Crosslinking of an oesophagus acellular matrix tissue scaffold

Abstract

The oesophagus acellular matrix (EAM) tissue-scaffold has the potential to serve as the foundation for a tissue-engineered oesophagus for repair of ablative defects. Similar to all collagen-based biomaterials, the EAM is subject to enzymatic degradation in vivo. The introduction of exogenous crosslinks to collagen molecules via glutaraldehyde (Glu) is the most accepted method of stabilizing collagen biomaterials, but fixation with Glu incurs adverse effects. Genipin (Gp), a naturally occurring crosslinking agent, has shown to be effective at improving the stability of collagen-based biomaterials with less cytotoxicity and reduced in vivo inflammatory responses than Glu. The aim of this study was to show that crosslinking with Gp improves the stability of the EAM while maintaining minimal biological reactivity and preserving EAM regeneration potential in a rat model. EAMs were crosslinked with Gp and Glu. Uncrosslinked EAMs served as controls. Denaturation temperature measurement and burst-pressure measurement after enzymatic degradation assays were used to determine the effectiveness of crosslinking on in vitro stability. Subcutaneous allograft implantation and oesophageal epithelial cell-seeding studies assessed the crosslinking effects on biological reactivity and regeneration potential, respectively. Both Gp and Glu improved EAM stability. After 30 days of implantation, the EAM elicited a minimal inflammatory response and crosslinking did not increase inflammation. Gp-crosslinked EAMs supported epithelial adhesion and proliferation while Glu-crosslinked EAMs did not. Gp improves the stability of the EAM while maintaining minimal biological reactivity and preserving EAM epithelial proliferation capacity, yielding a tissue scaffold that may form the basis of a durable and biocompatible tissue-engineered oesophagus.

Genipin, a Cross-linking Agent, Promotes Odontogenic Differentiation of Human Dental Pulp Cells

“…It also has been found to be an effective collagen cross-linking agent (16). In previous studies, genipin showed significantly lower cytotoxicity and inflammatory reaction than did glutaraldehyde (17)(18)(19), and there have been many trials to use genipin as a collagen cross-linking agent in tissue engineering fields (20)(21)(22)(23)(24).…”

Genipin crosslinking elevates the strength of electrochemically aligned collagen to the level of tendons

“…However, use of these enzymes in cultured tissue mimics is expensive. Chemical agents like glutaraledehyde (GA) have been widely used to stabilize collagenous matrices (Sung et al, 1999b); yet, toxicity is a real concern (Bhrany et al, 2008;Chang et al, 2002;Sung et al, 1999a). Irradiation with ultraviolet (UV) light has been used to crosslink collagen (Cornwell et al, 2007;Dong et al, 2005;Menter et al, 2001), but it has limited use to crosslink cellular tissues and tissue equivalents because of the potential of UV-mediated DNA degradation.…”

Esophageal tissue engineering

“…[74,75]. Les MD sont donc extrêmement employées dans l'ingénierie tissulaire vasculaire, valvulaire cardiaque, cutanée, et urologique [76].…”

“…La même équipe a par la suite stabilisé leurs scaffolds par « cross-linking » avec du glutaraldéhyde ou de la génipine afin de leurs conférer des forces mécaniques satisfaisantes. Le glutaraldéhyde engendrait une toxicité cellulaire non retrouvée avec la génipine [83]. Les DM oesophagiens permettent également la croissance de tissus musculaires lisses.…”

“…Les cellules utilisées peuvent être différenciées ou « à potentiel souches ». En ingénierie tissulaire oesophagienne [133], les cellules utilisées étaient principalement différenciées et représentées par des cellules épithéliales [78,82,83], des cellules musculaires lisses [84,89], des cellules musculaires striées [86,134] et des fibroblastes [64]. L'utilisation de cellules souches a été décrite dans l'ingénierie tissulaire oesophagienne [93].…”