Tunable 3D Agarose-Well to enhance structural integrity of a reconstructed human skin equivalent

•3D Agarose-Wells are prepared to enhance structural integrity of skin equivalents.•3D Agarose-Wells show the enhancement of flux increases in culturing.•Agarose are hydrophobically modified by a top-down synthesis from agarose.•3D Agarose-Wells possess various hydrophilic/hydrophobic balances. The successful reconstruction of the skin equivalents that closely resemble normal human skin tissue greatly relies on maintaining the structural integrity throughout the air-liquid culture. Therefore, this study presents a novel strategy to prepare 3D Agarose-Well which enhances structural integrity of reconstructed skin equivalents. Hydrogel forming of agarose with agarose-g-C18, which synthesized a hydrophobically modified agarose, generated the 3D Agarose-Wells that possess various hydrophilic/hydrophobic balances. As a result, the agarose-fibrils in the well, assembled with penetrated collagen in dermis layer were proved to enhance the structural integrity and further regulate the mechanical strength from 6.8 kPa to 20.9 kPa. Overall, this new strategy to prepare the skin equivalents in the 3D Agarose-Well would be actively used in tissue engineering, drug delivery, and cosmetics, etc.