Applying In Situ Ionic Crosslinking in Bioprinting Using Algae Cells

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Abstract
Bioprinting using algae cells has many potential applications including tissue engineering, environmental engineering, contaminant removal from water, and establishing space habitats. In extrusion-based bioprinting, bioink needs to be crosslinked after being extruded from the nozzle for printed constructs to first achieve and then maintain adequate shape fidelity. Crosslinking methods used in reported studies on algae-contained bioinks include both photo-crosslinking and ionic crosslinking. This paper reports a preliminary study where the coaxial nozzle-based in situ ionic crosslinking method was used in bioprinting of algae cells without additional crosslinking of printed samples for the first time. In comparison with photo-crosslinking, in situ ionic crosslinking can minimize bioink preparation time and complexity, eliminate cells’ exposure to ultraviolet radiation, and reduce the number of post-printing steps. In this preliminary study, the bioink was an alginate solution containing algae (Chlorella vulgaris) cells, and the crosslinking solution was a calcium chloride solution. The coaxial nozzle had two nozzles: inner and outer nozzles. In printing, the bioink was delivered through the outer nozzle while the crosslinking solution was delivered through the inner nozzle. The shape of the printed samples was a square block with dimensions of 30 × 30 × 10 mm. It was observed that, 9 days after printing, the algae cells grew within the printed samples, and the samples could keep their shapes relatively well. Many knowledge gaps exist regarding the effects of input variables in bioprinting of algae cells using this method. This paper discusses future research directions to fill these knowledge gaps.

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