Antimicrobial Zn-Carboxymethyl Chitosan Cryogel for Controlled Loading and Release of Ciprofloxacin via Coordination Bonds.

The local application of broad-spectrum antibiotics via polymeric drug delivery systems is a promising alternative to their systemic administration in wound healing, prevention and treatment of infections associated with surgical implants. However, low and poorly controlled loading efficiency and 100% burst release are common problems for the materials with weak physical interaction between antibiotics and polymeric matrices. Here, we report a new multifunctional carboxymethyl chitosan (CMC) cryogel, which efficiently prevents bacterial adhesion to the surface, kills bacteria in the solution via controlled release of ciprofloxacin (CIP), and promotes fibroblast proliferation.

Tuning Mechanical Properties, Swelling, and Enzymatic Degradation of Chitosan Cryogels Using Diglycidyl Ethers of Glycols with Different Chain Length as Cross-Linkers.

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.

Membrane vectorial lipidomic features of coral host cells' plasma membrane and lipid profiles of their endosymbionts Cladocopium.

The symbiotic relationships between coral animal host and autotrophic dinoflagellates are based on the mutual exchange and tight control of nutritional inputs supporting successful growth. The corals Sinularia heterospiculata and Acropora aspera were cultivated using a flow-through circulation system supplying seawater during cold and warm seasons of the year, then sorted into host cells and symbionts and subjected to phylogenetic, morphological, and advanced lipid analyses. Here we show, that the lipidomes of the dinoflagellates Cladocopium C1/C3 and acroporide-specific Cladocopium hosted by the corals, are determined by lipidomic features of different thermosensitivity and unique betaine- and phospholipid molecular species.

Chitosan Cryogels Cross-Linked with 1,1,3-Triglycidyloxypropane: Mechanical Properties and Cytotoxicity for Cancer Cell 3D Cultures.

Here, we have presented a new method of 1,1,3-triglycidyloxypropane (TGP) synthesis and investigated how cross-linker branching affects mechanical properties and cytotoxicity of chitosan scaffolds in comparison with those cross-linked using diglycidyl ethers of 1,4-butandiol (BDDGE) and poly(ethylene glycol) (PEGDGE). We have demonstrated that TGP is an efficient cross-linker for chitosan at a subzero temperature at TGP:chitosan molar ratios from 1:1 to 1:20. Although the elasticity of chitosan scaffolds increased in the following order of the cross-linkers PEGDGE > TGP > BDDGE, TGP provided cryogels with the highest compressive strength.

Chitosan versus Carboxymethyl Chitosan Cryogels: Bacterial Colonization, Human Embryonic Kidney 293T Cell Culturing and Co-Culturing.

The potential of chitosan and carboxymethyl chitosan (CMC) cryogels cross-linked with diglycidyl ether of 1,4-butandiol (BDDGE) and poly(ethylene glycol) (PEGDGE) have been compared in terms of 3D culturing HEK-293T cell line and preventing the bacterial colonization of the scaffolds. The first attempts to apply cryogels for the 3D co-culturing of bacteria and human cells have been undertaken toward the development of new models of host-pathogen interactions and bioimplant-associated infections. Using a combination of scanning electron microscopy, confocal laser scanning microscopy, and flow cytometry, we have demonstrated that CMC cryogels provided microenvironment stimulating cell-cell interactions and the growth of tightly packed multicellular spheroids, while cell-substrate interactions dominated in both chitosan cryogels, despite a significant difference in swelling capacities and Young's modulus of BDDGE- and PEGDGE-cross-linked scaffolds.

Sponge-like Scaffolds for Colorectal Cancer 3D Models: Substrate-Driven Difference in Micro-Tumors Morphology.

Macroporous scaffolds (cryogels) for the 3D cell culturing of colorectal cancer micro-tumors have been fabricated by cross-linking chitosan and carboxymethyl chitosan (CMC) with 1,4-butandiol diglycidyl ether (BDDGE) under subzero temperature. Due to the different intrinsic properties and reactivity of CMC and chitosan under the same cross-linking conditions, Young's moduli and swelling of the permeable for HCT 116 cells cryogels varied in the broad range 3-41 kPa and 3500-6000%, respectively. We have demonstrated that the morphology of micro-tumors can be controlled via selection of the polymer for the scaffold fabrication.

Chitosan Cross-Linking with Acetaldehyde Acetals.

Here we demonstrate the possibility of using acyclic diethylacetal of acetaldehyde (ADA) with low cytotoxicity for the fabrication of hydrogels via Schiff bases formation between chitosan and acetaldehyde generated in situ from acetals in chitosan acetate solution. This approach is more convenient than a direct reaction between chitosan and acetaldehyde due to the better commercial availability and higher boiling point of the acetals. Rheological data confirmed the formation of intermolecular bonds in chitosan solution after the addition of acetaldehyde diethyl acetal at an equimolar NH: acetal ratio.

Stimuli-Responsive Dual Cross-Linked -Carboxyethylchitosan Hydrogels with Tunable Dissolution Rate.

Here, we discuss the applicability of (methylenebis(salicylaldehyde)-MbSA) for the fabrication of the stimuli-responsive -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.

Physiological processes and lipidome dynamics in the soft coral Sinularia heterospiculata under experimental bleaching.

Coral polyps host intracellular symbiotic dinoflagellates (SD). The loss of SD (referred as bleaching) under stressful environmental conditions is the main reason of coral reef destruction, and therefore, intensively studied over the world. Lipids are the structural base of biomembranes and energy reserve of corals and are directly involved in the coral bleaching.

Isolation, characterization, and ecotoxicological application of marine mammal skin fibroblast cultures.

Marine mammal cell cultures are a multifunctional instrument for acquiring knowledge about life in the world's oceans in physiological, biochemical, genetic, and ecotoxicological aspects. We succeeded in isolation, cultivation, and characterization of skin fibroblast cultures from five marine mammal species. The cells of the spotted seal (Phoca largha), the sea lion (Eumetopias jubatus), and the walrus (Odobenus rosmarus) are unpretentious to the isolation procedure.

The effects of cold stress on Mytilus species in the natural environment.

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.

Chemical modulation of apoptosis in molluscan cell cultures.

This study focused on the alterations that occur in larval molluscan cells after administration of apoptotic inducers and inhibitors used in mammalian cells in response to cold stress. This is the first report on apoptosis modulation in molluscan cells assessed by flow cytometry. Mitochondrial activity, general caspase activation, and membrane integrity of control molluscan cells were compared to those processes in frozen-thawed molluscan cells, primary mouse embryonic fibroblasts, and human colon tumor cells prior to treatment and after incubation with apoptotic inducers or inhibitors.

The death pathways in mussel larval cells after a freeze-thaw cycle.

We analyzed cell viability, caspase activity, plasma membrane alterations and cell ultrastructure morphology to estimate the morphological and biochemical alterations that occur in bivalve molluscan cell cultures during cryopreservation. The use of 5% dymethyl sulfoxide as a cryoprotectant resulted in greater cell survival and a scarcity of destroyed cells lacking cytosol among dead cells. In this case, almost all cells died through necrosis or apoptosis, which appeared to increase in mussel cell cultures after a freeze-thaw cycle.

The contribution of apoptosis and necrosis in freezing injury of sea urchin embryonic cells.

Sea urchins have recently been reported to be a promising tool for investigations of oxidative stress, UV light perturbations and senescence. However, few available data describe the pathway of cell death that occurs in sea urchin embryonic cells after cryopreservation. Our study is focused on the morphological and functional alterations that occur in cells of these animals during the induction of different cell death pathways in response to cold injury.

Freezing tolerance of sea urchin embryonic cells: Differentiation commitment and cytoskeletal disturbances in culture.

This study focuses on the freezing tolerance of sea urchin embryonic cells. To significantly reduce the loss of physiological activity of these cells that occurs after cryopreservation and to study the effects of ultra-low temperatures on sea urchin embryonic cells, we tested the ability of the cells to differentiate into spiculogenic or pigment directions in culture, including an evaluation of the expression of some genes involved in pigment differentiation. A morphological analysis of cytoskeletal disturbances after freezing in a combination of penetrating (dimethyl sulfoxide and ethylene glycol) and non-penetrating (trehalose and polyvinylpyrrolidone) cryoprotectants revealed that the distribution pattern of filamentous actin and tubulin was similar to that in the control cultures.