Hyaluronic Acid Synthesis Research Articles

The effect of Salix alba L. bark extract on dark circles in vitro and in vivo.

Dark circles in the infraorbital area are a common cosmetic concern among individuals because they exhibit fatigue and are undesirable across all ages. Of the dark circle etiologies, blood stasis by poor vascular integrity can cause darkening of the lower eyelid skin, which might be alleviated by reduced endothelial permeability. In this study, we investigated the effects of Salix alba bark extract (SABE) on the synthesis of hyaluronic acid (HA) in fibroblasts and vascular integrity protection from inflammatory cytokine. We also performed a clinical trial investigating the effect of SABE on dark circles. To confirm the effect of SABE on HA synthesis in human dermal fibroblasts (HDF), we performed ELISA and real-time PCR. We investigated interaction HDF-secreted substance with vascular integrity, and human dermal microvascular endothelial cells (HMEC-1) were treated with conditioned medium (CM) from HDF treated with or without SABE. Subsequently, we conducted a clinical study on 29 subjects by having them apply SABE containing cream for 8 weeks. SABE treatment increased HA synthesis and regulated HMW-HA related gene expressions in HDF. CM from SABE-treated HDF alleviated endothelial permeability and led to improved vascular integrity in HMEC-1 cells. Treatment with the cream containing 2% SABE for 8 weeks improved the parameters measuring dark circles, skin microcirculation, and elasticity. Our results showed that SABE could protect against dark circles in vitro, and that topical treatment of SABE improved the clinical indexes of dark circles in a clinical study. Therefore, SABE can be used as an active ingredient for improving dark circles.

Complement protein C1q stimulates hyaluronic acid degradation via gC1qR/HABP1/p32 in malignant pleural mesothelioma.

Complement component C1q can act as a pro-tumorigenic factor in the tumor microenvironment (TME). The TME in malignant pleural mesothelioma (MPM) is rich in C1q and hyaluronic acid (HA), whose interaction enhances adhesion, migration and proliferation of malignant cells. HA-bound C1q is also capable of modulating HA synthesis. Thus, we investigated whether HA-C1q interaction would affect HA degradation, analyzing the main degradation enzymes, hyaluronidase (HYAL)1 and HYAL2, and a C1q receptor candidate. We first proceeded with the characterization of HYALs in MPM cells, especially HYAL2, since bioinformatics survival analysis revealed that higher HYAL2 mRNA levels have an unfavorable prognostic index in MPM patients. Interestingly, Real-Time quantitative PCR, flow cytometry and Western blot highlighted an upregulation of HYAL2 after seeding of primary MPM cells onto HA-bound C1q. In an attempt to unveil the receptors potentially involved in HA-C1q signaling, a striking co-localization between HYAL2 and globular C1q receptor/HABP1/p32 (gC1qR) was found by immunofluorescence, surface biotinylation and proximity ligation assays. RNA interference experiments revealed a potentially regulatory function exerted by gC1qR on HYAL2 expression, since C1QBP (gene for gC1qR) silencing unexpectedly caused HYAL2 downregulation. In addition, the functional blockage of gC1qR by a specific antibody hindered HA-C1q signaling and prevented HYAL2 upregulation. Thus, C1q-HA interplay is responsible for enhanced HYAL2 expression, suggesting an increased rate of HA catabolism and the release of pro-inflammatory and pro-tumorigenic HA fragments in the MPM TME. Our data support the notion of an overall tumor-promoting property of C1q. Moreover, the overlapping localization and physical interaction between HYAL2 and gC1qR suggests a potential regulatory effect of gC1qR within a putative HA-C1q macromolecular complex.

Modulation of lncRNA links endothelial glycocalyx to vascular dysfunction of tyrosine kinase inhibitor.

Novel cancer therapies leading to increased survivorship of cancer patients have been negated by a concomitant rise in cancer therapies-related cardiovascular toxicities. Sunitinib, a first line multi-receptor tyrosine kinase inhibitor, has been reported to cause vascular dysfunction although the initiating mechanisms contributing to this side effect remain unknown. Long non-coding RNAs (lncRNAs) are emerging regulators of biological processes in endothelial cells (ECs); however, their roles in cancer therapies-related vascular toxicities remain underexplored. We performed lncRNA expression profiling to identify potential lncRNAs that are dysregulated in human-induced pluripotent stem cell-derived ECs (iPSC-ECs) treated with sunitinib. We show that the lncRNA hyaluronan synthase 2 antisense 1 (HAS2-AS1) is significantly diminished in sunitinib-treated iPSC-ECs. Sunitinib was found to down-regulate HAS2-AS1 by an epigenetic mechanism involving hypermethylation. Depletion of HAS2-AS1 recapitulated sunitinib-induced detrimental effects on iPSC-ECs, whereas CRISPR-mediated activation of HAS2-AS1 reversed sunitinib-induced dysfunction. We confirmed that HAS2-AS1 stabilizes the expression of its sense gene HAS2 via an RNA/mRNA heteroduplex formation. Knockdown of HAS2-AS1 led to reduced synthesis of hyaluronic acid (HA) and up-regulation of ADAMTS5, an enzyme involved in extracellular matrix degradation, resulting in disruption of the endothelial glycocalyx which is critical for ECs. In vivo, sunitinib-treated mice showed reduced coronary flow reserve, accompanied by a reduction in Has2os and degradation of the endothelial glycocalyx. Finally, we identified that treatment with high molecular-weight HA can prevent the deleterious effects of sunitinib both in vitro and in vivo by preserving the endothelial glycocalyx. Our findings highlight the importance of lncRNA-mediated regulation of the endothelial glycocalyx as an important determinant of sunitinib-induced vascular toxicity and reveal potential novel therapeutic avenues to attenuate sunitinib-induced vascular dysfunction.

Heterologous Biosynthesis of Hyaluronic Acid Using a New Hyaluronic Acid Synthase Derived from the Probiotic Streptococcus thermophilus

Hyaluronic acid (HA) is a natural linear polysaccharide extensively used in many fields, including the food, medicine, and cosmetics industries. Currently, species that produce HA synthetase (HAS) from microbial sources are relatively small and mainly pathogenic, such as Streptococcus pyogenes and Pasteurella multicide. Moreover, there is limited research on the safe microbial sources of HAS. Thus, we characterized SthasA, a HAS derived from the probiotic Streptococcus thermophilus, and used it for the de novo synthesis of HA in a chassis strain of Bacillus amyloliquefaciens. Metabolic engineering of the precursor supply modules suggested that hasB (encoding UDPG dehydrogenase), which was derived from Corynebacterium glutamicum ATCC 13032, effectively promoted the accumulation of HA products. Furthermore, by combining the expression of the global regulatory factor CcpA, HA yield from the recombinant strain reached 3.20 g/L. Finally, we obtained a yield of 5.57 g/L HA with a molecular weight of 1.7 × 106 Da using various process optimization strategies in a 5 L bioreactor. This study enriches our understanding of obtaining HAS from non-pathogenic bacteria and provides a safe and effective process for producing HA, which has the potential to promote the industrial applications of HA further.

Downregulation of HAS‑2 regulates the chondrocyte cytoskeleton and induces cartilage degeneration byactivating the RhoA/ROCK signaling pathway.

Osteoarthritis (OA) is a progressive joint disorder, which is principally characterized by the degeneration and destruction of articular cartilage. The cytoskeleton is a vital structure that maintains the morphology and function of chondrocytes, and its destruction is a crucial risk factor leading to chondrocyte degeneration and OA. Hyaluronan synthase‑2 (HAS‑2) is a key enzyme in synthesizing hyaluronic acid (HA) invivo. The synthesis of high molecular weight HA catalyzed by HAS‑2 serves a vital role in joint movement and homeostasis; however, it is unclear what important role HAS‑2 plays in maintaining chondrocyte cytoskeleton morphology and in cartilage degeneration. The present study downregulated the expression of HAS‑2 by employing 4‑methylumbelliferone (4‑MU) and RNA interference. Invitro experiments, including reverse transcription‑quantitative PCR, western blotting, laser scanning confocal microscopy and flow cytometry were subsequently performed. The results revealed that downregulation of HAS‑2 could activate the RhoA/ROCK signaling pathway, cause morphological abnormalities, decrease expression of the chondrocyte cytoskeleton proteins and promote chondrocyte apoptosis. Invivo experiments, including immunohistochemistry and Mankin's scoring, were performed to verify the effect of HAS‑2 on the chondrocyte cytoskeleton, and it was revealed that inhibition of HAS‑2 could cause cartilage degeneration. In conclusion, the present results revealed that downregulation of HAS‑2 could activate the RhoA/ROCK pathway, cause abnormal morphology and decrease chondrocyte cytoskeleton protein expression, leading to changes in the signal transduction and biomechanical properties of chondrocytes, promotion of chondrocyte apoptosis and the induction of cartilage degeneration. Moreover, the clinical application of 4‑MU may cause cartilage degeneration. Therefore, targeting HAS‑2 may provide a novel therapeutic strategy for delaying chondrocyte degeneration, and the early prevention and treatment of OA.

Regulation of hyaluronic acid synthesis and breakdown can prevent significant damage of the lung epithelium in response to SARS-CoV-2 infection

COVID-19 remains an ongoing global pandemic causing significant respiratory distress and endothelial dysfunction in individuals susceptible to severe disease. Increased production of hyaluronic acid has been observed in patients with severe COVID-19, however its significance in COVID-19 pathogenesis is currently unknown. Hymecromone is a hyaluronic acid synthase inhibitor FDA approved for use in humans as a choleretic. We hypothesized that excessive production of hyaluronic acid by the airway epithelium, in response to SARS-CoV-2 infection, may increase hyaluronan fragments that augment epithelial dysfunction in COVID-19. We therefore evaluated a hyaluronic acid synthase inhibitor, hymecromone, for its potential to regulate the infection of respiratory epithelia by SARS-CoV-2. Using air-liquid interface models we evaluated the impact of SARS-CoV-2 infection on differentiated primary human airway epithelium in the BSL3 facility. At 100μg/ml hymecromone significantly inhibited the cytopathic effects of SARS-CoV-2 on airway epithelium (p<0.001). After 24 hours, SARS-CoV-2 virus accumulated in the basolateral compartment after apical infection of the airway cultures. Hymecromone also significantly inhibited this basolateral accumulation of viral particles versus untreated controls (p<0.05). Additionally, hymecromone alters the cytokine response to SARS-CoV-2 infection, dose-dependently decreasing apical secretion of cytokines including IL-17A/B/D, IL-21 and IL-5, and dose-dependently increasing the basolateral secretion of IL-17A/D and IL-1RA. All experiments were performed on three independent donor cells with three experimental repeats per condition. In conclusion, our data highlight the importance of the regulation of hyaluronic acid production in the response of the airway epithelium to SARS-CoV-2 infection and indicate hymecromone as a potential therapeutic option. Funding for this study was provided by the Rainwater Foundation. This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Interaction between epidermal growth factor receptor and C-C motif chemokine receptor 2 in the ovulatory cascade.

The epidermal growth factor receptor (EGFR) signaling pathway is one of the main pathways responsible for propagating the luteinizing hormone (LH) signal throughout the cumulus cells and the oocyte. Recently, we have proposed the C-C motif chemokine receptor 2 (CCR2) and its main ligand (monocyte chemoattractant protein-1, MCP1) as novel mediators of the ovulatory cascade. Our previous results demonstrate that the gonadotropins (GNT), amphiregulin (AREG), and prostaglandin E2 (PGE2) stimulation of periovulatory gene mRNA levels occurs, at least in part, through the CCR2/MCP1 pathway, proposing the CCR2 receptor as a novel mediator of the ovulatory cascade in a feline model. For that purpose, feline cumulus-oocyte complexes (COCs) were cultured in the presence or absence of an EGFR inhibitor, recombinant chemokine MCP1, and gonadotropins [as an inducer of cumulus-oocyte expansion (C-OE), and oocyte maturation] to further assess the mRNA expression of periovulatory key genes, C-OE, oocyte nuclear maturation, and steroid hormone production. We observed that MCP1 was able to revert the inhibition of AREG mRNA expression by an EGFR inhibitor within the feline COC. In accordance, the confocal analysis showed that the GNT-stimulated hyaluronic acid (HA) synthesis, blocked by the EGFR inhibitor, was recovered by the addition of recombinant MCP1 in the C-OE culture media. Also, MCP1 was able to revert the inhibition of progesterone (P4) production by EGFR inhibitor in the C-OE culture media. Regarding oocyte nuclear maturation, recombinant MCP1 could also revert the inhibition triggered by the EGFR inhibitor, leading to a recovery in the percentage of metaphase II (MII)-stage oocytes. In conclusion, our results confirm the chemokine receptor CCR2 as a novel intermediate in the ovulatory cascade and demonstrate that the EGFR/AREG and the CCR2/MCP1 signaling pathways play critical roles in regulating feline C-OE and oocyte nuclear maturation, with CCR2/MCP1 signaling pathway being downstream EGFR/AREG pathway within the ovulatory cascade.

A Safe-by-Design Approach for the Synthesis of a Novel Cross-Linked Hyaluronic Acid with Improved Biological and Physical Properties.

Hyaluronic acid (HA) is a polymer with unique biological properties that has gained in interest over the years, with applications in pharmaceutical, cosmetic, and biomedical fields; however, its widespread use has been limited by its short half-life. Therefore, a new cross-linked hyaluronic acid was designed and characterized using a natural and safe cross-linking agent, such as arginine methyl ester, which provided improved resistance to enzymatic action, as compared to the corresponding linear polymer. The antibacterial profile of the new derivative was shown to be effective against S. aureus and P. acnes, making it a promising candidate for use in cosmetic formulations and skin applications. Its effect on S. pneumoniae, combined with its excellent tolerability profile on lung cells, also makes this new product suitable for applications involving the respiratory tract.

4-Methylumbelliferone Targets Revealed by Public Data Analysis and Liver Transcriptome Sequencing.

4-methylumbelliferone (4MU) is a well-known hyaluronic acid synthesis inhibitor and an approved drug for the treatment of cholestasis. In animal models, 4MU decreases inflammation, reduces fibrosis, and lowers body weight, serum cholesterol, and insulin resistance. It also inhibits tumor progression and metastasis. The broad spectrum of effects suggests multiple and yet unknown targets of 4MU. Aiming at 4MU target deconvolution, we have analyzed publicly available data bases, including: 1. Small molecule library Bio Assay screening (PubChemBioAssay); 2. GO pathway databases screening; 3. Protein Atlas Database. We also performed comparative liver transcriptome analysis of mice on normal diet and mice fed with 4MU for two weeks. Potential targets of 4MU public data base analysis fall into two big groups, enzymes and transcription factors (TFs), including 13 members of the nuclear receptor superfamily regulating lipid and carbohydrate metabolism. Transcriptome analysis revealed changes in the expression of genes involved in bile acid metabolism, gluconeogenesis, and immune response. It was found that 4MU feeding decreased the accumulation of the glycogen granules in the liver. Thus, 4MU has multiple targets and can regulate cell metabolism by modulating signaling via nuclear receptors.

Evaluation of saffron extract bioactivities relevant to skin resilience

IntroductionSaffron, Crocus sativus L, is a perennial spice herb. It has been extensively studied for its antioxidant, antidepressant, and anti-inflammatory properties, and has recently gained new interest for use in high-end cosmetics. The present work aims to elucidate the skin-protective properties of saffron in human dermal fibroblasts. MethodsThe skin-protective properties of saffron extract were evaluated in terms of tyrosinase and collagenase inhibition activities, antioxidant activity in mouse macrophage cells, collagen synthesis and hyaluronic acid synthesis and cell migration activity in primary dermal fibroblast normal human neonatal cells (HDFn). ResultsSaffron’s main phytoactive constituents - crocins, picrocrocin, safranal, and crocetin - were quantified by LC-MS at 91.0, 61.5, 3.6, and 1.9 mg/g, respectively. Saffron extract inhibited tyrosinase and collagenase with IC50 0.78 mg/mL and 0.1 mg/mL, respectively. Saffron extracts (100–200 μg/mg) suppressed reactive oxygen species and nitric oxide generation in lipopolysaccharide-stimulated RAW 264.7 murine macrophage cells and promoted collagen and hyaluronic acid synthesis in HDFn. Saffron extract at 25 μg/mg significantly promoted migration of HDFn cells (wound healing capacity) compared to control (no treatment). ConclusionThe findings highlighted the potential benefits of saffron extract relevant to skin resilience.

Pure Platelet and Leukocyte-Platelet-Rich Plasma for Regenerative Medicine in Orthopedics-Time- and Preparation-Dependent Release of Growth Factors and Effects on Synovial Fibroblasts: A Comparative Analysis.

Intra-articular injections of autologous platelet concentrates are considered capable to enhance the healing of cartilage lesions, alleviate joint inflammation, and relieve other musculoskeletal pathological conditions. The aim of this study was to analyze the soluble fractions obtained from platelet-rich plasma (pure- and leukocyte-PRP) to compare time- and preparation-dependent modifications of growth factor concentrations and the supporting activity of the two preparations on synovial fibroblast growth and hyaluronic acid (HA) production in vitro. The release kinetics of FGF-2, SDF-1, VEGF, HGF, EGF, PD GF-AB/BB, IGF-1, VCAM-1, and TGF-β isoforms were followed up to 168 h after PRP activation, and their amounts were determined by multiplex-beads immunoassay. Synovial cell growth and supernatant HA production were respectively analyzed by Alamar Blue assay and ELISA. Time-dependent modifications grouped molecules in three peculiar patterns: one reaching the highest concentrations within 18 h and decreasing afterwards, another progressively increasing up to 168 h, and the last peaking at the central time points. Synovial fibroblast growth in response to L-PRP and P-PRP revealed differences over time and among added concentrations. Both preparations displayed a preserved supporting capacity of HA synthesis.

Green synthesis of hyaluronic acid coated, thiolated chitosan nanoparticles for CD44 targeted delivery and sustained release of Cisplatin in cervical carcinoma.

Cervical carcinoma is one of the most prevalent gynecological cancers throughout the world. Cisplatin is used as first line chemotherapy for treatment of cervical cancer, but it comes with plethora of side effects. The aim of this study was to develop hyaluronic acid coated, thiolated chitosan nanocarriers using green synthesis approach, for CD44 targeted delivery and sustained release of Cisplatin in cervical cancer cells. After synthesis through ionic gelation method, Zeta analysis showed that the nanoparticle size was 265.9nm with a zeta potential of +22.3mV and .226 PDI. SEM and TEM analysis confirmed the spherical shape and smooth surface of nanoparticles. FTIR and XRD showed the presence of characteristic functional groups, successful encapsulation of drug, and crystalline nature of nanoparticles respectively. Drug loading and entrapment efficiency were calculated to be 70.1% ± 1.2% and 45% ± .28% respectively. Analysis of in vitro drug release kinetics showed that drug release followed the Higuchi model at pH 6.8 and 7.4 and Cisplatin release for up to 72h confirmed sustained release. In vitro analysis on cervical cancer cells HeLa and normal cervical epithelial cells HCK1T was done through cell morphology analysis, trypan blue assay (concentration range of 10-80μg/ml), and MTT cytotoxic assay (concentration range of 10-90μg/ml). The results showed a higher cytotoxic potential of HA coated, thiolated chitosan encapsulated Cisplatin (HA-ThCs-Cis NP) nanoformulation as compared to pure Cisplatin in HeLa while in HCK1T, pure Cisplatin showed much higher toxicity as compared to HA-ThCs-Cis nanoformulation. These findings suggest that CD44 targeted delivery system can be a useful approach to minimize offtarget toxicities, give sustained release and better cellular uptake in cancer cells.

Brain inflammation induces alterations in glycosaminoglycan metabolism and subsequent changes in CS-4S and hyaluronic acid

It remains uncertain how brain glycosaminoglycans (GAGs) contribute to the progression of inflammatory disorders like multiple sclerosis (MS). We investigated here neuroinflammation-mediated changes in GAG composition and metabolism using the mouse model of experimental autoimmune encephalomyelitis (EAE) and sham-immunized mice as controls. Cerebellum, mid- and forebrain at different EAE phases were investigated using gene expression analysis (microarray and RT-qPCR) as well as HPLC quantification of CS and hyaluronic acid (HA). The cerebellum was the most affected brain region showing a downregulation of Bcan, Cspg5, and an upregulation of Dse, Gusb, Hexb, Dcn and Has2 at peak EAE. Upregulation of genes involved in GAG degradation as well as synthesis of HA and decorin persisted from onset to peak, and diminished at remission, suggesting a severity-related decrease in CS and increments in HA. Relative disaccharide quantification confirmed a 3.6 % reduction of CS-4S at peak and a normalization during remission, while HA increased in both phases by 26.1 % and 17.6 %, respectively. Early inflammatory processes led to altered GAG metabolism in early EAE stages and subsequent partially reversible changes in CS-4S and in HA. Targeting early modifications in CS could potentially mitigate progression of EAE/MS.

Fine-tuning the cell morphology of Corynebacterium glutamicum via dual-valve regulation for enhanced hyaluronic acid production

Enhanced synthesis of hyaluronic acid (HA) with recombinant Corynebacterium glutamicum as production host is achieved in this work. Hyaluronan synthase (HAS), which is a membrane protein acting as a key enzyme in HA biosynthesis, impacts both HA yield and its molecular weight. Cell morphology, which includes size, shape, and surface area, has a large impact on the expression and activity of HAS. Therefore, deliberate regulation of cell morphology holds the potential to enhance HA production. Here, we constructed three modules, namely the transporter module, the morphology tuning module and the HA synthesis module. The transporter module contains a strong constitutive promoter Ptuf and arabinose transport protein used to control the maximum amount of inducer entering the cell, thus reducing excessive cell deformation. The morphology tuning module contains an arabinose-inducible weak promoter PBAD and a cell-division-relevant gene used to sense intracellular inducer concentrations and achieve different degrees of change in cell size. These two modules work together, described as a dual-valve regulation, to achieve fine-tuning of cell morphology, resulting in a 1.87-fold increase in cell length and a 2.08-fold increase in cell membrane. When combined with the HA synthesis module, the HA titer reached 16.0 g/L, which is 1.6 times the yield reported in the previous morphology-engineered strain. Hence, for the first time, a morphologically engineered strain resulting in both high cell density and HA titer was constructed.

Protection Impacts of Coix lachryma-jobi L. Seed Lactobacillus reuteri Fermentation Broth on Hydrogen Peroxide-Induced Oxidative Stress in Human Skin Fibroblasts

This study researches the active ingredients in the fermented and aqueous extracts of three types of Coix seed. The results show that the active contents of total sugars, total phenols and total proteins of Coix seed fermented with Lactobacillus reuteri, Saccharomyces cerevisiae and Lactobacillus bulgaricus are significantly higher compared to unfermented Coix seed aqueous extract (CW), and the free radical scavenging ability of Coix seed Lactobacillus reuteri fermented extract (CLRF) is significant. The protective effects of CLRF on hydrogen peroxide (H2O2)-induced oxidative stress in human skin fibroblasts (HSF) are investigated. The results show that cell viability, total antioxidant capacity, superoxide dismutase (SOD) activity, catalase (CAT) activity, collagen type I (COL-I) content and synthesis of hyaluronic acid (HA) significantly increased, and reactive oxygen species (ROS), matrix metalloproteinase-1 (MMP-1) content are decreased in CLRF-treated HSF compared to CW and damage model groups, providing effective protection to skin structure. The results show that CLRF can stimulate the PI3K/AKT signaling pathway, inhibiting intracellular ROS levels, positively regulating COL-I genes and significantly reducing MMP-1 expression, with anti-oxidative damage effects. As such, this study provides a theoretical basis for applying CLRF as a novel anti-oxidative agent in the cosmetics industry.

Effects of pharmacological inhibition of hyaluronic acid synthesis on experimental endometriosis.

Dysregulated hyaluronic acid (HA) metabolism has been shown to be implicated in several pathologies including endometriosis. 4-Methylumbelliferone (4MU) is an HA synthesis inhibitor with proven antitumour activity. In this study, we aim to evaluate the effect of 4MU on endometriosis development both in vivo and in vitro. Endometriosis was surgically induced by uterine tissue auto-transplantation in 32 two-month-old BALB/c mice. Animals were designated into the early or late starting treatment group, which initiated on day 2 or day 15 after surgery, respectively. Within each group, 4MU 200 mg/kg/day or vehicle (Control) were administered by oesophageal gavage for 28 days. After sacrifice, the percentage of developed lesions, lesion size, cell proliferation, vascularization and HA deposition within the endometriotic-like lesions were evaluated. Cell viability was assessed in endometrial epithelial cells (ECC-1) and in endometrial stromal cells (t-HESC); and migration was evaluated in t-HESC. There was a significant reduction in the percentage of developed lesions in mice that started the 4MU treatment on day 2 compared with its respective control group, and compared with those that started treatment on day 15. However, no significant changes were found when analysing endometriotic-like lesion's cell proliferation, vascularization and HA deposition. In vitro, both cell viability and migration were inhibited by 4MU treatment. The inhibition of HA synthesis could be a beneficial and alternative option to treat endometriosis at the early stage of the disease. Further research is necessary to elucidate 4MU's mechanism of action and better strategies for delivering this promising drug.

Benefits of topical hyaluronic acid for skin quality and signs of skin aging: From literature review to clinical evidence.

Skin aging goes beyond a chronological process and also results from extrinsic factors referred to as the exposome. Hyaluronic acid (HA) is an important component of the extracellular matrix, with loss starting at 25 years old. While many studies of HA concern topical use, few literature reviews only address the use of topical HA in dermatology. This review describes the different characteristics of HA-containing cosmeceuticals, with a focus on skin aging and the impact of exposome factors on HA synthesis and degradation. A review was performed using the terms HA, hyaluronan, topical, dermatology, cosmetic, aging treatment, exposome, and cosmeceuticals. Results are also presented from a recent randomized controlled trial (RCT), which investigated the additional benefit of using a HA epidermic filler (HA-filler serum) combined with Botulinum toxin type A (BoNTA) to treat signs of skin aging. Subjects were randomized to two groups: HA-filler serum starting 24 h after the BoNTA injection then twice daily for 24 weeks, or the control group, which received BoNTA. HA is a key ingredient used in cosmeceuticals for its hydration/antiaging properties (hygroscopic, rheological, and viscoelastic). Several clinical studies indicate that HA is both well tolerated and effective, adjuvant to both post-surgical and facial rejuvenation procedures. In the RCT, one of few studies to combine BoNTA and HA with a 6-month follow-up, the HA-filler serum lengthened the duration of BoNTA's effect in reducing wrinkles. Numerous studies support HA-based cosmeceuticals as a noninvasive, effective solution for improving skin hydration and rejuvenation.

Prospects of Hyaluronidase Therapy in Novel COVID-19 Infection with Damage to Lungs

INTRODUCTION: In recent years, the production and regulation of synthesis of hyaluronic acid in COVID-19 has been actively studied. Hyaluronan plays a significant role in development of severe lung damage in COVID-19 and is a potential therapeutic target the action on which will probably improve prognosis for patients with COVID-19.
 AIM: To study prospects of using bovhyaluronidaze azoximer in complex treatment of patients with COVID-19 with lung damage at the inpatient stage.
 MATERIALS AND METHODS: Thirty five patients (6 men and 29 women) aged 58.9 12.9 years hospitalized with COVID-19 infection, were examined. Capillary blood saturation (SpO2) was 80.1 8.6%, the volume of lung damage in X-ray computed tomography (X-ray CT) was 45.1 19.4% on the right and 40.0 19.5% on the left. All the patients received treatment according to the Temporary Guidelines: prevention, diagnosis and treatment of novel coronavirus infection. Ver. 14 (27.12.2021). Besides, as part of complex treatment for COVID-19, bovhyaluronidaze azoximer was administered intramuscularly on the 21.9 6.8th day of illness with a course of 10 injections (once in 3 days).
 RESULTS: In the course of comprehensive treatment including bovhyaluronidaze azoximer, increase in SpO2 was recorded: in 7 patients ― after 1 injection (4.2 1.7%), in 24 ― after 2 injections (5.4 0.6%), another 4 patients did not show any significant increase in SpO2 after the first two injections. Increase in SpO2 after the 1st injection inversely correlated with age (r = -0.34; p 0.05) and the initial saturation (r = -0.38; p 0.05). Increase in SpO2 after the second injection correlated with the day of illness on which treatment with bovhyaluronidaze azoximer began (r = -0.36; p 0.05).
 CONCLUSION: Use of bovhyaluronidaze azoximer in complex treatment for COVID-19 with the lung damage at the inpatient stage can be effective in younger patients with more expressed initial reduction of SpO2, and also in case of administration of the drug in the early stages of the disease. The data obtained in the pilot study, dictate the necessity of studying the level of hyaluronic acid in blood of patients with COVID-19 and lung damage and its role in risk stratification of such patients.

Clinically Relevant Biology of Hyaluronic Acid in the Desmoplastic Stroma of Pancreatic Ductal Adenocarcinoma.

Pancreatic ductal adenocarcinoma (PDAC) is notorious for its poor outcome. The presence of a dense desmoplastic stroma is a hallmark of this malignancy, and abundant hyaluronic acid (HA) within this stroma is a common feature of PDAC. At the end of 2019, an HA-targeting drug, after initial promise, failed phase 3 clinical trials in PDAC. This failure in the face of such strong evidence for biological importance forces us to turn back to the research and seek a better understanding of HA biology in PDAC. Therefore, in this review, we reexamine what is known about HA biology, the methods used to detect and quantify HA, and the ability of the biological models in which HA has been investigated to recapitulate an HA-rich desmoplastic tumor stroma. The role of HA in PDAC relies on its complex interplay with a range of HA-associated molecules, which have not been as extensively investigated as HA itself. Therefore, using large genomic data sets, we cataloged the abundance and activity in PDAC of molecules that modulate HA synthesis, degradation, protein interactions, and receptor binding. Based on their association with clinical characteristics and individual patient outcomes, we suggest a small number of HA-associated molecules that warrant further investigation as biomarkers and drug targets.

Anti-aging efficacy of solid-state fermented ginseng with Aspergillus cristatus and its active metabolites.

Aspergillus cristatus is a beneficial fungus of microbial fermented teas such as China’s Fuzhuan brick tea and Pu-erh tea, and is commonly called golden flower fungus (GFF) because its cleistothecium has a yellow millet or sand grain shape. Since natural materials fermented with GFF exhibit various physiological activities, a new active cosmeceutical ingredient was developed by solid-state fermentation of ginseng, a famous active material for healthy skin, with GFF. The extract of solid-state fermented ginseng with GFF (GFFG) exhibited potent anti-aging efficacy on the skin such as the increase of hyaluronic acid synthesis, aquaporin expression, and mRNA level of filaggrin in HaCaT keratinocyte. GFFG also inhibited the expression of MMP-1 increased by TNF-α in human dermal fibroblast. Sophisticated chromatographic and spectroscopic studies have elucidated isodihydroauroglaucin and flavoglaucin as the metabolites which were not present in ginseng extract nor GFF extract alone. Bioassay of these metabolites revealed that these compounds were part of active principles of GFFG. These results suggest that GFFG would be a potential active ingredient in anti-aging cosmeceutical products.