Vladimír Velebný

This prospective, single-arm, exploratory postmarketing study preliminarily evaluated the clinical response and plasma biomarker changes in 18 client-owned dogs with naturally occurring osteoarthritis (OA) treated with sodium hyaluronate (Bonharen). Patients received intravenous injections of Bonharen Intravenous at a dose of 0.15 mL/kg (1.3–1.6 mg/kg hyaluronic acid once a week for consecutive five weeks). Clinical parameters (lameness, joint pain, mobility, swelling) were assessed weekly and two weeks after the final dose was given via standardized scoring. The plasma concentrations of selected inflammatory, cartilage-related, and oxidative stress biomarkers were measured before treatment and two weeks after the final dose. Clinical improvement in lameness and/or joint pain on palpation was observed in nearly half of the patients. No clinical deterioration was recorded at any time point. Physical activity increased in all patients with reduced baseline activity. Significant decreases in the plasma levels of prostaglandin E₂, Δ17-6-keto prostaglandin F1α, malondialdehyde, and hyaluronan were detected, indicating reduced systemic inflammation and oxidative stress. In addition, an increase in plasma hydroxybutyrate and decrease in the collagen-breakdown marker prolyl-hydroxyproline were observed. No adverse effects were reported. These findings suggest that intravenous hyaluronic acid (Bonharen) may represent a safe and promising component to multimodal OA management in dogs and demonstrate the feasibility of integrating plasma biomarkers in canine OA studies.

Hyaluronic acid (HA), a natural polymer produced through biotechnological processes, is initially synthesized with a high molecular weight, which is subsequently reduced for specific applications. This work aims to develop an experimental laboratory device that enables the controlled depolymerization of HA to achieve a targeted molecular weight. This is accomplished by applying precisely regulated temperature and pressure conditions that facilitate the cleavage of HA polymer chains as they pass through a narrow capillary. The process also allows for a controlled exposure time to elevate the temperature and pressure, with the rapid transition between standard and elevated conditions ensuring temporal stability. A key innovation of this approach is the selective application of an elevated temperature to a designated capillary section, adjustable from 0.5 to 5 m in 0.5 m increments, allowing for a broad range of reaction times. Numerical simulations verified temperature distribution along the capillary during heating and cooling. The device’s effectiveness is demonstrated by cleaving HA chains (initial Mw = 2150 kDa) dissolved in an aqueous solution at a 0.1 mg/mL concentration. Results from a factorial experiment that evaluated the extremes of three main variables show cleavage across a wide molecular weight range, reaching values as low as 8 kDa, with a variance of less than 5%. This study presents a viable device for the selective depolymerization of HA via physical parameters alone, eliminating the need for additional substances such as acids, hydroxides, or enzymes.

Nanofibrous materials represent a very promising form of advanced carrier systems that can be used industrially, especially in regenerative medicine as highly functional bandages, or advanced wound dressings. By incorporation of antimicrobial additives directly into the structure of the nanofiber carrier, the functionality of the layer is upgraded, depending on the final requirement—bactericidal, bacteriostatic, antiseptic, or a generally antimicrobial effect. Such highly functional nanofibrous layers can be prepared mostly by electrospinning technology from both synthetic and natural polymers. The presence of a natural polymer in the composition is very advantageous. Especially in medical applications where, due to the presence of the material close to the human body, the healing process is more efficient and without the occurrence of an unwanted inflammatory response. However, converting natural polymers into nanofibrous form, with a homogeneously distributed and stable additive, is a great challenge. Thus, a combination of natural and synthetic materials is often used. This review clearly summarizes the issue of the incorporation and effectiveness of different types of antimicrobial substances, such as nanoparticles, antibiotics, common antiseptics, or substances of natural origin, into electrospun nanofibrous layers made of mostly natural polymer materials. A section describing the problematic aspects of antimicrobial polymers is also included.

In the realm of surgical and dental applications, hyaluronic acid (HA) braided threads show significant therapeutic potential due to their incorporation of pharmaceutical active ingredients. This study primarily focuses on resolving the crucial challenge of devising a deposition method that can ensure both precision and uniformity in the content of the active ingredient Octenidine dihydrochloride (OCT) within each segment of the threads. Our objective in this study was to develop a continuous deposition method for OCT onto a braided thread composed of 24 hyaluronic acid-based fibers, aiming for a specific OCT content of 0.125 µg/mm, while maintaining a maximum allowable deviation of ±15% in OCT content. The motivation behind designing this novel method stemmed from the necessity of employing a volatile solvent for the active agent. Conventional wetting methods proved unsuitable due to fluctuations in the solution’s concentration during deposition, and alternative methods known to us demanded intricate technical implementations. The newly introduced method offers distinct advantages, including its online processing speed, scalability potential, and cost-efficiency of the active agent solution. Additionally, it minimizes the impact on the natural polymer thread, preserving energy by obviating the need for complete thread saturation. Our research and precise apparatus development resulted in achieving the desired thread properties, with an OCT content of (1.51 ± 0.09) µg per 12 mm thread piece. These findings not only validate the suitability of this innovative method for depositing active agents but also extend its potential applicability beyond dental care.

The physical stresses during cryopreservation affect stem cell survival and further proliferation. To minimize or prevent cryoinjury, cryoprotective agents (CPAs) are indispensable. Despite the widespread use of 10% dimethyl sulfoxide (DMSO), there are concerns about its potential adverse effects. To bypass those effects, combinations of CPAs have been investigated. This study aimed to verify whether high-molecular-hyaluronic acid (HMW-HA) serves as a cryoprotectant when preserving human mesenchymal stem cells (hMSCs) to reduce the DMSO concentration in the cryopreservation medium. We studied how 0.1% or 0.2% HMW-HA combined with reduced DMSO concentrations (from 10% to 5%, and 3%) affected total cell count, viability, immunophenotype, and differentiation potential post-cryopreservation. Immediately after cell revival, the highest total cell count was observed in 10% DMSO-stored hMSC. However, two weeks after cell cultivation an increased cell count was seen in the HMW-HA-stored groups along with a continued increase in hMSCs stored using 3% DMSO and 0.1% HMW-HA. The increased total cell count corresponded to elevated expression of stemness marker CD49f. The HA-supplemented cryomedium did not affect the differential potential of hMSC. Our results will participate in producing a ready-to-use product for cryopreservation of mesenchymal stem cells.

Hyaluronan (HA) comprises a fundamental component of the extracellular matrix and participates in a variety of biological processes. Half of the total amount of HA in the human body is present in the skin. HA exhibits a dynamic turnover; its half-life in the skin is less than one day. Nevertheless, the specific participants in the catabolism of HA in the skin have not yet been described in detail, despite the essential role of HA in cutaneous biology. A deeper knowledge of the processes involved will act to support the development of HA-based topical and implantable materials and enhance the understanding of the various related pathological cutaneous conditions. This study aimed to characterize the distribution and activity of hyaluronidases and the other proteins involved in the degradation of HA in healthy human full-thickness skin, the epidermis and the dermis. Hyaluronidase activity was detected for the first time in healthy human skin. The degradation of HA occurred in lysates at an acidic pH. HA gel zymography revealed a single band corresponding to approximately 50 kDa. This study provided the first comprehensive view of the distribution of canonic HA-degrading proteins (HYAL1 and HYAL2) in human skin employing IHF and IHC. Furthermore, contrary to previous assumptions TMEM2, a novel hyaluronidase, as well as CEMIP, a protein involved in HA degradation, were localized in the human epidermis, as well as in the dermis.

All-trans-retinoic acid (atRA) is a potent ligand that regulates gene expression and is used to treat several skin disorders. Hyaluronic acid (HA) was previously conjugated with atRA (HA-atRA) to obtain a novel amphiphilic compound. HA-atRA forms micelles that incorporate hydrophobic molecules and facilitate their transport through the skin. The aim of this study was to determine the influence of HA-atRA on gene expression in skin cells and to compare it with that of unbound atRA. Gene expression was investigated using microarrays and a luciferase system with a canonical atRA promoter. HA-atRA upregulated gene expression similarly to atRA. However, HA-atRA activated the expression of cholesterol metabolism genes, unlike atRA. Further investigation using HPLC and filipin III staining suggested that the treated cells induced cholesterol synthesis to replenish the cholesterol removed from the cells by HA-atRA. HA modified with oleate (HA-C18:1) removed cholesterol from the cells similarly to HA-atRA, suggesting that the cholesterol removal stemmed from the amphiphilic nature of the two derivatives. HA-atRA induces retinoid signaling. Thus, HA-atRA could be used to treat skin diseases, such as acne and psoriasis, where the combined action of atRA signaling and anti-inflammatory cholesterol removal may be potentially beneficial.

Formation of peritoneal adhesions (PA) is one of the major complications following intra-abdominal surgery. It is primarily caused by activation of the mesothelial layer and underlying tissues in the peritoneal membrane resulting in the transition of mesothelial cells (MCs) and fibroblasts to a pro-fibrotic phenotype. Pro-fibrotic transition of MCs—mesothelial-to-mesenchymal transition (MMT), and fibroblasts activation to myofibroblasts are interconnected to changes in cellular metabolism and culminate in the deposition of extracellular matrix (ECM) in the form of fibrotic tissue between injured sides in the abdominal cavity. However, ECM is not only a mechanical scaffold of the newly synthetized tissue but reciprocally affects fibrosis development. Hyaluronan (HA), an important component of ECM, is a non-sulfated glycosaminoglycan consisting of N-acetyl-D-glucosamine (GlcNAc) and D-glucuronic acid (GlcUA) that can affect the majority of processes involved in PA formation. This review considers the role of endogenously produced HA in the context of different fibrosis-related pathologies and its overlap in the development of PA.