Enhanced critical-size calvarial bone healing by ASCs engineered with Cre/loxP-based hybrid baculovirus.
Calvarial bone repair remains challenging for adults. Although adipose-derived stem cells (ASCs) hold promise to heal bone defects, use of ASCs for critical-size calvarial bone repair is ineffective. Stromal cell-derived factor 1 (SDF-1) is a chemokine capable of triggering stem cell migration. Although recombinant SDF-1 protein is co-delivered with other molecules including BMP-2 to facilitate calvarial bone repair, these approaches did not yield satisfactory healing. This study aimed to exploit a newly developed Cre/loxP-based hybrid baculovirus for efficient gene delivery and prolonged transgene expression in ASCs. We demonstrated that transduction of rat ASCs with the hybrid Cre/loxP-based baculovirus enabled robust and sustained expression of functional BMP-2 and SDF-1. Expression of BMP-2 or SDF-1 alone failed to effectively induce rat ASCs osteogenesis and healing of critical-size calvarial bone defects. Nonetheless, prolonged BMP-2/SDF-1 co-expression in ASCs synergistically activated both Smad and ERK1/2 pathways and hence potentiated the osteogenesis. Consequently, transplantation of the hybrid baculovirus-engineered, BMP-2/SDF-1-expressing ASCs/scaffold constructs potently healed the critical-size (6 mm) calvarial bone defects (filling ≈70% of defect volume), which considerably outperformed the calvarial bone repair using BMP-2/SDF-1 delivered with biomaterial-based scaffolds. These data implicated the potential of Cre/loxP-based hybrid baculovirus vector for ASCs engineering and calvarial bone healing.
Department of Chemical Engineering, National Tsing Hua University, Hsinchu, 300, Taiwan.
Shih-Chun
SC
Lo
Department of Chemical Engineering, National Tsing Hua University, Hsinchu, 300, Taiwan.
Kuei-Chang
KC
Li
Center for Tissue Engineering, Chang Gung Memorial Hospital, Taoyuan, 333, Taiwan; Department of Orthopaedic, Chang Gung Memorial Hospital, Taoyuan, 333, Taiwan.
Yu-Han
YH
Chang
Department of Chemical Engineering, National Tsing Hua University, Hsinchu, 300, Taiwan.
Mu-Nung
MN
Hsu
Department of Chemical Engineering, National Tsing Hua University, Hsinchu, 300, Taiwan.
Li-Yu
LY
Sung
Department of Chemical Engineering, National Tsing Hua University, Hsinchu, 300, Taiwan.
Truong Anh
TA
Vu
Department of Chemical Engineering, National Tsing Hua University, Hsinchu, 300, Taiwan. Electronic address: ychu@mx.nthu.edu.tw.
Yu-Chen
YC
Hu
D053687:Adult Stem Cells / Q000502:physiology* / Q000637:transplantation* / Q000821:virology; D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics / Q000378:metabolism*; D001861:Bone Regeneration / Q000502:physiology; D060846:Cell Engineering / Q000379:methods*; D002478:Cells, Cultured; D054377:Chemokine CXCL12 / Q000235:genetics / Q000378:metabolism*; D005260:Female; D051381:Rats; D017207:Rats, Sprague-Dawley; D012887:Skull Fractures / Q000473:pathology / Q000503:physiopathology / Q000628:therapy*; D014161:Transduction, Genetic / Q000379:methods*; D016896:Treatment Outcome
C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; C515255:CXCL12 protein, human; D054377:Chemokine CXCL12
Adipose-derived stem cells; BMP-2; Baculovirus; Calvarial bone healing; Cre/loxP; SDF-1