Development of a bioink using exopolysaccharide from Rhizobium sp. PRIM17

Abstract

Biopolymers play a significant role in tissue engineering, including in the formulation of bioinks that require careful selection of the biopolymers having properties ideal for printability and supporting biological entities such as cells. Alginate is one of the most widely explored natural biopolymers for tissue engineering applications due to its biocompatibility, cross-linking ability, hydrophilic nature, and easy incorporation with other polymers. Here, a succinoglycan-like exopolysaccharide (EPS-R17) produced by a bacterial strain Rhizobium sp. PRIM17 was incorporated with alginate for the development of a bioink. The physicochemical characterization of EPS-R17 was performed before formulating the bioink with alginate. The bioink formulation was prepared by mixing different concentrations of EPS with an alginate solution at room temperature under sterile atmosphere. The prepared bioink was characterized for rheological properties, biocompatibility, and a bioplotting experiment was also conducted to mimick the extrusion bioprinting. The EPS-R17 was composed of glucose, galactose, and rhamnose with a molecular weight of 69.98 kDa. It was thermally stable up to 260 °C and showed characteristic FT-IR peaks (1723.3 cm^-1) for succinyl groups. The EPS-R17 showed biocompatibility with keratinocytes (HaCaT), and fibroblasts (HDF) in vitro. The rheological properties of EPS-R17-alginate bioink at different combinations showed shear thinning behavior at 25 and 37 °C. Amplitude sweep measurements showed the gel-like nature of the polymer combinations in the solution system superior to alginate or EPS-R17 alone. The combination of 1 % EPS-R17 and 1.5 % alginate showed good compressive strength and swelling behavior. Extrusion bioprinting mimicked using a bioplotting experiment showed the sustained cell viability in the polymer matrix of EPS-R17-alginate bioink. The results indicate that the EPS-R17 can be used in combination with alginate for bioinks for bioprinting applications for providing physical properties and favorable bioactivities.

Keywords: Alginate; Bioink; Exopolysaccharide; Extrusion bioprinting; Rheology; Rhizobium.