Small molecule inhibitor of TGF-β signaling enables robust osteogenesis of autologous GMSCs to successfully repair minipig severe maxillofacial bone defects

Autor: Aerali Heinayati, Dongyu Bao, Bin Wang, Haiyan Qin, Liudi Wang, Huifen Liu, Xin Tong, Xiaochen Ding, Anyuan Shi
Rok vydání: 2019
Předmět:
0301 basic medicine
Male
Bone Regeneration
Swine
Gingiva
Medicine (miscellaneous)
Bone Morphogenetic Protein 2
Bone Morphogenetic Protein 4
TGF-β signaling
Mice
0302 clinical medicine
Osteogenesis
Transforming Growth Factor beta
Osteogenic differentiation
Medicine
lcsh:QD415-436
Cells
Cultured
lcsh:R5-920
Mice
Inbred BALB C
Communicable disease
Cell Differentiation
Bone defect
030220 oncology & carcinogenesis
Benzamides
Molecular Medicine
Stem cell
lcsh:Medicine (General)
Mice
Nude
Dioxoles
Biochemistry
Genetics and Molecular Biology (miscellaneous)
Bone morphogenetic protein 2
lcsh:Biochemistry
03 medical and health sciences
In vivo
SB431542
Animals
Humans
BMP
Smad3 Protein
Bone regeneration
Cell Proliferation
business.industry
Regeneration (biology)
Research
Mesenchymal stem cell
Mesenchymal Stem Cells
Cell Biology
GMSCs
030104 developmental biology
Apoptosis
Cancer research
business
Zdroj: Stem Cell Research & Therapy
Stem Cell Research & Therapy, Vol 10, Iss 1, Pp 1-15 (2019)
ISSN: 1757-6512
Popis: Background Clinically, for stem cell-based therapy (SCBT), autologous stem cells are considered better than allogenic stem cells because of little immune rejection and no risk of communicable disease infection. However, severe maxillofacial bone defects restoration needs sufficient autologous stem cells, and this remains a challenge worldwide. Human gingival mesenchymal stem cells (hGMSCs) derived from clinically discarded, easily obtainable, and self-healing autologous gingival tissues, have higher proliferation rate compared with autologous bone marrow mesenchymal stem cells (hBMSCs). But for clinical bone regeneration purpose, GMSCs have inferior osteogenic differentiation capability. In this study, a TGF-β signaling inhibitor SB431542 was used to enhance GMSCs osteogenesis in vitro and to repair minipig severe maxillofacial bone defects. Methods hGMSCs were isolated and cultured from clinically discarded gingival tissues. The effects of SB431542 on proliferation, apoptosis, and osteogenic differentiation of hGMSCs were analyzed in vitro, and then, SB431542-treated hGMSCs composited with Bio-Oss® were transplanted into immunocompromised mice subcutaneously to explore osteogenic differentiation in vivo. After that, SB431542-treated autologous pig GMSCs (pGMSCs) composited with Bio-Oss® were transplanted into circular confined defects (5 mm × 12 mm) in minipigs maxillary to investigate severe bone defect regeneration. Minipigs were sacrificed at 2 months and nude mice at 8 weeks to retrieve specimens for histological or micro-CT or CBCT analysis. Effects of SB431542 on TGF-β and BMP signaling in hGMSCs were investigated by Western Blot or qRT-PCR. Results One micromolar of SB431542 treatment induced a robust osteogenesis of hGMSCs in vitro, without adverse effect on apoptosis and growth. In vivo, 1 μM SB431542 treatment also enabled striking osteogenesis of hGMSCs subcutaneously in nude mice and advanced new bone formation of pGMSCs in minipig maxillary bone defect model. In addition, SB431542-treated hGMSCs markedly increased bone-related proteins expression, and BMP2 and BMP4 gene expression. Conversely, SMAD3 protein-dependent TGF-β signal pathway phosphorylation was decreased. Conclusions Our study show that osteogenic differentiation of GMSCs treated with TGF-β signaling inhibitor SB431542 was increased, and SB431542-treated autologous pig GMSCs could successfully repair minipig severe maxillofacial bone defects. This preclinical study brings about a promising large bone regeneration therapeutic potential of autologous GMSCs induced by SB431542 in clinic settings. Electronic supplementary material The online version of this article (10.1186/s13287-019-1281-2) contains supplementary material, which is available to authorized users.
Databáze: OpenAIRE
Externí odkaz:
Nepřihlášeným uživatelům se plný text nezobrazuje