In vitro generation of a scaffold-free tissue-engineered construct (TEC) derived from human synovial mesenchymal stem cells: biological and mechanical properties and further chondrogenic potential.
The purpose of this study was to characterize a tissue-engineered construct (TEC) generated with human synovial mesenchymal stem cells (MSCs). MSCs were cultured in medium with ascorbic acid 2-phosphate (Asc-2P) and were subsequently detached from the substratum. The detached cell/matrix complex spontaneously contracted to develop a basic TEC. The volume of the TEC assessed by varying initial cell density showed that it was proportional to initial cell densities up to 4 x 10(5) cells/cm(2). Assessment of the mechanical properties of TEC using a custom device showed that the load at failure and stiffness of the constructs significantly increased with time of culture in the presence of Asc-2P, while in the absence of Asc-2P, the constructs were mechanically weak. Thus, the basic TEC possesses sufficiently self-supporting mechanical properties in spite of not containing artificial scaffolding. TEC further cultured in chondrogenic media exhibited positive alcian blue staining with elevated expression of chondrogenic marker genes. Based on these findings, such human TEC may be a promising method to promote cartilage repair for future clinical application.
Ando
Wataru
W
Department of Orthopaedics, Osaka University Graduate School of Medicine, Osaka, Japan.
Tateishi
Kosuke
K
Katakai
Daisuke
D
Hart
David A
DA
Higuchi
Chikahisa
C
Nakata
Ken
K
Hashimoto
Jun
J
Fujie
Hiromichi
H
Shino
Konsei
K
Yoshikawa
Hideki
H
Nakamura
Norimasa
N
D000328:Adult; D001205:Ascorbic Acid / Q000031:analogs & derivatives / Q000378:metabolism; D001696:Biomechanical Phenomena; D055396:Bone Morphogenetic Protein 2 / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D020219:Chondrogenesis / Q000187:drug effects; D003094:Collagen / Q000096:biosynthesis; D003470:Culture Media; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D008297:Male; D008422:Materials Testing; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects; D013583:Synovial Membrane / Q000166:cytology*; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds
D055396:Bone Morphogenetic Protein 2; D003470:Culture Media; C011669:ascorbate-2-phosphate; D003094:Collagen; D001205:Ascorbic Acid