The transitions and water state in three types of hydrogels have been examinated by differential scanning calorimetry (DSC) in dried and hydrated states correlated with dynamical mechanical thermal analysis (DMTA), namely substituted anhydride modified collagens and their corresponding hydrogels, hybrid hydrogels of collagen and poly (N-isopropyl acrylamide) (pNIPAM) with various nanoclays and hydroxyapatite and polyvinyl alcohol (PVA) gel and mixed hyaluronic acid cryogels have been investigated. The denaturation temperature (Td), denaturation enthalpy, and glass transition temperature (Tg) and water states (free or bonded) in the hydrated samples have been determined, and they were found to be dependent on chemical composition, thermal history, and moisture content. It was established that the denaturation of collagen is an irreversible kinetic process that is governed by the temperature history, nanoparticle type in hybrid hydrogels, specimen hydration, and cross-linking agent amount, amongst other variables. The interaction of water with hybrid collagen and water state in semi-interpenetrated polymer network depends on hydrogel composition and thermal history. The hybrid hydrogels keep the temperature responsiveness. Contact angle and swelling measurements demonstrated the correlation between hydrophobicity/hydrophilicity balance and state of water in the polymer network and the denaturation temperature values. Thermal characterization of the HA/PVA hydrogels unloaded and loaded with methotrexate and ciprofloxacin was shown that Tg of PVA is strongly affected by the water content, the water induced a plasticizing effect on the polymer, lowering its glass transition temperature. The processing conditions e.g. number of cycles of freeze/thaw process or time of thaw cure affected both, crystallinity and physical network showing variations in the Tg values of the systems. The evidenced low-temperature peaks indicate the presence of freezable bound water within the polymeric network. Transition temperatures determined at higher temperature were assigned to the relaxation mechanism of the glycosidic bonds and structural rearrangements due to the loss of the residual water. Crystallinity degree increased with number of the FT cycles and by the loading of drugs into cryrogel network because of a good compatibility between the two constituents.