Introduction3D-bioprinted preclinical tumour model enables the fabrication of spatially defined tumour microenvironment (TME) mimicking the in-vivo situation. It is challenging to opt a bioink that is biocompatible, with minimum to no cytotoxicity & inflammatory-response. Here, we compare tunicate-derived nanocellulose (NC) against widely used semi-synthetic, gelatin methacrylate (GelMA) biomaterial for best mirroring HNSCC TME via bioprinting technology.

MethodsUsing HNSCC-lines (UM-SCC-14C, 11B & 22B), we bioprinted HNSCC cell-laden bioinks in NC-based hydrogels (TEMPO-oxidized NC (TEMPO-NC) & carboxymethylated NC (Carboxy-NC)) comparing to GelMA in alginate (GelMAA). Culturing bioconstructs for 21days in tumour media, we investigated bioink compatibility subjecting to a luminescence-based viability assay & immunofluorescence imaging for E-cadherin (n-3) within the 3D architecture.

ResultsOver 21 days, UM-SCC-22B (metastatic) cell-laden constructs showed significantly higher viability than 11B & 14C constructs. HNSCC cells in Carboxy-NC displayed significantly high viability than cells in TEMPO-NC. Over this period, viability in Carboxy-NC were comparable to GelMAA. HNSCC cells in Carboxy-NC constructs formed cell clusters indicative of cell proliferation. Overall, all HNSCC constructs expressed E-cadherin, an epithelial marker.

DiscussionDifferent cell lines showed differing behaviour reflecting their origin. The differences in bio-chemical & mechanical properties resulted in differing viability & proliferation, demonstrating carboxymethyl backbone within Carboxy-NC supports viability to a similar level as GelMAA. Given, NC bioinks endure minimum to no inflammatory-response, Carboxy-NC is preferred over GelMAA, while establishing a multicellular HNSCC TME model.

The authors declare that there is no conflict of interest