Abstract
Objective To investigate the biological characteristics of human umbilical cord mesenchymal stem cells (hUCMSC) transfected with.bone morphogenetic protein-2 (BMP-2) gene in vitro and in vivo.Methods hUCMSC were cultured in vitro.Based on the different culture media added,three groups were designed:blank group (10% FCS/DMEM),control group (10% FCS/DMEM + empty lentiviral particles),experimental group (10% FCS/DMEM + BMP-2 lentiviral particles).Cell differentiation was examined by alkaline phosphatase (ALP) measurement kit.The expression of osteopontin ( OPN ),type 1 collagen ( COL1 ) and BMP-2 was detected by using Western boltting.Von kossa staining method was used to study the calcification effects. In the rat muscle bag, the complex containing BMP-2-transfected hUCMSCs/COL1 was transplanted.The ectopic bone in the implanted area was observed by the CT 3D reconstruction and hematoxylin and eosin (HE) staining.Results The positive rate of hUCMSC transfected with BMP-2 gene was (90.95 ± 4.35 ) %.As compared with the blank group and control group,the proliferation in experimental group was promoted at 5th and 7th day after culture (P < 0.05 ).The ALP activity was increased from (26.492 ±7.105) U/g·prot at 3rd day to (38.625 ± 11.592) U/g·prot at 5th day,even to (49.732 ± 13.068) U/g·prot at 14th day in experimental group as compared with other two groups (P < 0.05).The COL1,OPN and BMP-2 proteins were overexpressed in experimental group,but weakly expressed in the blank group and control group ( P < 0.05 ).Many mineralized nodes were observed 18 days after transfection of hUCMSC with BMP-2 gene.The ectopic bone formed in the rat muscle bag implanted with BMP-2-transfected hUCMSCs/COL1.Conclusion Transfection of BMP-2 gene can promote osteogenic potential of hUCMSCs. Key words: Bone morphogenetic protein-2; Umbilical cord mesenchymal stem cell; Osteogenic differntiation; Transfection
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