Production and Characterization of Electrospun Chitosan, Nanochitosan and Hyaluronic Acid Membranes for Skin Wound Healing.

The development of new wound dressings made from biomaterials, which offer a better cost-benefit ratio and accelerate the healing process, is increasing nowadays. Various biopolymers can be electrospun to form functional membranes for wound healing. Therefore, in this study, chitosan and nanochitosan membranes with or without hyaluronic acid were prepared using the electrospinning technique, characterized and evaluated in the healing of skin wounds in rats. Chitosan and nanochitosan solutions, with or without hyaluronic acid, were prepared at concentrations of 1%-4% using PEO (polyethylene oxide) and subjected to the electrospinning process to obtain membranes characterized by scanning electron microscopy (SEM), mechanical tests, and antimicrobial activity. The healing effect of the membranes was evaluated by monitoring the area of the lesions, contraction of the wounds, histologic analysis, and induction of pro-inflammatory cytokine (IL-1 α and TNF-α) production in rats. The nanochitosan and nanochitosan membranes with hyaluronic acid achieved greater fiber diameter and uniformity, resistance, elasticity, and thermal stability, in addition to good adhesion to the wound bed and permeation capacity. Despite not presenting antimicrobial activity in vitro, they contributed to the production of pro-inflammatory interleukins in the animals tested, provided physical protection, reduced the wound area more markedly until the seventh day of the evaluation, with an acceleration of the healing process and especially when functionalized with hyaluronic acid. These results indicate that the membranes may be promising for accelerating the healing process of chronic wounds in humans.

Effect of hyaluronic acid-stabilized silver nanoparticles on lettuce (Lactuca sativa L.) seed germination.

Nanotechnology has brought significant advancements to agriculture through the development of engineered nanomaterials (ENPs). Silver nanoparticles (AgNPs) capped with polysaccharides have been applied in agricultural diagnostics, crop pest management, and seed priming. Hyaluronic acid (HA), a natural polysaccharide with bactericidal properties, has been considered a growth regulator for plant tissues and an inducer of systemic resistance against plant diseases. Additionally, HA has been employed as a stabilizing agent for AgNPs. This study investigated the synthesis and effects of hyaluronic acid-stabilized silver nanoparticles (HA-AgNPs) as a seed priming agent on lettuce (Lactuca sativa L.) seed germination. HA-AgNPs were characterized using several techniques, exhibiting spherical morphology and good colloidal stability. Germination assays conducted with 0.1, 0.04, and 0.02 g/L of HA-AgNPs showed a concentration-dependent reduction in seed germination. Conversely, lower concentrations of HA-AgNPs significantly increased germination rates, survival, tolerance indices, and seed water absorption compared to silver ions (Ag+). SEM/EDS indicated more significant potential for HA-AgNPs internalization compared to Ag+. Therefore, these findings are innovative and open new avenues for understanding the impact of Ag+ and HA-AgNPs on seed germination.

Hyaluronic acid-mediated epithelial-mesenchymal transition of human lens epithelial cells via CD44 and TGF-ß2.

PURPOSE: The epithelial-mesenchymal transition of human lens epithelial cells plays a role in posterior capsule opacification, a fibrotic process that leads to a common type of cataract. Hyaluronic acid has been implicated in this fibrosis. Studies have investigated the role of transforming growth factor (TGF)-ß2 in epithelial-mesenchymal transition. However, the role of TGF-ß2 in hyaluronic acid-mediated fibrosis of lens epithelial cell remains unknown. We here examined the role of TGF-ß2 in the hyaluronic acid-mediated epithelial-mesenchymal transition of lens epithelial cells. METHODS: Cultured human lens epithelial cells (HLEB3) were infected with CD44-siRNA by using the Lipofectamine 3000 transfection reagent. The CCK-8 kit was used to measure cell viability, and the scratch assay was used to determine cell migration. Cell oxidative stress was analyzed in a dichloro-dihydro-fluorescein diacetate assay and by using a flow cytometer. The TGF-ß2 level in HLEB3 cells was examined through immunohistochemical staining. The TGF-ß2 protein level was determined through western blotting. mRNA expression levels were determined through quantitative real-time polymerase chain reaction. RESULTS: Treatment with hyaluronic acid (1.0 µM, 24 h) increased the epithelial-mesenchymal transition of HLEB3 cells. The increase in TGF-ß2 levels corresponded to an increase in CD44 levels in the culture medium. However, blocking the CD44 function significantly reduced the TGF-ß2-mediated epithelial-mesenchymal transition response of HLEB3 cells. CONCLUSIONS: Our study showed that both CD44 and TGF-ß2 are critical contributors to the hyaluronic acid-mediated epithelial-mesenchymal transition of lens epithelial cells, and that TGF-ß2 in epithelial-mesenchymal transition is regulated by CD44. These results suggest that CD44 could be used as a target for preventing hyaluronic acid-induced posterior capsule opacification. Our findings suggest that CD44/TGF-ß2 is crucial for the hyaluronic acid-induced epithelial-mesenchymal transition of lens epithelial cells.

Analysis of the Morphology, Rheology, and Clinical Applicability of a Hybrid Injectable with Hyaluronic Acid and Hydroxyapatite.

BACKGROUND: Treating signs of aging requires skeletal restructuring, facial contour redefinition, and sagging skin treatment. Hence, HarmonyCa® was recently introduced in the market, combining hyaluronic acid and hydroxyapatite. However, there are no studies evaluating the physicochemical properties of this material. PURPOSE: To analyze the morphology, rheology, and clinical applicability of HarmonyCa® over a 180-day follow-up. METHODS: A morphological analysis with scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDS) and rheology measurement with a rheometer. The clinical applicability analysis included a tissue ptosis patient who received the product and was reassessed 30 and 180 days after applying HarmonyCa®. RESULTS: SEM and EDS analyses showed two distinct morphologies related to hydroxyapatite and hyaluronic acid. The rheological measurements of HarmonyCa® showed G' and G″ of 51.56 and 44.01 Pa, respectively. The tan δ value was 0.8503, and G* was 108.1 Pa. The clinical outcomes were satisfactory. CONCLUSION: HarmonyCa® presents a good synergy between hyaluronic acid and hydroxyapatite. Also, the rheological analysis showed moderate viscosity for the product, with a clinical indication for subcutaneous application. LEVEL OF EVIDENCE IV: 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 .

Novel approach to soft tissue regeneration: in vitro study of compound hyaluronic acid and horizontal platelet-rich fibrin combination.

OBJECTIVE: This study aims to develop a compound biomaterial to achieve effective soft tissue regeneration. METHODOLOGY: Compound hyaluronic acid (CHA) and liquid horizontal-platelet-rich fibrin (H-PRF) were mixed at a ratio of 1:1 to form a CHA-PRF gel. Human gingival fibroblasts (HGFs) were used in this study. The effect of CHA, H-PRF, and the CHA-PRF gel on cell viability was evaluated by CCK-8 assays. Then, the effect of CHA, H-PRF, and the CHA-PRF gel on collagen formation and deposition was evaluated by qRT‒PCR and immunofluorescence analysis. Finally, qRT‒PCR, immunofluorescence analysis, Transwell assays, and scratch wound-healing assays were performed to determine how CHA, H-PRF, and the CHA-PRF gel affect the migration of HGFs. RESULTS: The combination of CHA and H-PRF shortened the coagulation time of liquid H-PRF. Compared to the pure CHA and H-PRF group, the CHA-PRF group exhibited the highest cell proliferation at all time points, as shown by the CCK-8 assay. Col1a and FAK were expressed at the highest levels in the CHA-PRF group, as shown by qRT‒PCR. CHA and PRF could stimulate collagen formation and HGF migration, as observed by fluorescence microscopy analysis of COL1 and F-actin and Transwell and scratch healing assays. CONCLUSION: The CHA-PRF group exhibited greater potential to promote soft tissue regeneration by inducing cell proliferation, collagen synthesis, and migration in HGFs than the pure CHA or H-PRF group. CHA-PRF can serve as a great candidate for use alone or in combination with autografts in periodontal or peri-implant soft tissue regeneration.

Crosslinked Collagen-Hyaluronic Acid Scaffold Enhances Interleukin-10 Under Co-Culture of Macrophages And Adipose-Derived Stem Cells.

The skin, the human body's largest organ, possesses a protective barrier that renders it susceptible to various injuries, including burns. Following burn trauma, the inflammatory process triggers both innate and adaptive immune responses, leading to the polarization of macrophages into two distinct phenotypes: the pro-inflammatory M1 and the anti-inflammatory M2. This dual response sets the stage for wound healing and subsequent tissue regeneration. Contributing to this transition from M1 to M2 polarization are human adipose-derived stem cells (ASCs), which employ paracrine signaling and inflammation suppression to enhance the remodeling phase. ASCs, when combined with biocompatible polymers, can be integrated into functional scaffolds. This study introduces an 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide-crosslinked (EDC-crosslinked) collagen-hyaluronic acid (Col-HA) scaffold assembled with ASCs, designed as a natural biomaterial device to modulate macrophage behavior in vitro under co-culture conditions. This innovation aims to improve wound healing processes. The EDC-crosslinked Col-HA scaffold favored the release of anti-inflammatory cytokines by ASCs, which indicated the M2 prevalence. In tissue engineering, a critical objective lies in the development of functional biomaterials capable of guiding specific tissue responses, notably the control of inflammatory processes. Thus, this research not only presents original findings but also points toward a promising avenue within regenerative medicine.

Aplicação de ácido hialurônico e corticoide através de uma punção guiada por ultrassonografia no tratamento de disfunção da articulação temporomandibular: relato de caso / Hyaluronic and corticoid application through an ultrasound-guided puncture in the treatment of temporomandibular joint dysfunctions: case report / Aplicación de ácido hialurónico y corticoide a través de una punción guiada por ecografía en el tratamiento de la disfunción de la articulación temporomandibular: informe de un caso

Introdução: As Disfunções temporomandibulares (DTM) incluem desordens dos músculos da mastigação, das articulações temporomandibulares e da inervação local, frequentemente associadas a dor orofacial e que resultam em mioartropatias do Sistema Mastigatório. A tendência atual tende a começar com tratamento conservador e progredir a procedimentos mais invasivos na falha dos tratamentos iniciais. Relato de caso: O presente relato visa mostrar o resultado de uma técnica invasiva para o tratamento de uma DTM grave, com a aplicação do ácido hialurônico e de corticoide através de uma punção guiado por ultrassonografia. A paciente apresentava dor crônica e perda importante de peso devido a limitação da abertura da boca. A RM demonstrou disfunção das ATMs, com sinais de deslocamento parcial do disco direito anteromedialmente. Foi realizada a aplicação bilateral intra-articular de ácido hialurônico e de corticoide através de uma punção guiado por ultrassonografia. Considerações Finais: A associação destas classes na punção de ATMs ainda não está bem estabelecida havendo necessidade de estudos complementares para avaliar eficácia, como este relato de caso, que se mostrou favorável com grande melhora clínica da paciente... (AU)

Introduction: Temporomandibular dysfunctions (TMD) include disorders of the masticatory muscles, temporomandibular joints, and local innervation, often associated with orofacial pain and resulting in myoarthropathies of the masticatory system. The current trend tends to begin with conservative treatment and progress to more invasive procedures if the initial treatments fail. Case Report: The present report aims to show the result of an invasive technique for the treatment of a severe TMD, with the application of hyaluronic acid and corticoid through an ultrasound-guided puncture. The patient presented with chronic pain and significant weight loss due to limited mouth opening. MRI demonstrated TMJ dysfunction, with signs of partial anteromedial dislocation of the right disc. Bilateral intra-articular application of hyaluronic acid and corticoid was performed through an ultrasound guided puncture. Final considerations: The association of these classes in TMJ puncture is still not well established, and further studies are needed to evaluate efficacy, as in this case report, which proved favorable, with great clinical improvement for the patient... (AU)

Introducción: Los trastornos temporomandibulares (TTM) incluyen trastornos de los músculos masticatorios, de las articulaciones temporomandibulares y de la inervación local, a menudo asociados a dolor orofacial y que dan lugar a mioartropatías del sistema masticatorio. La tendencia actual es comenzar con un tratamiento conservador y progresar hacia procedimientos más invasivos al fracasar los tratamientos iniciales. Informe de un caso: El presente informe pretende mostrar el resultado de una técnica invasiva para el tratamiento de un TTM severo, con la aplicación de ácido hialurónico y corticoide a través de una punción guiada por ecografía. El paciente presentaba dolor crónico y una importante pérdida de peso debido a la limitación de la apertura bucal. La RMN demostró una disfunción de la ATM, con signos de dislocación parcial del disco derecho anteromedialmente. Se realizó la aplicación intraarticular bilateral de ácido hialurónico y corticoide mediante una punción guiada por ecografía. Consideraciones finales: La asociación de estas clases en la punción de la ATM aún no está bien establecida y se necesitan más estudios para evaluar la eficacia, como en el reporte de este caso, que resultó favorable con gran mejoría clínica del paciente... (AU)

Clinical efficacy of hyaluronic acid in the treatment of periodontal intrabony defect: a systematic review and meta-analysis.

OBJECTIVE: This systematic review aimed to evaluate the effects of hyaluronic acid (HA) alone or in combination with any bone substitute for the treatment of intrabony defects (IBDs). MATERIAL AND METHODS: Six databases were searched up to April 2022 to find randomized clinical trials comparing the clinical effects of open flap debridement (OFD) + HA versus OFD alone (first group) or OFD + HA + bone substitutes versus OFD + bone substitutes (second group) in the treatment of IBDs with a follow-up of at least 3 months. Random effects models of mean differences were used to determine the clinical attachment level (CAL) gain, probing depth (PD) reduction, and radiographic bone fill (RBF). RESULTS: Of the 276 studies identified, 6 were included in the qualitative synthesis, and 5 in the meta-analyses. The meta-analyses in the first group showed a statistically significant differences for CAL gain (mean difference [MD]:1.00; 95% confidence interval [CI]:0.65 - 1.35; n = 2) and PD reduction (MD: 0.76; 95%CI: 0.34 - 1.17; n = 2) favoring HA + OFD at 6 months. However, in the second group, the meta-analyses did no show additional effect of HA in association with bone substitute was demonstrated for either CAL gain (MD: 0.57; 95%CI: - 0.30 - 1.43; n = 2) or PD reduction (MD: 1.05; 95%CI: - 0.38 - 2.47; n = 2) but did show significant differences for RBF (MD: 0.57; 95%CI: 0.15 - 0.99; n = 2) at 12 months. CONCLUSION: Compared with OFD alone, local application of HA in the treatment of IBDs provided a significant CAL gain and PD reduction at 6 months. However, its combination with bone substitutes showed no statistically significant differences at 12 months. CLINICAL RELEVANCE: The use of OFD + HA improves the CAL and PD in the treatment of IBDs compared to OFD only after 6 months of follow-up. These results are not maintained after 12 months.

In Vivo Models for the Study of Hyaluronic Acid Fillers: A Review.

BACKGROUND: Hyaluronic acid (HA) filler application is one of the most frequent minimally invasive aesthetic procedures used worldwide. Its properties and characterization, performance, effects in other tissues, and response to complication treatments have been studied in several animal models. This review aims to categorize animal models considering the advantages and disadvantages regarding the purpose of the study. METHODS: Literature research was made using MEDLINE via PubMed by two reviewers using keywords "hyaluronic acid" "filler" and "animal model". Full-text articles published in English and with an in vivo animal model were included for data extraction. RESULTS: The rat model was the most common animal used to evaluate properties or characteristics and degradation of HA fillers. Rabbits were preferred for evaluating HA embolism treatments; however, anatomical names of the arteries differ in some studies. Mice and rats used as vascular occlusion model are challenging due to the size of the vessels and viscosity of the filler. CONCLUSION: There is a wide variability of options of in vivo animal models to evaluate HA fillers. The animal characteristics, laboratory resources, and HA properties should be considered in accordance with the objective of the study, when choosing the ideal model. LEVEL OF EVIDENCE III: This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. 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 .

Effect of 0.15% sodium hyaluronate on tear film breakup time in healthy anesthetized cats.

OBJECTIVES: This study aimed to evaluate the effect of 0.15% sodium hyaluronate (SH) on tear film breakup time (TFBT) in healthy anesthetized cats. PROCEDURES: Forty cats undergoing elective surgery were enrolled. TFBT was assessed before anesthesia to obtain baseline values. As a preanesthetic medication, cats received midazolam, tramadol, and cetamine combined in the same syringe. For anesthetic induction and maintenance, propofol and isoflurane were used. After a 15 min stabilization period to achieve the surgical anesthetic plane, one eye was treated with one drop of SH, while the other eye received saline and served as a control. TFBT was measured at the end of the general anesthesia (T40) and 35 (T75) and 80 min (T120) after the termination of the anesthesia. TFBT values were compared between the control and SH-treated eyes; both values were also compared with the baseline values (p < .05). RESULTS: In the control eyes, TFBT significantly decreased from baseline at all time points (p < .001), while in SH-treated eyes, TFBT significantly increased from baseline only at T40 (p < .0001). In SH-treated eyes, TFBT was significantly higher than that in the control eyes at all time points (p < .001). CONCLUSIONS: In healthy cats, TFBT decreases significantly after 40 min of general anesthesia, and one drop of 0.15% sodium hyaluronate was able to maintain the stability of the tear film for up to 75 min in treated eyes. However, the isolated effect of each drug used in our anesthetic protocol on TFBT should be executed in further studies.

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

BACKGROUND: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.

Hyaluronic acid nanoemulsions improve piplartine cytotoxicity in 2D and 3D breast cancer models and reduce tumor development after intraductal administration.

Nanoemulsions modified with chitosan (NE-Q) or hyaluronic acid (NE-HA), developed for intraductal administration of piplartine (piperlongumine) and local breast cancer treatment, were evaluated for cytotoxic effects in vitro in 2D and 3D breast cancer models and in vivo in a chemically induced carcinogenesis model. Droplet size was lower than 100 nm, and zeta potential varied from +17.9 to -25.5 mV for NE-Q and NE-HA, respectively. Piplartine nanoencapsulation reduced its IC50 up to 3.6-fold in T-47D and MCF-7 monolayers without differences between NE-Q and NE-HA, and up to 6.6-fold in cancer spheroids. Cytotoxicity improvement may result from a more efficient NE-mediated delivery, as suggested by stronger fluorescent staining of cells and spheroids. In 1-methyl-1-nitrosourea -induced breast cancer models, intraductal administration of piplartine-loaded NE-HA inhibited breast tumor development and histological alterations. These results support the potential applicability of piplartine-loaded NE-HA for intraductal treatment of breast cancer.

Effects of the Application of Decellularized Amniotic Membrane Solubilized with Hyaluronic Acid on Wound Healing.

A perfect graft for wound care must be readily available without affecting the immune response, covering and protecting the wound bed. Considering previous studies have already established the use of hyaluronic acid (HA) for the treatment of wounds but the data presented on the amniotic membrane (AM) and its promising effects on healing still requires further investigation, this study aimed to evaluate the effects of the application of a decellularized amniotic membrane solubilized with hyaluronic acid on the healing process of cutaneous wounds on the 7th and 14th day, to evaluate the evolution of the wound and the inflammatory phases in these two times. Cutaneous lesions were excised from the dorsal region and 96 Wistar rats were divided into four groups: I-Excisional wound (EW); II-EW + AM; III-EW + HA; IV-EW + AM + HA. The present study demonstrated that the proposed combined therapy favors the tissue repair process of the epithelial lesion. Results showed a reduction in pro-inflammatory cytokines, an increase in anti-inflammatory cytokines, an increase in TGF-ß, and attenuation of oxidative stress, reducing the acute inflammatory response and promoting the beginning of tissue repair. We concluded that the proposed therapies accelerated the inflammatory process with anticipation of the repair phase.

Photobiomodulation associated with lipid nanoparticles and hyaluronic acid accelerate the healing of excisional wounds.

Objectives: This article aimed to investigate the effects of the association between photobiomodulation and hyaluronic acid incorporated in lipid nanoparticles in an epithelial lesion model in inflammatory parameters and oxidative stress. Methods: Eighty Wistar rats were randomly assigned to the following groups: epithelial lesion group (EL); EL+PBM; EL+HA; EL+SLNs; EL+SLNs-HA; EL+PBM+HA; EL+PBM+SLNs; EL+PBM+SLNs-HA. The animals were anesthetized with 4% isofluorane after shaving and induced to an epithelial lesion. Topical treatment with a gel containing HA (0.9%) and/or SLNs (10 mg/mL) and with laser irradiation occurred daily for 1 week. Results: The results showed an increase in wound contraction on the seventh day in the LE + LBM + AH-NPL group, a reduction in pro-inflammatory cytokines (IL-6, IL-1ß, and TNF-α), an increase in anti-inflammatory cytokines (IL- 4 and IL-10) and TGF-ß. The levels of pro-inflammatory cytokine IL-4 and TGF-ß also showed an increase in the LE + NPL-AH, LE + FBM + AH, LE + FBM + NPL and LE + FBM + NPL-AH groups. Regarding oxidative stress parameters, the levels of DCF and nitrite decreased in the combined therapy group when compared to the control group, as well as oxidative damage (carbonyl and sulfhydryl). In the antioxidant defense, there was an increase in GSH and SOD in the combination therapy group. Histological analysis showed a reduction in inflammatory infiltrate in the combination therapy group. The number of fibroblasts and the compaction of collagen fibers did not obtain significant responses. Conclusions: Results analyzed together showed that the combined therapy favored the repair process, and that studies can be carried out to enhance the histological analysis therapy favored the tissue repair process and that studies can be carried out to enhance the histological analysis.

The effects of sulfated hyaluronan in breast, lung and colorectal carcinoma and monocytes/macrophages cells: Its role in angiogenesis and tumor progression.

Hyaluronan (HA) is a component of the extracellular matrix (ECM) it is the main non-sulfated glycosaminoglycan able to modulate cell behavior in the healthy and tumor context. Sulfated hyaluronan (sHA) is a biomaterial derived from chemical modifications of HA, since this molecule is not naturally sulfated. The HA sulfation modifies several properties of the native molecule, acquiring antitumor properties in different cancers. In this study, we evaluated the action of sHA of ~30-60 kDa with different degrees of sulfation (0.7 sHA1 and 2.5 sHA3) on tumor cells of a breast, lung, and colorectal cancer model and its action on other cells of the tumor microenvironment, such as endothelial and monocytes/macrophage cells. Our data showed that in breast and lung tumor cells, sHA3 is able to modulate cell viability, cytotoxicity, and proliferation, but no effects were observed on colorectal cancer cells. In 3D cultures of breast and lung cancer cells, sHA3 diminished the size of the tumorsphere and modulated total HA levels. In these tumor models, treatment of monocytes/macrophages with sHA3 showed a downregulation of the expression of angiogenic factors. We also observed a decrease in endothelial cell migration and modulation of the hyaluronan-binding protein TSG-6. In the breast in vivo xenograft model, monocytes/macrophages preincubated with sHA1 or sHA3 decreased tumor vasculature, TSG-6 and HA levels. Besides, in silico analysis showed an association of TSG-6, HAS2, and IL-8 with biological processes implicated in the progression of the tumor. Taken together, our data indicate that sHA in a breast and lung tumor context is able to induce an antiangiogenic action on tumor cells as well as in monocytes/macrophages (Mo/MØ) by modulation of endothelial migration, angiogenic factors, and vessel formation.

Effects of a collagen hyaluronic acid silk-fibroin patch with the electroconductive element polyaniline on left ventricular remodeling in an infarct heart model.

Biodegradable cardiac patches have been able to induce improvement in left ventricular (LV) remodeling. A novel scaffold patch made with collagen and silk-fibroin (COL-SF) was further associated to polyaniline (PANi) to increase conductivity. Thus, this study investigated the safety of the association of PANi to a patch, and the improvement in LV remodeling in a myocardial infarct (MI) rat model. Wistar rats underwent MI induction. MI was confirmed with echocardiographic and after 2 weeks, animals (n = 10/group) were randomized into: (a) COL-SF hyaluronic acid patch, (b) PANi hyaluronic acid patch, (c) MI Control (just repeat thoracotomy). Healthy animals were also followed. Echocardiography was performed at pre-treatment, and at 2-, 4-, and 8-weeks post-treatment. Hearts underwent hemodynamic evaluation on Langendorff apparatus and histology for LV thickness and percent of infarct size. Liver, kidneys, and blood samples were evaluated for biochemical, hematological, oxidative stress, and histology. There was a tendency of lower %infarct size in patched animals. LV thickness was higher in the patched animals than controls. Functional echocardiographic indices %Fractional shortening and %LV ejection fraction decreased in the MI control group, but not in the patched animals. PANi presented higher %LVEF versus MI control. PANi presented higher liver transaminases; no morphological changes were observed in histology. Elevation of antioxidant markers was observed. COL-SF and PANi patches were able to induce better remodeling features compared to MI controls on %infarct size and LV thickness and have not presented echocardiographic worsening. Polyaniline may present a slight improvement on LV remodeling, despite associated to signs of hepatotoxicity and pro-oxidant effect.

Physicochemical characterization and in vitro antioxidant activity of hyaluronic acid produced by Streptococcus zooepidemicus CCT 7546.

Hyaluronic acid (HA) is a biopolymer with applications in different areas such as medicine and cosmetics. HA is currently either isolated from animal sources or produced by microbial fermentation. Animal HA presents some disadvantages such as high cost and risk of viral cross-species or another infectious agent. In the present study, we evaluated the physicochemical characteristics and in vitro antioxidant capacity of HA produced by Streptococcus zooepidemicus CCT 7546. In addition, commercial sodium hyaluronate (SH) from an animal source was used as control. The microbial HA yield after purification was 69.8 mg/L. According to Fourier transform infrared spectroscopy, it was seen that bacterial and animal HA spectra are overlapped. The thermogravimetric analysis revealed that microbial HA was more stable than its equivalent from the animal source. However, scanning electron microscopy indicates that the purification method used in the animal product was more effective. Microbial HA showed activity in total antioxidant capacity (14.02 ± 0.38%), reducing power (18.18 ± 6.43%), DPPH radical-scavenging (5.57 ± 0.23 kmol TE/g), and hydroxyl radical-scavenging (28.39 ± 2.40%) tests. Therefore, in vitro antioxidant tests demonstrated that the antioxidant action mechanism occurs through scavenging reactive oxygen species (ROS) and donating electrons/hydrogen atoms.

Hyaluronic Acid In Vitro Response: Viability and Proliferation Profile of Human Chondrocytes in 3D-Based Culture.

OBJECTIVES: This study aimed to evaluate the efficacy of hyaluronic acid in the viability and proliferation profile of human femoral-tibial joint cartilage affected by osteoarthritis using in vitro models of chondrocytes in a 2-dimensional (2D)- and 3-dimensional (3D)-based culture model by spheroids. DESIGN: In vitro study of knee cartilage affected by osteoarthritis that required surgical treatment. Samples were cultured and exposed to hyaluronic acid (100 and 500 µM; intervention group) or vehicle solution. In monolayer or 2D culture, proliferation and cell viability were measured, and nuclear morphometry was analyzed by 4',6'-diamino-2-fenil-indol (DAPI) staining. The 3D-based culture established from the culture of articular cartilage of patients submitted to total knee arthroplasty evaluated the diameter, viability, and fusion ability of the chondrospheres created. RESULTS: Samples from 3 patients resulted in viable cultures, with chondrocyte cells exhibiting a potential for cell proliferation and viability to establish a culture. Hyaluronic acid (100 and 500 µM) improved chondrocyte viability and proliferation up to 72 hours in contact when compared with the control group, and no nuclear irregularities in morphology cell characteristics were observed by DAPI. In the 3D evaluation, hyaluronic acid (500 µM) improved the cellular feedback mechanisms, increasing the survival and maintenance of the chondrospheres after 7 days of analysis, showing the intrinsic capacity of chondrospheres grouped in the attempt to rearrange and reestablish new articular tissue. CONCLUSIONS: The 2D- and 3D-based culture models with hyaluronic acid improved chondrocyte viability and proliferation and demonstrated the ability of freshly formed chondrospheres to undergo fusion when placed together in the presence of hyaluronic acid.

The effect of hyaluronic acid on the periodontium in spontaneous periodontitis in rats / El efecto del ácido hialurónico sobre el periodonto en periodontitis espontánea en ratas

SUMMARY: The aim of the article was to study changes in periodontal tissues in rats with spontaneous periodontitis (SP) and to evaluate the effect of hyaluronic acid (HA) on the state of the periodontium. Wistar rats with signs of SP were divided into 6 groups: 1) intact group; 2) intact animals with HA "HD-1,0 MDa"; 3) SP group; 4) SP with HA "S-2,4 MDa"; 5) SP with HA "ST-2,4 MDa"; 6) SP with HA "HD-1,0 MDa". The study of the periodontium rats with SP noted the main structural changes (collagen reduction, resorption of alveolar bone, dilatation and stasis of the vessels of the periodontium, gingival papilla and tooth pulp), which were assessed as moderate. Morphological evidence of inflammation was infiltration of neutrophils into the connective tissue of the gums, without the formation of abscesses. Local administration of HA did not cause additional structural damage in periodontal tissues of rats with SP, but also did not affect changes in the microvascular system of periodontium and tooth pulp, periodontal ligaments, only a tendency to inhibit alveolar bone resorption in rats was noted. One can consider the tendency to improve the condition of periodontal tissues in the group of rats injected with high molecular HA and HA with mannitol (2.4 MDa).

RESUMEN: El objetivo del artículo fue estudiar los cambios en los tejidos periodontales en ratas con periodontitis espontánea (PE) y evaluar el efecto del ácido hialurónico (HA) sobre el estado del periodonto. Las ratas Wistar con signos de PE se dividieron en 6 grupos: 1) grupo intacto; 2) animales intactos con HA "HD-1,0 MDa"; 3) grupo PE; 4) PE con HA "S-2,4 MDa"; 5) PE con HA "ST-2,4 MDa"; 6) PE con HA "HD-1,0 MDa". En las ratas con PS se observaron los principales cambios estructurales (reducción de colágeno, reabsorción del hueso alveolar, dilatación y estasis de los vasos del periodonto, papila gingival y pulpa dentaria), que fueron evaluados como moderados. La evidencia morfológica de inflamación fue la infiltración de neutrófilos en el tejido conectivo de las encías, sin la formación de abscesos. La administración local de HA no causó daño estructural adicional en los tejidos periodontales de las ratas con PE, pero tampoco se produjo cambios en el sistema microvascular del periodonto y en la pulpa dental y ligamentos periodontales.Se observó una tendencia a inhibir la resorción del hueso alveolar. Se puede considerar la tendencia a mejorar el estado de los tejidos periodontales en el grupo de ratas inyectadas con HA de alto peso molecular y HA con manitol (2,4 MDa).

Syndecan-1 Depletion Has a Differential Impact on Hyaluronic Acid Metabolism and Tumor Cell Behavior in Luminal and Triple-Negative Breast Cancer Cells.

Glycosaminoglycans (GAGs) and proteoglycans (PGs) are major components of the glycocalyx. The secreted GAG and CD44 ligand hyaluronic acid (HA), and the cell surface PG syndecan-1 (Sdc-1) modulate the expression and activity of cytokines, chemokines, growth factors, and adhesion molecules, acting as critical regulators of tumor cell behavior. Here, we studied the effect of Sdc-1 siRNA depletion and HA treatment on hallmark processes of cancer in breast cancer cell lines of different levels of aggressiveness. We analyzed HA synthesis, and parameters relevant to tumor progression, including the stem cell phenotype, Wnt signaling constituents, cell cycle progression and apoptosis, and angiogenic markers in luminal MCF-7 and triple-negative MDA-MB-231 cells. Sdc-1 knockdown enhanced HAS-2 synthesis and HA binding in MCF-7, but not in MDA-MB-231 cells. Sdc-1-depleted MDA-MB-231 cells showed a reduced CD24-/CD44+ population. Furthermore, Sdc-1 depletion was associated with survival signals in both cell lines, affecting cell cycle progression and apoptosis evasion. These changes were linked to the altered expression of KLF4, MSI2, and miR-10b and differential changes in Erk, Akt, and PTEN signaling. We conclude that Sdc-1 knockdown differentially affects HA metabolism in luminal and triple-negative breast cancer model cell lines and impacts the stem phenotype, cell survival, and angiogenic factors.