RESUMO
Biological cryopreservation often involves using a cryoprotective agent (CPA) to mitigate lethal physical stressors cells endure during freezing and thawing, but effective CPA concentrations are cytotoxic. Hence, natural polysaccharides have been studied as biocompatible alternatives. Here, a subset of 26 natural polysaccharides of various chemical composition was probed for their potential in enhancing the metabolic post-thaw viability (PTV) of cryopreserved Vero cells. The best performing cryoprotective polysaccharides contained significant fucose amounts, resulting in average PTV 2.8-fold (up to 3.1-fold) compared to 0.8-fold and 2.2-fold for all non-cryoprotective and cryoprotective polysaccharides, respectively, outperforming the optimized commercial CryoStor™ CS5 formulation (2.6-fold). Stoichiometrically, a balance between fucose (18-35.7 mol%), uronic acids (UA) (13.5-26 mol%) and high molecular weight (MW > 1 MDa) generated optimal PTV. Principal component analysis (PCA) revealed that fucose enhances cell survival by a charge-independent, MW-scaling mechanism (PC1), drastically different from the charge-dominated ice growth disruption of UA (PC2). Its neutral nature and unique properties distinguishable from other neutral monomers suggest fucose may play a passive role in conformational adaptability of polysaccharide to ice growth inhibition, or an active role in cell membrane stabilization through binding. Ultimately, fucose-rich anionic polysaccharides may indulge in polymer-ice and polymer-cell interactions that actively disrupt ice and minimize lethal volumetric fluctuations due to a balanced hydrophobic-hydrophilic character. Our research showed the critical role neutral fucose plays in enhancing cellular cryopreservation outcomes, disputing previous assumptions of polyanionicity being the sole governing predictor of cryoprotection.
Assuntos
Fucose , Gelo , Animais , Chlorocebus aethiops , Fucose/metabolismo , Células Vero , Congelamento , Crioprotetores/farmacologia , Crioprotetores/química , Criopreservação/métodos , Polissacarídeos/farmacologia , Polímeros/farmacologia , Sobrevivência CelularRESUMO
Recently, polysaccharide-based hydrogels crosslinked with the trivalent iron cation have attracted interest due to their remarkable properties that include high mechanical stability, stimuli-responsiveness, and enhanced absorptivity. In this study, a Fe3+ crosslinked hydrogel was prepared using the biocompatible extracellular polysaccharide (EPS) secreted by the marine bacterium Alteromonas macleodii Mo169. Hydrogels with mechanical strengths (G') ranging from 0.3 kPa to 44.5 kPa were obtained as a result of the combination of different Fe3+ (0.05-9.95 g L-1) and EPS (0.3-1.7 %) concentrations. All the hydrogels had a water content above 98 %. Three different hydrogels, named HA, HB, and HC, were chosen for further characterization. With strength values (G') of 3.2, 28.9, and 44.5 kPa, respectively, these hydrogels might meet the strength requirements for several specific applications. Their mechanical resistance increased as higher Fe3+ and polymer concentrations were used in their preparation (the compressive hardness increased from 8.7 to 192.1 kPa for hydrogel HA and HC, respectively). In addition, a tighter mesh was noticed for HC, which was correlated to its lower swelling ratio value compared to HA and HB. Overall, this preliminary study highlighted the potential of these hydrogels for tissue engineering, drug delivery, or wound healing applications.
Assuntos
Alteromonas , Hidrogéis , Ferro , Polissacarídeos Bacterianos , Hidrogéis/química , Alteromonas/química , Polissacarídeos Bacterianos/química , Ferro/química , Materiais Biocompatíveis/química , Reagentes de Ligações Cruzadas/química , Força CompressivaRESUMO
Microalgae are photosynthetic organisms that can produce biomolecules with industrial interest, including exopolysaccharides (EPS). Due to their structural and compositional diversity, microalgae EPS present interesting properties that can be considered in cosmetic and/or therapeutic areas. Seven microalgae strains from three different lineages, namely Dinophyceae (phylum Miozoa), Haptophyta, and Chlorophyta, were investigated as EPS producers. All strains were found to be EPS producers, though the highest EPS yield was obtained for Tisochrysis lutea, followed by Heterocapsa sp. (126.8 and 75.8 mg L−1, respectively). Upon assessment of the polymers chemical composition, significant contents of unusual sugars, including fucose, rhamnose, and ribose, were found. Heterocapsa sp. EPS stood out due to its high content of fucose (40.9 mol%), a sugar known to confer biological properties to polysaccharides. The presence of sulfate groups (10.633.5 wt%) was also noticed in the EPS produced by all microalgae strains, thus contributing to the possibility that these EPS might have biological activities worth exploring.(AU)