Fish scale containing alginate dialdehyde-gelatin bioink for bone tissue engineering

Abstract

The development of biomaterial inks suitable for biofabrication and mimicking the physicochemical properties of the extracellular matrix is essential for bioprinting technology in tissue engineering applications. The use of animal-derived proteinous materials, such as jellyfish collagen, or fish scale gelatin, has become an important pillar in biomaterial ink design to increase the bioactivity of hydrogels. However, besides the extraction of proteinous structures, the use of structurally intact fish scales as an additive could increase biocompatibility and bioactivity of hydrogels due to its organic (collagen and gelatin) and inorganic (hydroxyapatite) contents, while simultaneously enhancing mechanical strength in 3D printing applications. To test this hypothesis, we present here a composite biomaterial ink composed of fish scale (FS) and alginate dialdehyde (ADA)-gelatin (GEL) for three-dimensional (3D) bioprinting applications. We fabricate 3D cell-laden hydrogels using mouse pre-osteoblast MC3T3-E1 cells. We evaluate the physicochemical and mechanical properties of FS incorporated ADA-GEL biomaterial inks as well as the bioactivity and cytocompatibility of cell-laden hydrogels. Due to the distinctive collagen orientation of the FS, the compressive strength of the hydrogels significantly increases with increasing FS particle content. Addition of FS also provides a tool to tune hydrogel stiffness. FS particles were homogeneously incorporated into the hydrogels. Particle-matrix integration was confirmed via scanning electron microscopy. FS incorporation in the ADA-GEL matrix increased the osteogenic differentiation of MC3T3-E1 cells in comparison to pristine ADA-GEL, as FS incorporation led to increased ALP activity and osteocalcin secretion of MC3T3-E1 cells. Due to the significantly increased stiffness and supported osteoinductivity of the hydrogels, FS structure as a natural collagen and hydroxyapatite source contributed to the biomaterial ink properties for bone engineering applications. Our findings indicate that ADA-GEL/FS represents a new biomaterial ink formulation with great potential for 3D bioprinting applications, and FS is confirmed as promising additive for bone tissue engineering applications.

Biopolymers for Tissue Engineering: Crosslinking, Printing Techniques, and Applications

“…In addition, a composite biomaterial ink composed of Fish Scale and alginate dialdehyde (ADA)-Gel for the fabrication of 3D cell-laden hydrogels using mouse pre-osteoblast MC3T3-E1 cells was revealed as a suitable biomaterial ink formulation with great potential for 3D bioprinting for bone TE applications [62].…”

3D bioprinting of mouse pre-osteoblasts and human MSCs using bioinks consisting of gelatin and decellularized bone particles

“…With high surface area and unique structural shape, inorganic additives can significantly influence the properties of 3D scaffolds with tailored functionalities. In bone tissue engineering, inorganic materials-reinforced bioink formulations have been reported with successful outcomes in many studies using different materials and designs [4,[24][25][26][27][28]. Utilizing both organic and inorganic materials in a composite structure is a suitable approach to mimic the bone tissue extracellular matrix (ECM).…”

Three-Dimensional Artificial Skin Construct Bioprinted with a Marine-Based Biocomposite

“…However, further in vitro cellular investigation using keratinocytes must be conducted to outline the potential for a tissue-engineered skin graft. Additionally, Özenler et al have demonstrated that the incorporation of fish scale into alginate dialdehyde-bioink can significantly enhance the mechanical properties of the 3D graft and evoke a favorable biological response to cells …”