Generation of genipin cross-linked fibrin-agarose hydrogel tissue-like models for tissue engineering applications

Abstract

Generation of biomimetic and biocompatible artificial tissues is the basic research objective for tissue engineering (TE). In this sense, the biofabrication of scaffolds that resemble the tissues' extracellular matrix (ECM) is an essential aim in this field. Uncompressed and nanostructured fibrin-agarose hydrogels (FAH and NFAH respectively) emerged as promising scaffold in TE, but its structure and biomechanical properties must be improved in order to broad their TE applications. Here we generated and characterized novel membrane-like models with increased structural and biomechanical properties based on the chemical cross-linking of FAH and NFAH with genipin (GP at 0.1, 0.25, 0.5 and 0.75%). Furthermore, scaffolds were subjected to rheological (G, G', G" modulus), ultrastructural and ex vivo biocompatibility analyses. Results showed that all GP concentrations increased the stiffness (G) and especially the elasticity (G') of FAH and NFAH. Ultrastructural analyses demonstrated that GP and nanostructuration of FAH allowed controlling the porosity of FAH. In addition, biological studies revealed that higher concentration of GP (0.75%) started to compromise the cell function and viability. Finally, this study demonstrated the possibility to generate natural and biocompatible FAH and NFAH with improved structural and biomechanical properties by using 0.1% to 0.5% of GP. However, further in vivo studies are needed in order to demonstrate the biocompatibility, biodegradability and regeneration capability of these cross-linked scaffolds.

Plant-Derived Biomaterials: A Review of 3D Bioprinting and Biomedical Applications

“…Agarose is derived from red seaweed and has garnered interest in the plant derived 3D bioprinting sphere due to its ability to be prepared as a thermal-reversible gel (Zarrintaj et al, 2018). It exhibits many of the properties of alginate, emulating the extracellular matrix with high water uptake, but with the advantage of hydrogen bond-mediated self-gelation without the need for potentially toxic crosslinking agents such as genipin (Campos et al, 2018). It has however been noted to be adversely mechanically affected by the presence of cells: cells within agarose diminish the gel strength due to interference with the hydrogen bonding required for crosslinking and gelation (Shoichet et al, 1996).…”

Fibrin Matrices as (Injectable) Biomaterials: Formation, Clinical Use, and Molecular Engineering

“…Genipin derives from a naturally occurring compound (geniposide, a glycoside present in various fruits) via β‐glucosidase treatment, and can react with amines at two sites of its molecular structure and then further cross‐link a material through oxidative oligomerization (Figure C). Due to the unselective nature of these reactions, genipin crosslinking can increase both fibrin modulus and its resistance to degradation, but can also allow its covalent mixing with other materials, for example, silk, alginate, and decellularized ECM particles, thereby minimizing phase separation phenomena. Poly(methyl methacrylate) (PMMA) beads have also been incorporated in fibrin prior to the addition of genipin; after cross‐linking, and dissolution of the beads in acetone, this allows to introduce a controlled microporosity in fibrin gels …”

Magneto-Responsive Hydrogels: Preparation, Characterization, Biotechnological and Environmental Applications

“…They are also promising materials in the development of bio-artificial tissue of corneas, oral mucosa, skin, cartilage and abdominal-wall. 117 However, one should note that the viscoelastic modulus of scaffolds plays a crucial role on the cell response. For instance, for osteogenic differentiation, rigid (modulus ca.…”