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Demineralized Bone Matrix Combined Bone Marrow Mesenchymal Stem Cells, Bone Morphogenetic Protein-2 and Transforming Growth Factor-[beta]3 Gene Promoted Pig Cartilage Defect Repair.(Research Article)

Objectives 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-[beta]3 (Ad-TGF-[beta]3) promotes the repair of the full-thickness... Full description

Journal Title: PLoS ONE Dec 29, 2014, Vol.9(12)
Main Author: Wang, Xin
Other Authors: Li, Yanlin , Han, Rui , He, Chuan , Wang, Guoliang , Wang, Jianwei , Zheng, Jiali , Pei, Mei , Wei, Lei
Format: Electronic Article Electronic Article
Language: English
Subjects:
ID: ISSN: 1932-6203 ; DOI: 10.1371/journal.pone.0116061
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title: Demineralized Bone Matrix Combined Bone Marrow Mesenchymal Stem Cells, Bone Morphogenetic Protein-2 and Transforming Growth Factor-[beta]3 Gene Promoted Pig Cartilage Defect Repair.(Research Article)
format: Article
creator:
  • Wang, Xin
  • Li, Yanlin
  • Han, Rui
  • He, Chuan
  • Wang, Guoliang
  • Wang, Jianwei
  • Zheng, Jiali
  • Pei, Mei
  • Wei, Lei
subjects:
  • Tissue Engineering
  • Collagen
  • Bone Morphogenetic Proteins
  • Adenoviruses
  • Enzyme-Linked Immunosorbent Assay
  • Transforming Growth Factors
  • Swine
ispartof: PLoS ONE, Dec 29, 2014, Vol.9(12)
description: Objectives 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-[beta]3 (Ad-TGF-[beta]3) 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-[beta]3 (T group), Ad-BMP-2 + Ad-TGF-[beta]3(BT group), cells infected with empty Ad served as a negative group(N group), the expression of the BMP-2 and TGF-[beta]3 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-[beta]3 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, HE, safranin O staining and Odriscoll score. Results Ad-BMP-2 and Ad-TGF-[beta]3 (BT group) infected cells acquired strong type II collagen staining compared with Ad-BMP-2 (B group) and Ad-TGF-[beta]3 (T group) along. The Ad-BMP-2 and Ad-TGF-[beta]3 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-[beta]3 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.
language: English
source:
identifier: ISSN: 1932-6203 ; DOI: 10.1371/journal.pone.0116061
fulltext: fulltext
issn:
  • 1932-6203
  • 19326203
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titleDemineralized Bone Matrix Combined Bone Marrow Mesenchymal Stem Cells, Bone Morphogenetic Protein-2 and Transforming Growth Factor-[beta]3 Gene Promoted Pig Cartilage Defect Repair.(Research Article)
creatorWang, Xin ; Li, Yanlin ; Han, Rui ; He, Chuan ; Wang, Guoliang ; Wang, Jianwei ; Zheng, Jiali ; Pei, Mei ; Wei, Lei
ispartofPLoS ONE, Dec 29, 2014, Vol.9(12)
identifierISSN: 1932-6203 ; DOI: 10.1371/journal.pone.0116061
subjectTissue Engineering ; Collagen ; Bone Morphogenetic Proteins ; Adenoviruses ; Enzyme-Linked Immunosorbent Assay ; Transforming Growth Factors ; Swine
descriptionObjectives 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-[beta]3 (Ad-TGF-[beta]3) 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-[beta]3 (T group), Ad-BMP-2 + Ad-TGF-[beta]3(BT group), cells infected with empty Ad served as a negative group(N group), the expression of the BMP-2 and TGF-[beta]3 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-[beta]3 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, HE, safranin O staining and Odriscoll score. Results Ad-BMP-2 and Ad-TGF-[beta]3 (BT group) infected cells acquired strong type II collagen staining compared with Ad-BMP-2 (B group) and Ad-TGF-[beta]3 (T group) along. The Ad-BMP-2 and Ad-TGF-[beta]3 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-[beta]3 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.
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titleDemineralized Bone Matrix Combined Bone Marrow Mesenchymal Stem Cells, Bone Morphogenetic Protein-2 and Transforming Growth Factor-[beta]3 Gene Promoted Pig Cartilage Defect Repair.(Research Article)
descriptionObjectives 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-[beta]3 (Ad-TGF-[beta]3) 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-[beta]3 (T group), Ad-BMP-2 + Ad-TGF-[beta]3(BT group), cells infected with empty Ad served as a negative group(N group), the expression of the BMP-2 and TGF-[beta]3 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-[beta]3 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, HE, safranin O staining and Odriscoll score. Results Ad-BMP-2 and Ad-TGF-[beta]3 (BT group) infected cells acquired strong type II collagen staining compared with Ad-BMP-2 (B group) and Ad-TGF-[beta]3 (T group) along. The Ad-BMP-2 and Ad-TGF-[beta]3 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-[beta]3 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.
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titleDemineralized Bone Matrix Combined Bone Marrow Mesenchymal Stem Cells, Bone Morphogenetic Protein-2 and Transforming Growth Factor-[beta]3 Gene Promoted Pig Cartilage Defect Repair.(Research Article)
authorWang, Xin ; Li, Yanlin ; Han, Rui ; He, Chuan ; Wang, Guoliang ; Wang, Jianwei ; Zheng, Jiali ; Pei, Mei ; Wei, Lei
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atitleDemineralized Bone Matrix Combined Bone Marrow Mesenchymal Stem Cells, Bone Morphogenetic Protein-2 and Transforming Growth Factor-[beta]3 Gene Promoted Pig Cartilage Defect Repair
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abstractObjectives 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-[beta]3 (Ad-TGF-[beta]3) 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-[beta]3 (T group), Ad-BMP-2 + Ad-TGF-[beta]3(BT group), cells infected with empty Ad served as a negative group(N group), the expression of the BMP-2 and TGF-[beta]3 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-[beta]3 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, HE, safranin O staining and Odriscoll score. Results Ad-BMP-2 and Ad-TGF-[beta]3 (BT group) infected cells acquired strong type II collagen staining compared with Ad-BMP-2 (B group) and Ad-TGF-[beta]3 (T group) along. The Ad-BMP-2 and Ad-TGF-[beta]3 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-[beta]3 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.
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date2014-12-29