Using Stem Cell Screening-enriching-combining Technique For Promoting Bone Regeneration

BECKGROUND:Bone defect caused by various reasons is a common clinical problem which demands graft treatment.However,there are many defects in current bone repair materials.Bone marrow mesenchymal stem cells(MSC)are regarded as the seed cells of tissue engineering and have received more and more attention.In this study,we used the stem cell screening-enriching-combining technique to construct graft material for promoting bone regeneration.OBJECTIVES:The objectives were to explore the rapid-adhesion characteristic of bone marrow MSC;to analyze the effect of stem cell screening-enriching-combining technique on screening and enriching MSCs;and to observe the curative effect of the technique.METHODS:Human bone marrow was processed by stem cell screening-enriching-combining technique: bone marrow filtered through beta-tricalcium phosphate(?-TCP)granules as it circulated in a circulation pipeline,MSC/?-TCP composites were grafted into bone defect sites of patients.Before and after filtration,MSC number and nucleated cells number were counted;nucleated cells vitality was measured by trypan blue excluding rate;bone marrow samples were tested by bacteriological examination to assess the safety.Confocal microscopy was performed to observe MSC adhesion on the ?-TCP.Forty-two patients with bone defects were followed up to observe the curative effect of the technique.RESULTS:By stem cell screening-enriching-combining technique,85.53% bone marrow MSC was screened and enriched on the ?-TCP granules.There is no significant change in the number of nucleated cells(P > 0.05).There was no significant change in the activity of nucleated cells before and after filtration(P > 0.05).Only two bone marrow samples of one patient were contaminated,but treatment outcome was good.Confocal microscopy showed that MSC adhered,proliferated,and formed colonies on the ?-TCP.All patients with bone defects were effectively repaired;the average healing time was 5.04±2.11 months.CONCLUSIONS:Bone marrow MSCs have the characteristic rapid adhesion.By stem cell screening-enriching-combining technique,most of bone marrow MSC could be effectively screened,enriched and combined with ?-TCP simultaneously.The MSC/?-TCP composites prepared by the technique can effectively promote human bone regeneration.