Use of hyaluronic acid associated with triamcinolone acetonide or ozone gas in the treatment of induced osteoarthritis in rabbits

Purpose: To evaluate the effects of the intra-articular application of hyaluronic acid associated with triamcinolone acetonide, and ozone gas in the treatment of induced osteoarthritis in rabbit’s stifles.Methods: Twenty-one Norfolk rabbits were submitted to cranial cruciate ligament transection of the left stifle. After six weeks of the surgery, the rabbits were randomized assigned into three groups: G1 (control) – saline solution (0.9%); G2 – hyaluronic acid associated with triamcinolone; G3 – ozone gas, submitted to three intra-articular applications every seven days. Results: Significant differences occurred: osteophytes at medial femoral condyle (G2 > G1, G2 > G3) on radiography exam; thickening of the medial condyle (G1 > G3, G2 > G3) on ultrasound exam; osteophytes at lateral tibial condyle (G2 > G1, G2 > G3), and medial femoral condyle (G1 > G2, G3 > G1) on computed tomography. Histologically, mean values of chondrocytes in the femur and tibia in G3 and G2 were statistically lower. Conclusions: The intra-articular injection of hyaluronic acid associated with triamcinolone accentuated degenerative joint disease by imaging and macroscopic evaluations, and by histological findings, this treatment and the ozone gas treatment showed similar effects and were inferior to the saline solution (0.9%).

Development and characterization of hyaluronic acid-lysine nanoparticles with potential as innovative dermal filling

ABSTRACT Skin aging causes changes such as wrinkles and flaccidity leading to a large demand for aesthetic procedures, including dermal filling. A key agent in dermal filling is hyaluronic acid (HA), which is a naturally occurring glycosaminoglycan. However, it is a hydrophilic macromolecule that experiences great difficulty in crossing the skin barrier causing most commercial formulations containing it to be injectable, which in turn brings risks since they involve an invasive technique. In that sense, the aim of this study was to develop and characterize nanoparticles obtained from ionic interaction between HA and lysine (Lys) for use as a potential agent of dermal filling for topical application, increasing and improving its applicability and safety. To this end, nanoparticles were obtained by dripping of Lys over HA under magnetic stirring. A nanometric size was confirmed and a suitable surface charge was obtained by zeta potential. Nanoparticles were almost spherical in shape with a smooth surface. Interaction between raw materials for preparing nanoparticles was studied by FTIR and NMR spectroscopy and an ionic interaction was confirmed. These physicochemical features suggest that obtained nanoparticles can be further used as a topical dermal filling.

Expression of heparanase in basal cell carcinoma and squamous cell carcinoma

Abstract: Background: Heparanase is an enzyme that cleaves heparan sulfate chains. Oligosaccharides generated by heparanase induce tumor progression. Basal cell carcinoma and squamous cell carcinoma comprise types of nonmelanoma skin cancer. Objectives: Evaluate the glycosaminoglycans profile and expression of heparanase in two human cell lines established in culture, immortalized skin keratinocyte (HaCaT) and squamous cell carcinoma (A431) and also investigate the expression of heparanase in basal cell carcinoma, squamous cell carcinoma and eyelid skin of individuals not affected by the disease (control). Methods: Glycosaminoglycans were quantified by electrophoresis and indirect ELISA method. The heparanase expression was analyzed by quantitative RT-PCR (qRTPCR). Results: The A431 strain showed significant increase in the sulfated glycosaminoglycans, increased heparanase expression and decreased hyaluronic acid, comparing to the HaCaT lineage. The mRNA expression of heparanase was significantly higher in Basal cell carcinoma and squamous cell carcinoma compared with control skin samples. It was also observed increased heparanase expression in squamous cell carcinoma compared to the Basal cell carcinoma. Conclusion: The glycosaminoglycans profile, as well as heparanase expression are different between HaCaT and A431 cell lines. The increased expression of heparanase in Basal cell carcinoma and squamous cell carcinoma suggests that this enzyme could be a marker for the diagnosis of such types of non-melanoma cancers, and may be useful as a target molecule for future alternative treatment.

Remodeling of the middle third of the face with fillers / Remodelamento do terço médio da face com preenchedores

Facial aging is a consequence of multiple intrinsic and extrinsic interactive factors, leading to loss of volume and repositioning of facial fat and bone remodeling. Breakthroughs have been seen in recent decades in the ability to reduce the signs of aging. Filling techniques and numerous products available on the market have generated new interest in the study of facial anatomy. Fillers are able to restore volume in limited areas and are suitable for treating facial wrinkles and loss of subcutaneous volume. The gold standard remains hyaluronic acid because it is more biocompatible and has greater permanence in the skin, without being definitive. The objective of this report is to review the anatomy of the middle third of the face and the main indications for filling and the techniques used for rejuvenation.

O envelhecimento facial é consequência de múltiplos fatores intrínsecos e extrínsecos que interagem entre si, levando à perda de volume e reposicionamento da gordura facial, assim como o remodelamento ósseo. Nas últimas décadas, houve um grande avanço na Medicina em reduzir os sinais do envelhecimento. As técnicas de preenchimento e os inúmeros produtos disponíveis no mercado trouxeram novo interesse no estudo da anatomia da face. Os preenchedores têm como função restaurar o volume de áreas restritas, portanto, são adequados para tratar rugas faciais e perda de volume subcutâneo. O padrão ouro atualmente é o ácido hialurônico, por ser mais biocompatível e ter uma permanência maior na pele, sem ser definitivo. O objetivo desse artigo é revisar a anatomia do terço médio da face, assim como as principais indicações de preenchimento e técnicas utilizadas para o rejuvenescimento.

Tissue hyaluronan expression, as reflected in the sputum of lung cancer patients, is an indicator of malignancy

Hyaluronan (HA) shows promise for detecting cancerous change in pleural effusion and urine. However, there is uncertainty about the localization of HA in tumor tissue and its relationship with different histological types and other components of the extracellular matrix, such as angiogenesis. We evaluated the association between HA and degree of malignancy through expression in lung tumor tissue and sputum. Tumoral tissue had significantly increased HA compared to normal tissue. Strong HA staining intensity associated with cancer cells was significant in squamous cell carcinoma compared to adenocarcinoma and large cell carcinoma. A significant direct association was found between tumors with a high percentage of HA and MVD (microvessel density) in tumoral stroma. Similarly significant was the direct association between N1 tumors and high levels of HA in cancer cells. Cox multivariate analysis showed significant association between better survival and low HA. HA increased in sputum from lung cancer patients compared to cancer-free and healthy volunteers and a significant correlation was found between HA in sputum and HA in cancer tissue. Localization of HA in tumor tissue was related to malignancy and reflected in sputum, making this an emerging factor for an important diagnostic procedure in patients suspected to have lung cancer. Further study in additional patients in a randomized prospective trial is required to finalize these results and to validate our quantitative assessment of HA, as well as to couple it to gold standard sputum cytology.

Glucosamine and chondroitin sulfate association increases tibial epiphyseal growth plate proliferation and bone formation in ovariectomized rats

OBJECTIVE: The growth plate consists of organized hyaline cartilage and serves as a scaffold for endochondral ossification, a process that mediates longitudinal bone growth. Based on evidence showing that the oral administration of glucosamine sulfate (GS) and/or chondroitin sulfate (CS) is clinically valuable for the treatment of compromised articular cartilage, the current study evaluated the effects of these molecules on the tibial epiphyseal growth plate in female rats. METHOD: The animals were divided into two control groups, including vehicle treatment for 45 days (GC45) and 60 days (GC60) and six ovariectomized (OVX) groups, including vehicle treatment for 45 days (GV45), GS for 45 days (GE45GS), GS+CS for 45 days (GE45GS+CS), vehicle for 60 days (GV60), GS for 60 days (GE60GS) and GS+CS for 60 days (GE60GS+CS). At the end of treatment, the tibias were dissected, decalcified and processed for paraffin embedding. Morphological and morphometric methods were employed for analyzing the distal tibial growth plates using picrosirius red staining and the samples were processed for histochemical hyaluronan detection. Morphometric analyses were performed using the 6.0ProPlus¯ Image system. RESULTS: Notably, after 60 days of treatment, the number of proliferative chondrocytes increased two-fold, the percentage of remaining cartilage increased ...

Optimization of flask culture medium and conditions for hyaluronic acid production by a streptococcus equisimilis mutant nc2168

A mutant designated NC2168, which was selected from wild-type Streptococcus equisimilis CVCC55116by ultraviolet ray combined with60Co-γ ray treatment and does not produce streptolysin, was employed to produce hyaluronic acid (HA). In order to increase the output of HA in a flask, the culture medium and conditions for NC2168 were optimized in this study. The influence of culture medium ingredients including carbon sources, nitrogen sources and metal ions on HA production was evaluated using factional factorial design. The mathematical model, which represented the effect of each medium component and their interaction on the yield of HA, was established by the quadratic rotary combination design and response surface method. The model estimated that, a maximal yield of HA could be obtained when the concentrations of yeast extract, peptone, glucose, and MgSO4 were set at 3 g/100 mL, 2 g/100 mL, 0.5 g/100 mL and 0.15 g/100 mL, respectively. Compared with the values obtained by other runs in the experimental design, the optimized medium resulted in a remarkable increase in the output of HA and the maximum of the predicted HA production was 174.76 mg/L. The model developed was accurate and reliable for predicting the production of HA by NC2168.Cultivation conditions were optimized by an orthogonal experimental design and the optimal conditions were as follows: temperature 33ºC, pH 7.8, agitation speed 200 rpm, medium volume 20 mL.

Influência do envelhecimento na concentração de ácido hialurônico nas pregas vocais de ratas fêmeas / Influence of aging on hyaluronic acid concentration in the vocal folds of female rats

A vibração das pregas vocais é um importante fator envolvido na produção vocal e o envelhecimento pode alterar a quantidade de ácido hialurônico da prega vocal levando a disfonia. OBJETIVO: Este estudo compara a concentração de ácido hialurônico nas pregas vocais de ratas fêmeas idosas e jovens. Desenho do estudo: estudo experimental. MATERIAL E MÉTODO: Foram utilizadas pregas vocais de 13 ratas fêmeas divididas em dois grupos: cinco ratas idosas e oito ratas jovens. A concentração tecidual do ácido hialurônico foi determinada por meio de método fluorimétrico utilizando a proteína de ligação ao ácido hialurônico imobilizada em placas de enzyme-linked immunosorbent assay (ELISA) e também conjugada à biotina. Estreptavidina marcada com európio foi adicionada e, depois de európio ter sido liberado com o uso de solução de enhancement; a fluorescência final foi medida em um fluorímetro. RESULTADOS: Foram encontradas as seguintes concentrações de ácido hialurônico nas pregas vocais de acordo com os grupos: 581,7 ng/mg em ratas idosas e 1275,6 ng/mg em ratas jovens. A análise estatística mostrou diferença entre os grupos. CONCLUSÃO: A prega vocal de ratas idosas tem uma menor concentração de ácido hialurônico do que a concentração da prega vocal de ratas jovens.

The vibration of the vocal fold lamina propria is an important factor involved in vocal production and aging may change the amount of hyaluronic acid in the vocal fold leading to dysphonia. AIMS: This study compares the concentration of hyaluronic acid in vocal folds of aged and young female rats. Study design: experimental. MATERIALS AND METHODS: We used the vocal cords of 13 female rats divided into two groups: five aged rats and eight young ones. The tissue concentration of hyaluronic acid was determined using the fluorimetric method with the hyaluronic acid binding-protein coated on plates of enzyme-linked immunosorbent assay (ELISA), conjugated with biotin. Europium-labeled streptavidin was added and, after europium release with the use of enhancement solution, the final fluorescence was measured in a fluorometer. RESULTS: We found the following concentrations of hyaluronic acid in vocal fold according to the group: 581.7 ng/mg in old female rats and 1275.6 ng/mg in young female rats. Statistical analysis showed differences between groups. CONCLUSIONS: The vocal folds of old female rats have a lower concentration of hyaluronic acid when compared to such concentration on the vocal folds of young female rats.

Ascaris lumbricoides: Estudio de la cinética de captación de ácido hialurónico / Ascaris lumbricoides: Study of kinetics of hyaluronic acid capture

El ácido hialurónico (AH) participa de la respuesta inmune porque es un constituyente de las barreras mecánicas naturales, un estimulador de la inflamación y un ligando de receptores involucrados en la inmunidad. En experiencias previas se demostró que adultos y larvas de Ascaris lumbricoides pueden unir AH. El objetivo de este trabajo fue estudiar la cinética de captación de AH por este nematodo. Se trabajó con un extracto de parásito adulto y un concentrado de larvas. Se utilizó la técnica modificada de Inhibición de la Agregación por Adhesión para detección del receptor CD44 soluble de hialuronato. Se evaluó la cinética de captación de AH por el parásito, variando el tiempo de contacto entre ambos desde un tiempo inicial hasta 90 minutos, con intervalos de 10 minutos. Para todos los tiempos se calcularon los valores de CexpAdhE AH e IexpCP AH que fueron analizados por las pruebas de Wilcoxon y de Friedman. Los resultados mostraron que al tiempo inicial de contacto, el parásito adulto y las larvas captan una pequeña cantidad de AH. A los 10 minutos se produjo la mayor captación que se mantuvo sin diferencias significativas durante 50 minutos. A partir de ese tiempo, hubo un leve incremento en la captación que permaneció sin diferencia significativa hasta el tiempo final. Esta experiencia demuestra que in vitro, A. lumbricoides realiza la mayor captación a los 10 minutos de contacto, por lo que se sugiere que in vivo sería capaz de unir AH rápidamente a los fines de modular la respuesta inmune.

Hyaluronic acid (HA) particípales in the immune response because it is a constituent of natural mechanical barriers, an inflammation stimulator and a ligand for receptors involved in immunity. Previous experiences have shown that adult specimens and larval concéntrales oí A. lumbricoides have hyaluronan binding capacity. The aim ofthis work was to study the kinetics ofhyaluronic acid capture by this parasite. Work was carried out with an extract of adult specimen and a larval concéntrate. The modlfled test of serum soluble CD44 Detectlon by Aggregatlon Inhlbltlon was used. Kinetics hyaluronlc acid capture by the nematode was assessed by varylng the contad time between HA and parasite, from an initial time to 90 minutes, with 10-minute intervals. CexpAdhE HA ana lexpCP HA were calculated for all the times and they were analysed by Wilcoxon and Friedman Tests The results showed that at initial time of contad, the adult paraslte and the larvae had captured a small amount of HA. The major capture was made at 10 minutes and it was maintained for 50 minutes without significad differences. From that time there was a slight increase in capture and it remaíned so untíi the end of the experiment without considerable variations. This experience demonstrates that at 10 minutes of in vitro contad, A. lumbricoides captures most of the HA, whích suggests that in vivo, the parasite would be able to capture HA quickly for the purpose of modulating the immune response.

Glycosaminoglycan distribution in the rat uterine cervix during the estrous cycle

OBJECTIVE: To analyze the amount of glycosaminoglycans in the uterine cervix during each phase of the rat estrous cycle. DESIGN: Based on vaginal smears, forty female, regularly cycling rats were divided into four groups (n = 10 for each group): GI - proestrous, GII - estrous, GIII - metaestrous and GIV - diestrous. Animals were sacrificed at each phase of the cycle, and the cervix was immediately removed and submitted to biochemical extraction and determination of sulfated glycosaminoglycans and hyaluronic acid. The results were analyzed by ANOVA followed by the Bonferroni post-hoc test. RESULTS: The uterine cervix had the highest amount of total sulfated glycosaminoglycans and dermatan sulfate during the estrous phase (8.90 ± 0.55 mg/g of cetonic extract, p<0.001; and 8.86 ± 0.57 mg/g of cetonic extract, p<0.001). In addition, there was more heparan sulfate at the cervix during the proestrous phase (0.185 ± 0.03 mg/g of cetonic extract) than during any other phase (p<0.001). There were no significant changes in the concentration of hyaluronic acid in the uterine cervix during the estrous cycle. CONCLUSION: Our data suggest that the amount of total sulfated glycosaminoglycans may be influenced by hormonal fluctuations related to the estrous cycle, with dermatan sulfate and heparan sulfate being the glycosaminoglycans most sensitive to hormonal change.

Analysis of hyaluronic acid concentration in rat vocal folds during estral and gravidic puerperal cycles / Análise da concentração do ácido hialurônico nas pregas vocais de ratas durante o ciclo estral e ciclo gravídico-puerperal

Hormone plays an important role in the larynx. Among other substances, vocal folds contain hyaluronic acid, which tissue concentration may vary according to hormone action. AIM: the objective of this study is to analyze hyaluronic acid concentration in the vocal folds during estral and gravidic-puerperal cycles. MATERIALS AND METHODS: Experimental study. 40 adult rats were divided into two groups. In the first group we used 20 rats to establish the concentration of hyaluronic acid during the estral cycle and in the second group, 20 animals were submitted to the same procedure but during the gravidic-puerperal cycle. RESULTS: Variations in hyaluronic acid concentration was not observed during the estral cycle. In the gravidic puerperal cycle group, an increase in hyaluronic acid concentration was observed in the puerperal subgroup. Comparing the two groups of estral and gravidic-puerperal cycles, no difference was observed. CONCLUSIONS: In comparing all subgroups of estral and gravidic-puerperal cycles, an increase in hyaluronic acid concentration was noticed only in the puerperal phase.

Os hormônios exercem importante influência sobre a laringe. A prega vocal contém, entre outras substâncias, o ácido hialurônico, cuja concentração nos tecidos pode variar com a ação dos hormônios. OBJETIVO: O objetivo deste trabalho é analisar comparativamente a concentração do ácido hialurônico nas pregas vocais de ratas durante o ciclo estral e ciclo gravídico-puerperal. FORMA DE ESTUDO: Experimental. MATERIAL E MÉTODO: Foram utilizadas 40 ratas adultas, divididas em dois grupos, no primeiro grupo utilizamos 20 ratas para determinação da concentração do ácido hialurônico no ciclo estral, no segundo grupo, também de 20 animais, foi realizado o mesmo experimento no ciclo gravídico-puerperal. RESULTADOS: No grupo do ciclo estral não observou-se variação da concentração do ácido hialurônico. No grupo do ciclo gravídico-puerperal houve aumento da concentração do ácido hialurônico no subgrupo do puerpério. Na comparação entre os dois grupos do ciclo estral e gravídico-puerperal não houve diferença. Quando comparamos todos os subgrupos há diferença no grupo do puerpério. CONCLUSÕES: Comparando-se todos os subgrupos do ciclo estral e ciclo gravídico-puerperal, só no puerpério houve aumento da concentração do ácido hialurônico.

Valor preditivo de marcadores séricos de fibrose hepática em pacientes portadores de hepatite crônica viral C / Predictive value of serum markers of hepatic fibrosis in patients with chronic hepatitis C

INTRODUÇÃO: Os marcadores séricos têm sido empregados na avaliação da fibrose hepática em pacientes portadores de hepatite crônica C (HCC). OBJETIVOS: Avaliar a capacidade do índice aspartato aminotransferase (AST)/alanina aminotransferase (ALT), dos níveis séricos de gama-glutamiltransferase (GGT), contagem de plaquetas, do índice AST/plaquetas (APRI) e do ácido hialurônico (AH) em predizer a intensidade da fibrose hepática na HCC e a variação desses marcadores após tratamento com interferon. PACIENTES E MÉTODOS: Em 72 pacientes portadores de hepatite C determinamos no soro o índice AST/ALT, GGT, plaquetas, índice APRI (obtido pelo quociente AST/plaquetas) e o AH, que foram comparados ao estadiamento histológico, segundo os critérios de METAVIR. Receberam tratamento com interferon e ribavirina 65 pacientes. Os indivíduos que concluíram o tratamento (n = 33) realizaram nova dosagem dos marcadores séricos de fibrose para comparar com os níveis pré-tratamento. RESULTADOS: Observamos que a GGT, a contagem de plaquetas, o índice APRI e o AH se correlacionaram com estádio de doença hepática (p < 0,01), exceto o índice AST/ALT. A análise das áreas sob as curvas ROC (AUC) evidenciaram que a melhor associação com estadiamento hepático foi para o índice APRI e a dosagem sérica do AH: AUC (APRI) = 0,85 e AUC (AH) = 0,86. Na avaliação pós-terapia com interferon, apenas a GGT e o índice APRI apresentaram redução de seus níveis (p < 0,05). CONCLUSÃO: O AH e o índice APRI apresentaram maior acurácia no estadiamento da fibrose, podendo ser aplicados como métodos diagnósticos alternativos na HCC.

INTRODUCTION: Serum markers have been used in the assessment of liver fibrosis in patients with chronic hepatitis C (CHC). AIMS: We evaluated the capacity of aspartate aminotransferase (AST)/alanine aminotransferase (ALT) ratio, gama-glutamyltransferase (GGT) levels, platelet count, the AST to platelet ratio index (APRI) and serum hyaluronic acid (HA) to predict the intensity of hepatic fibrosis in patients with CHC and the variation of these markers after therapy with interferon. PATIENTS AND METHODS: In 72 patients with hepatitis C, AST/ALT ratio, GGT levels, platelet count, the APRI index (calculated as the ratio of AST to platelets) and serum HA concentration were determined and compared to histological staging according to the scoring system of METAVIR. Sixty-five patients received interferon and ribavirin therapy. The individuals that completed the treatment (n = 33) underwent a new test for serum marker of fibrosis in order to compare it with pre-treatment levels. RESULTS: GGT levels, platelet count, the APRI index and serum HA were correlated with the stage of hepatic fibrosis (p < 0.01), except AST/ALT ratio. The analysis of the areas under the ROC curve (AUC) evidenced that APRI and HA levels were the markers with the best association with hepatic staging: AUC (APRI) = 0.85 and AUC (HA) = 0.86. After therapy with interferon, only GGT and the APRI showed reduction of their levels (p < 0.05). CONCLUSION: HA and the APRI index were the most accurate approaches to liver fibrosis staging and they may be used as alternative diagnostic methods in patients with CHC.

Clinical Application of Non-invasive Diagnosis for Hepatic Fibrosis / 대한간학회지

Progressive hepatic fibrosis with development of cirrosis is a feature of chronic liver disease. Assessing fibrosis is important for predicting disease progression and patient management. Liver biopsy is the current gold standard for the diagnosis of liver fibrosis. However, liver biopsy is an invasive procedure. Alternative non-invasive methods have been developed. Serum markers are useful in predicting liver cirrhosis, but accuracy of serum markers is not satisfactory in the assessment of fibrosis. Newly developed transient elastography (Fibroscan) is a non-invasive method of measuring liver stiffness. Fibroscan has been reported to be superior in early detection of cirrhosis to serum markers. Factors influencing it's performance are not fully investigated. The evaluation of new tests should be continued to perform.

Efectos del ácido hialurónico de alto peso molecular en lesiones del cartílago articular: experiencia en gonartrosis / Effects of hyaluronic acid on the articute cartilage: experience in gonarthrosis

Se realizó un estudio prospectivo en el Hospital Domingo Luciani, para evaluar la eficacia del ácido hialurónico (AH) sobre las lesiones del cartilago articular utilizando para ello 25 pacientes (30 rodillas), de ambos sexos y edades entre 40 y 60 años, con diagnóstico de gonartrosis incipiente (grados I y II) según la clasificación radiológica de Ahlbãck, en los cuales se evaluaron los siguientes parámetros: dolor, limitación de la flexión, limitación de la extensión y lesión del cartilago articular utilizando la artroscopia antes y despúes del tratamiento. El estudio se dividió en tres etapas: diagnóstico, tratamiento (15 rodillas con AH y 15 rodillas sin AH o control) y evaluación y seguimiento (4 meses posterior a tratamiento). Los resultados observados al comparar ambos grupos reportaron una evidente mejoría en los parámetros de evaluación de los pacientes tratados con AH, lo cual fue demostrado estadísticamente (análisis de varianza p<0,001), excepto en el parámetro de limitación de la extensión, donde a pesar de evidenciar mejoría clínica, estadísticamente no pudo ser demostrado. Del estudio se puede concluir que el protocolo de trabajo demostró ser ineficiente para el manejo de gonartrosis incipiente, y que el Ácido Hialurónico no sólo produce mejoría clínica del cuadro, sino que también promueve la cicatrización de las lesiones del cartílago articular