Dynamic–Mechanical Properties of Bioartificial Polymeric Materials

Blends of natural and synthetic polymers were studied for potential applications in the biomedical field. Collagen and hyaluronic acid were mixed in aqueous solution with poly(vinyl alcohol) and poly(acrylic acid). The properties of the blends were studied by differential scanning calorimetry and dynamic mechanical thermal analysis. Some methods were also investigated to enhance the miscibility of the polymers in these blends.

Films from blends of hyaluronic acid with poly(vinyl alcohol) and with ethylene-vinyl alcohol copolymers were prepared from solutions in water and dimethylsulfoxide by a casting method. The study of the interactions between the natural and synthetic component has been carried out through thermal and morphological analysis. The results show that the crystallinity of poly(vinyl alcohol) and its copolymers decreases with increasing hyaluronic acid content and these crystallizable polymers exhibit no tendency to crystallize when hyaluronic acid content is more than 80 wt%. The relatively good miscibility in these blends may be due to the capacity of the natural and synthetic component, each with abundant hydroxyl groups, to interact mutually through hydrogen bonding. The presence of such interactions was supported by the observation of the melting point depression of the synthetic component in thermal analysis of the blends. Melting point depression was more accentuated in melt recrystallized blends. These results suggest that the interactions between the two polymers are enhanced if the blends are brought close to the melting point of the synthetic component.

Poly (vinyl alcohol) and hyaluronic acid hydrogels as potential biomaterial systems - A comprehensive review

Bulls with varying freezability exhibit substantial variation in semen characteristics after cryopreservation. Sperm freezability is positively correlated with membrane cholesterol content, membrane integrity, mitochondrial activityand antioxidant content. The purpose of this study was to determine the optimal concentration of hyaluronic acid (HA) in bull sperm with different cryotolerances. Simmental bulls (n = 10) semen samples were taken and categorized based on their progressive motility (PM) after freeze-thawing: Group I, consisting of bulls (n = 5) with progressive sperm motility ≥45%, was considered good freezability ejaculates (GF), and Group II, including bulls (n = 5) with progressive sperm motility ≤30%, was considered poor freezability ejaculates (PF) bulls. Semen samples were diluted with a Tris-egg-yolk-glycerol (TEYG) extender containing various concentrations of HA: without HA (control), 1 mM HA, 2 mM HAand 4 mM HA. After the freeze-thaw process, sperm kinematics, plasma membrane and acrosome integrity, mitochondrial activityand apoptotic status were evaluated. The addition of 1 mM HA to the diluent of bulls with GF increased PM and linearity (LIN) compared to the control group (p < 0.05). Normal morphology was improved after thawing in the samples treated with 1and 2 mM HA in the GF and PF bullsrespectively. The membrane and acrosome integrity of GF bulls treated with 1 mM HA was significantly (p < 0.05) greater than that of the control groups. Adding 1 mM HA to the extender of bulls with GF and PF improved the proportion of viable cells compared with the highest concentration (4 mM) of HA. The mitochondrial activity of PF bulls treated with 1and 2 mM HA was significantly (p < 0.05) greater than that of the controls and 4 mM HA. Finally, it can be concluded that adding low doses of HA (1 mM) to the TEYG extender of GF and PF bulls ameliorated the post-thaw semen quality.

Biological lubricants are essential for the effective functioning of biomedical systems that involve relative motion. Hyaluronic acid (HA) is often used for its excellent lubrication properties; however, its long-term effectiveness is compromised by its tendency to accelerate metal corrosion. Incorporating cellulose nanocrystals (CNCs) into HA offers a promising solution due to CNCs' corrosion-inhibition properties, tunable rheological behavior, and tribological properties. This study examines the impact of varying HA and CNC concentrations on friction, wear, and corrosion in HA/CNC suspensions. Optimal performance was achieved with 2wt% CNC in 1mg/mL HA, corresponding to the onset of gelation. Below this CNC concentration, lubrication improves through the formation of a protective film and the mitigation of corrosion. Increasing CNC beyond 2wt% yields minimal additional benefit. HA concentration also plays a key role: at low HA levels, corrosion predominates, leading to increased friction and wear. In contrast, HA levels above 1mg/mL in 2wt% CNC suspensions enhance lubricant film formation due to improved rheology, resulting in reduced wear, coefficient of friction, and corrosion. These results underscore the importance of optimizing HA and CNC concentrations to maximize their synergistic effects in advanced bio-tribological lubrication applications.

This work was aimed to explore the potential effect of hyaluronic acid (HA) initial concentration (7.0 - 14.0 % w/v) on cross-linking efficiency of HA hydrogels cross-linked with 1,4-butanediol diglycidyl ether (BDDE). The results revealed that the hydrogel prepared at 10.0 % HA concentration exhibited a slower degradation rate, a lower swelling ability and more regular porosity than those prepared at either lower or higher HA concentration. After four days incubating with hyaluronidase, the content of NAG (N-acetyl glucosamine) remaining in the 10.0 HA hydrogel was 25.1±1.9 % with respect to the total NAG content found in the original mass. In contrast, the hydrogels prepared at 7.0 % and 14.0 % HA concentration showed a less remaining content of NAG equaled to approximately 15.9±5.4 % and 19.5±2.6 % respectively. On the other hand, the swelling ability of tested hydrogels was steadily decreased with the increase of HA initial concentration until the 10.0 % HA hydrogel and then showed an opposite trend. Based on this finding, the 10.0 % HA hydrogel exhibited the lowest swelling ratio which was observed at 129±3.2 g/g in distilled water and at 116±2.4 g/g in phosphate buffer saline (PBS). The SEM images showed various morphologies within the entire range of tested hydrogels. However, the hydrogel prepared at 10.0 % HA concentration was more homogenous and appeared with narrower pore-size distribution ranged in diameter from less than 50 μm to approximately 300 μm. Finally, the effect of HA initial concentration was investigated by FTIR which confirmed that the 10.0 % HA hydrogel was subject to a greater loss of (- OH) at 3343 cm-1 than other hydrogels except the 11.0 % HA hydrogel. This phenomenon was probably attributed to the formation of pendants that allowed the 11.0 % HA hydrogel to appear with a lower peak intensity than the 10.0 % HA hydrogel in the FTIR spectra. In conclusion, the HA initial concentration plays a crucial role in determining the cross-linking efficiency of HA hydrogels cross-linked with BDDE.

Histological analysis of hyaluronic acid (HA) applied in the nasolabial fold (NLF) studied gel distribution into the dermis and subcutaneous fat, correlating density of NLF tissues, with cohesivity, concentration and degradation of HA. Ten patients received two injection points on each NLF. They were intradermal near the nasal ala and subcutaneous near the oral commissure. Injection points received 0.1ml of 24mg/ml HA cross-linked by BDDE 2.0ppm through a bolus technique injection without retrograde backflow. Biopsies were taken 72h and 9months after application to histological analysis. Dermis density fragmented the gel into small nodules, spreading it into the reticular dermis. Seventy-two hours after application, average diameter of the small nodules was 0.30mm and at month nine 0.05mm. Softness of the subcutaneous fat and high concentration of HA created a single and compact nodule. Seventy-two hours after application, average of nodule diameters was 1.05mm and at month nine 0.49mm. HA biodegradation occurred on the external surface of the gel. The single nodule offered small contact surface to enzymes, slowing gel degradation. The sum average of the diameters of the small nodules offered a large contact surface to enzymes, accelerating gel degradation. Permanence of the HA into the tissues depended on the relationship between the gel distribution and its degradation. Due to the small contact surface of the single nodule to enzymes, permanence of the HA into the subcutaneous fat was more extended than into the dermis. This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

IntroductionOsteoarthritis (OA) is the most common musculoskeletal disease. Current treatments for OA are mainly symptomatic and inadequate since none results in restoration of fully functional cartilage. Hyaluronic Acid (HA) intra-articular injections are widely accepted for the treatment of pain associated to OA. The goal of HA viscosupplementation is to reduce pain and improve viscoelasticity of synovial fluid. Platelet-rich plasma (PRP) has been also employed to treat OA to possibly induce cartilage regeneration. The combination of HA and PRP could supply many advantages for tissue repair. Indeed, it conjugates HA viscosupplementation with PRP regenerative properties. The aim of this study was to evaluate the rheological and biological properties of different HA compositions in combination with PRP in order to identify (i) the viscoelastic features of the HA-PRP blends, (ii) their biological effect on osteoarthritic chondrocytes and (iii) HA formulations suitable for use in combination with PRP.Materials and MethodsHA/PRP blends have been obtained mixing human PRP and three different HA at different concentrations: 1) Sinovial, 0.8% (SN); 2) Sinovial Forte 1.6% (SF); 3) Sinovial HL 3.2% (HL); 4) Hyalubrix 1.5% (HX). Combinations of phosphate buffered saline (PBS) and the four HA types were used as control. Rheological measurements were performed on an Anton PaarMCR-302 rheometer. Amplitude sweep, frequency sweep and rotational measurements were performed and viscoelastic properties were evaluated. The rheological data were validated performing the tests in presence of Bovine Serum Albumin (BSA) up to ultra-physiological concentration (7%). Primary osteoarthritic chondrocytes were cultured in vitro with the HA and PRP blends in the culture medium for one week. Cell viability, proliferation and glycosaminoglycan (GAG) content were assessed.ResultsPRP addition to HA leads to a decrease of viscoelastic shear moduli and increase of the crossover point, due to a pure dilution effect. For viscosupplements with HA concentration below 1% the viscoelasticity is mostly lost. Results were validated also in presence of proteins, which in synovial fluid are more abundant than HA.Chondrocytes proliferated overtime in all different culture conditions. The proliferation rate was higher in chondrocytes cultured in the media containing PRP compared to the cultures with different HA alone. GAG content was significantly higher in chondrocytes cultured in PRP and HL blend.DiscussionWe investigated the rheological and biological properties of four different HA concentrations when combined with PRP giving insights on viscoelastic and biological properties of a promising approach for future OA therapy. Our data demonstrate that PRP addition is not detrimental to the viscosupplementation effect of HA. Viscosupplements containing low HA concentration are not indicated for combination with PRP, as the viscoelastic properties are lost. Although having the same rheological behavior of SF and HX, HL was superior in stimulating extracellular matrix production in vitro.

The role of cytokines in cervical ripening: Correlations between the concentrations of cytokines and hyaluronic acid in cervical mucus and the induction of hyaluronic acid production by inflammatory cytokines by human cervical fibroblasts

Biomaterials are substances designed for biomedical purposes. They have to be biocompatible, biodegradable, and non-toxic. Biomaterials can be obtained from natural and synthetic polymers. Natural high molecular weight compounds have to be isolated from natural sources. The management of food industry wastes is important because they are biologically active and can be hazardous for human and environment. Fish eyeballs can be a potential source of glycosaminoglycans (GAGs) as for instance hyaluronic acid (HA) or chondroitin sulfate (CS). In the experimental studies, GAGs were isolated from vitreous of Cyprinus Carpio and Salmonidae eyeballs. The concentration of HA and CS was determined by the spectrophotometric method at 520 and 525 nm, respectively, with the use of a standard curve. The results showed higher content of HA and CS in the Salmonidae than in Cyprinus Carpio. However, in each isolated GAGs mixture, both polymers were detected. It can be assumed that fish vitreous eyeballs can be managed as the natural source of GAGs.

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.

Hyaluronic acid (HA) plays a vital role in many tissues, influencing water content and mechanical function, and has been shown to have positive biological effects on cell behavior in vitro. To begin to determine whether these benefits can be accessed if HA is incorporated into collagen-based scaffolds for tissue engineering, HA-collagen composite matrices were prepared and selected properties evaluated. HA-collagen scaffolds were cross-linked with carbodiimide and loss rates of HA in culture medium assessed. Scaffold pore structures were evaluated by light and electron microscopy. Adult canine chondrocytes were grown in selected HA-collagen scaffolds to assess the effects of HA on cell behavior. Homogenous HA-collagen slurries were achieved when polyionic complexes were suppressed. HA was uniformly distributed through the scaffolds, which demonstrated honeycomb-like pores with interconnectivity among pores increasing as HA content increased. Virtually all of the HA added to the collagen slurry was incorporated into the composite scaffolds that underwent a 7-day cross-linking protocol. After 5 days in culture medium, the HA content in the scaffolds was 5-7% regardless of initial HA loading. After only 2 weeks in culture cartilaginous tissue was found in the chondrocyte-seeded HA-collagen scaffolds. This study contributes to the understanding of the effects of HA content, pH, and cross-link treatment on pore characteristics and degradation behavior essential for the design of HA-collagen scaffolds. The demonstration that these scaffolds can be populated by chondrocytes and support in vitro formation of cartilaginous tissue warrants further investigation of this material system for tissue engineering.

Hyaluronic acid (HA), known for diverse properties, was investigated for its potential in dental pulp therapy. This study investigated the potential of HA in dental pulp therapy by examining the physical properties and effects of zinc oxide eugenol (ZOE) pulpotomy materials containing varying HA concentrations on rat molar teeth. In vitro tests assessed compressive strength and hardness of ZOE materials blended with HA (0.5%, 1%, 3%) and HA gels (0.54%, 0.8%). 120 samples, encompassing the control group, underwent compressive strength testing, while 60 samples were designated for hardness assessment. In vivo experiments on rat molars studied histological effects of HA-containing ZOE on dental pulp over 1 week and 1 month. Gels with HA concentrations of 0.5%, 1%, and 0.54% were used in pulpotomy on 22 rats. Each rat underwent the procedure on four teeth, with one tooth serving as a control, totaling 88 teeth subjected to the intervention. In the analyses, SPSS 22.0 was used and the significance level was set at P = 0.05. Findings showed that HA at 0.5% maintained compressive strength, but higher concentrations decreased mechanical properties significantly (P = 0.001). Histological assessments indicated better outcomes with lower HA concentrations in terms of odontoblast layer continuity (P = 0.005 at 1 month) and pulp vitality (P = 0.001 at 1 week and P = 0.018 at 1 month). The study suggests HA holds promise for pulpotomy and regenerative endodontic treatments, but further research is needed to understand long-term clinical implications.

Hyaluronic acid metabolism in keloid fibroblasts

We examined the effect of hyaluronic acid in promoting proliferation of undifferentiated progenitor cells through the CD44 receptor during eosinopoiesis in vitro. Undifferentiated umbilical cord blood cells were purified on the first day to isolate primitive progenitor cells expressing the CD34 hemopoietic surface marker. Culture in wells coated with 100 micrograms/ml hyaluronic acid caused a 198 +/- 28.7% augmentation of proliferation of CD34+ progenitor cells at 3 wk (p < 0.01). By contrast, concentrations of hyaluronic acid > 10 micrograms/ml inhibited proliferation of unfractionated cord blood mononuclear cells. The augmented proliferation of precursor cells caused by hyaluronic acid was associated with complete (93.0 +/- 5.12%) differentiation to eosinophil morphology. By contrast, concentrations of hyaluronic acid > or = 10 micrograms/ml inhibited eosinophilic differentiation of unfractionated mononuclear cells. Wright-Giemsa staining demonstrated 95.4 +/- 2.92% eosinophils for CD34+ cells cultured for 3 wk without hyaluronic acid (control) and 93.8 +/- 5.11% for CD34+ cells cultured in hyaluronic acid-coated wells (100 micrograms/ml); for unfractionated cells, 94.0 +/- 3.02% demonstrated eosinophilic morphology in control wells at 3 wk vs 55.4 +/- 8.34% in hyaluronic acid-coated (100 micrograms/ml) wells (p < 0.05). Augmented proliferation caused by hyaluronic acid was attenuated completely by the anti-CD44 mAbs, 212.3 and IM7.8.1. Pretreatment of CD34+ cells with 5 micrograms/ml 212.3 inhibited the augmented proliferation caused by the optimal concentration of hyaluronic acid (100 micrograms/ml) from 260 +/- 39.2% of control growth to 114 +/- 16.4% of control growth (p = 0.02). Inhibition was comparable for IM7.8.1. Control mAb (LM2) to the beta 2 integrin subunit CD11b had no effect on proliferation induced by hyaluronic acid. We demonstrate that hyaluronic acid stimulates the growth of CD34+ selected umbilical cord blood cells into specifically differentiated mature eosinophils. This process is modulated by the CD44 receptor on the progenitor cell population.