Interpenetrating Polymer Network of Hydroxyethyl Methacrylate and Chitosan as Cryogels for Tissue Engineering Applications

Tissue engineering is a vast expanding field with applications in areas such as tissue/organ transplantation, drug delivery, in vitro models, and so on. Biomaterials form an essential component in tissue engineering by acting as a template for cellular activity, therefore, novel tissue-engineered biomaterials with innovative properties are in high demand. Hence, this work proposes an interpenetrating polymer network cryogel of chitosan and hydroxyethyl methacrylate (HEMA) as a tissue-engineered biomaterial with uniform and high cell seeding throughout the cryogel matrix. The physical analysis of the cryogels demonstrated a highly macroporous structure exhibiting uniform pore size distribution and overall porosity through field emission-scanning electron microscopy (FE-SEM) analysis with pore sizes lying in the range of 50–200 μm and 150–400 μm in horizontal and transverse plane, respectively. The cryogels were also found to be degradable with an average percent degradation of 17.28 ± 1.47% in 4 weeks, and their mechanical properties revealed an average compressive strength of 0.05 MPa and an elastic modulus of 3 MPa. Further, biological characterization of the cryogel through direct contact test depicted an excellent biocompatibility with L929 mouse fibroblast and MC3T3-E1 preosteoblasts with negligible presence of dead cells in and around the cryogel. Uniform and high cell seeding with an increasing proliferation trend of MC3T3-E1 cells was observed on these cryogels through live-dead staining and MTT assay at day 1, 3, and 7-time point. Cell adherence studies via FE-SEM analysis also depicted a similar trend with uniform MC3T3-E1 cell distribution along with highly flattened morphology and extracellular matrix production. Therefore, based on their promising physico-chemical and biological properties, HEMA-Chitosan cryogels exhibit a strong potential application as tissue engineered biomaterials.