title: Optimal combination of soluble factors for tissue engineering of permanent cartilage from cultured human chondrocytes.
abstract: Since permanent cartilage has poor self-regenerative capacity, its regeneration from autologous human chondrocytes using a tissue engineering technique may greatly benefit the treatment of various skeletal disorders. However, the conventional autologous chondrocyte implantation is insufficient both in quantity and in quality due to two major limitations: dedifferentiation during a long term culture for multiplication and hypertrophic differentiation by stimulation for the redifferentiation. To overcome the limitations, this study attempted to determine the optimal combination in primary human chondrocyte cultures under a serum-free condition, from among 12 putative chondrocyte regulators. From the exhaustive 2(12) = 4,096 combinations, 256 were selected by fractional factorial design, and bone morphogenetic protein-2 and insulin (BI) were statistically determined to be the most effective combination causing redifferentiation of the dedifferentiated cells after repeated passaging. We further found that the addition of triiodothyronine (T3) prevented the BI-induced hypertrophic differentiation of redifferentiated chondrocytes via the suppression of Akt signaling. The implant formed by the human chondrocytes cultured in atelocollagen and poly(l-latic acid) scaffold under the BI + T3 stimulation consisted of sufficient hyaline cartilage with mechanical properties comparable with native cartilage after transplantation in nude mice, indicating that BI + T3 is the optimal combination to regenerate a clinically practical permanent cartilage from autologous chondrocytes.
authors.affiliation: Departments of Cartilage and Bone Regeneration (Fujisoft), and Sensory and Motor System Medicine, Faculty of Medicine, The University of Tokyo, Hongo 7-3-1, Bunkyo-ku, Tokyo 113-8655, Japan.
authors.forename: Guangyao
authors.initials: G
authors.lastname: Liu
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kawaguchi
authors.forename: Toru
authors.initials: T
authors.lastname: Ogasawara
authors.forename: Yukiyo
authors.initials: Y
authors.lastname: Asawa
authors.forename: Junji
authors.initials: J
authors.lastname: Kishimoto
authors.forename: Tsuguharu
authors.initials: T
authors.lastname: Takahashi
authors.forename: Ung-Il
authors.initials: UI
authors.lastname: Chung
authors.forename: Hisayo
authors.initials: H
authors.lastname: Yamaoka
authors.forename: Hirotaka
authors.initials: H
authors.lastname: Asato
authors.forename: Kozo
authors.initials: K
authors.lastname: Nakamura
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Takato
authors.forename: Kazuto
authors.initials: K
authors.lastname: Hoshi
mesh_terms: D000293:Adolescent; D001705:Bioprosthesis; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000494:pharmacology*; D002356:Cartilage / Q000166:cytology* / Q000378:metabolism; D002357:Cartilage Diseases / Q000628:therapy; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D002648:Child; D019902:Chondrocytes / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation; D004338:Drug Combinations; D005260:Female; D006801:Humans; D007004:Hypoglycemic Agents / Q000494:pharmacology*; D007328:Insulin / Q000494:pharmacology*; D008297:Male; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D012038:Regeneration / Q000187:drug effects; D015398:Signal Transduction / Q000187:drug effects; D023822:Tissue Engineering; D016212:Transforming Growth Factor beta / Q000494:pharmacology*; D014284:Triiodothyronine / Q000494:pharmacology*
chemical_list: C525718:BMP2 protein, human; C525719:Bmp2 protein, mouse; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D004338:Drug Combinations; D007004:Hypoglycemic Agents; D007328:Insulin; D016212:Transforming Growth Factor beta; D014284:Triiodothyronine; D051057:Proto-Oncogene Proteins c-akt