title: Type II collagen and glycosaminoglycan expression induction in primary human chondrocyte by TGF-β1.
abstract: BACKGROUND
  A localized non-surgical delivery of allogeneic human chondrocytes (hChonJ) with irradiated genetically modified chondrocytes (hChonJb#7) expressing transforming growth factor-β1 (TGF-β1) showed efficacy in regenerating cartilage tissue in our pre-clinical studies and human Phase I and II clinical trials. These previous observations led us to investigate the molecular mechanisms of the cartilage regeneration.
  
  
  METHODS
  Genetically modified TGF-β1preprotein was evaluated by monitoring cell proliferation inhibition activity. The effect of modified TGF-β1 on chondrocytes was evaluated based on the type II collagen mRNA levels and the amount of glycosaminoclycan (GAG) formed around chondrocytes, which are indicative markers of redifferentiated chondrocytes. Among the cartilage matrix components produced by hChonJb#7 cells, type II collagen and proteoglycan, in addition to TGF-β1, were also tested to see if they could induce hChonJ redifferentiation. The ability of chondrocytes to attach to artificially induced defects in rabbit cartilage was tested using fluorescent markers.
  
  
  RESULTS
  Throughout these experiments, the TGF-β1 produced from hChonJb#7 was shown to be equally as active as the recombinant human TGF-β1. Type II collagen and GAG production were induced in hChonJ cells by TGF-β1 secreted from the irradiated hChonJb#7 cells when the cells were co-cultured in micro-masses. Both hChonJ and hChonJb#7 cells could attach efficiently to the defect area in the rabbit cartilage.
  
  
  CONCLUSIONS
  This study suggests that the mixture (TG-C) of allogeneic human chondrocytes (hChonJ) and irradiated genetically modified human chondrocytes expressing TGF-β1 (hChonJb#7) attach to the damaged cartilage area to produce type II collagen-GAG matrices by providing a continuous supply of active TGF-β1.
authors.affiliation: TissueGene Inc., 9605 Medical Center Dr. Suite 200, Rockville, MD, 20850, USA. hyoon@tissuegene.com.
authors.forename: Hyun Joo
authors.initials: HJ
authors.lastname: Yoon
authors.affiliation: Department of Rehabilitation and Personal training, Konyang University, 158, Gwanjeodong-ro, Daejeon, Seo-gu, Korea. bum3340@hanmail.net.
authors.forename: Suk Bum
authors.initials: SB
authors.lastname: Kim
authors.affiliation: TissueGene Inc., 9605 Medical Center Dr. Suite 200, Rockville, MD, 20850, USA. dsomaiya@tissuegene.com.
authors.forename: Dhara
authors.initials: D
authors.lastname: Somaiya
authors.affiliation: TissueGene Inc., 9605 Medical Center Dr. Suite 200, Rockville, MD, 20850, USA. mnoh@tissuegene.com.
authors.forename: Moon Jong
authors.initials: MJ
authors.lastname: Noh
authors.affiliation: Kolon Life Science, 13 Kolon-ro, Gwacheon-si, Gyeonggi-do, Korea. klschoi@kolon.com.
authors.forename: Kyoung-Baek
authors.initials: KB
authors.lastname: Choi
authors.affiliation: Kolon Life Science, 13 Kolon-ro, Gwacheon-si, Gyeonggi-do, Korea. cllim@kolon.com.
authors.forename: Chae-Lyul
authors.initials: CL
authors.lastname: Lim
authors.affiliation: Kolon Life Science, 13 Kolon-ro, Gwacheon-si, Gyeonggi-do, Korea. yalli74@kolon.com.
authors.forename: Hyeon-Youl
authors.initials: HY
authors.lastname: Lee
authors.affiliation: Kolon Life Science, 13 Kolon-ro, Gwacheon-si, Gyeonggi-do, Korea. csgr@kolon.com.
authors.forename: Yeon-Ju
authors.initials: YJ
authors.lastname: Lee
authors.affiliation: TissueGene Inc., 9605 Medical Center Dr. Suite 200, Rockville, MD, 20850, USA. yiyoungsuk@yahoo.com.
authors.forename: Youngsuk
authors.initials: Y
authors.lastname: Yi
authors.affiliation: TissueGene Inc., 9605 Medical Center Dr. Suite 200, Rockville, MD, 20850, USA. ortholee@tissuegene.com.
authors.forename: Kwan Hee
authors.initials: KH
authors.lastname: Lee
mesh_terms: D000818:Animals; D019556:COS Cells; D002357:Cartilage Diseases / Q000378:metabolism / Q000473:pathology / Q000628:therapy; D002358:Cartilage, Articular / Q000378:metabolism / Q000473:pathology / Q000601:surgery; D002448:Cell Adhesion; D002522:Chlorocebus aethiops; D019902:Chondrocytes / Q000187:drug effects / Q000473:pathology* / Q000637:transplantation; D020219:Chondrogenesis / Q000187:drug effects; D018920:Coculture Techniques; D024043:Collagen Type II / Q000378:metabolism*; D004195:Disease Models, Animal; D005269:Femur / Q000378:metabolism / Q000473:pathology / Q000601:surgery; D015316:Genetic Therapy / Q000379:methods; D006025:Glycosaminoglycans / Q000378:metabolism*; D056945:Hep G2 Cells; D006801:Humans; D007223:Infant; D009154:Mutation; D011817:Rabbits; D012038:Regeneration; D014162:Transfection; D053773:Transforming Growth Factor beta1 / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D015854:Up-Regulation
chemical_list: D024043:Collagen Type II; D006025:Glycosaminoglycans; C508960:TGFB1 protein, human; D053773:Transforming Growth Factor beta1