title: Effect of BMPs and Wnt3a co-expression on the osteogenetic capacity of osteoblasts.
abstract: In the present study, third‑generation autologous‑inactivated bone morphogenic protein 2 (BMP2), BMP4, BMP6, BMP7, BMP9 and Wnt3a lentiviral vectors were constructed and integrated into the genome of MC3T3‑E1 murine mesenchymal stem cells (MMSCs) to produce osteoinductive factor gene‑modified MMSCs. The transfection efficiency of each osteoinductive factor was then determined by detecting the expression levels of runt related transcription factor 2 (Runx2) mRNA. The cotransfection with combinations of two lentiviruses was performed, and the expression levels of bone γ‑carboxyglutamate protein and alkaline phosphatase in the MC3T3‑E1 cell culture supernatant were detected. The expression level of Runx2 mRNA was detected by reverse transcription‑polymerase chain reaction, and western blotting was performed to detect the protein expression levels of BMP2, BMP4, BMP6, BMP7, BMP9 and Wnt3a. The results demonstrated that the recombinant lentiviruses were successfully transfected into MC3T3‑E1 cells. The relative expression levels of Runx2 mRNA were greatest in the BMP2 group, sequentially followed by the BMP4, BMP9, BMP7, Wnt3a and BMP6 groups. The results of cotransfection of MC3T3‑E1 cells (a total of 8 groups) demonstrated that BMP‑2 and BMP‑7 exhibited the highest cotransfection efficiency. Western blot analysis demonstrated that following BMP2 and BMP7 cotransfection of MC3T3‑E1 cells, the protein expression levels of BMP2, BMP4, BMP6, BMP7, BMP9 and Wnt3a were increased compared with control cells. In conclusion, the third‑generation lentiviral vectors effectively improved the osteogenic efficiencies of MC3T3‑E1 cells, which provided an important theoretical basis and therapeutic strategy for bone reconstruction and tissue engineering.
authors.affiliation: Department of Orthopaedics, The General Hospital of Tianjin Medical University, Tianjin 300052, P.R. China.
authors.forename: Wendong
authors.initials: W
authors.lastname: Ruan
authors.affiliation: Department of Orthopaedics, The General Hospital of Tianjin Medical University, Tianjin 300052, P.R. China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Xue
authors.affiliation: Department of Orthopaedics, The General Hospital of Tianjin Medical University, Tianjin 300052, P.R. China.
authors.forename: Yaqi
authors.initials: Y
authors.lastname: Zong
authors.affiliation: Department of Orthopaedics, The General Hospital of Tianjin Medical University, Tianjin 300052, P.R. China.
authors.forename: Chao
authors.initials: C
authors.lastname: Sun
mesh_terms: D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000096:biosynthesis / Q000235:genetics*; D055415:Bone Morphogenetic Protein 4 / Q000096:biosynthesis / Q000235:genetics*; D055418:Bone Morphogenetic Protein 6 / Q000096:biosynthesis / Q000235:genetics*; D055419:Bone Morphogenetic Protein 7 / Q000096:biosynthesis / Q000235:genetics*; D001861:Bone Regeneration / Q000235:genetics*; D002454:Cell Differentiation / Q000235:genetics*; D050659:Core Binding Factor Alpha 1 Subunit / Q000096:biosynthesis; D018507:Gene Expression Regulation, Developmental; D055427:Growth Differentiation Factor 2 / Q000096:biosynthesis / Q000235:genetics*; D016086:Lentivirus / Q000235:genetics; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism; D051379:Mice; D010006:Osteoblasts / Q000166:cytology; D010012:Osteogenesis / Q000235:genetics*; D023822:Tissue Engineering; D014162:Transfection; D060509:Wnt3A Protein / Q000096:biosynthesis / Q000235:genetics*
chemical_list: C525719:Bmp2 protein, mouse; C525810:Bmp4 protein, mouse; C525817:Bmp6 protein, mouse; D055396:Bone Morphogenetic Protein 2; D055415:Bone Morphogenetic Protein 4; D055418:Bone Morphogenetic Protein 6; D055419:Bone Morphogenetic Protein 7; D050659:Core Binding Factor Alpha 1 Subunit; C526108:Gdf2 protein, mouse; D055427:Growth Differentiation Factor 2; C493716:Runx2 protein, mouse; D060509:Wnt3A Protein; C555511:Wnt3a protein, mouse; C525820:bmp7 protein, mouse