Abstract
The insufficient healing capacity of articular cartilage necessitates mechanically functional biologic tissue replacements. Using cells to form biomimetic cartilage implants is met with the challenges of cell scarcity and donor site morbidity, requiring expanded cells that possess the ability to generate robust neocartilage. To address this, this study assesses the effects of expansion medium supplementation (bFGF, TFP, FBS) and self-assembled construct seeding density (2, 3, 4 million cells/5 mm dia. construct) on the ability of costochondral cells to generate biochemically and biomechanically robust neocartilage. Results show TFP (1 ng/mL TGF-β1, 5 ng/mL bFGF, 10 ng/mL PDGF) supplementation of serum-free chondrogenic expansion medium enhances the post-expansion chondrogenic potential of costochondral cells, evidenced by increased glycosaminoglycan content, decreased type I/II collagen ratio, and enhanced compressive properties. Low density (2 million cells/construct) enhances matrix synthesis and tensile and compressive mechanical properties. Combined, TFP and Low density interact to further enhance construct properties. That is, with TFP, Low density increases type II collagen content by over 100%, tensile stiffness by over 300%, and compressive moduli by over 140%, compared with High density. In conclusion, the interaction of TFP and Low density seeding enhances construct material properties, allowing for a mechanically functional, biomimetic cartilage to be formed using clinically relevant costochondral cells.
Highlights
Articular cartilage injury manifests in joint pain and dysfunction as repair tissue is unable to recapitulate native biochemical and biomechanical properties [1]
Tissue engineering efforts are met with the challenges of 1) identifying a source of healthy cells associated with minimal donor site morbidity and 2) developing conditions to expand these cells while maintaining their ability to generate biomechanically functional tissue
Monolayer Cell Morphology and Proliferation Cell morphology in adherent monolayer culture is illustrated in Morphology, Histology, and Immunohistochemistry After 4 wks culture in non-adherent agarose wells, costal chondrocytes generated robust neocartilage
Summary
Articular cartilage injury manifests in joint pain and dysfunction as repair tissue is unable to recapitulate native biochemical and biomechanical properties [1]. Disorders of the temporomandibular joint (TMJ), including disc perforation and displacement, have been associated with progressive articular cartilage degeneration and the development of osteoarthritis. Tissue engineering may present a more conservative approach to replace the damaged articular surface with functional, autologous neocartilage. Tissue engineering efforts are met with the challenges of 1) identifying a source of healthy cells associated with minimal donor site morbidity and 2) developing conditions to expand these cells while maintaining their ability to generate biomechanically functional tissue. Addressing these challenges may lead to a therapeutic approach with long-term regenerative potential
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