Abstract
Abstract Background: Recombinant human bone morphogenetic proteins (rhBMPs) have been characterized especially chondrogenic and osteogenic activity both in vitro and in vivo studies. However, delivery of more than one growth factor by sustained release carrier to orthopedic site has yet been questionable in terms of efficacy and synergism. Objective: Evaluate osteoinductivity and synergistic effect of rhBMP-2 and -7 using absorbable collagen sponge (ACS) carrier system in vivo. Methods: cDNA of BMP-2 and -7 active domains were cloned and expressed in Escherichia coli BL21 StarTM (DE3) using pRSETc expression system. Then, the purified rhBMPs were loaded onto ACS and evaluated by in vivo rat subcutaneous bioassay. Two and eight weeks postoperatively, all treated groups were histologically verified for evidence of new bone formation and neovascularization by hematoxylin-eosin staining and light microscopy. Results: The Wistar rat treated with rhBMP-2 or -7/ACS exhibited new bone formation, compared to ACS control. The group treated with ACS supplemented with both rhBMP-2 and -7 significantly showed the osteoid matrix very well-organized into trabeculae-like structure with significant blood vessel invasion. Conclusion: The osteogenic induction of rhBMPs was combined with ACS carrier in the in vivo bioassay. In addition, the combination of both two potent recombinant osteoinductive cytokines, rhBMP-2 and -7, with ACS carrier demonstrated synergistic effect and might be a more promising and effective choice for therapeutic applications.
Highlights
Recombinant human bone morphogenetic proteins have been characterized especially chondrogenic and osteogenic activity both in vitro and in vivo studies
We examined the osteogenic induction of absorbable collagen sponge (ACS) combined with either Recombinant human bone morphogenetic proteins (rhBMPs), compared to only ACS and ACS with either rhBMP alone in the in vivo rat subcutaneous implant model
After verifying inframe insertion by DNA sequencing, rhBMP cDNAs were subcloned from pGEM T-easy vector into pRSETc expression vector and induced by 0.5 mM isopropyl β-Dthiogalactopyranoside (IPTG) to express heterologous protein in E. coli BL21 StarTM (DE3) under T7 RNA polymerase/promoter system
Summary
Recombinant human bone morphogenetic proteins (rhBMPs) have been characterized especially chondrogenic and osteogenic activity both in vitro and in vivo studies. Objective: Evaluate osteoinductivity and synergistic effect of rhBMP-2 and -7 using absorbable collagen sponge (ACS) carrier system in vivo. The purified rhBMPs were loaded onto ACS and evaluated by in vivo rat subcutaneous bioassay. Results: The Wistar rat treated with rhBMP-2 or -7/ACS exhibited new bone formation, compared to ACS control. Conclusion: The osteogenic induction of rhBMPs was combined with ACS carrier in the in vivo bioassay. The combination of both two potent recombinant osteoinductive cytokines, rhBMP-2 and -7, with ACS carrier demonstrated synergistic effect and might be a more promising and effective choice for therapeutic applications
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