Effect of recombinant adeno-associated BMP-4/7 fusion gene on the biology of BMSCs

Abstract

The aim of the study was to investigate the effect of recombinant adeno-associated virus (AAV) with morphogenetic protein 4/7 fusion gene on the ossification of rabbit bone marrow stromal cells (BMSCs). The genes BMP-4 and BMP-7 were obtained through one-step reverse transcriptase polymerase chain reaction from human placental cells. The BMP-4/7 fusion gene was generated through recombination. The rabbit BMSCs were transfected with the recombinant AAV vectors carrying AAV-BMP-4/7 with various multiplicity of infection (MOI) values. Expression of fusion gene BMP-4/7 protein was determined by the ELISA method. The ossification of cells was evaluated by observing alkaline phosphatase (ALP) and osteocalcin (OC) activity after transfection for 7 and 14 days. We successfully constructed the recombinant AAV with the BMP-4/7 fusion gene. A 29-20 kDa protein was shown by SDS-PAGE electrophoresis following transfection with AAV-BMP-4/7. The fusion protein BMP-4/7 in BMSCs transferred by AAV showed a positive correlation with various MOI values. There was significantly higher ALP and OC activity in the AAV-BMP-4/7 transfection groups than in the AAV-EGFP groups (t(ALP)=896.88, P<0.001, t(OC)=543.24, P<0.01). The fusion gene BMP-4/7 is capable of enhancing the expression of BMPs and possesses significant ossification activity through AAV.

Dental pulp cells that express adeno-associated virus serotype 2-mediated BMP-7 gene enhanced odontoblastic differentiation

“…Hakki et al observed that BMP-7 stimulates both ALP activity and biomineralization in a dose-dependent manner to promote the osteogenic differentiation of human periodontal ligament stem cells 26) . The strong odontogenic effects that we observed in our study are consistent with previous reports that used adeno-associated viral vectors in a variety of cell types 27,28) . Additionally, DSPP and OCN have been used as indicators of odontoblastic differentiation.…”

Overexpression of Bone Morphogenetic Protein-1 Promotes Osteogenesis of Bone Marrow Mesenchymal Stem Cells In Vitro

“…Thus, in the present study, we used lentiviral vectors for transfection. Regarding the other transfection vectors, one study used the Adeno-associated virus (AAV) virus vector system to construct the AAV-BMP-4/7 vector, making the screening of recombinant vectors more straightforward, and they successfully built the recombinant AAV-BMP-4/7 virus that carried the BMP-4/7 fusion gene in a relatively short time [30]. This may provide a new tool for BMPs-related gene research.…”

Genetic Engineering of Mesenchymal Stem Cells for Regenerative Medicine

“…overexpress genes, such as BMP-4, -7 [125] and BMP-6, claimed to be even more potent proosteogenic factors than BMP-2 in the BMP-2-engineered BM-MSCs [126].…”

“…GDF-5 [102][103][104] TIMP-1 [106] Bcl-2 [101] BONE FORMATION BMP-2 [86][109-111] BMP-2/SDF-1β [113] BMP-2/HIF-1α [114] BMP-2/Ang-1 [47] BMP-2/VEGF [116][117][118] BMP-2/miR-148b [142] BMP-4/-6/-7 [125,126] VEGF [119,120] Epo [121] IGF-I [128,129] OGP [127] Leptin [130] PDGF [131] ephrinB2 [132] ITGA2 [133] NELL1 [134] β-catenin [135] HI Cb TA Fo Na LIVER REGENERATION VEGF [144] ATP7B [155] HN HN…”