Genipin Cross-Linked Fibrin Hydrogels for in vitro Human Articular Cartilage Tissue-Engineered Regeneration

Abstract

Our objective was to examine the potential of a genipin cross-linked human fibrin hydrogel system as a scaffold for articular cartilage tissue engineering. Human articular chondrocytes were incorporated into modified human fibrin gels and evaluated for mechanical properties, cell viability, gene expression, extracellular matrix production and subcutaneous biodegradation. Genipin, a naturally occurring compound used in the treatment of inflammation, was used as a cross-linker. Genipin cross-linking did not significantly affect cell viability, but significantly increased the dynamic compression and shear moduli of the hydrogel. The ratio of the change in collagen II versus collagen I expression increased more than 8-fold over 5 weeks as detected with real-time RT-PCR. Accumulation of collagen II and aggrecan in hydrogel extracellular matrix was observed after 5 weeks in cell culture. Overall, our results indicate that genipin appeared to inhibit the inflammatory reaction observed 3 weeks after subcutaneous implantation of the fibrin into rats. Therefore, genipin cross-linked fibrin hydrogels can be used as cell-compatible tissue engineering scaffolds for articular cartilage regeneration, for utility in autologous treatments that eliminate the risk of tissue rejection and viral infection.

Fibrin matrices: The versatile therapeutic delivery systems

“…Introduction of additional crosslinks in the fibrin gels with crosslinking agents like glutaraldehyde lowers pore size as well as susceptibility to proteolysis [33,66,67]. Genipin, isolated from Gardenia jasminoides fruits, was used successfully by Dare et al [68] as a less toxic alternative to glutaraldehyde for crosslinking of fibrin matrices and induce chondrogenic differentiation of human primary articular chondrocytes. Sell et al [69] crosslinked electrospun fibrinogen scaffolds with three bi-fuctional reagents-glutaraldehyde, 1-ethyl-3-(3, di-methyl aminopropyl) carbodiamide (EDC) and genipin and investigated their peak stress modulus and in vitro degradation.…”

Evaluating osteochondral defect repair potential of autologous rabbit bone marrow cells on type II collagen scaffold

“…Recently, Dare et al fabricated a genipin cross-linked human fibrin hydrogel system as a scaffold for articular cartilage tissue engineering. Their results showed that genipin appeared to inhibit the inflammatory reaction observed 3 weeks after subcutaneous implantation of the fibrin hydrogel into rats (Dare et al 2009). Our results were in agreement with their reports showing that the repair site did not result in the inflammation during experimental period.…”

Triple-Layered Cell Sheet for Tissue-Engineering the Synovial Membrane of the Temporomandibular Joint

“…Numerous studies have focused on tissue-engineering the condylar cartilage and the disc of the TMJ [Almarza and Athanasiou, 2004;Detamore and Athanasiou, 2005;Dare et al, 2009;Hillel et al, 2009;Morotomi et al, 2012]. This study, for the first time, focused on creating a tissueengineered SM of the TMJ with an original method, because we hold that an SM with abnormal function will cause continuous damage to the cartilage and disc rather than nourish them.…”