title: Demineralized bone matrix combined bone marrow mesenchymal stem cells, bone morphogenetic protein-2 and transforming growth factor-β3 gene promoted pig cartilage defect repair.
abstract: OBJECTIVE
  To investigate whether a combination of demineralized bone matrix (DBM) and bone marrow mesenchymal stem cells (BMSCs) infected with adenovirus-mediated- bone morphogenetic protein (Ad-BMP-2) and transforming growth factor-β3 (Ad-TGF-β3) promotes the repair of the full-thickness cartilage lesions in pig model.
  
  
  METHODS
  BMSCs isolated from pig were cultured and infected with Ad-BMP-2(B group), Ad-TGF-β3 (T group), Ad-BMP-2 + Ad-TGF-β3(BT group), cells infected with empty Ad served as a negative group(N group), the expression of the BMP-2 and TGF-β3 were confirmed by immunofluorescence, PCR, and ELISA, the expression of SOX-9, type II collagen(COL-2A), aggrecan (ACAN) in each group were evaluated by real-time PCR at 1w, 2w, 3w, respectively. The chondrogenic differentiation of BMSCs was evaluated by type II collagen at 21d with immunohistochemical staining. The third-passage BMSCs infected with Ad-BMP-2 and Ad-TGF-β3 were suspended and cultured with DBM for 6 days to construct a new type of tissue engineering scaffold to repair full-thickness cartilage lesions in the femur condyles of pig knee, the regenerated tissue was evaluated at 1,2 and 3 months after surgery by gross appearance, H&E, safranin O staining and O'driscoll score.
  
  
  RESULTS
  Ad-BMP-2 and Ad-TGF-β3 (BT group) infected cells acquired strong type II collagen staining compared with Ad-BMP-2 (B group) and Ad-TGF-β3 (T group) along. The Ad-BMP-2 and Ad-TGF-β3 infected BMSCs adhered and propagated well in DBM and the new type of tissue engineering scaffold produced hyaline cartilage morphology containing a stronger type II collagen and safranin O staining, the O'driscoll score was higher than other groups.
  
  
  CONCLUSIONS
  The DBM compound with Ad-BMP-2 and Ad-TGF-β3 infected BMSCs scaffold has a good biocompatibility and could well induce cartilage regeneration to repair the defects of joint cartilage. This technology may be efficiently employed for cartilage lesions repair in vivo.
authors.affiliation: Department of Sports Medicine, First Affiliated Hospital of Kunming Medical University, Kunming, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Sports Medicine, First Affiliated Hospital of Kunming Medical University, Kunming, China.
authors.forename: Yanlin
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Diabetology, First Affiliated Hospital of Kunming Medical University, Kunming, China.
authors.forename: Rui
authors.initials: R
authors.lastname: Han
authors.affiliation: Department of Sports Medicine, First Affiliated Hospital of Kunming Medical University, Kunming, China.
authors.forename: Chuan
authors.initials: C
authors.lastname: He
authors.affiliation: Department of Sports Medicine, First Affiliated Hospital of Kunming Medical University, Kunming, China.
authors.forename: Guoliang
authors.initials: G
authors.lastname: Wang
authors.affiliation: Department of Sports Medicine, First Affiliated Hospital of Kunming Medical University, Kunming, China.
authors.forename: Jianwei
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Sports Medicine, First Affiliated Hospital of Kunming Medical University, Kunming, China.
authors.forename: Jiali
authors.initials: J
authors.lastname: Zheng
authors.affiliation: Department of Orthopedics, West Virginia University, Morgantown, United States of America.
authors.forename: Ming
authors.initials: M
authors.lastname: Pei
authors.forename: Mei
authors.initials: M
authors.lastname: Pei
authors.affiliation: Department of Orthopedics, Warren Alpert Medical School of Brown University, Providence, United States of America.
authors.forename: Lei
authors.initials: L
authors.lastname: Wei
mesh_terms: D000256:Adenoviridae / Q000378:metabolism; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D018775:Bone Demineralization Technique; D001854:Bone Marrow Cells / Q000166:cytology; D001857:Bone Matrix / Q000737:chemistry* / Q000648:ultrastructure; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics* / Q000627:therapeutic use; D002358:Cartilage, Articular / Q000473:pathology*; D002454:Cell Differentiation; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D020219:Chondrogenesis; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation; D015316:Genetic Therapy; D006801:Humans; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology; D034421:Sus scrofa; D014161:Transduction, Genetic; D053782:Transforming Growth Factor beta3 / Q000235:genetics* / Q000627:therapeutic use
chemical_list: D015415:Biomarkers; D055396:Bone Morphogenetic Protein 2; D053782:Transforming Growth Factor beta3