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Comprehensive characterization of chondrocyte cultures in plasma and whole blood biomatrices for cartilage tissue engineering
Author(s) -
Schulz Ronny M.,
Haberhauer Marcus,
Zernia Göran,
Pösel Claudia,
Thümmler Christian,
Somerson Jeremy S.,
Huster Daniel
Publication year - 2014
Publication title -
journal of tissue engineering and regenerative medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.835
H-Index - 72
eISSN - 1932-7005
pISSN - 1932-6254
DOI - 10.1002/term.1561
Subject(s) - chondrocyte , aggrecan , cartilage , extracellular matrix , tissue engineering , chemistry , biomedical engineering , agarose , matrix (chemical analysis) , chondrogenesis , type ii collagen , hyaline cartilage , cartilage oligomeric matrix protein , pathology , articular cartilage , anatomy , biochemistry , biology , chromatography , osteoarthritis , medicine , alternative medicine
Many synthetic polymers and biomaterials have been used as matrices for 3D chondrocyte seeding and transplantation in the field of cartilage tissue engineering. To develop a fully autologous carrier for chondrocyte cultivation, we examined the feasibility of allogeneic plasma and whole blood‐based matrices and compared them to agarose constructs. Primary articular chondrocytes isolated from 12‐month‐old pigs were embedded into agarose, plasma and whole blood matrices and cultivated under static‐free swelling conditions for up to four weeks. To evaluate the quality of the synthesized extracellular matrix (ECM), constructs were subjected to weekly examinations using histological staining, spectrophotometry, immunohistochemistry and biochemical analysis. In addition, gene expression of cartilage‐specific markers such as aggrecan, Sox9 and collagen types I, II and X was determined by RT‐PCR. Chondrocyte morphology was assessed via scanning electron microscopy and viability staining, including proliferation and apoptosis assays. Finally, 13 C NMR spectroscopy provided further evidence of synthesis of ECM components. It was shown that chondrocyte cultivation in allogeneic plasma and whole‐blood matrices promoted sufficient chondrocyte viability and differentiation behaviour, resulting in neo‐formation of a hyaline‐like cartilage matrix. Copyright © 2012 John Wiley & Sons, Ltd.