Anti-Inflammatory Effects of Boric Acid in Treating Knee Osteoarthritis: Biochemical and Histopathological Evaluation in Rat Model.

This study aimed to examine the anti-inflammatory properties of boric acid (BA) in treating knee osteoarthritis (KOA) in rats, evaluating its biochemical and histopathological therapeutic effects. A KOA rat model was induced by injecting monosodium iodoacetate into the knee joint. Random assignment was performed for the experimental groups as follows: group-1(control), group-2(KOA control), group-3 (BA:4 mg/kg, orally), group-4(BA:10 mg/kg, orally), group-5(BA:4 mg/kg, intra-articularly), and group-6(BA:10 mg/kg, intra-articularly). The rats received 100 µL of BA intra-articularly on days 1, 7, 14, and 21 or 1 mL orally once a day (5 days/week) for 4 weeks. Serum levels of interleukin-1ß (IL-1ß), tumor necrosis factor-α (TNF-α), and activity of matrix metalloproteinase-13 (MMP-13) were measured. Histopathological and immunohistochemical analyses were performed on knee joint samples using specific antibodies for IL-1ß, TNF-α, MMP-13, and nitric oxide synthase-2 (NOS-2). Group-2 exhibited higher serum IL-1ß and TNF-α levels and MMP-13 activity than group-1 (P < 0.05). However, IL-1ß and TNF-α levels and MMP-13 activity were lower in all treatment groups than in group-2, with statistically significant reductions observed in groups-4, 5, and 6. Histopathologically, group-2 displayed joint space narrowing, cartilage degeneration, and deep fissures. Groups-5 and 6 demonstrated significant joint space enlargement, articular cartilage tissue regeneration, and immunostaining patterns similar to those in group-1. Immunohistochemically, group-2 showed significant increases in IL-1ß, TNF-α, MMP-13, and NOS-2 expression. However, all treatment groups exhibited reductions in these expression levels compared to group-2, with statistically significant decreases observed in groups-5 and 6 (P < 0.01). BA shows potential efficacy in reducing inflammation in experimental KOA model in rats. It may be a promising therapeutic agent for KOA, warranting further clinical studies for validation.

The Evaluation of the Effects of Nanoemulsion Formulations Containing Boron and/or Zinc on the Wound Healing in Diabetic Rats.

Wound healing remains a challenging clinical problem, especially in the presence of diabetes. Diabetic patients have the impaired ability to fight infection and insufficient inflammatory response. The aim of this study was to evaluate the effects of boronophenylalanine (BFA) and/or Zn-containing nanoemulsion (NE) formulations on wound healing in diabetic rats. MTT and scratch assays were performed to evaluate the proliferative effects of BFA and/or Zn on human dermal fibroblast (HDF) cells and the migration of these cells, respectively. The BFA and/or Zn-NE were prepared, and the effects of NEs on wound healing in diabetic rats were evaluated by applying once a day for 14 days. MTT assay showed that 10 to 25 µM BFA and/or 50 µM Zn had very significant positive effects on cell proliferation. In the scratch assay, 10 µM BFA significantly increased the migration of HDF cell compared with control. The droplet sizes of all the NEs were <115 nm and their zeta potential values were in range of (-) 23.9 ± 2.356 to (-) 33.1 ± 1.438 mV. There was a significant reduction in the wound contraction values (%) of the groups treated with the BFA and/or Zn-NE on the 14th day compared with the untreated diabetic rats group. According to histopathological findings, wound healing was nearly complete in BFA and/or Zn-NE compared with untreated diabetic rats. Especially, the group treated with the NE containing the low concentration of BFA showed highly promising results in wound healing of diabetic rats within 14 days with complete epithelialization and the completely closed wound area.

Evaluation of cytokines in protective effect of docosahexaenoic acid in experimental achilles tendinopathy rat model induced with type-1 collagenase.

BACKGROUND: We aimed to investigate the effectiveness of docosahexaenoic acid (DHA) as a treatment for Achilles tendinopathy (AT) induced with type-I collagenase in rats and compare it with collagen. METHODS: The AT model was induced with type I collagenase, and animals were randomly assigned to groups. Group 1:AT, Group 2: Collagen (7.2 mg/kg/day), Group 3:DHA (300 mg/kg/day), and Group 4:DHA (100 mg/kg/day). Right tendons of Group1 were used as a healthy control (HC). Oral treatments were applied for eight weeks. Serum tumor necrosis factor-alpha(TNF-α), matrix metalloproteinase-13 (MMP-13), and interleukin-1 beta(IL-1ß) concentrations were determined by ELISA. Tendon samples were taken for histopathological evaluation and examined immunohistochemically with antibodies specific for Col1A1, TNF-α, MMP-13, IL-1ß, and nitric oxide synthase-2(NOS-2). The ultimate tensile force (UTF) yield force(YF) and stiffness were measured by biomechanical assessments. RESULTS: UTF,YF and stiffness values were increased in all treatment groups compared to the AT control, a significant increase was found in Group 2 (p < 0.05). There was severe degeneration of tendon cells in the AT control. The tendon cells in samples from Groups 2-3 were less degraded, and this was statistically significant (p < 0.05). TNF-α, MMP-13, IL-1ß, and NOS-2 expressions were significantly higher in the AT control compared to the HC. In all treatment groups, their concentrations were lower than in the AT control. Serum TNF-α, MMP-13, and IL-1ß levels were lower in all treatment groups (Especially in Group3 (p < 0.001)) compared to Group1. CONCLUSION: The efficacy of high-dose DHA as a treatment for AT was investigated from biochemical, histopathological, and biomechanical perspectives. The results showed that DHA could be an alternative treatment compound to collagen.

Effects of Bisphenol A Released From Composite Fillings on Reproductive Hormone Levels in Men.

OBJECTIVES: Composite resins are the most preferred filling material because of their excellent aesthetic qualities. However, a filling material should also be biocompatible as well as aesthetic. The aim of this study was to determine the serum and saliva bisphenol-A (BPA) levels and to examine the effects of serum BPA on reproductive hormone levels after healthy men were treated with composite fillings. METHODS: Eighteen healthy males each received 2 composite restorations. Saliva and blood samples of subjects were collected before resin application and 1 day and 1, 3, and 5 weeks after the resin was applied. BPA amounts in samples were detected using high-performance liquid chromatography (HPLC). Serum gonadotropins, testosterone, sex hormone binding globulin, free androgen index, and oestrogen levels were measured with radioimmunological assay kits. Statistical analysis of data was made using Friedman, Wilcoxon signed ranks and Mann-Whitney U tests (α = 0.05). RESULTS: The amount of BPA released from composite resins over time was not significantly elevated in either saliva or serum (P > 0.5). In addition, serum BPA levels were significantly higher than saliva BPA levels for both composites (P < .05), but saliva and serum BPA levels were not statistically different when comparing the 2 composites (P > .05). CONCLUSIONS: BPA from composite resins used in this study did not significantly alter serum hormone levels.

Investigation of the efficacy of daidzein in experimental knee osteoarthritis-induced with monosodium iodoacetate in rats.

BACKGROUND: Osteoarthritis (OA) is a degenerative chronic illness that most frequently occurs in the knee joint. Daidzein (DZ) an isoflavone has anti-inflammatory and antioxidant activity. The aim of this study was to evaluate the effectiveness of DZ as a treatment for experimental knee OA (KOA) in rats. METHOD: An experimental KOA model was induced by monosodium iodoacetate (MIA) in rats. Thereafter, 49 Wistar albino male rats (250-300 g, 12-16 weeks old) were randomly divided into 7 groups: C (healthy control); DC (KOA + saline); hyaluronic acid (HA); HA+ intraarticular (ia) DZ; oral (po) DZ; ia DZ; HA + po DZ groups. DZ and/or HA were administered intraarticularly to the rats as 50 µL on days 1, 7, 14, and 21. Alternatively, the DZ was administered orally as 0.5 mL twice daily for 21 days. After the treatment, rats were sacrificed by decapitation under general anesthesia. Serum samples were analyzed to determine the total oxidant status (TOS) and total antioxidant status (TAS) and the levels of TNF-α, IL-1ß, MMP-13, and DZ. Knee joint samples underwent histopathological examination, and TNF-α, IL-1ß, NOS2, and MMP-13 were analyzed with immunohistochemical methods. RESULTS: HA, DZ, and DZ + HA effectively reduced the levels of TNF-α, IL-1ß, and MMP-13 in the serum of the DC group (p < 0.001). In groups that received HA, DZ, or DZ + HA, the serum TAS increased compared with the DC group (p < 0.05). When the DZ + HA combination was used, a more pronounced reduction in the levels of TNFα, NOS2, IL-1ß, and MMP-13 was observed in knee joints. In addition, the cracks on the cartilage surface and fibrillation were completely improved in the groups that received HA, DZ, or DZ + HA compared with the DC group. CONCLUSION: DZ had anti-inflammatory and antioxidant effects in a rat OA model. Therefore, DZ, as monotherapy or especially in combination with HA, may be a promising and beneficial therapy for OA. Key Points •DZ has been shown to reduce TNF-α, IL-1ß, and MMP-13 both in serum and in tissue samples taken from the knee-joints. •The cracks on the cartilage surface and fibrillation in KOA were completely improved by using DZ and DZ + HA combination. •DZ may be useful to eliminate/reduce/ameliorate inflammation and oxidative damage in the pathogenesis of KOA. •DZ, alone or in combination with HA, may be a promising natural compound with beneficial effects in the treatment of KOA.

Serum metabolite profiling of ST-segment elevation myocardial infarction using liquid chromatography quadrupole time-of-flight mass spectrometry.

ST segment elevation myocardial infarction (STEMI) is one of the most common global causes of cardiovascular disease-related death. Several metabolites may change during STEMI. Hence, analysis of metabolites in body fluid may be considered as a rapid and accurate test for initial diagnosis. This study has therefore attempted to determine the variation in metabolites identified in the serum of STEMI patients (n = 20) and 15 controls. Samples collected from the Cardiology Department, Medical Faculty, Ataturk University, were extracted by liquid-liquid extraction and analysed using liquid chromatography quadrupole time-of-flight mass spectrometry. The METLIN database was used for the identification and characterization of metabolites. According to Q-TOF/MS measurements, 231 m/z values, which were significantly different between groups (P < 0.01 and fold analysis >1.5) were detected. Metabolite identification was achieved via the Human Metabolome database. According to the multivariate data analysis, leucine, isoleucine, l-proline, l-alanine, glycine, fumaric acid, citrate, succinate and carnitine levels were decreased, whereas levels of propionic acid, maleic acid, butyric acid, urea, oleic acid, palmitic acid, lysoPC [18:2(9Z)], glycerol, phoshpatidylethanolamine, caffeine and l-lactic acid were increased in STEMI patients compared with controls. In conclusion, malonic acid, maleic acid, fumaric acid and palmitic acid can be used as biomarkers for early risk stratification of patients with STEMI.

The Effect of Parietin Isolated From Rheum ribes L on In Vitro Wound Model Using Human Dermal Fibroblast Cells.

Parietin is one of the well-known anthraquinone compounds that can be extracted from Rheum ribes L. In this study, we aimed to investigate the effects of parietin isolated from Rheum ribes L on an in vitro wound model using human dermal fibroblast cells and compare its effectiveness against zinc. The antioxidant effect of parietin was determined by using the 1,1-diphenyl-2-picrylhydrazine (DPPH) method. Human dermal fibroblast cells were cultured in proculture medium and were kept until 100% confluence was achieved. The wound model was created by using a pipette tip. After that, different concentrations of parietin and zinc (final concentrations in the well to be 5-250 µM and 25-200 µM, respectively) were added into the medium. The proliferation-inducing effect on cell viability was determined by using MTT assay. Images of cells were taken at 0, 12, and 24 hours. According to the DPPH method, parietin exhibited have antioxidant activity. According to the MTT results, parietin exhibited significant proliferation-inducing effect on cell viability in a dose range of 5 to 10 M, and zinc showed significant proliferation-inducing effect on cell viability at dose 50 µM ( P < .05). In addition, the image of cell proliferation was also shown at the same doses at 24 hours. In this study, we claim that parietin induces cell proliferation at low doses in cases of dermal fibroblast loss. In conclusion, parietin as an alternative to zinc in wound healing could be used by clinicians in the future with more extensive studies.

Design and Optimization of a New Voltammetric Method for Determination of Isoniazid by Using PEDOT Modified Gold Electrode in Pharmaceuticls.

Isoniazid (INH) was studied with regard to its electrochemical treatment on a strongly alkaline solution of a poly (3,4-ethylenedioxythiophene)-modified gold electrode, using both cyclic voltammetric and controlled potential techniques. Electrocatalytic performance measurements of this composite electrode toward oxidation of INH exhibited an increase of 4-folds in oxidation peak densities compared to the bare gold electrode. Central composite design method was used to obtain optimum experimental conditions, and used critical parameters (pH (A), scan rate (mV/s, B) and temperature (C, C) to assess the effects on amperometric response. Optimum experimental conditions were achieved by using a pH of 9.2 with a scan rate of 260 mV/s and a temperature of 30 C. Under these circumstances, a good linear relationship was observed between peak current densities and INH concentration in the range of 0.05-2 µM, with correlation coefficient of 0.9998. Furthermore, the method was very sensitive (limit of quantitation, 0.0043 µM), accurate (relative error, -5.65 to 4.03) and precise (relative standard deviation %, ≤ 7.97). The method was also applied to determine INH in pharmaceutical formulations, and statistically compared the results with the official method using the two one-sided equivalence test; the results were in good agreement with those obtained by the official and reported methods.

A new spectrofluorimetric method for determination of losartan potassium in rabbit plasma and its application to pharmacokinetic study.

A new spectrofluorimetric method to determine losartan potassium (LP) in rabbit plasma is described. The method was based on measuring the native fluorescence of LP in acidic medium. Optimum excitation and emission wavelengths were found to be 248 nm and 410 nm, respectively, in methanol that was diluted with a sulfurous acid solution LP was extracted from rabbit plasma by methyl-tertiary-butyl-ether in acidic media and then back extracted with NaOH. The calibration curves were linear between 0.025 and 0.5 µg/mL with a lower limit of detection 0.004 µg/mL. Precision and accuracy values of the method were calculated as lower than 4.97% and ± 5.68, respectively and the recovery of LP from rabbit plasma was higher than 91.1%. In addition, stability studies of LP in rabbit plasma were carried out and demonstrated its good stability at - 20 °C and at room temperature. The developed and validated method was successfully applied for estimating the pharmacokinetic parameters of LP following oral administrations of a single 10 mg LP/kg to rabbits and it could be concluded that the method can be applied to clinical trials.

GC-FID and HPLC-DAD Methods for the Determination of Menadione Sodium Bisulphite Directly and by Converting Menadione Sodium Bisulphite to Menadione in Pharmaceutical Preparation.

was performed in both direct analysis of MSB and analysis of MN by converting MSB to MN with sodium carbonate. GC-FID method was carried out on the HP-5 capillary column GC-FID and HPLC-DAD methods were developed for determination of menadione (MN) and menadione sodium bisulphite (MSB). By means of each method, quantitative analysis of MSB in commercial pharmaceutical using nitrogen gas. HPLC-DAD method was achieved on the reversed phase C8 column by using a mobile phase consisting methanol and water. The calibration curves of GC-FID and HPLC-DAD for both analytes were linear in the same concentration range (0.5-20 µg/mL). Both methods were validated in terms of precision, accuracy, recovery and limits of detection (LOD) and quantitation (LOQ). Although LOD values of HPLC-DAD method (0.010 µg/mL for MN and 0.005 µg/mL for MSB) is lower than obtained values with GC-FID method (0.04 µg/mL for MN and 0.06 µg/mL for MSB), both methods gave similar and favorable results in terms of precision and accuracy. The Student's t-test was applied to investigate the significant of the different between the results of MSB determination with direct analysis of MSB and analysis of MN by converting MSB to MN by means of GC-FID and HPLC-DAD method in dosage form.

Optimization of a spectrofluorimetric method based on a central composite design for the determination of potassium losartan in pharmaceutical products

Here, a spectrofluorimetric method for the determination of potassium losartan (PL) in pharmaceutical products is described. The effects of critical parameters, pH, acid molarity, and temperature, on the fluorescence intensity of PL were analyzed, and these parameters were optimized using a central composite design (CCD). The highest fluorescent intensity at excitation (λex) and emission (λem) wavelengths of 248 nm and 410 nm, respectively, was achieved using 0.01 M sulfurous acid (pH 2) at 21.6 °C. Under optimum conditions, the method was linear from 0.025-0.5 µg/mL, with a reasonably high correlation coefficient (0.9993). Furthermore, the method was very sensitive (LOQ, 0.006), accurate (RE, ≤7.06), and precise (%RSD, ≤6.51). After development and validation of the method, samples containing PL were analyzed with this method, and the obtained data were statistically compared with those obtained with a previously published reference method using a two one-sided equivalence test (TOST). According to the data, the results from the proposed and reference assays were equivalent.

Descreve-se método espectrofluorométrico para a determinação de losartana potássica (PL) em produtos farmacêuticos. Os efeitos de parâmetros críticos (pH, molaridade ácida e temperatura) na intensidade da fluorecência foram otimizados usando o planejamento de componente central (DCC). A mais alta intensidade fluorescente com λex=248 nm e λem= 410 nm foi obtida usando ácido sulfúrico 0.01 M (pH 2) e 21.6 ºC. Nas condições ideais, a linearidade do método foi estabelecida na faixa de concentração de 0.025-0.5 µg/mL com coeficiente de correlação bastante elevado (0.9993). Além disso, o método foi muito sensível com valor de LOQ 0.006, exato (RE≤7.06) e preciso (RSD%≤6.51). Depois do desenvolvimento e validação do método, amostras de medicamentos contendo PL foram analisadas com este método e os resultados obtidos foram comparados estatisticamente com método de referência, publicado anteriormente, usando o Teste de equivalência TOST (Teste de Equivalência Unilateral). De acordo com os dados estatísticos, os resultados do ensaio de referência e do método proposto foram equivalentes.

Spectrofluorimetric Determination of α-Tocopherol in Capsules and Human Plasma.

A simple, sensitive and rapid spectrofluorimetric method for determination of α-tocopherol in pharmaceutical capsule and human plasma was developed and validated. The native fluorescence of α-tocopherol was measured at 334 nm with excitation at 291 nm, after extraction of α-tocopherol from human plasma hexane:dichloromethane mixture. The calibration curves were linear (R≥0.9993) in the concentration range of 0.25-2.5 µg/ml of α-tocopherol in both standard solutions and plasma samples. The developed method was directly and easily applied for determination of α-tocopherol in the plasma of healthy volunteers and different type of bladder cancer and stomach cancer patients and also pharmaceutical capsule.