Environmental stressors induce changes in marine mussels from molecular (e.g., neurotransmitter and chaperone concentration, and expression of immune- and heat-shock protein-related genes) to physiological (e.g., filtration and heart rates, the number of circulating hemocytes) levels. Temperature directly affects the biogeographic distribution of mussels. Chaperones might form an essential part of endogenous protective mechanisms for the adaptation of these animals to low temperatures in nature. Here, we review the available studies dealing with cold stress responses of Mytilidae family members in their natural environment.

Here, we discuss the applicability of (methylenebis(salicylaldehyde)—MbSA) for the fabrication of the stimuli-responsive N-carboxyethylchitosan (CEC) hydrogels with a tunable dissolution rate under physiological conditions. In comparison with non-covalent salicylimine hydrogels, MbSA cross-linking via covalent bis(‘imine clip’) and non-covalent hydrophobic interactions allowed the fabrication of hydrogels with storage moduli > 1 kPa at ten-fold lower aldehyde/CEC molar ratio with the preservation of pH- and amino-acid responsive behavior. Although MbSA-cross-linked CEC hydrogels were stable at neutral and weakly alkaline pH, their disassembly in cell growth medium (Dulbecco’s modified Eagle’s medium, DMEM) under physiological conditions was feasible due to transimination reaction with amino acids contained in DMEM. Depending on the cross-linking density, the complete dissolution time of the fabricated hydrogels varied from 28 h to 11 days. The cytotoxicity of MbSA cross-linked CEC hydrogels toward a human colon carcinoma cell line (HCT 116) and primary human dermal fibroblasts (HDF) was remarkably lower in comparison with CEC-salicylimine hydrogels. Fast gelation, relatively low cytotoxicity, and tunable stimuli-induced disassembly under physiological conditions make MbSA cross-linked CEC hydrogels promising for drug encapsulation and release, 3D printing, cell culturing, and other biomedical applications.

Cross-linking chitosan at room and subzero temperature using a series of diglycidyl ethers of glycols (DEs)—ethylene glycol (EGDE), 1,4-butanediol (BDDE), and poly(ethylene glycol) (PEGDE) has been investigated to demonstrate that DEs can be a more powerful alternative to glutaraldehyde (GA) for fabrication of biocompatible chitosan cryogels with tunable properties. Gelation of chitosan with DEs was significantly slower than with GA, allowing formation of cryogels with larger pores and higher permeability, more suitable for flow-through applications and cell culturing. Increased hydration of the cross-links with increased DE chain length weakened intermolecular hydrogen bonding in chitosan and improved cryogel elasticity. At high cross-linking ratios (DE:chitosan 1:4), the toughness and compressive strength of the cryogels decreased in the order EGDE > BDDE > PEGDE. By varying the DE chain length and concentration, permeable chitosan cryogels with elasticity moduli from 10.4 ± 0.8 to 41 ± 3 kPa, toughness from 2.68 ± 0.5 to 8.3 ± 0.1 kJ/m³, and compressive strength at 75% strain from 11 ± 2 to 33 ± 4 kPa were fabricated. Susceptibility of cryogels to enzymatic hydrolysis was identified as the parameter most sensitive to cross-linking conditions. Weight loss of cryogels increased with increased DE chain length, and degradation rate of PEGDE-cross-linked chitosan decreased 612-fold, when the cross-linker concentration increased 20-fold.

This paper reviews original and literature data on the cryoresistance of the cells of marine organisms. The technology for the cryopreservation of these objects includes the selection of freezing conditions and the use, in addition to traditional cryoprotectants, of combinations of exogenous lipids, antioxidants, and disaccharide trehalose as a membrane stabilizing agent. We propose an approach for the preservation of marine invertebrate cells. The approach is based on the use of biologically active substances obtained from the tissues of marine organisms. Our results demonstrated the synergistic activity of these components of cryoprotective mixtures, and, at the same time, the specificity of antioxidant effects. An analysis of the factors that determine the choice of cryoprotectors was performed for various cell types. The development of the cryopreservation methods of marine organisms provides an opportunity for their wide application in both developmental biology and in marine biotechnology and serves as an important prerequisite for the cryobank creation.