Scaffold‐ and serum‐free hypertrophic cartilage tissue engineering as an alternative approach for bone repair. Issue 7 (16th February 2020)

• Record Type: Journal Article
• Authors: Kronemberger, Gabriela S.; Dalmônico, Gisele M. L.; Rossi, André L.; Leite, Paulo Emílio Correa; Saraiva, Antonio M.; Beatrici, Anderson; Silva, Karina Ribeiro; Granjeiro, José Mauro; Baptista, Leandra Santos
• Is Part Of: Artificial organs. Volume 44:Issue 7(2020)
• Page Start: E288
• Page End: E299
• Publication Date: 2020-02-16
• Subjects: adipose stem/stromal cells -- bone -- hypertrophic cartilage -- scaffold‐free -- serum‐free -- spheroids -- tissue engineering; Artificial organs -- Periodicals; 617.956
• DOI: 10.1111/aor.13637
• Languages: English
• ISSNs: 0160-564X

Abstract

Abstract: Human adipose stem/stromal cell (ASC) spheroids were used as a serum‐free in vitro model to recapitulate the molecular events and extracellular matrix organization that orchestrate a hypertrophic cartilage phenotype. Induced‐ASC spheroids (ø = 450 µm) showed high cell viability throughout the period of culture. The expression of collagen type X alpha 1 chain ( COLXA1 ) and matrix metallopeptidase 13 ( MMP‐13 ) was upregulated at week 2 in induced‐ASC spheroids compared with week 5 ( P < .001) evaluated by quantitative real‐time PCR. In accordance, secreted levels of IL‐6 ( P < .0001), IL‐8 ( P < .0001), IL‐10 ( P < .0001), bFGF ( P < .001), VEGF ( P < .0001), and RANTES ( P < .0001) were the highest at week 2. Strong in situ staining for collagen type X and low staining for TSP‐1 was associated with the increase of hypertrophic genes expression at week 2 in induced‐ASC spheroids. Collagen type I, osteocalcin, biglycan, and tenascin C were detected at week 5 by in situ staining, in accordance with the highest expression of alkaline phosphatase ( ALPL ) gene and the presence of calcium deposits as evaluated by Alizarin Red O staining. Induced‐ASC spheroids showed a higher force required to compression at week 2 ( P < .0001). The human ASC spheroids under serum‐free inducer medium and normoxic culture conditions were induced to a hypertrophic cartilage phenotype, opening a new perspective to recapitulate endochondral ossification in vivo.