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A preliminary comparison of two different polyacrylamide hydrogel fabrication methods demonstrate differences in stiffness measurements and adhesion abilities of osteosarcoma cells

The purpose of this research was to compare two different polyacrylamide hydrogel fabrication methods described in the literature and assess their use in mechanotransduction studies in osteosarcoma. Both methods employ succinimide chemistry to functionalize the hydrogel surface for cell response studies, one in the form of NHS, while the other in the form of Sulfo-SANPAH. Six hydrogels of two different stiffness were created for each method and were evaluated on their receptiveness to cell seeding and Young’s moduli with atomic force microscopy. Both hydrogel fabrication methods lack reproducibility as significant differences were observed in stiffness measurements between six hydrogels at both 0.5 kPa and 50 kPa stiffnesses. Despite the Sulfo- SANPAH method of preparation being more receptive to cell seeding and generating the expected effects on known mechanotransducers, it appeared to have larger variabilities in stiffness measurements for both 0.5 kPa and 50 kPa prepared hydrogels. Researchers may employ the Sulfo-SANPAH method to study the impact of ECM of varying stiffness on osteosarcoma cell mechanotransduction, but should remain cautious when interpreting results as a function of the expected stiffness. Studies should be accompanied by measurements of Young's modulus whenever possible.