Abstract
BackgroundChondrocyte transplantation to address cartilage damage is an established solution. Because hyaluronic acid (HA) is an essential component for homeostasis of the cartilage, in order to arrive at methodologies to utilize its advantages in cell-based therapies, we compared the HA retention capability of a thermoreversible gelation polymer scaffold-based environment (3D-TGP) with conventional in vitro cell culture methodologies. MethodsChondrocytes derived from osteoarthritis-affected knee joint cartilage of elderly patients were used and accomplished in three phases. In Phase I, the levels of HA secreted by chondrocytes were measured in culture supernatant. In Phase II, retention capacity of externally added HA was quantified indirectly by measuring the HA released in culture supernatant, and in Phase III, the expression of CD44 on cells was analysed by immunohistochemistry. ResultsIn Phase I, the average HA in the 3D supernatant was 3% that of 2D. In phase II, 80% of externally added HA was detected in the 2D on day 7, while in 3D-TGP, only 0.1% was released until day 21. In Phase III, 2D yielded individual cells that started degenerating from the third week; in 3D-TGP cells grew for a longer duration, formed a tissue-like architecture with extracellular matrix with significantly intense staining of CD44 than 2D. ConclusionThe capability of the 3D-TGP culture environment to retain HA and support chondrocytes to grow with a tissue-like architecture expressing higher HA content is considered advantageous as it serves as an in vitro culture platform that enables tissue engineering of cartilage tissue with native hyaline phenotype and higher HA expression. The in vitro environment being conducive, based on this data, we also recommend that the TGP be tried as an encapsulation material in clinical studies of chondrocyte implantation for optimal clinical outcome.
Highlights
Cell-based therapies with and without scaffolds, in vitro engineered cartilage tissue, gene-enhanced cartilage regeneration and physical modulation in vivo to direct stem cells present in the body to become chondrogenic cells are the different approaches in regenerative therapies for cartilage repair [1].The biomaterials used for tissue engineering of articular cartilage support the chondrocytes by providing the optimal physical and biochemical conditions for developing and maintaining the hyaline phenotype of the chondrocytes, which influences the quality of repair after transplantation
A thermo-reversible gelation polymer (TGP)-hydrogel based in vitro culture system has been previously employed for cartilage tissue engineering of bovine [4], rabbit [5], and human chondrocytes, [6] which has helped to firmly maintain the hyaline phenotype for a longer period than in cartilage tissue engineered by other methods, both in vitro [4,5] and in vivo [5], apart from enabling osteoarthritis (OA)-derived chondrocytes to express pluripotency markers, such as UEA-1 in vitro [7]
We evaluated whether the TGP scaffold is capable of retaining hyaluronic acid (HA) and its implications on HA secretion and expression during in vitro culture of chondrocytes derived from osteoarthritic knees for evaluating its potentials as both an in vitro culture scaffold as well as an encapsulation material during transplantation for cartilage regeneration
Summary
Cell-based therapies with and without scaffolds, in vitro engineered cartilage tissue, gene-enhanced cartilage regeneration and physical modulation in vivo to direct stem cells present in the body to become chondrogenic cells are the different approaches in regenerative therapies for cartilage repair [1].The biomaterials used for tissue engineering of articular cartilage support the chondrocytes by providing the optimal physical and biochemical conditions for developing and maintaining the hyaline phenotype of the chondrocytes, which influences the quality of repair after transplantation. A thermo-reversible gelation polymer (TGP)-hydrogel based in vitro culture system has been previously employed for cartilage tissue engineering of bovine [4], rabbit [5], and human chondrocytes, [6] which has helped to firmly maintain the hyaline phenotype for a longer period than in cartilage tissue engineered by other methods, both in vitro [4,5] and in vivo [5], apart from enabling osteoarthritis (OA)-derived chondrocytes to express pluripotency markers, such as UEA-1 in vitro [7]. Conclusion: The capability of the 3D-TGP culture environment to retain HA and support chondrocytes to grow with a tissue-like architecture expressing higher HA content is considered advantageous as it serves as an in vitro culture platform that enables tissue engineering of cartilage tissue with native hyaline phenotype and higher HA expression.
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