title: MicroRNA-26a-modified adipose-derived stem cells incorporated with a porous hydroxyapatite scaffold improve the repair of bone defects.
abstract: Tissue-engineered bone substitutes are frequently used to repair bone defects. Adipose-derived stem cells (ASCs) are a promising source of cells for repairing bone tissue, however, insufficient osteogenic potency remains the main obstacle for their application. The present study aimed to enhance the osteogenic potency of ASCs by transfection of microRNA (miR)‑26a, a novel osteogenic and angiogenic promoting miRNA. An inverted fluorescence microscope was used to observe transfection efficiency, while a scanning electron microscope was used to detect morphological alterations. Cell proliferation was monitored continuously for 7 days using a Cell Counting kit‑8 assay. Osteogenic differentiation was determined by reverse transcription quantitative polymerase chain reaction, alkaline phosphatase (ALP) staining, collagen secretion and extracellular matrix (ECM) mineralization. ASCs were incorporated with a porous hydroxyapatite (HA) scaffold to create a novel tissue‑engineered bone substitute and inserted into the critical tibia defect of rats. New bone formation was evaluated by hematoxylin and eosin and Masson's trichrome staining. The results demonstrated that miR‑26a was successfully delivered into the cytoplasm, while the morphology and proliferation of ASCs were not significantly altered. Osteogenic‑associated genes were markedly upregulated and ALP production, collagen secretion and ECM mineralization were all increased following transfection of miR‑26a. Histological evaluation demonstrated that the modified cells accompanied with a porous HA scaffold markedly promoted new bone formation within the defective area. In conclusion, miR‑26a transfection significantly improved the osteogenic potency of ASCs suggesting that modified ASCs incorporated with a HA scaffold may be used as a potential bone substitute.
authors.affiliation: Department of Orthopaedics, No. 113 Hospital of PLA, Ningbo, Zhejiang 315000, P.R. China.
authors.forename: Zhenlin
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Department of Orthopaedics, Xijing Hospital of PLA, Xi'an, Shaanxi 710032, P.R. China.
authors.forename: Dawei
authors.initials: D
authors.lastname: Zhang
authors.affiliation: Department of Otorhinolaryngology, No. 113 Hospital of PLA, Ningbo, Zhejiang 315000, P.R. China.
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Hu
authors.affiliation: Department of Orthopaedics, No. 113 Hospital of PLA, Ningbo, Zhejiang 315000, P.R. China.
authors.forename: Jiwei
authors.initials: J
authors.lastname: Cheng
authors.affiliation: Department of Orthopaedics, No. 113 Hospital of PLA, Ningbo, Zhejiang 315000, P.R. China.
authors.forename: Chuanmeng
authors.initials: C
authors.lastname: Zhuo
authors.affiliation: Department of Orthopaedics, No. 113 Hospital of PLA, Ningbo, Zhejiang 315000, P.R. China.
authors.forename: Xiancong
authors.initials: X
authors.lastname: Fang
authors.affiliation: Department of Orthopaedics, No. 113 Hospital of PLA, Ningbo, Zhejiang 315000, P.R. China.
authors.forename: Yongming
authors.initials: Y
authors.lastname: Xing
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000818:Animals; D018786:Bone Substitutes / Q000737:chemistry; D002478:Cells, Cultured; D017886:Durapatite / Q000737:chemistry; D035683:MicroRNAs / Q000235:genetics* / Q000627:therapeutic use; D010012:Osteogenesis; D016062:Porosity; D051381:Rats; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D013977:Tibia / Q000293:injuries* / Q000502:physiology; D023822:Tissue Engineering; D054457:Tissue Scaffolds / Q000737:chemistry*; D014162:Transfection
chemical_list: D018786:Bone Substitutes; C556274:MIRN26 microRNA, rat; D035683:MicroRNAs; D017886:Durapatite