Hyaluronic Acid Concentration Research Articles

Development and Characterization of n-Propyl Gallate Encapsulated Solid Lipid Nanoparticles-Loaded Hydrogel for Intranasal Delivery.

The objective of the present study was to develop n-propyl gallate-loaded solid lipid nanoparticles (PG-SLNs) in a hydrogel (HG) formulation using Transcutol-P (TC-P) as a permeation enhancer. Modified solvent injection technique was applied to produce optimized PG-SLNs via the Quality by Design approach and central composite design. The in vitro mucoadhesion, scavenging activity, drug release, permeation studies of PG from PG-SLNs-loaded HG were evaluated under simulated nasal conditions. Compared with in vitro release behavior of PG from SLNs, the drug release from the PG-SLNs-loaded HG showed a lower burst effect and sustained release profile. The cumulative permeation of PG from PG-SLNs-loaded HG with TC-P was 600 μg/cm2 within 60 min, which is 3–60-fold higher than PG-SLNs and native PG, respectively. Raman mapping showed that the distribution of PG-SLNs was more concentrated in HG having lower concentrations of hyaluronic acid. The scavenging assay demonstrated increased antioxidant activity at higher concentrations of HG. Due to enhanced stability and mucoadhesive properties, the developed HG-based SLNs can improve nasal absorption by increasing residence time on nasal mucosa. This study provides in vitro proof of the potential of combining the advantages of SLNs and HG for the intranasal delivery of antioxidants.

Repetitive Synthesis of High-Molecular-Weight Hyaluronic Acid with Immobilized Enzyme Cascades.

Industrial hyaluronic acid (HA) production comprises either fermentation with Streptococcus strains or extraction from rooster combs. The hard‐to‐control product quality is an obstacle to these processes. Enzymatic syntheses of HA were developed to produce high‐molecular‐weight HA with low dispersity. To facilitate enzyme recovery and biocatalyst re‐use, here the immobilization of cascade enzymes onto magnetic beads was used for the synthesis of uridine‐5’‐diphosphate‐α‐d‐N‐acetyl‐glucosamine (UDP‐GlcNAc), UDP‐glucuronic acid (UDP‐GlcA), and HA. The combination of six enzymes in the UDP‐sugar cascades with integrated adenosine‐5'‐triphosphate‐regeneration reached yields between 60 and 100 % for 5 repetitive batches, proving the productivity. Immobilized HA synthase from Pasteurella multocida produced HA in repetitive batches for three days. Combining all seven immobilized enzymes in a one‐pot synthesis, HA production was demonstrated for three days with a HA concentration of up to 0.37 g L−1, an average M W of 2.7–3.6 MDa, and a dispersity of 1.02–1.03.

Concentrations of tissue inhibitor of matrix metalloproteinase-1 and hyaluronic acid in canine amniotic membranes cryopreserved for different time points and its effects in dogs with complicated corneal ulcers.

To determine the concentrations of total protein (TP), tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) and hyaluronic acid (HA) in amniotic membranes (AMs) harvested from placentas of bitches of different ages and cryopreserved for different time points. The outcomes of complicated corneal defects of dogs repaired with AMs stored for the same time points were also evaluated. Ten cryopreserved canine AMs were stored for short term (2-50days), middle term (92-210days), or long term (256-357days). TP was quantified by Bradford's test, whereas TIMP-1 and HA were quantified by ELISA. Twenty-one dogs that had an AM transplantation to restore deep or perforating corneal wounds were selected. TIMP-1levels were lower in AMs cryopreserved for middle term (p=.02) and long term (p=.0009), when compared to AMs stored for short term. TP (p=.39) and HA (p=.18) concentrations in AMs did not differ among the storage time. TIMP-1 concentration in AMs correlated with storage time (R=-.65, p<.0001), while TP (R=-.33, p=.07) and HA concentrations did not (R=-.15, p=.41). The age of donors did not correlate with the components evaluated in the AMs. Corneal defects repaired with AMs stored for short term healed sooner than the ones repaired with AMs stored for middle (p<.01) and long term (p=.02). Additionally, TIMP-1levels in AMs correlated negatively with the epithelization time (R=-.62, p=.002). Graft opacity was severe in 55% of cases. However, the HA levels in AMs correlated negatively with the opacification score (R=-.47, p=.03). Vision was observed in more patients who presented deep ulcers and descemetoceles, than in the ones with perforations (p=.004). TIMP-1 concentration in canine AMs significantly decreased over a year storage time, while TP and HA concentrations did not change during the same period. The age of donors did not correlate with the components evaluated in the AMs. Complicated corneal defects repaired with AMs cryopreserved for short term healed sooner and tended to be less opaque; however, satisfactory to optimal outcomes were achieved even in the eyes repaired with AMs stored for up to a year.

Use of serum hyaluronic acid as a biomarker of endothelial glycocalyx degradation in dogs with septic peritonitis.

To describe daily changes in serum concentrations of hyaluronic acid (HA), a biomarker of endothelial glycocalyx degradation, in dogs with septic peritonitis and to determine whether relationships exist among serum concentrations of HA and biomarkers of inflammation and patient fluid status. 8 client-owned dogs. Serum samples that had been collected for a previous study and stored at -80°C were used. Blood samples were collected at admission and daily thereafter during hospitalization and were analyzed for concentrations of HA and interleukins 6, 8, and 10. Patient data including acute patient physiologic and laboratory evaluation score, type and amount of fluids administered daily, and daily CBC and lactate concentration results were recorded. To determine the significant predictors of HA concentration, a general linear mixed model for repeated measures was developed. All dogs survived to discharge. Concentrations of HA ranged from 18 to 1,050 ng/mL (interquartile [25th to 75th percentile] range, 49 to 119 ng/mL) throughout hospitalization. Interleukin-6 concentration was a significant predictor of HA concentration as was total administered daily fluid volume when accounting for interleukin-6 concentration. When fluid volume was analyzed independent of inflammatory status, fluid volume was not a significant predictor. Concentrations of HA did not significantly change over time but tended to increase on day 2 or 3 of hospitalization. Results supported the theory that inflammation is associated with endothelial glycocalyx degradation. Dogs recovering from septic peritonitis may become more susceptible to further endothelial glycocalyx damage as increasing fluid volumes are administered.

Network Pharmacological Analysis and Experimental Validation of the Mechanisms of Action of Si-Ni-San Against Liver Fibrosis.

Background: Si-Ni-San (SNS), a commonly used traditional Chinese medicine (TCM) formula, has potency against liver diseases, such as hepatitis and non-alcoholic fatty liver disease (NAFLD). However, the therapeutic efficacy and pharmacological mechanisms of action of SNS against liver fibrosis remain largely unclear. Methods: A carbon tetrachloride (CCl4)-induced liver fibrosis mouse model was adopted for the first time to investigate the beneficial effects of SNS on liver fibrosis. The potential mechanisms of action of SNS were explored using the network pharmacology-based strategy and validated with the aid of diverse assays. Results: SNS treatment reduced collagen and ECM deposition, downregulated fibrosis-related factor (hyaluronic acid and laminin) contents in serum, maintained the morphological structure of liver tissue, and improved liver function in the liver fibrosis model. Based on network pharmacology results, apoptosis, inflammation and angiogenesis, together with the associated pathways (including VEGF, TNF, caspase, PPAR-γ and NF-κB), were identified as the mechanisms underlying the effects of SNS on liver fibrosis. Further in vivo experiments validated the significant mitigatory effects of SNS on inflammatory infiltration and pro-inflammatory cytokine contents (IFNγ, IL-1β and TGF-β1) in liver tissues of mice with liver fibrosis. SNS suppressed pathologic neovascularization as well as levels of VEGFR1, VEGF and VEGFR2 in liver tissues. SNS treatment additionally inhibited hepatic parenchyma cell apoptosis in liver tissues of mice with liver fibrosis and regulated apoptin expression while protecting L02 cells against apoptosis induced by TNF-α and Act D in vitro. Activation of hepatic stellate cells was suppressed and the balance between MMP13 and TIMP1 maintained in vitro by SNS. These activities may be associated with SNS-induced NF-κB suppression and PPAR-γ activation. Conclusion: SNS effectively impedes liver fibrosis progression through alleviating inflammation, ECM accumulation, aberrant angiogenesis and apoptosis of hepatic parenchymal cells along with inhibiting activation of hepatic stellate cells through effects on multiple targets and may thus serve as a novel therapeutic regimen for this condition.

Comparative Investigation of Collagen-Based Hybrid 3D Structures for Potential Biomedical Applications.

Collagen is a key component for devices envisaging biomedical applications; however, current increasing requirements impose the use of multicomponent materials. Here, a series of hybrid collagen-based 3D materials, comprising also poly(ε-caprolactone) (PCL) and different concentrations of hyaluronic acid (HA)—in dense, porous or macroporous form—were characterized in comparison with a commercially available collagen sponge, used as control. Properties, such as water uptake ability, water vapour sorption, drug loading and delivery, were investigated in correlation with the material structural characteristics (composition and morphology). Methylene blue (MB) and curcumin (CU) were used as model drugs. For spongeous matrices, it was evidenced that, in contrast to the control sample, the multicomponent materials favor improved sustained release, the kinetics being controlled by composition and cross-linking degree. The other characteristics were within an acceptable range for the intended purpose of use. The obtained results demonstrate that such materials are promising for future biomedical applications (wound dressings and lab models).

Changes in visual quality with soft contact lenses after the instillation of hyaluronic acid eye drops

PurposeTo evaluate the changes in visual function and anterior surface aberrations during soft contact lens (SCL) wear after the instillation of hyaluronic acid (HA) eye drops with different viscosity. MethodsA prospective, randomized, and participant-masked study was performed. Twenty healthy participants (25.4 ± 2.6 years) were evaluated. Hydrogel (Ocufilcon D) and silicone-hydrogel (Somofilcon A) SCL were randomly assigned to both eyes of the same participant. Visual function in terms of high- and low-contrast corrected distance visual acuity (CDVA) and anterior contact lens surface aberrations (RMS HOA) were measured before and after the instillation, at different times, of different eye drops: saline (control) and 0.1%, 0.2%, and 0.3% HA. ResultsCompared with the saline solution, during hydrogel SCL wear, there was an improvement (P < 0.05) in high-contrast CDVA after 3 and 10 min with 0.1% HA, and after 5 and 20 min with 0.2% HA. During silicone-hydrogel SCL wear, there was a deterioration (P < 0.05) in high-contrast CDVA after 1 and 30 min with 0.3% HA. Additionally, during silicone-hydrogel SCL wear, there was also a deterioration (P < 0.05) in low-contrast CDVA after 5 and 20 min with 0.3% HA. In terms of RMS HOA, there were no clinically relevant changes with both SCL. ConclusionsThe instillation of HA eye drops could have a different effect on visual quality depending on their concentration of HA, the contact lens material, its surface ionicity, or other physicochemical properties that should be studied in future studies.

Micellar Hyaluronidase and Spiperone as a Potential Treatment for Pulmonary Fibrosis.

Concentration of hyaluronic acid (HA) in the lungs increases in idiopathic pulmonary fibrosis (IPF). HA is involved in the organization of fibrin, fibronectin, and collagen. HA has been proposed to be a biomarker of fibrosis and a potential target for antifibrotic therapy. Hyaluronidase (HD) breaks down HA into fragments, but is a subject of rapid hydrolysis. A conjugate of poloxamer hyaluronidase (pHD) was prepared using protein immobilization with ionizing radiation. In a model of bleomycin-induced pulmonary fibrosis, pHD decreased the level of tissue IL-1β and TGF-β, prevented the infiltration of the lung parenchyma by CD16+ cells, and reduced perivascular and peribronchial inflammation. Simultaneously, a decrease in the concentrations of HA, hydroxyproline, collagen 1, total soluble collagen, and the area of connective tissue in the lungs was observed. The effects of pHD were significantly stronger compared to native HD which can be attributed to the higher stability of pHD. Additional spiperone administration increased the anti-inflammatory and antifibrotic effects of pHD and accelerated the regeneration of the damaged lung. The potentiating effects of spiperone can be explained by the disruption of the dopamine-induced mobilization and migration of fibroblast progenitor cells into the lungs and differentiation of lung mesenchymal stem cells (MSC) into cells of stromal lines. Thus, a combination of pHD and spiperone may represent a promising approach for the treatment of IPF and lung regeneration.

Optimization of a floating poloxamer 407-based hydrogel using the Box-Behnken design: in vitro characterization and in vivo buoyancy evaluation for intravesical instillation

Intravesical instillation of a poloxamer 407 (PLX)-based hydrogel offers advantages such as thermo-sensitivity and sol-to-gel transition, but its utility is limited by urinary obstruction and insufficient bladder residence time. To overcome these obstacles, a floating PLX-hydrogel (FPH) was developed using sodium bicarbonate (BC) as a floating agent and hyaluronic acid (HA) as a gel strength modulator. The FPH composition was optimized using the Box-Behnken design with three independent variables: X1 [PLX concentration, 23.91%], X2 [BC concentration, 5.15%], and X3 [HA concentration, 3.49%]. The quadratic model was the best fit (desirability function, 0.623), resulting in response parameters of Y1 [floating time, 53.7 s], Y2 [gelation temperature gap, 20.3°C], and Y3 [duration time of gel, 396.7 min]. Rheological observations revealed the mechanical rigidity (storage modulus > loss modulus at elevated temperature) of the optimized FPH (phase transition temperature, 15.08°C). Gel erosion and drug release studies were performed using the gravimetric method; the remaining FPH fraction decreased exponentially with time, and gemcitabine release was biphasic and surface erosion-controlled. In vivo buoyancy was evaluated in rats using ultrasonography; these results were similar to those of the in vitro floating behavior. Thus, optimized FPH for intravesical instillation is a prospective option for bladder cancer treatment.

Hyaluronic Acid Concentration in Female Vocal Folds With Reinke's Edema.

The aim of the present study was to investigate hyaluronic acid (HA) concentrations in vocal folds among patients with Reinke's edema. Prospective and experimental study. Single tertiary center. An HA binding protein isolated from bovine nasal cartilage was used to identify and isolate the HA from samples. Plates coated with biotin-conjugated binding protein and streptavidin-europium conjugate were sequentially incubated with 18 Reinke's edema samples and 11 female vocal fold cover samples from cadavers (the superficial layer of the lamina propria; control group). After the release of europium from streptavidin in enhancement solution, final fluorescence was measured in a fluorometer. The mean HA concentration in Reinke's edema vocal folds was significantly higher than that in the control vocal folds (9.2 × 103 vs 0.9 × 103µg/g). Vocal fold covers affected by Reinke's edema present a higher concentration of HA than do vocal fold covers with no edema.

Formulation, Optimization, and In-Vitro Evaluation of Hyaluronic Acid Buccal Films Containing Benzydamine Hydrochloride

Aim: Mucoadhesive buccal films provide drug delivery for both local and systemic circulation with increased patient compliance. The objective of this study was to develop a buccal film containing benzydamine hydrochloride (BNZ) by using hyaluronic acid (HA) as a film-forming agent and glycerin (Gly) as a plasticizer for the anti-inflammatory effect on the mucosal ulcer. Material and Methods: The effects of different concentrations of HA and Gly on responses were analyzed using 23 factorial design. The HA and Gly ratio were selected as independent factors, and work of mucoadhesion, tensile strength, elongation at break, and contact angle values were chosen as dependent factors. Nine different film formulations were prepared at constant BNZ concentration (0.15%) by solvent casting method. Mechanical and mucoadhesion properties of film formulations were evaluated, and also contact angles were calculated. The BNZ release profiles were evaluated in simulated saliva solution (pH 6.8). Results: All film formulations were successfully prepared by solvent casting method. Increasing HA concentration from 0.5 to 2 % raised the mechanical and mucoadhesion properties of film formulations. Although the addition of Gly decreased the tensile strength values, it increased the elongation at break values about ten times. All formulations released more than 60 % BNZ in 60 mins. Conclusion: It is concluded that HA oral mucoadhesive film formulations containing BNZ are promising for treating oral ulcers.

Clinical effectiveness and safety of intra-articular injection of HYALGO in the management of knee osteoarthritis symptoms: A multicenter prospective study

BackgroundThe reduced concentration of hyaluronic acid in the synovial fluid, leading to impairment of joint function and painful symptomatology during knee osteoarthritis (OA), can be restored by using injectable formulations of hyaluronic acid (HA) and chondroitin sulfate (CS), variable for relative composition, HA/CS molecular modifications, and injection protocols. The present study aims to assess the safety and performance of the intra-articular (IA) viscosupplementing agent HYALGO, a formulation combining 40 mg/mL HA (>1700 kDa) and 40 mg/mL CS, in the treatment of patients suffering from knee OA. Methods74 patients affected by knee lesions classified as grade II and III according to Kellgren and Lawrence classification were prospectively recruited and treated with three HYALGO injections (2 mL) given one week apart. Visual analogue scale (VAS) pain changes were monitored at each injection and over-time at 6, 14, and 26 weeks of follow-up. Secondary endpoints were: Western Ontario McMaster University Osteoarthritis index (WOMAC), Patient's Global Assessment (PGA) score, Clinical Observer Global Assessment (COGA) score, Outcome Measures in Rheumatology Committee (OMERACT) and Osteoarthritis Research Society International (OARSI) responders rates. Patients were also assessed for changes in their ultrasound joint scores according to the criteria of the OMERACT US Task Force Group. ResultsPain reduction was statistically significant starting from the first IA injection. Mean pain reduction from baseline to week 26 was −90.6%. At 26 weeks, WOMAC Pain was reduced by −62.7%, WOMAC Stiffness by −47.2%, WOMAC Physical Function by −54.1%; Total WOMAC by −53.8%. The VAS PGA change from baseline was −48.0 [mm] and VAS COGA -41.0 [mm]. Responders at week 26 were 78.4%. Ultrasound parameters (joint effusion, synovial thickness, and popliteal cysts) improved or remained stable from baseline to week 6. ConclusionsThree injections of HYALGO were safe and effective to manage symptomatic knee OA, with a beneficial effect that increased progressively over time, peaking 6 months after injection.

Investigation of synovial fluid lubricants and inflammatory cytokines in the horse: a comparison of recombinant equine interleukin 1 beta-induced synovitis and joint lavage models

BackgroundLameness is a debilitating condition in equine athletes that leads to more performance limitation and loss of use than any other medical condition. There are a limited number of non-terminal experimental models that can be used to study early inflammatory and synovial fluid biophysical changes that occur in the equine joint. Here, we compare the well-established carpal IL-1β-induced synovitis model to a tarsal intra-articular lavage model, focusing on serial changes in synovial fluid inflammatory cytokines/chemokines and the synovial fluid lubricating molecules lubricin/proteoglycan 4 and hyaluronic acid. The objectives of this study were to evaluate clinical signs; synovial membrane and synovial fluid inflammation; and synovial fluid lubricants and biophysical properties in response to carpal IL-1β synovitis and tarsal intra-articular lavage.ResultsHyaluronic acid (HA) concentrations, especially high molecular weight HA, and synovial fluid viscosity decreased after both synovitis and lavage interventions. Synovial fluid lubricin concentrations increased 17–20-fold for both synovitis and lavage models, with similar changes in both affected and contralateral joints, suggesting that repeated arthrocentesis alone resulted in elevated synovial fluid lubricin concentrations. Synovitis resulted in a more severe inflammatory response based on clinical signs (temperature, heart rate, respiratory rate, lameness and joint effusion) and clinicopathological and biochemical parameters (white blood cell count, total protein, prostaglandin E2, sulfated glycosaminoglycans, tumor necrosis factor-α and CC chemokine ligands − 2, − 3, − 5 and − 11) as compared to lavage.ConclusionsSynovial fluid lubricin increased in response to IL-1β synovitis and joint lavage but also as a result of repeated arthrocentesis. Frequent repeated arthrocentesis is associated with inflammatory changes, including increased sulfated glycosaminoglycan concentrations and decreased hyaluronic acid concentrations. Synovitis results in more significant inflammatory changes than joint lavage. Our data suggests that synovial fluid lubricin, TNF-α, CCL2, CCL3, CCL5, CCL11 and sGAG may be useful biomarkers for synovitis and post-lavage joint inflammation. Caution should be exercised when performing repeated arthrocentesis clinically or in experimental studies due to the inflammatory response and loss of HA and synovial fluid viscosity.

Integration of a Nucleoside Triphosphate Regeneration System in the One‐pot Synthesis of UDP‐sugars and Hyaluronic Acid

AbstractEnzymatic synthesis is a promising approach to produce hyaluronic acid (HA) products with defined molecular weight (MW) and dispersity. We here report on the evaluation of nucleoside triphosphate (NTP) regeneration system for the synthesis of the precursors uridine‐5'‐diphosphate α‐d‐N‐acetylglucosamine (UDP‐GlcNAc) and uridine‐5'‐diphosphate α‐d‐glucuronic acid (UDP‐GlcA) and integration in the one‐pot synthesis of HA. By integration of polyphosphate kinase from Ruegeria pomeroyi (RpPPK2‐3) into the enzyme cascade to synthesize UDP‐GlcA we reached 89 % yield after 1 h with a regeneration cycle number (RCN) of 17. With the UDP‐GlcNAc cascade, the ATP concentration was lowered 250‐fold and reached 94 % yield after 1 h with an RCN of 234. Integration of the RpPPK2‐3 regeneration system in one‐pot HA synthesis afforded optimization of MgCl2 and ATP starting concentrations. With 25 mM MgCl2 and 0.1 mM ATP, a HA concentration of 0.81 g L−1 with an average MW of 1.17 MDa and dispersity of about 1.08 and an RCN of 75 was obtained. We conclude that integration of NTP regeneration with RpPPK2‐3 expanded the enzymatic possibilities to produce HA with lower range MW.

Melatonin Enhances Chondrocytes Survival Through Hyaluronic Acid Production and Mitochondria Protection

IntroductionMelatonin, an indoleamine synthesized mainly in the pineal gland, plays a key role in regulating the homeostasis of bone and cartilage. Reactive oxygen species (ROS) are released during osteoarthritis (OA) and associated with cartilage degradation. Melatonin is a potent scavenger of ROS.Material and methodsWe used rat chondrocytes as a research objective in our study and investigated the effect of melatonin. We evaluated cell viability by MTS, cell apoptosis and mitochondrial transmembrane potential (MMP) by flow cytometry, hyaluronic acid (HA) concentrations by ELISA assay, relative gene and protein expression by RT-qPCR and WB assay, the subcellular location of nuclear factor κB (NF-κB) p65 and Rh-123 dye by immunofluorescence, oxygen consumption rate (OCR) by Seahorse equipment.ResultsMTS and flow cytometry assay indicated that melatonin prevented H2O2-induced cell death. Melatonin suppressed the expression of HAS1 both from mRNA and protein level in time- and dose-dependent manners. Furthermore, melatonin treatment attenuated the decrease in HA production in response to H2O2 stimulation and repressed the expression of cyclooxygenase-2 (COX-2). Melatonin significantly decreased the nuclear volume of p65 and inhibited the expression of p50 induced by H2O2. In addition, melatonin restored the MMP and OCR of mitochondria, exerted mitochondrial protection function.ConclusionsMelatonin has a protective effect on chondrocytes, possibly by upregulating HA synthesis and inhibiting the NF-kB signaling.

Physicochemical Properties of Injectable Hyaluronic Acid: Skin Quality Boosters

AbstractIn addition to clinical research on the efficacy and compatibility of injectable hyaluronic acid (HA) based fillers, ample in vitro research on the physicochemical properties of these materials is published. Different HA‐based product ranges are also available as skin quality boosters (SQBs) to improve skin quality. The aim of this study is to investigate the physicochemical properties of specific HAs for improving skin quality, as, to the best of the authors' knowledge, no data is published on this topic to date. This paper may provide a better understanding of clinical performance and differentiation between these SQBs. Five HA injectable hydrogels (Belotero Revive, Juvederm Volite, Restylane Skinbooster Vital, Viscoderm Hydrobooster, and Profilho) from the SQB product ranges of different companies and manufacturing technologies are investigated for their extrusion force, swelling degree, rheological performance, and cohesivity. There are significant differences in extrusion force, swelling degree, rheological performance, and cohesivity between the assessed SQBs. HA concentration (mg mL−1) exhibits statistically significant positive correlations with extrusion force, swelling degree, tan delta, and cohesivity. This study provides a physicochemical characterization of different SQBs and information to improve understanding of this type of product.

Hyaluronic acid and its use in dentistry.

The purpose of this paper is to evaluate the role of hyaluronic acid in bio-revitalization (HABR) by testing several extracellular matrix biological parameters in cultured dermal fibroblasts. To this aim, fibroblastic expressed genes after exposition to three HABR medical devices were evaluated. Cells were seeded on a layer of three different medical devices containing 6.2, 10 and 20 mg/ml of HABR for 24 h. Real Time Polymerase Chain Reaction was performed to investigate gene expressions. Genes encoding HABR synthesis and degradation, Metalloproteinases 2 and 3 and Desmoplakin production as well as GDF6, and IGF1 were activated by hyaluronic acid products. The in vitro study showed similar effects on tested genes despite a different concentration of hyaluronic acid contained in the medical devices and the simultaneous presence of other additives. Based on the reported data, gene activations are an aspect of metabolic modulation of signalling pathways rather than the proportional production of a specific connective tissue molecule. Indeed, different HABR concentration and the presence of other additives did not change the overall effect on the studied genes. We believe that the optimization of extracellular matrix micro-environment, obtained by enhanced structural support with HABR, leads to functional and metabolic improvement.

Influence of high-molecular hyaluronic acid on MCF-7 breast cancer stem cells

In recent years, more and more data have been accumulating on the high radio- and chemo-resistance of tumor stem cells (CSCs), confirming the assumption that these cells play an important role in the recurrence of the tumor process in some cancer patients. Therefore it is of considerable interest to elucidate the patterns and mechanisms of the formation of the CSC pool under the influence of various factors, including one of the main components of the extracellular matrix, hyaluronic acid (HA). The purpose of the study was to elucidate the effects of HA with a molecular weight of (1-3) x 106 Dа on the amount of CSCs in MCF-7 breast cancer cell culture in vitro. Material and methods. Using the MTT test, the viability of MCF-7 cancer cells was determined after incubation with HA at a concentration of 0.3-5.0 mg/ml. Using flow cytometry, we evaluated the change in the relative and absolute amount of CSCs under the influence of HA (0.6 mg/ml). CSCs were detected by CD44+CD24-/low immunophenotype, as well as by their ability to efficiently efflux Hoechst 33342 fluorescent dye from cells and form so-called side population (SP). Results. Viability of cancer cells in the general population decreased with increasing concentration of HA (R=-0.94, p=0.006). At the same time the incubation of cells with HA led to an increase in both the relative and absolute amounts of CSCs (SP) compared with the control. Thus, the relative number of SP cells increased by 2.0 times (p = 0.002), CD44 + CD24- / low cells - by 6.1 times (p = 0.005). The absolute number of these cells changed to a lesser extent compared to the control: the increase was 1.6 (p = 0.004) and 2.9 times (p = 0.02), respectively. Conclusion. Thus, the effect of high-molecular weight HA on breast cancer cells of MCF-7 line leads to an increase in amount of CSCs, while the viability of non-stem cancer cells decreases under the same conditions.

Effects of Oral Hyaluronic Acid Administration in Dogs Following Tibial Tuberosity Advancement Surgery for Cranial Cruciate Ligament Injury.

Simple SummaryHyaluronic acid injections into the stifle are often used for managing osteoarthritis in dogs. Giving hyaluronic acid orally would be easier, but to date we do not have enough information regarding its effects when administered by this route. For this reason, in this study we evaluated the effects of oral administration of hyaluronic acid in dogs with cranial cruciate ligament rupture after surgical resolution. Dogs were divided into two groups that received either oral hyaluronic acid or a placebo. We measured changes in several biomarkers of osteoarthritis before surgery, and at ten weeks after surgery. Results showed significant improvements in some of these biomarkers, namely synovial fluid levels of hyaluronic acid and paraoxonase-1. These changes indicate that post-op oral administration of hyaluronic acid may be effective for the management of stifle osteoarthritis in the dog.Hyaluronic acid (HA) intraarticular injection is used in the management of osteoarthritis in veterinary medicine. However, HA oral administration is less common given the scarce currently available scientific evidence. This study was aimed at evaluating the effects of oral HA administration on synovial fluid concentrations of several selected biomarkers in dogs with cranial cruciate ligament (CCL) injury operated on using the tibial tuberosity advancement (TTA) technique. Fifty-five dogs were included in this prospective, randomized, double-blind, clinical study; they were randomly assigned to receive either a placebo (group A; n = 25) or HA (group B; n = 30) orally for 10 weeks. Synovial fluid samples were obtained before surgery, and at 10 weeks postoperatively to measure concentrations of HA, haptoglobin, nitric oxide, and paraoxonase-1. After 10 weeks, group HA showed a significant increase in HA concentration (p = 0.0016) and a significant decrease in PON-1 concentration (p = 0.011) compared to baseline. In conclusion, post-op oral HA administration in canine patients with CCL injury leads to improvements in osteoarthritis biomarkers, namely higher synovial fluid HA concentrations and reduced synovial fluid paraoxonase-1 concentrations. These findings support the bioavailability of orally-administered HA and its usefulness in improving biomarkers of osteoarthritis.

Koncentrace kyseliny hyaluronové v synoviální tekutině jako ukazatel závažnosti a stupně postižení u pacientů s gonartrózou

Koncentrace kyseliny hyaluronové v synoviální tekutině jako ukazatel závažnosti a stupně postižení u pacientů s gonartrózou