Redox Properties of Mn(III)porphyrins in Hyaluronan Oxidative Degradation: Single Electron Transfer Mechanism.

Cationic Mn(III)-meso-tetraarylporphyrin derivatives, substituted in para position with different size alkyl chains, were investigated to function as antioxidants in free-radical degradation of high-molar-mass hyaluronan by the methods of rotational viscometry and oximetry. The results of rotational viscometry showed that MnTM-4-PyP5+, MnTE-4-PyP5+, MnTPr-4-PyP5+, MnTPen-4-PyP5+ and MnTHep-4-PyP5+ showed high efficiency in decomposing H2O2, and reducing of peroxidized hyaluronan. When using oxygen electrode, MnTE-4-PyP5+, MnTPr-4-PyP5+, MnTPen-4-PyP5+, and MnTHep-4-PyP5+ applied to function as protective antioxidants in hyaluronan degradation, the uptake of dissolved oxygen from the reaction milieu was rapid, followed by continual increase in oxygen concentration up to the end of the measurement. However, when especially MnTE-4-PyP5+, MnTPr-4-PyP5+, and MnTPen-4-PyP5+ were examined as hyaluronan chain-breaking antioxidants, after short-term dissolved oxygen uptake, almost no increase in oxygen concentration was shown.

Hyaluronan: Sources, Structure, Features and Applications.

Hyaluronan (HA) is a non-sulfated glycosaminoglycan that is present in a variety of body tissues and organs. Hyaluronan has a wide range of biological activities that are frequently influenced by molar mass; however, they also depend greatly on the source, purity, and kind of impurities in hyaluronan. High-molar-mass HA has anti-inflammatory, immunosuppressive, and antiangiogenic properties, while low-molar-mass HA has opposite properties. A number of chemical modifications have been performed to enhance the stability of HA and its applications in medical practice. Hyaluronan is widely applied in medicine, such as viscosupplementation, ophthalmology, otolaryngology, wound healing, cosmetics, and drug delivery. In this review, we summarized several medical applications of polymers based on the hyaluronan backbone.

Taxifolin Reduces Blood Pressure via Improvement of Vascular Function and Mitigating the Vascular Inflammatory Response in Spontaneously Hypertensive Rats.

The effect of a 10-day-long treatment with taxifolin (TAX, 20 mg/kg/day p.o.) was investigated on spontaneously hypertensive rats (SHRs) with a focus on the vascular functions of isolated femoral arteries and thoracic aortas. TAX reduced blood pressure in SHRs. In femoral arteries, TAX increased acetylcholine-induced relaxation, reduced the maximal NA-induced contraction, and reduced acetylcholine-induced endothelium-dependent contraction (EDC); however, TAX had no effect on the vascular reactivity of isolated thoracic aortas. In addition, TAX elevated the total nitric oxide synthase (NOS) activity and iNOS protein expression but reduced cyclooxygenase-2 (COX2) protein expression in the tissue of the abdominal aorta without changes in Nos2 and Ptgs2 gene expressions. TAX also increased the gene expression of the anti-inflammatory interleukin-10 (Il10). In addition, in vitro studies showed that TAX has both electron donor and H atom donor properties. However, TAX failed to reduce superoxide production in the tissue of the abdominal aorta after oral administration. In conclusion, our results show that a decrease in the blood pressure in TAX-treated SHRs might be attributed to improved endothelium-dependent relaxation and reduced endothelium-dependent contraction. In addition, the results suggest that the effect of TAX on blood pressure regulation also involves the attenuation of COX2-mediated pro-inflammation and elevation of anti-inflammatory pathways.

Preparation and Physicochemical Characterization of Gelatin-Aldehyde Derivatives.

The present study aimed at preparing novel free-radical scavenging and water-soluble compounds derived from gelatin. Specifically, gelatin−syringaldehyde, gelatin−anisaldehyde, and gelatin−vanillin were synthesized and thoroughly studied for their physicochemical properties. In particular, the compounds were characterized by UV-Vis spectroscopy, Fourier-transform infrared spectroscopy, and scanning electron microscopy. Notably, as demonstrated by thermogravimetry and differential scanning calorimetry, all three derivatives exhibited higher thermal stability than gelatin itself. Free-radical scavenging activities of the examined compounds were explored by (i) a standard spectrophotometric ABTS assay and (ii) an assay of oxidative degradation of hyaluronic acid monitored by rotational viscometry. We found that gelatin and gelatin−syringaldehyde demonstrated the highest efficacy in scavenging •OH radicals, whereas gelatin−anisaldehyde was the least effective. The efficacy of scavenging alkyloxy- and alkylperoxy-type free radicals via hydrogen-atom-transferring property was in the following order: gelatin > gelatin−vanillin > gelatin−syringaldehyde > gelatin−anisaldehyde. Electron-donor properties determined using the ABTS assay revealed the following order in one-electron reduction of ABTS•+: gelatin > gelatin−anisaldehyde > gelatin−vanillin > gelatin−syringaldehyde.

Reaction of N-Acetylcysteine with Cu2+: Appearance of Intermediates with High Free Radical Scavenging Activity: Implications for Anti-/Pro-Oxidant Properties of Thiols.

We studied the kinetics of the reaction of N-acetyl-l-cysteine (NAC or RSH) with cupric ions at an equimolar ratio of the reactants in aqueous acid solution (pH 1.4−2) using UV/Vis absorption and circular dichroism (CD) spectroscopies. Cu2+ showed a strong catalytic effect on the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) radical (ABTSr) consumption and autoxidation of NAC. Difference spectra revealed the formation of intermediates with absorption maxima at 233 and 302 nm (ε302/Cu > 8 × 103 M−1 cm−1) and two positive Cotton effects centered at 284 and 302 nm. These intermediates accumulate during the first, O2-independent, phase of the NAC autoxidation. The autocatalytic production of another chiral intermediate, characterized by two positive Cotton effects at 280 and 333 nm and an intense negative one at 305 nm, was observed in the second reaction phase. The intermediates are rapidly oxidized by added ABTSr; otherwise, they are stable for hours in the reaction solution, undergoing a slow pH- and O2-dependent photosensitive decay. The kinetic and spectral data are consistent with proposed structures of the intermediates as disulfide-bridged dicopper(I) complexes of types cis-/trans-CuI2(RS)2(RSSR) and CuI2(RSSR)2. The electronic transitions observed in the UV/Vis and CD spectra are tentatively attributed to Cu(I) → disulfide charge transfer with an interaction of the transition dipole moments (exciton coupling). The catalytic activity of the intermediates as potential O2 activators via Cu(II) peroxo-complexes is discussed. A mechanism for autocatalytic oxidation of Cu(I)−thiolates promoted by a growing electronically coupled −[CuI2(RSSR)]n− polymer is suggested. The obtained results are in line with other reported observations regarding copper-catalyzed autoxidation of thiols and provide new insight into these complicated, not yet fully understood systems. The proposed hypotheses point to the importance of the Cu(I)−disulfide interaction, which may have a profound impact on biological systems.

Hyaluronic acid and chitosan-based electrospun wound dressings: Problems and solutions.

To date, available review papers related to the electrospinning of biopolymers including polysaccharides for wound healing were focused on summarizing the process conditions for two candidates, namely chitosan and hyaluronic acid. However, most reviews lack the discussion of problems of hyaluronan and chitosan electrospun nanofibers for wound dressing applications. For this reason, it is required to update information by providing a comprehensive overview of all factors which may play a role in the electrospinning of hyaluronic acid and chitosan for applications of wound dressings. This review summarizes the fabricated chitosan and hyaluronic acid electrospun nanofibers as wound dressings in the last years, including methods of preparations of nanofibers and challenges for the electrospinning of both pure chitosan and hyaluronic acid and strategies how to overcome the existing difficulties. Moreover, in this review the biological roles and mechanisms of chitosan and hyaluronic acid in the wound healing process are explained including the advantages of nanofibers for ideal wound management using the common solvents, copolymers enhancing spinning process, and the most biologically active incorporated substances thereby providing drug delivery in wound healing.

Assessment of the Substance Antioxidative Profile by Hyaluronan, Cu(II) and Ascorbate.

In the minireview presented here, the authors discuss the evaluation of inhibitory effect of substances in the phases of initiation and propagation of high-molar-mass hyaluronan oxidative degradation. The experimental approach should be considered as original since on using a simple experimental assay it is possible to prove both the so-called "preventive" and "chain-breaking" antioxidant activity of investigated water-soluble endo- or exogenous substances.

Ortho Isomeric Mn(III) N-Alkyl- and Alkoxyalkylpyridylporphyrins-Enhancers of Hyaluronan Degradation Induced by Ascorbate and Cupric Ions.

High levels of hyaluronic acid (HA) in tumors correlate with poor outcomes with several types of cancers due to HA-driven support of adhesion, migration and proliferation of cells. In this study we explored how to enhance the degradation of HA into low-molecular fragments, which cannot prevent the immune system to fight tumor proliferation and metastases. The physiological solution of HA was exposed to oxidative degradation by ascorbate and cupric ions in the presence of either one of three ortho isomeric Mn(III) substituted N-alkyl- and alkoxyalkylpyridylporphyrins or para isomeric Mn(III) N-methylpyridyl analog, commonly known as mimics of superoxide dismutase. The changes in hyaluronan degradation kinetics by four Mn(III) porphyrins were monitored by measuring the alteration in the dynamic viscosity of the HA solution. The ortho compounds MnTE-2-PyP5+ (BMX-010, AEOL10113), MnTnBuOE-2-PyP5+ (BMX-001) and MnTnHex-2-PyP5+ are able to redox cycle with ascorbate whereby producing H2O2 which is subsequently coupled with Cu(I) to produce the •OH radical essential for HA degradation. Conversely, with the para analog, MnTM-4-PyP5+, no catalysis of HA degradation was demonstrated, due to its inertness towards redox cycling with ascorbate. The impact of different Mn(III)-porphyrins on the HA decay was further clarified by electron paramagnetic resonance spectrometry. The ability to catalyze the degradation of HA in a biological milieu, in the presence of cupric ions and ascorbate under the conditions of high tumor oxidative stress provides further insight into the anticancer potential of redox-active ortho isomeric Mn(III) porphyrins.

Hyaluronan as a Prominent Biomolecule with Numerous Applications in Medicine.

Hyaluronan (HA) is a natural glycosaminoglycan present in many tissues of all vertebrates. HA has various biological functions, which are dependent on its molar mass. High-molar-mass HA has anti-angiogenic, immunosuppressive and anti-inflammatory properties, while low-molar-mass HA has opposite effects. HA has also antioxidative properties, however on the other hand it can be readily degraded by reactive oxygen species. For many years it has been used in treatment of osteoarthritis, cosmetics and in ophthalmology. In the last years there has been a growing interest of HA to also be applied in other fields of medicine such as skin wound healing, tissue engineering, dentistry and gene delivery. In this review we summarize information on modes of HA administration, properties and effects of HA in various fields of medicine including recent progress in the investigation of HA.

Self-Associating Polymers Chitosan and Hyaluronan for Constructing Composite Membranes as Skin-Wound Dressings Carrying Therapeutics.

Chitosan, industrially acquired by the alkaline N-deacetylation of chitin, belongs to ß-N-acetyl-glucosamine polymers. Another ß-polymer is hyaluronan. Chitosan, a biodegradable, non-toxic, bacteriostatic, and fungistatic biopolymer, has numerous applications in medicine. Hyaluronan, one of the major structural components of the extracellular matrix in vertebrate tissues, is broadly exploited in medicine as well. This review summarizes that these two biopolymers have a mutual impact on skin wound healing as skin wound dressings and carriers of remedies.

Versatile Use of Chitosan and Hyaluronan in Medicine.

Chitosan is industrially acquired by the alkaline N-deacetylation of chitin. Chitin belongs to the ß-N-acetyl-glucosamine polymers, providing structure, contrary to α-polymers, which provide food and energy. Another ß-polymer providing structure is hyaluronan. A lot of studies have been performed on chitosan to explore its industrial use. Since chitosan is biodegradable, non-toxic, bacteriostatic, and fungistatic, it has numerous applications in medicine. Hyaluronan, one of the major structural components of the extracellular matrix in vertebrate tissues, is broadly exploited in medicine as well. This review summarizes the main areas where these two biopolymers have an impact. The reviewed areas mostly cover most medical applications, along with non-medical applications, such as cosmetics.

Antioxidant and antibacterial polyelectrolyte wound dressing based on chitosan/hyaluronan/phosphatidylcholine dihydroquercetin.

Antioxidant and antimicrobial wound dressings are the most favorable for acute and chronic wounds treatment. Herein, we formulated a multifunctional polyelectrolyte wound dressing membrane on the basis of chitosan (Ch) and hyaluronan (HA) enhanced by phosphatidylcholine dihydroquercetin (PCDQ). Physicochemical properties and microstructures of fabricated films were investigated adopting Fourier transform infrared (FT-IR) spectroscopy, thermogravimetric analysis (TGA) and scanning electron microscope (SEM). Furthermore, water uptakes, wettability profiles, surface roughness, and mechanical characteristics of the developed membranes were studied. The developed wound dressing revealed free radical scavenging potency, hemocompatibility with a tendency to enhance blood clotting. Furthermore, incorporation of PCDQ significantly promoted the antibacterial and anti-inflammatory activities of Ch/HA/PCDQ. Moreover, Ch/HA/PCDQ films exhibited cellular compatibility towards mouse fibroblast cells. The capability of Ch/HA/PCDQ to promote wound healing was evaluated using adult Wistar albino female rats. The in vivo findings demonstrated that Ch/HA/PCDQ films significantly ameliorated mouse full-thickness wounds as evidenced by a reduction in the wound area. Moreover, histological examinations of wounds dressed with Ch/HA/PCDQ illustrated a prominent re-epithelialization compared with wounds handled with the cotton gauze and Ch/HA dressings, exposing the efficiency of PCDQ. These findings emphasized that a Ch/HA/PCDQ membrane has outstanding potential for wound healing and skin regeneration.

Oxidative Degradation of High-Molar-Mass Hyaluronan: Effects of Some Indole Derivatives to Hyaluronan Decay.

Indole derivatives such as isatin (a natural compound), cemtirestat, stobadine, and its derivatives (synthetic compounds) are known to have numerous positive effects on human health due to regulation of oxidative status. The aim of the study was to assess radical scavenging capacities of these compounds and explore their potential protective effects against reactive oxygen species formed during Cu(II) ions and ascorbate-induced degradation of high-molar-mass hyaluronan. Based on the IC50 values determined by the ABTS assay, the most effective compound was SM1M3EC2·HCl reaching the value ≈ 11 µmol/L. The lowest IC50 value reached in the DPPH assay was reported for cemtirestat ≈ 3 µmol/L. Great potency of inhibition of hyaluronan degradation was shown by cemtirestat, followed by isatin even at low concentration 10 µmol/L. On the other hand, stobadine·2HCl had also a protective effect on hyaluronan degradation, however at greater concentrations compared to cemtirestat or isatin. SME1i-ProC2·HCl reported to be a less effective compound and SM1M3EC2·HCl can be considered almost ineffective compared to stobadine·2HCl. In conclusion, our results showed that both isatin and cemtirestat were capable of attenuating the degradation of high-molar-mass hyaluronan due to their ability to complex/sequester cupric ions.

Enhancement of wound healing by chitosan/hyaluronan polyelectrolyte membrane loaded with glutathione: in vitro and in vivo evaluations.

The inflammation of chronic wounds generally causes delaying their healing process. The present work aims to formulate a wound dressing polyelectrolyte membrane based on chitosan (Ch) and sodium hyaluronate (HA) loaded with glutathione (GSH). The membrane types (Ch/HA and Ch/HA/GSH) were examined by Fourier transform infrared spectroscopy (FT-IR). The material properties were further investigated using thermal gravimetric analysis (TGA) and scanning electron microscopy (SEM). Physical characteristics of the prepared membranes, such as wettability, surface roughness, and mechanical properties were determined by standard experimental methods. In vitro assays were used to evaluate the haemocompatibility, thrombogenicity, and cytotoxicity of the membranes. The wound healing examined using a standard rat model exhibited a progress at exploiting the Ch/HA/GSH-type membranes compared to a bicomponent Ch/HA membrane or a "dry" healing wound. Histological examination of the recovered skin confirmed the visual observations. In conclusion, in vivo study results assert that Ch/HA/GSH is a proper wound-dressing for healing the chronic skin wounds.

Skin wound healing with composite biomembranes loaded by tiopronin or captopril.

Novel wound dressings composed of chitosan (Ch) and hyaluronan (HA) loaded with tiopronin or captopril as antiinflammatory drugs were prepared. Composite biomembranes were examined in skin wounds of ischemic rabbits with the aim to accelerate the process of healing. The results proved that the biomembranes composed of Ch/HA/tiopronin or Ch/HA/captopril facilitated healing of skin wounds compared to untreated animals and animals treated with Ch/HA membranes. These results were confirmed by histology. Cu(II) ions and ascorbate-induced high-molar-mass HA degradation by means of rotational viscometry was performed and the ability of the both drugs to scavenge reactive oxygen species was evaluated. The results showed that captopril as well as tiopronin decreased the rate of HA degradation exclusively at higher concentrations.

Impact of Ergothioneine, Hercynine, and Histidine on Oxidative Degradation of Hyaluronan and Wound Healing.

A high-molecular weight hyaluronan is oxidatively degraded by Cu(II) ions and ascorbate-the so called Weissberger biogenic oxidative system-which is one of the most potent generators of reactive oxygen species, namely •OH radicals. Ergothioneine, hercynine, or histidine were loaded into chitosan/hyaluronan composite membranes to examine their effect on skin wound healing in ischemic rabbits. We also explored the ability of ergothioneine, hercynine, or histidine to inhibit hyaluronan degradation. Rotational viscometry showed that ergothioneine decreased the degree of hyaluronan radical degradation in a dose-dependent manner. While histidine was shown to be potent in scavenging •OH radicals, however, hercynine was ineffective. In vivo results showed that the addition of each investigated agent to chitosan/hyaluronan membranes contributed to a more potent treatment of ischemic skin wounds in rabbits compared to untreated animals and animals treated only with chitosan/hyaluronan membranes.

The importance of ergothioneine synthesis in ancient time by organisms living in oxygen free atmosphere.

The paper published by Ruczyszky and Liu (2017) reports on the biosynthesis of ergothioneine under both aerobic and anaerobic conditions. We would like to suggest a hypothesis as to what could be the reason that microorganisms on the Earth synthesized ergothioneine under anaerobic conditions.

Molecular hydrogen: potential in mitigating oxidative-stress-induced radiation injury 1.

Uncontrolled production of oxygen and nitrogen radicals results in oxidative and nitrosative stresses that impair cellular functions and have been regarded as causative common denominators of many pathological processes. In this review, we report on the beneficial effects of molecular hydrogen in scavenging radicals in an artificial system of •OH formation. As a proof of principle, we also demonstrate that in rat hearts in vivo, administration of molecular hydrogen led to a significant increase in superoxide dismutase as well as pAKT, a cell survival signaling molecule. Irradiation of the rats caused a significant increase in lipid peroxidation, which was mitigated by pre-treatment of the animals with molecular hydrogen. The nuclear factor erythroid 2-related factor 2 is regarded as an important regulator of oxyradical homeostasis, as well as it supports the functional integrity of cells, particularly under conditions of oxidative stress. We suggest that the beneficial effects of molecular hydrogen may be through the activation of nuclear factor erythroid 2-related factor 2 pathway that promotes innate antioxidants and reduction of apoptosis, as well as inflammation.

Chitosan/hyaluronan/edaravone membranes for anti-inflammatory wound dressing: In vitro and in vivo evaluation studies.

A novel wound healing material composed of chitosan (Ch) and hyaluronan (HA) boosted with edaravone (Ed) as an anti-inflammatory drug was developed. The fabricated membranes were verified using FT-IR, and the thermal properties were estimated employing TGA instrument. Moreover, Physical characterizations of the prepared membranes demonstrated a decrease in the membrane wettability, whereas an increase in membrane roughness was monitored due to the effect of edaravone supplementation. A comparative study of free-radical scavenging activity of edaravone itself was carried out by two in vitro approaches: uninhibited/inhibited hyaluronan degradation and decolorization of ABTS methods in normal and simulated inflammation condition (acidic condition). Accordingly, the scavenging activity of edaravone was significantly diminished to OH and peroxy-/alkoxy-type radicals in acidic conditions in compared to the neutral reactions. The biochemical studies evidenced the haemocompatibility of the examined membranes. The consequence of membranes composed of Ch/HA/Ed on the wound healing of the rat's skin was studied, and the macroscopic and microscopic investigations revealed remarkable healing at 21st day post-surgery compared with injuries treated with cotton gauze as a negative control in addition to Ch/HA membrane without edaravone. For these reasons, the Ch/HA/Ed membrane could be implemented as wound dressing material.

MitoQ Loaded Chitosan-Hyaluronan Composite Membranes for Wound Healing.

Two self-associating biopolymers, namely chitosan (Ch) and a high-molar-mass hyaluronan (HA), were used to prepare membranes with the aim to protect and to enhance the healing of injured skin. A mitochondrially-targeted antioxidant-MitoQ-was incorporated into the mixture of biopolymers prior to their self-association. These three-component membranes were evaluated in detail utilising surface roughness measurements, contact angle measurements, hemocompatibility, and thrombogenicity analyses. Furthermore, in vivo application of Ch/HA/MitoQ membranes was assessed on injured rabbit and rat skin utilizing histological methods. The results showed that the prepared thrombogenic Ch/HA/MitoQ membranes had higher roughness, which allowed for greater surface area for tissue membrane interaction during the healing processes, and lower cytotoxicity levels than controls. MitoQ-loaded composite membranes displayed superior healing properties in these animal models compared to control membranes.