Fibroblast ve keratinosit hücrelerinin in vitro üç boyutlu ko-kültüründe kolajen/keratin/aljinat hidrojeli

Due to the high demand for advanced biomaterials in tissue engineering, research has focused on biomimetic hydrogels and three-dimensional (3D) co-culture systems to enhance tissue formation. The aim of this study was to examine the potential of Collagen/Keratin/Alginate (Col/Ker/Alg) hydrogel as a tissue scaffold for fibroblast and keratinocyte 3D co-culture. Collagen and keratin were extracted from bovine Achilles tendons and human hair waste, respectively, and characterized using FTIR and SDS-PAGE. FTIR confirmed the presence of key functional groups, while SDS-PAGE identified distinct collagen and keratin bands. Hydrogels were cross-linked using calcium chloride at 0.10 M, 0.50 M, and 1.0 M concentrations and analyzed for morphology, swelling, degradation, protein release, and porosity. The 0.50 M cross-linked hydrogel exhibited the highest water uptake, a highly porous and interconnected structure, and the most significant protein release after Day 2. Degradation studies showed that 0.10 M hydrogels degraded rapidly, while 1.0 M hydrogels were more stable. Cell culture experiments confirmed that all hydrogel formulations supported fibroblast viability for 9 days, with 0.50 M proving optimal. Micro-patterned 3D-printed stamps were used to create channel formations mimicking natural tissue structures. The hydrogel matrix, seeded with human dermal fibroblasts (HDF-a) and keratinocytes (HaCaT), was assessed over 21 days for viability and proliferation using resazurin reduction and fluorescence imaging. Results demonstrated that the Col/Ker/Alg hydrogel provides a favorable 3D microenvironment for cell proliferation, indicating its potential as a promising scaffold for skin tissue engineering applications.