The healing of critical-size calvarial bone defects in rat with rhPDGF-BB, BMSCs, and β-TCP scaffolds.
Platelet-derived growth factor-BB (PDGF-BB) plays important roles in regenerating damaged tissue. In this study we investigated the effects of a tissue-engineered bone combined with recombinant human PDGF-BB (rhPDGF-BB), bone marrow stem cells (BMSCs) and β-tricalcium phosphate (β-TCP) to repair critical-size calvarial bone defects in rat. Proliferation and osteogenic differentiation of BMSCs treated with different concentration rhPDGF-BB (0, 10, and 50 ng/ml) was evaluated by MTT, alkaline phosphatase (ALP) activity, alizarin red staining and real-time quantitative PCR (RT-qPCR) analysis of osteogenic gene. BMSCs were then combined with rhPDGF-BB-loaded β-TCP and transplanted into 5 mm calvarial bone defects. The new bone formation and mineralization was evaluated by micro-computerized tomography (Micro-CT) and histological analysis at week 8 after operation. It was observed that the proliferation of BMSCs treated with rhPDGF-BB was enhanced with a time- and dose- dependent manner. There were increased ALP activity, mineralized deposition and elevated mRNA levels of osteogenic gene for BMSCs treated with rhPDGF-BB, particularly in the 50 ng/ml group. Histological analysis showed new bone formation and mineralization in the rhPDGF-BB/BMSCs/β-TCP group was significantly higher than BMSCs/β-TCP, rhPDGF-BB/β-TCP, and β-TCP alone group (P < 0.05). In conclusion, rhPDGF-BB/BMSCs/β-TCP is a promising tissue-engineered bone for craniofacial bone regeneration.
Department of Prosthodontics, School of Stomatology, Ninth People's Hospital, Shanghai Key Laboratory of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Ling
L
Xu
Kaige
K
Lv
Wenjie
W
Zhang
Xiuli
X
Zhang
Xinquan
X
Jiang
Fuqiang
F
Zhang
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D000077214:Becaplermin; D001842:Bone and Bones / Q000187:drug effects* / Q000473:pathology; D002130:Calcium Phosphates; D002478:Cells, Cultured; D017931:DNA Primers; D005456:Fluorescent Dyes; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D006801:Humans; D008297:Male; D020574:Proto-Oncogene Proteins c-sis / Q000494:pharmacology*; D051381:Rats; D011916:Rats, Inbred F344; D060888:Real-Time Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000494:pharmacology; D014057:Tomography, X-Ray Computed; D014945:Wound Healing / Q000187:drug effects*
D002130:Calcium Phosphates; D017931:DNA Primers; D005456:Fluorescent Dyes; D020574:Proto-Oncogene Proteins c-sis; D011994:Recombinant Proteins; C485817:beta-tricalcium phosphate; D000077214:Becaplermin; D000469:Alkaline Phosphatase