Improved calvarial bone repair by hASCs engineered with Cre/loxP-based baculovirus conferring prolonged BMP-2 and MiR-148b co-expression.
Repairing large calvarial bone defects remains a challenging task. Previously, it was discovered that that miR-148b, when acting in concert with bone morphogenetic protein 2 (BMP-2), enhanced the osteogenesis of human adipose-derived stem cells (hASCs) and improved calvarial bone healing in nude mice. However, the molecular target of miR-148b remained elusive. Here it is revealed that miR-148b directly targets NOG, whose gene product (noggin) is an antagonist to BMPs and negatively regulates BMP-induced osteogenic differentiation and bone formation. A new Cre/loxP-based baculovirus system was employed to drive prolonged BMP-2 and miR-148b overexpression in hASCs, wherein the BMP-2 overexpression induced noggin expression but the concurrent miR-148b expression downregulated noggin, thus relieving the negative regulatory loop and ameliorating hASC osteogenesis without hindering hASC proliferation or triggering appreciable cytotoxicity. Implantation of the engineered hASCs coexpressing BMP-2 and miR-148b into nude mice enabled substantial repair of critical-size calvarial bone defects (4 mm diameter) at 12 weeks post-transplantation, filling 83% of the defect area, 75% of bone volume and restoring the bone density to 89% of the original bone density. Such superior healing effects indicate the potential of the Cre/loxP-based baculovirus-mediated BMP-2/miR-148b expression for calvarial bone repair. Copyright © 2016 John Wiley & Sons, Ltd.
Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
Kuei-Chang
KC
Li
Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
Shih-Chun
SC
Lo
Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
Li-Yu
LY
Sung
Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
Ya-Hsin
YH
Liao
Center for Tissue Engineering, Chang Gung Memorial Hospital, Taoyuan, Taiwan.|Department of Orthopaedic, Chang Gung Memorial Hospital, Taoyuan, Taiwan.
Yu-Han
YH
Chang
Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
Yu-Chen
YC
Hu
D000273:Adipose Tissue / Q000378:metabolism* / Q000473:pathology; D000818:Animals; D016367:Baculoviridae; D055396:Bone Morphogenetic Protein 2 / Q000096:biosynthesis* / Q000235:genetics; D001861:Bone Regeneration; D005260:Female; D015870:Gene Expression; D064593:Heterografts; D006801:Humans; D019426:Integrases / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D035683:MicroRNAs / Q000096:biosynthesis* / Q000235:genetics; D012886:Skull / Q000293:injuries / Q000378:metabolism / Q000473:pathology; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000378:metabolism* / Q000473:pathology; D014161:Transduction, Genetic
C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; C550570:MIRN148 microRNA, human; D035683:MicroRNAs; C045073:Cre recombinase; D019426:Integrases
BMP-2; Cre/loxP; adipose-derived stem cells; baculovirus; calvarial bone defect; miR-148b; noggin