Boronic acid and diol-containing polymers: how to choose the correct couple to form "strong" hydrogels at physiological pH.

Dynamic covalent hydrogels crosslinked by boronate ester bonds are promising materials for biomedical applications. However, little is known about the impact of the crosslink structure on the mechanical behaviour of the resulting network. Herein, we provide a mechanistic study on boronate ester crosslinking upon mixing hyaluronic acid (HA) backbones modified, on the one hand, with two different arylboronic acids, and on the other hand, with three different saccharide units. Combining rheology, NMR and computational analysis, we demonstrate that carefully selecting the arylboronic-polyol couple allows for tuning the thermodynamics and molecular exchange kinetics of the boronate ester bond, thereby controlling the rheological properties of the gel. In particular, we report the formation of "strong" gels (i.e. featuring slow relaxation dynamics) through the formation of original complex structures (tridentate or bidentate complexes). These findings offer new prospects for the rational design of hydrogel scaffolds with tailored mechanical response.

Rheology of partially and totally oxidized red seaweed galactans.

Agarose and κ-carrageenan were oxidized using (2,2,6,6-tetramethylpiperidinyl)oxy (TEMPO) in the presence of NaOCl and NaBr. Products with several degrees of oxidation were structurally characterized. The mechanical spectra were determined: derivatives with a medium to high degree of oxidation give rise to polysaccharides that behave like dilute solutions in water, whereas those with a degree of oxidation close to 20 % keep the gelling properties with a different thermo-rheological response towards pH (6.5 or 4.0) and counterions (K+ or Ca2+) in comparison with the native polysaccharides. For instance, they showed a marked dependence on the presence of calcium ions, observed in the increase of thermal stability and dynamic elastic component (G') value, due to the known interaction of this divalent cation with the carboxylate groups. In this sense, these derivatives with low oxidation degrees have proven to be not only thermosensitive, like the native polysaccharide, but also pH- and calcium-sensitive.

Design of Soft Nanocarriers Combining Hyaluronic Acid with Another Functional Polymer for Cancer Therapy and Other Biomedical Applications.

The rapid advancement in medicine requires the search for new drugs, but also for new carrier systems for more efficient and targeted delivery of the bioactive molecules. Among the latter, polymeric nanocarriers have an increasingly growing potential for clinical applications due to their unique physical and chemical characteristics. In this regard, nanosystems based on hyaluronic acid (HA), a polysaccharide which is ubiquitous in the body, have attracted particular interest because of the biocompatibility, biodegradability and nonimmunogenic property provided by HA. Furthermore, the fact that hyaluronic acid can be recognized by cell surface receptors in tumor cells, makes it an ideal candidate for the targeted delivery of anticancer drugs. In this review, we compile a comprehensive overview of the different types of soft nanocarriers based on HA conjugated or complexed with another polymer: micelles, nanoparticles, nanogels and polymersomes. Emphasis is made on the properties of the polymers used as well as the synthetic approaches for obtaining the different HA-polymer systems. Fabrication, characterization and potential biomedical applications of the nanocarriers will also be described.

Disaccharides obtained from carrageenans as potential antitumor agents.

Carrageenans are sulfated galactans found in certain red seaweeds with proven biological activities. In this work, we have prepared purified native and degraded κ-, ι-; and λ-carrageenans, including the disaccharides (carrabioses) and disaccharide-alditols (carrabiitols) from seaweed extracts as potential antitumor compounds and identified the active principle of the cytotoxic and potential antitumor properties of these compounds. Both κ and ι-carrageenan, as well as carrageenan oligosaccharides showed cytotoxic effect over LM2 tumor cells. Characterized disaccharides (carrabioses) and the reduced product carrabiitols, were also tested. Only carrabioses were cytotoxic, and among them, κ-carrabiose was the most effective, showing high cytotoxic properties, killing the cells through an apoptotic pathway. In addition, the cells surviving treatment with κ-carrabiose, showed a decreased metastatic ability in vitro, together with a decreased cell-cell and cell-matrix interactions, thus suggesting possible antitumor potential. Overall, our results indicate that most cytotoxic compounds derived from carrageenans have lower molecular weights and sulfate content. Potential applications of the results emerging from the present work include the use of disaccharide units such as carrabioses coupled to antineoplasics in order to improve its cytotoxicity and antimetastatic properties, and the use of ι-carrageenan as adjuvant or carrier in anticancer treatments.

Gracilariopsis hommersandii, a red seaweed, source of agar and sulfated polysaccharides with unusual structures.

Red seaweed Gracilariopsis hommersandii produces important amounts of non-gelling galactans, which were extracted with hot water (GrC, yield, 37%, viscosity average molecular weight, Mv 109 kDa), comprising agarose and sulfated galactan structures. The alkali modified derivative, GrCTr (Mv 95 kDa), gave a galactose:3,6-anhydrogalactose molar ratio of 1.0:0.9, and a more regular structure, favouring gelation (melting and gelling temperatures 64 and 14 °C, respectively). The rheological properties of this product suggest possible applications as hydrocolloid. G. hommersandii also biosynthesizes non gelling sulfated galactan fractions with diads constituted by ß-d-galactose and partially cyclized α-l-galactose units or non-cyclized α-d-galactose residues. Sulfation was mainly detected on C6 or C4 of the ß-d-galactose units, and on C6 and, in minor amounts, on C3 of the α-l-galactose units. The presence of ß-apiuronic acid was demonstrated for these fractions as side chains of the galactan backbone. Carrageenan structures were found for the first time in an agarophyte of the Gracilariales.

Minor polysaccharidic constituents from the red seaweed Hypnea musciformis. Appearance of a novel branched uronic acid.

Two polysaccharide fractions isolated from Hypnea musciformis after room temperature- and hot water extraction, soluble after KCl precipitation of the more abundant carrageenans, were subfractionated by ion-exchange chromatography eluting with increasing concentrations of NaCl. The lowest NaCl concentration (0.2M) eluted agarans. The dl-hybrids (or mixtures) eluted at intermediate concentrations of NaCl. The d/l-galactose ratio and the sulfate proportion increased with the NaCl concentration. Different types of substitution were present, mainly at C-3 with sulfate, Xyl and methylated Gal stubs, as well as low amounts of 3,6-AnGal. A novel constituent, identified as 3-C-carboxy-d-erythrose1 in its ß-furanosic form, was found linked to C-6 of ß-Gal units. A search carried out in other species like Iridaea undulosa and Kappaphycus alvarezii also revealed the same constituent. Finally, the late-eluting fractions were mostly carrageenans, with a structure consistent with that of a κ/ι/ν-carrageenan hybrid.

DFT/PCM theoretical study of the conversion of methyl 4-O-methyl-α-d-galactopyranoside 6-sulfate and its 2-sulfated derivative into their 3,6-anhydro counterparts.

Modeling of the conversion of methyl 4-O-methyl-α-d-galactopyranoside 6-sulfate (2) and 2,6-disulfate (1) into methyl 3,6-anhydro-4-O-methyl-α-d-galactopyranoside (4) and its 2-sulfate (3), respectively (Scheme 1) has been carried out using DFT at the M06-2X/6-311 + G(d,p)//M06-2X/6-31 + G(d,p) level with the polarizable continuum model (PCM) in water. The three steps necessary for the alkaline transformation of 6-sulfated (and 2,6-disulfated) galactose units into 3,6-anhydro derivatives were evaluated. The final substitution step appears to be the rate limiting, involving an activation energy of ca. 23 kcal/mol. The other two steps (deprotonation and chair inversion) combined involve lower activation energies (9-12 kcal/mol). Comparison of the thermodynamics and kinetics of the reactions suggest that if the deprotonation step precedes the chair inversion, the reaction should be faster for both compounds. No major differences in reaction rate can be theoretically predicted to be caused by the presence of sulfate on O-2, although one experimental result suggested that O-2 sulfation should increase the reaction rate. The conformational pathways are complex, given the large number of rotamers available for each compound, and the way that some of these rotamers combine into some of the pathways. In any case, the conformation (O)S2 appears as a common intermediate for the chair inversion processes.

Partial and total C-6 oxidation of gelling carrageenans. Modulation of the antiviral activity with the anionic character.

The optimal conditions for the full C-6 oxidation of κ- and ι-carrageenans using (2,2,6,6-tetramethylpiperidinyl)oxy (TEMPO) in the presence of sodium hypochlorite and sodium bromide were assessed. The fully oxidized products were characterized by NMR spectroscopy. Partially oxidized products were also obtained and analyzed by chemical and spectroscopical methods. The antiviral activity of carrageenans against herpes simplex virus HSV-1 and HSV-2 determined by plaque reduction assay, was not largely affected by full oxidation of the polysaccharides, but an increase in activity was detected by partial oxidation. A specific overoxidation on C-2 of the 3,6-anhydrogalactose moiety of κ-carrageenan was identified, solved experimentally and rationalized through the application of molecular modeling.

Chemical and rheological characterization of the carrageenans from Hypnea musciformis (Wulfen) Lamoroux.

This work reports a chemical and rheological study of the carrageenans from Hypnea musciformis, a red seaweed commercially known for its production of κ-carrageenan. The polysaccharides were extracted with water both at room temperature and at 90 °C: the yield of the latter was about six times larger than the former. Fractionation with KCl yielded a large proportion (50-67%) of a precipitate with 0.125 M of this salt for both extracts, with characteristics of a nearly pure κ-carrageenan, as determined by methylation analysis and NMR spectroscopy. Smaller amounts of fractions precipitating at higher concentrations showed a basic κ-carrageenan structure, but included some ι-carrageenan diads. The KCl-soluble polysaccharides showed a larger complexity, containing d- and l-galactans or d/l-hybrids. Some differences in the rheological properties of these carrageenans have been found. Although all KCl-precipitating polysaccharides form true gels at 10 °C in presence of KCl, those extracted with hot water form stronger gels than those extracted at room temperature. Both purified κ-carrageenans show lower gelling and melting temperature than the whole polysaccharides from which they were originated.