2.0) months, P < 0.05]. ③ Adverse events and side effects: Except 4 patients had iliac bone pain, all patients during the treatment had no infection and other complications, there were no re-fracture occurred at the follow-up of 8 months.CONCLUSION: The autologous MSC transplant treatment of exhibits a short duration and good effect for bone non-union, has obvious advantages over traditional bone graft.
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@#Objective To investigate the clinical effect of percutaneous autogenous bone marrow stem cell transplant on bone nonunion. Methods 140 patient with bones nonunion were divided into the autogenous bone transplants (A) group and the autogenous bone marrow stem cell transplant (B) group, with 70 cases in each group. They were assessed 2 and 5 months after operation. Results A few calluses could be observed at the end of fracture in 42 cases in group A 2 months after operation, and massive callus formation could be observed in 46 cases 5 months after operation. However, massive callus formation could be observed in 60 cases in group B 2 months after operation, and continual callus formation could be observed in 45 cases 5 months after operation. The mean healing time was (8.4±1.8) months in group A, and (6.5±2.0) months in group B (P<0.05). No side-effects have been observed during the treatment. Conclusion The percutaneous autogenous bone marrow stem cell transplant is more effective on bone nonunion compared with the traditional treatment.
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AIM:To construct recombinant adenovirus vector carrying human miR-133a and study its expression in human mesenchymal stem cells(hMSCs).METHODS:The PCR product containing miR-133a was amplified from human genomic DNA and inserted into the adenoviral shuttle vector pAdTrack-CMV.Then the recombinant shuttle plasmid linearized by pmeⅠwas cotransformed into competent E.coli.BJ5183 with the adenoviral backbone plasmid pAdEasy-1 to generate the recombinant adenovirus vector rAd-mir-133a.rAd-mir-133a was then packaged and amplified in human embryonic kidney 293(HEK293) cells.The purified rAd-miR-133a was used to infect the hMSCs and the expression of miR-133a was detected by non-quantitative RT-PCR and real-time PCR.RESULTS:The recombinant adenovirus shuttle vector pAdTrack-CMV-miR-133a was constructed and verified by restriction endonuclease analysis and DNA sequence analysis.rAd-miR-133a was successfully packaged and amplified in HEK293 cells.The transcriptions of primary miR-133a and mature miR-133a were over-expressed in the hMSCs infected with rAd-miR-133a.CONCLUSION:The recombinant adenovirus vector carrying human miR-133a is successfully constructed,which lay a foundation for miR-133a function study.