Betula pendula leaf extract and to investigate basic parameters influencing the drug release pattern. To fully assess the statistical significance of the influence of the individual factors in the tablet formulation development as well as the combination of them, Tukey’s tests and a complete 33 factor analysis of variance (ANOVA) were applied. The following three factors were studied at three levels : influence of the hydrophobic/hydrophilic polymer ratio Ethylcellulose (EC)/Hydroxypropyl methylcellulose (HPMC) , influence of HPMC molecular weight , and influence of the compression force applied . The effect of these varied parameters on the drug release parameter t80 was evaluated statistically. Twenty-seven tablet models were formulated, including all possible combinations of the variables. The obtained drug release profiles demonstrated that a 25/75 (EC/HPMC) ratio was the most suitable for prolonging the release process. Increasing the molecular weight of HMPC from 500 kDa to 750–1150 kDa and applying higher compression force significantly influenced the studied t80 values and caused sustained drug release (t80 up to 7.97 h). The combination of the hydrophilic HPMC polymer with the hydrophobic EC can result in the formation of a promising drug-carrying matrix, offering effective control of the drug release process.The aim of the study was to develop polymer matrix tablets with modified release of dry  Modern phytotherapy is a successful combination of traditional knowledge about the therapeutic use of medicinal plants and novel pharmacological, pharmacognostic and technological approaches based on an in-depth knowledge of the herbal chemical composition, mechanisms of action of the biologically active substances, and formulating the most appropriate pharmaceutical form . The maiBetula pendula Roth, Betulaceae) are known and used in traditional medicine and phytotherapy in various forms [Betulaceae is a large family containing more than 150 species of trees and shrubs, primarily distributed in the Northern Hemisphere ,5. Produus forms ,7. In ouus forms ,9. DespiModified-release pharmaceutical forms are widely used due to the numerous advantages they offer. The use of different types of polymers to control the drug release process represents one of the main aspects of the development of different types of matrix systems. Hydrophilic matrices are often the first choice in the development of modified-release pharmaceutical forms ,11,12. TAn increasing number of reports on the use of experiments in the development and optimization of various drug delivery systems can be found in the literature. Different statistical design approaches are often used as a powerful tool to optimize process parameters and pharmaceutical form characteristics ,17,18,19Betula pendula leaf extract and offering improved biopharmaceutical characteristics. The work is focused on the precise optimization of various factors such as compression parameters, type, concentration and molecular weight of the polymers used, which affect the characteristics of the formulation and the drug release process. The influence of the varied parameters, both individually and in combinations, was thoroughly evaluated using statistical DOE (design of experiments) approaches.Therefore, the aim of the research was to design polymer tablet matrix systems as a platform providing delayed release of the included dry w/w) ethoxyl basis), magnesium stearate and talc  were purchased from Sigma Aldrich . Hydroxypropyl methylcellulose of different molecular weights  was purchased from The Dow Chemical Company . Other chemicals and reagents used were of analytical grade. Birch leaves  were collected from the Rhodopes Floristic Region of Bulgaria in June–August 2019. The leaves were identified by their macroscopic and microscopic characteristics according to the requirements of the European Pharmacopoeia 9. They were dried in the shade without using high temperatures to preserve biologically active substances. Dried leaves were ground to 5 mm in size.Hyperoside , ethylcellulose . Ethanol extracts and dispersed colloidal silica were homogenized prior to and throughout the spray drying process by stirring with an electromagnetic stirrer at 300 rpm. Solutions were fed to a 0.7 mm spray nozzle. Nitrogen pressure was kept constant during the process—6 bar at a gas flow rate of 600 L/h, a peristaltic pump speed of 15%, an inlet temperature of 140 °C and aspiration of 95%.Dry 3 full factorial designs. Tablets were prepared by the wet granulation method with an average weight of 0.700 g. Different concentrations and ratios of the hydrophobic polymer ethylcellulose (EC) and the hydrophilic polymer hydroxypropyl methylcellulose (HPMC)  were used, varying the viscosity of the HPMC used  and the applied compression force  (The development of the model tablet formulations containing dry extract was performed using 3and 2 t) .3 factor analysis of variance (ANOVA) was applied. Three factors (independent variables) were determined and studied in experimental design at three levels: low (−1), medium (0) and high (+1), as follows: influence of the hydrophobic/hydrophilic polymer ratio (EC/HPMC), influence of HPMC molecular weight, and influence of the compression force applied, and their effect was evaluated on the dependent variable t80, indicating the time required to release 80% of the extract included in the tablet. To fully assess the statistical significance of the influence of the individual factors as well as the combination of them, a complete 3The required amounts of ethylcellulose, hydroxypropyl methylcellulose and dry extract were mixed, ground and sieved through a 0.5 mm sieve. The wet mixing was carried out with the gradual addition of 96% ethanol and continuous homogenization. Granulation was performed on a granulator ERWEKA AR 403 FGS  through a 2 mm sieve. The wet granular mass was dried at 45 °C to a moisture content of 3.5%. The classification of the granules was performed through a 1.6 mm sieve. The granular mass was dusted with the required amounts of talc and magnesium stearate for 5 min. The tablet compression was performed on a single-punch eccentric tablet press, the ERWEKA EP-1 with 13 mm punches at three different compression forces.The rheological characteristics of the 9 models of granular mixtures obtained were determined according to the pharmacopoeial methods for evaluation: angle of repose, flow rate through an orifice, compressibility index (Carr’s index) and Hausner ratio. The control parameters of the prepared 27 tablet models were determined according to the tests described in the European Pharmacopoeia 9, respectively: uniformity of mass, disintegration time, resistance to crushing and friability. Analyses were performed with laboratory equipment: tapped density tester SVM Erweka, disintegration tester Erweka—ZT 121, tablet hardness tester Erweka—TBH 125 and friabilator Erweka—TA 120.2O, pH 3.7): B (CH3CN) was used in a gradient mode from 90(A):10(B) to 10(A):90(B) with a flow rate of 0.9 mL/min. Detection was performed at 335 nm. The quantitative determination was performed by the method of the external standard. Results were presented as mean ± SD values, provided from 3 independent measurements. The drug release profile was monitored for a period of 8 h. The dissolution profiles of all tested models were fitted to zero-order, first-order, Higuchi and Korsmeyer–Peppas kinetic models.Dissolution tester Sotax CH 4123, Apparatus 2 (paddle), was used at the following operating parameters: test medium: 900 mL (0.1 N HCl buffer solution of pH 1.2 as Simulated Gastric Fluid (SGF), temperature 37 ± 0.5 °C, speed 150 rpm). The release of the incorporated plant extract from the polymer systems was evaluated by quantitative analysis of the flavanoid hyperoside using high-performance liquid chromatography (HPLC). HPLC system Varian Prostar with a column Hitachi C18 AQ  and PDA detector was used. A mobile phase A . The varied parameters of the 27 model tablet compositions—uniformity of mass, disintegration time, friability and hardness—are presented in The model tablets showed good uniformity of mass and relatively high hardness, varying in the range from 88 N to 165 N (which correlated with the applied compression force). All tested tablets showed very low friability, ranging from 0.01% to 0.126%.Our preliminary investigations have shown that a satisfactory delay in the release process from tablet models containing only hydrophilic polymers (HPMC) could not be achieved. The combination of hydrophobic ethylcellulose with the hydrophilic hydroxypropyl methylcellulose, as well as the change in the hydrophobic-hydrophilic polymer ratio, offers ample opportunities to control the release process. There is also evidence in the literature that combining a hydrophilic matrix of HPMC with a small amount of ethylcellulose results in a delay in the initial intensive release of well-soluble drugs and more effective control of the release process .Usually, the presence of a highly soluble substance in a matrix of HPMC generates an additional osmotic gradient, resulting in a faster rate of polymer swelling and a large increase in gel thickness. In the presence of water, the mobility of the polymer chains is enhanced, resulting in the gradual transformation of a glassy matrix into a rubbery, swollen gel. At higher polymer concentrations, the viscosity of the gel matrix is increased, which results in a decrease in the effective diffusion coefficient of the drug . In a stOther factors that could significantly affect the release process are the viscosity of the polymer used as well as the compression force applied in the preparation of the tablets. 2 values obtained from different mathematical models were evaluated and presented in To determine the release kinetic mechanism, in vitro drug release data were fitted to different kinetic models: zero-order, first-order, Higuchi and Korsmeyer–Peppas. The R2 (from 0.992 to 0.999). The Korsmeyer–Peppas mathematical model is often used to analyze the release of drugs from polymer systems and reflects the linear relationship between the logarithmic amount of substance released over time t (Log Mt/M0 and Logt).Korsmeyer–Peppas was determined to be the most suitable kinetic model, describing the process of in vitro drug release from all tested tablet models and showing the highest values of RIn most of the studied models , the established values for the diffusion exponent n were in the range of 0.45 to 0.95. These results confirmed the assumption that the mechanisms of diffusion and erosion were involved simultaneously and influenced the process of releasing the substance included in the matrix system. Values of n less than or equal to 0.45 are an indicator for a Fickian diffusion transport mechanism (Case I transport). Values of n between 0.45 and 0.95 usually indicate non-Fickian diffusion , which includes a combination of diffusion and erosion in the release mechanism. Values above 0.95 indicate a dominant erosion release mechanism (Case II transport and Super Case II transport). The relative complexity of the matrix system containing a combination of hydrophobic and hydrophilic polymers is a prerequisite for the release mechanism to be influenced by more than one process ,28,29.In a study investigating the influence of HPMC with different molecular weights in composite matrix tablets, authors found values for the diffusion exponent n ranging from 0.48 to 0.50, indicating that the release was governed by anomalous transport, i.e., the combination of the drug diffusion through the gel layer and the erosion of the polymer gel layer .To assess the effect of the EC concentration on the release process, model formulations were developed by varying the ratio between the hydrophobic and hydrophilic polymers . When comparing analogous models containing HPMC with the same molecular weight and obtained with the same compression force, the following trend is clearly observed: increasing the amount of the hydrophobic polymer from 10% (level −1) to 25% (level 0) leads to significant elongation of t80 . Model FThe release profiles of the experimental models showed that increasing the EC concentration led to a reduced initial release and prolongation of t80. This is probably due to the fact that the presence of the hydrophobic polymer in the matrix slows down the intensive water ingress in the initial moments, even before the formation of a viscous hydrogel, hence the dissolution of the included plant extract and effective control of the release process. It is noteworthy that more significant differences are observed in models containing low-molecular-weight HPMC. This was expected, as the high molecular weights of K15M and K100M create a more viscous hydrogel layer, which slows down the release process more efficiently. However, as the concentration of EC increased, the observed trend changed. Comparison of the models F10  and F1  showed a decrease in t80 from 5.26 h to 3.12 h. This pattern was preserved in other analogous models containing medium- and high-molecular-weight HPMC and 25% and 40% ethylcellulose, respectively. In each case, models containing 40% EC showed faster release of the included extract and correspondingly lower values for t80. This can be explained by the fact that the large hydrophobic ethylcellulose molecules create local disturbances in the integrity of the viscous hydrogel layer, leading to lower retention of the soluble substances and faster drug release.A similar pattern has been reported by other authors. Madgulkar et al. use response surface methodology for the formulation and optimization of sustained-release tablets of venlafaxine resinates. HPMC and EC were taken as independent variables in the central composite design for two factors at three levels. In vitro dissolution studies were performed to study the release kinetic parameters. In this case also, like in other matrix systems of similar type, the diffusion exponent n values are indicative of a coupling of diffusion and erosion mechanisms. Increased n values are observed with an increase in HPMC content, even at higher EC levels. The drug released was measured at the 2nd and 8th hours. A general tendency is clearly noticed when comparing the presented models: as the amount of HPMC increases, the release process slows down. However, the amount of EC does not affect the release process in the same way. Comparing the models F1–F3 containing the lowest concentration of HPMC (41.62 mg) and increasing concentrations of EC  shows that the double increase in the amount of EC (model F1 to model F2) leads to a delay in the release process and a correspondingly smaller amount of released drug at the 2nd and 8th hours. However, when the concentration is further increased (model F3), the release process does not slow down further, and even a greater amount of released substance is observed compared to model F2. In the models with increased HMPC concentration, the tendency is maintained—low and medium EC values lead to slower release, compared to the highest concentrations, which generally show faster release of the included drug substance .The results of Tukey’s test on the effect of the hydrophobic/hydrophilic polymer ratio (EC/HPMC) on the release time (t80) are presented in It can be concluded that the addition of EC to the hydrogel matrix improves the control of the release process, but only up to a certain polymer concentration. Furthermore, increasing concentration has the opposite effect and rather accelerates the release of the drug substances.The molecular weight of the hydrophilic polymer used is essential for the rate and extent of drug release. When comparing analogous models obtained with the same hydrophobic/hydrophilic polymer ratio of 40/60 and the same compression force of 1 t , the following trend is clearly outlined: increasing the molecular weight of the hydrophilic polymer used  leads to a delay in the release of the included plant extract. This difference is more significant when comparing the models prepared with the lowest viscosity polymer, K4M, and those with medium viscosity, K15M (models F1 and F4). When comparing the F4 and F7 models (including the medium molecular weight K15M and the high molecular weight K100M), the trend was preserved, but although statistically significant, it is much less pronounced.In a study from 2014, Jain and co-authors investigated the effect of HPMC molecular weight and concentration on the in vivo erosion behavior of HPMC matrix tablets. Different formulations were investigated, containing varying proportions of two HPMC grades with different molecular weights. The in vivo erosion behavior and gastrointestinal transit were investigated using magnetic marker monitoring. The authors concluded that the erosion is strongly dependent on the composition of the formulations. Models containing a larger amount of high molecular weight HPMC or a higher content of HPMC exhibit a relatively slower erosion rate, and vice versa. Therefore, the appropriate erosion characteristics of HPMC matrix tablets can be modulated by manipulating the amount of polymer and content of HPMC grades with different molecular weights .Gao et al. conducted a mechanistic study of the influence of formulation variables on matrix performance and drug release from HPMC matrix tablets. The effects of HPMC/lactose ratio and HPMC viscosity grade (molecular weight) on solute release and swelling of matrix tablets were investigated. Drug, lactose and HPMC releases were monitored simultaneously. The authors suggested that the strong dependence of HPMC release on viscosity grade can be explained on the basis of the concept of polymer disentanglement concentration. They consider that the HPMC/lactose ratio modulates drug release rate by altering drug diffusivity, while the HPMC viscosity grade impacts matrix dissolution and gel layer thickness development below a critical molecular weight. High-viscosity grade HPMC (>4000 cps), forming slowly dissolving matrices, demonstrates similar drug release rates, mainly due to the same drug diffusivity as a result of the identical gel composition and thickness. The higher apparent drug diffusivity and release rate in fast dissolving matrices (< or = 100 cps) are probably explained by swelling inhomogeneity .The post-hoc test showed that the molecular weight of 750 kDa led to 2.0637 (on average) higher values for t80 compared to 500 kDa, while the highest molecular weight of 1150 kDa only led to 0.3744 higher values for t80 compared to 750 kDa .The obtained results are in accordance with some data published by other authors  that theThe general trend observed among all analogous models in varying the compression force is that as the pressure increases, the release rate slows down, but statistically, the change in this independent variable has the lowest effect on the values of t80. This pattern was more pronounced when comparing the models obtained at 1 t and those at 1.5 t pressure. The differences in the models obtained at 1.5 t and 2 t were insignificant.As the compression force increases, the density of the obtained matrix system increases, as do the inner interparticle interactions. This leads to a stronger matrix structure and slower water entry in the initial moment, which can probably explain the intensive initial release in the models obtained at the lowest pressure of 1 t.In a study by Hirun and Kraisit, composite matrix tablets were developed, and the effect of HPMC k-series on porosity, compatibility and release behavior was studied. Scanning electron microscopy was used for the morphological analysis of the top view of the matrix tablets and the side view of the broken tablets. The tablet’s surface morphology was rough, with aggregated particles, small holes and cracks visible in the cross-sections. According to the type of HPMC used, a dense, continuous mass was observed, with some irregularities and rough-breaking pieces. The inner structures of all tablets were composed of solids and air, known as tablet porosity. It can be considered a feature that impacts liquid and drug transport through the solid dosage form. The authors emphasize that the compression force and tablet composition may affect the surface area of the pores and the number of distinct pore diameters in the tablets, thereby affecting the release process .As seen in To visualize the correlation between the three parameters, 3D surface plots are presented , explain2 indicates what part of the changes in the independent variable will lead to changes in the resultant (dependent) variable, i.e., R2 = 0.984 shows that with 98.4% certainty, the changes in the release time of 80% of the included dry plant extract can be explained by the factor variables of the model. The corrected coefficient of determination, R2Adj., is 0.977. Its value, which is close to 1, indicates that there is a significant linear correlation between the factor variables and the resulting variable. Moreover, the very small difference between the values of R2 and R2Adj. shows that the influence of additional conditions in the statistical model is insignificant.Betula pendula leaf extract can be successfully used as phytotherapeutic products, providing delayed release of the included plant extract. Such pharmaceutical formulations can offer improved biopharmaceutical characteristics, longer maintenance of drug therapeutic plasma concentrations without the need for frequent dosing, reduced risk of abrupt concentration fluctuations and limited drug side effects.The following conclusions can be summarized from the studies conducted on the drug release process and the influence of the studied variables: the concentration of EC significantly affects the release process, and the EC/HPMC ratio of 25/75 is most suitable for prolonging the release process from the formulated polymer matrices and eliminating the initial intensive release. The increase in the molecular weight of the HMPC used led to a delay in the release process, with more significant differences between K4M and K15M  and a less pronounced change in the highest molecular weight, K100M (1150 kDa). Although it had a less significant effect on the values of t80, the increase in the applied compression force led to a delay in the release process. F18 can be defined as the most promising model, obtained at a ratio of EC/HPMC 25/75, a molecular weight of HPMC 1150 kDa and a compression force of 2 t, showing the longest time to release 80% of the included active substance (7.97 h). The proposed polymer formulations with dry


Various weight fractions of the reinforcement materials, ranging from 0 to 0.5 wt.%, were incorporated into the epoxy. The aim is to enhance the characteristics and durability of the polymers for potential utilization in different mechanical applications. The addition of hybrid additives consisting of Al2O3 nanoparticles and graphite to the epoxy resin had a noticeable effect on the performance of the epoxy nanocomposites. The incorporation of these additives resulted in increased elasticity, strength, toughness, ductility, and hardness as the concentration of reinforcement increased. The enhancement in the stiffness, mechanical strength, toughness and ductility reached 33.9%, 25.97%, 25.3% and 16.7%, respectively. Furthermore, the frictional tests demonstrated a notable decrease in both the coefficient of friction and wear with the rise of the additives’ weight fraction. This improvement in the structural integrity of the epoxy nanocomposites led to enhanced mechanical properties and wear resistance. The SEM was utilized to assess the surfaces of tested samples and provide insights into the wear mechanism.The main target of this study is to evaluate the impact of hybrid reinforcement using Al Lately, numerous industries have been increasingly utilizing polymers as viable alternatives to metals and metal alloys due to their advantages that can be used in various applications. Polymers have many attractive characteristics, such as lightweightness, affordability, non-toxicity, corrosion resistance, and ease of manufacturing or production, which allow outstanding design flexibility . Among tRecently, polymer nanocomposites have gained significant attention from both researchers and industrial sectors as they present promising materials with outstanding behavior. This performance arises from the interconnection between the polymer matrix and incorporated nanoparticles ,7. ThrouTo improve polymer properties and mechanical, thermal, electrical, and tribological characteristics, a lot of researchers have focused on incorporating nano-additives (nano-fillers) into epoxy resins. In an investigation into the thermomechanical performance of epoxy/block-copolymer/core-shell rubber nanocomposites conducted by Bajpai et al. , the aut2O3) are ceramic fillers that are popular for their remarkable behavior such as superior resistance to wear, high stiffness, corrosion resistance, and scratch resistance. Additionally, they have outstanding mechanical performance, high toughness, and creep resistance [2O3 nanoparticles (high loading) was examined to evaluate its mechanical properties [2O3. They evaluated the tribological and mechanical performance of the epoxy for up to 0.4 wt.% weight fraction of Al2O3. The outcomes illustrated a considerable enhancement in the mechanical performance and the wear resistance.The inclusion of ceramic fillers is usually employed to preserve the mechanical performance of the composites such as elasticity modulus and strength . The utisistance . Epoxy coperties . The invoperties  tried toCarbon materials such as graphite, graphene nanoplatelets, and carbon nanotubes, are famous for their ability in the production of multifunctional engineering components. Such carbon materials possess the ability to minimize friction and improve wear behavior . In addiThe utilization of hybrid nanofillers in reinforcing polymer nanocomposites proved that the combination of different filler types is able to improve the physical, tribological and mechanical performance of the nanocomposite relative to the utilization of a single filler type . It has 2O3 and graphite with different loading fractions on the mechanical and tribological performance of the epoxy nanocomposite. The epoxy nanocomposite’s mechanical properties  were determined as a function in the nanoparticle loading fraction. Additionally, the coefficient of friction and the wear of the epoxy nanocomposites were assessed under the variation of the normal loads and sliding distances. In order to gain insights into the wear mechanism of the produced composites, the morphological properties of the nanocomposite surfaces were examined through scanning electron microscopy (SEM). Additionally, X-ray diffraction (XRD) analysis was employed to assess the phase composition of the hybrid epoxy nanocomposite.Based on the literature survey, the inclusion of hybrid nanoparticles in the epoxy can result in a significant impact on the produced polymer’s nanocomposite properties. Each type of nanoparticle can contribute to the enhancement of a specific property. Furthermore, the nanoparticle loading fraction has a crucial function in controlling the performance of the produced nanocomposite. The primary purpose of the current investigation is to study the influence of incorporating hybrid nanoparticles of Al2O3 nanoparticles were obtained from US Research Nanomaterials, Inc., Houston, TX, USA. The Al2O3 has the following specifications: purity of 99% or greater, 100% alpha phase, particle size 80 nm, and density of 3.97 g/cm3. The previous specifications were obtained from the company datasheet. On the other hand, the graphite (Gr) was obtained from the same company, with the following specifications: fixed carbon 99.9+%; moisture: <0.3%; ash: <0.5%; particle size 400 nm–1.2 µm diameter, and with a flaky morphology with a thickness of 80 nm and 8–6 m2/g surface area.For the current investigation, the epoxy was obtained from Graffiti Resin Company, Saudi Arabia. The package consisted of two components: a non-pigmented liquid epoxy which is the resin and a hardener which is used for curing the epoxy resin. These two compounds are typically mixed in specific ratios (1:1) to initiate the curing process and achieve the desired properties of the epoxy. The Al2O3 nanoparticles and graphite were subjoined into the epoxy. The epoxy/Al2O3/Gr mixture was mechanically stirred at room temperature for a duration time of 30 min, with a rotational speed of 150 rpm. The previous process aimed to distribute the Al2O3 nanoparticles and Gr uniformly inside the epoxy matrix. Secondly, the hardener was carefully blended, in a 1:1 ratio, with the epoxy/Al2O3/Gr mixture for 3 min. The previous process is crucial to achieve inclusive homogenization of the mixture components without creating air bubbles inside the composite and to ensure uniform nanoparticle distribution throughout the epoxy matrix. The weight fractions of both the Al2O3 nanoparticles and the graphite are provided in 2O3 nanoparticles, graphite, and the hardener, were poured into molds that were prepared beforehand. The molds were designed specifically for the production of samples used in compression and tribological tests. The mixtures were left for 24 h at room temperature to harden and be ready for tests. The curing time was identified based on the recommendations provided in the epoxy datasheet. The waiting time is allowed to give the mixture a proper crosslinking and hardening, and such a process leads to the formation of solid specimens and composite specimens ready for subsequent testing and characterization. Furthermore, a reference sample without any additives was also fabricated. This reference sample serves as a baseline for comparison purposes, and allows a direct evaluation of the impact of the Al2O3 and Gr nanoparticles on the mechanical and tribological performance of the epoxy composite. The sample production process was conducted based on standard precautions to ensure the accurate mixing and homogeneity of epoxy formulation. Firstly, Al2O3/Gr samples underwent compression, utilizing a computer-controlled servo-hydraulic universal testing machine. The machine had a maximum load capacity of 30 tons and the compression rate was adjusted to 1.5 mm/min. From the stress–strain curves achieved, various mechanical characteristics of the epoxy nanocomposites were derived. In accordance with ASTM D2240 [The compression specimens were designed following the standard of ISO 604 Plastics . The speTM D2240 , the har2O3 nanoparticles and graphite. The examination revealed that the Al2O3 nanoparticles displayed an approximately spherical shape, as shown in 2O3 nanoparticles and graphite aligned with the data provided on the data sheet obtained from the manufacturing company. These analyses confirmed the consistency between the observed properties of the nanoparticles and the specifications provided by the supplier.SEM and TEM analyses were utilized to investigate the morphology of the Al2O3 nanoparticles and graphite. Additionally, XRD was utilized to examine the chemical composition of the epoxy/Al2O3/Gr nanocomposites. 2O3 nanoparticles utilized in the current study. The analysis reveals that the alumina powder exclusively consisted of the α-Al2O3 phase. It is noteworthy that these Al2O3 peaks align with those documented in a prior investigation carried out by Mulpur et al. [2O3 nanoparticles and graphite in the epoxy resin is reflected in the XRD pattern of the epoxy composite. However, the analysis confirms that no chemical reaction occurred between the Al2O3, the graphite, and the epoxy matrix. The presence of the nano additives did not modify the crystal structure of the epoxy, and the epoxy retained its distinctive characteristics and identity within the resulting composite.XRD (X-ray diffraction) analysis was employed to explore the chemical compositions of the materials used in the current study: epoxy, as well as the Alr et al. . In contr et al. . Moreove2O3 and graphite, the hardness consistently and significantly rises. The D-index scale was used to measure the hardness of the nanocomposite samples, and the average hardness of the pure epoxy was recorded as 73.4. As the concentration of additives increased in the nanocomposite, the epoxy nanocomposite exhibited an increase in hardness. The highest level of hardness was observed at a weight fraction of 0.5 wt%, measuring 85.1 on the D-index scale. This represented a significant enhancement of 16%, compared to the hardness of the pure epoxy. The observed increase in composite hardness can be attributed to the favorable interface between the Al2O3 and graphite particles and epoxy resin. This interface facilitates the efficient load transfer, resulting in enhanced hardness. Moreover, the hardness of the composite relies on the intermolecular bonds formed among the matrix and the reinforcement material. The noticeable increase in hardness indicates that the additive particles are uniformly distributed within the epoxy resin, further supporting their role in reinforcing the composite structure.According to 2O3 and graphite particles within the epoxy resin. During the compression tests, the stress and strain values were carefully recorded for the epoxy nanocomposites containing various weight fractions of Al2O3 and graphite particles. During the investigation of the impact of adding Al2O3 and graphite particles onto the epoxy composites, several compressive properties were examined, including Young’s modulus, yield strength, ductility and toughness. By incorporating Al2O3 and graphite particles into the epoxy, the nanoparticles were effectively dispersed into the epoxy chains. This arrangement resulted in a reduction in the epoxy chain flexibility, leading to an improvement in the compressive properties of the epoxy nanocomposite.In order to examine the load-carrying capacity of the epoxy nanocomposites, a compression test was performed. The aim was to explore the compressive properties resulting from the addition of the Al2O3 and graphite weight fraction increased from 0.1 to 0.5 wt%. The pure epoxy modulus of elasticity was recorded as 2.77 GPa. However, at a weight fraction of 0.5 wt% for both additives, the Young’s modulus experienced a significant enhancement of 33.9%, reaching 3.71 GPa. Furthermore, the findings indicate that as the content of Al2O3 and graphite particles in the epoxy increased, there was a corresponding increase in the nanocomposite strength. Specifically, when the weight fraction of Al2O3 and graphite reached 0.5 wt%, the strength exhibited a significant improvement of 25.97% when compared to the pure epoxy.2O3. Aluminum oxide possesses a high Young’s modulus of approximately 1400 GPa, which contributes to the overall stiffness of the composite. During the test, the applied compressive force is distributed between the epoxy and the aluminum oxide. In the presence of nanoparticles, when a crack is initiated within the composite, the nanoparticles play a crucial role in healing the crack and preventing its propagation. This crack-healing mechanism effectively increases the overall strength of the composite [The enhanced strength of the epoxy nanocomposites can be attributed to the superior characteristics of Alomposite .2O3 and graphite particles, the plastic deformation area of the composite expanded. Consequently, the toughness of each epoxy nanocomposite was assessed by computing the area for each nanocomposite under the stress–strain diagram. To quantify the toughness enhancement, the relative rise in toughness for each nanocomposite was estimated in comparison to the neat epoxy, as presented in 2O3 and graphite. This interface promotes interfacial adhesive bonding, which facilitates the transfer of load from the epoxy to the Al2O3 and graphite particles. As the reinforcement content was increased in the composite, there was a steady increment in the toughness of the material. Specifically, when 0.5 wt.% for both Al2O3 and graphite were added to the epoxy, the toughness of the nanocomposites was improved by 25.3%. The recorded results are in line with the research conducted by Emiroglu et al. [2O3 and graphite content, reaching a relative ductility of 16.7% compared to the pure epoxy.With the increase in the concentration of the Alu et al. , where tu et al. . The rel2O3 and graphite particles on the epoxy/Al2O3/Gr nanocomposite coefficient of friction, the nanocomposite specimens were subjected to rubbing against a stainless-steel counterpart. The rubbing was performed in the existence of different loads, specifically between 10 and 40 N, at 0.4 m/s sliding speed. The coefficient of friction for each condition was recorded. In order to investigate the impact of loading fractions of Al2O3/Gr nanocomposites, friction tests were carried out for various durations: 10, 20, 30, and 40 min, at a constant load of 40 N. In order to investigate the effect of sliding distance/time on the tribological characteristics of epoxy/Al2O3 nanoparticles in the epoxy/Al2O3/Gr nanocomposites played a significant role in achieving a more uniform and consistent transferred film on the steel counterpart. This uniformity of the transferred film further facilitated a dropping in the coefficient of friction as the sliding duration time increased [The observed decrease in the coefficient of friction accompanied by increasing friction duration time can be attributed to several factors. Firstly, the long friction duration time and rubbing against the counterpart contribute to the smoothing of the composite surface, leading to a reduction in friction. Additionally, prolonged sliding duration generates sufficient heat to induce localized melting of the nanocomposites . This mencreased .2O3/Gr nanocomposites  were measured under different normal loads. The results, in 2O3 and graphite particles led to a decrease in weight loss. This decrease in weight loss occurred due to the improved mechanical characteristics of the nanocomposites. Specifically, the increased concentration of additive particles enhanced the strength of the epoxy nanocomposites. As a result, the strength of bonding among the epoxy resin and the additive particles prohibited the surface degradation during the friction test and improved the wear strength of the nanocomposites. However, with the rise in applied load, a significant amplification in the weight loss of the nanocomposite samples was noticed. This increase in wear may have occurred due to the generation of heat resulting from the kinetic energy which resulted from the relative motion between the rubbing surfaces. The increase in temperature affects the contact region, leading to surface softening. Consequently, high shear resistance is created, resulting in a notable rise in nanocomposite weight loss and the formation of some transferred layers. Additionally, the elevated temperature in the contact area during the sliding test can degrade the shear strength of the epoxy sample. This resulted in escaping reinforcement particles from the epoxy sample throughout the friction test, which may act as abrasive particles on the surface of the epoxy nanocomposite, consequently contributing to increased weight loss [2O3 may work as rolling balls, effectively functioning as a solid lubricant. This behavior helps to reduce the friction coefficient of the nanocomposites in comparison with the pure epoxy [2O3/Gr nanocomposites. It can be observed that increasing the sliding distance/time had a minimal impression on the wear rate of the nanocomposite samples. This limited effect can be attributed to the increased shear resistance among the nanocomposite sample surfaces and the counterpart, which occurs as layers of nanocomposite are transferred to the counterpart.To assess the wear performance of the fabricated nanocomposites, the weight loss of both the pure epoxy (Epoxy0) and epoxy/Alght loss . Neverthre epoxy . Figure 2O3/Gr nanocomposites, the surface of samples after the tribology tests were examined utilizing SEM (scanning electron microscopy), as illustrated in 2O3 and graphite particles exhibit a relatively smooth surface. This indicates that the presence of these additive particles improves the nanocomposite surface strength; consequently, the wear and the coefficient of friction for the nanocomposites decreased, even at low-loading content. Moreover, the graphite particles work as a solid lubricant, reducing the shear resistance throughout the tribology test. In 2O3 and graphite is a fatigue delamination mechanism.In order to gain further insights into the improved wear properties of the epoxy/Al2O3 and graphite particles up to 0.2 wt.% resulted in a relatively smooth sample surface with a decrease in microcracks. This change in the wear mode is attributed to micro-plowing, as shown in 2O3 and graphite particles to 0.3, 0.4, and 0.5 wt.% led to an improvement in the bonding strength and hardness of the proposed epoxy nanocomposites. This in turn enhanced the interfacial adhesion among the matrix and the additive particles. The improved interfacial adhesion facilitated the efficient stress–energy transfer and consequently enhanced the wearing out of the sample surface. As a result, Increasing the concentration of Al2O3 nanoparticles and graphite additives on the various properties of epoxy nanocomposites was investigated. The study outcomes can be concluded as follows:An enhancement of 16% in nanocomposite hardness was achieved, due to uniform dispersion of added particles.2O3 and graphite particle concentration from 0.1 to 0.5 wt.% resulted in 33.9% increase in stiffness, relative to pure epoxy.Increased Al2O3 led to 25.97% increase in mechanical strength compared to neat epoxy.Adding 0.5 wt.% of AlThe toughness is enhanced by 25.3% and the ductility by 16.7% in the 0.5 wt.% composite.An enhancement in the tribological performance was achieved where the coefficient of friction was reduced by 20.75%.A noticeable enhancement in the wear resistance under various frictional conditions is achieved.2O3 and graphite particle concentration reduced surface shear resistance, leading to decreased wear in the nanocomposites.The worn surface examination revealed that increased AlIn the presented experimental study, the impact of reinforcing epoxy with a com-bination of Al


The aim of the study was to develop a diagnostic method for the quantitative determination of the main components of cartilage tissue of various types based on multivariate IR spectral analysis and verification of data using classical chemical analysis.Ksp coefficient was determined as the ratio of the mass fractions of collagen and chondroitin sulfate. Its value was compared with the value of Kchem, equal to the ratio of the mass fractions of collagen and chondroitin sulfate, obtained using the classical chemical analysis of these substances.Cartilages of the nasal septum, knee joint, rib, and nucleus pulposus of the intervertebral disc, as well as trypsinized and defective cartilage samples, were examined as samples. The IR spectra of the cartilage samples, as well as calibration mixtures of collagen and chondroitin sulfate, were obtained. The IR spectra were collected using the attenuated total reflectance techniques, and their processing was performed using the TQ Analyst software and the principal component regression calibration technique. Based on calibration dependence, the Ksp increases from 0.88±0.05 (Kchem~0.8) to 4.55. The use of a calibration model with a complete analysis of the cartilage IR spectrum made it possible to determine the difference in the ratio of the main components in the matrix of different samples in the absence of obvious changes in the IR spectra. Thus, a statistically significant decrease in the content of chondroitin sulfate in degraded articular cartilage  was shown compared with intact samples .The IR spectra of cartilage tissues are a superposition of the IR spectra of collagen and chondroitin sulfate and qualitatively reflect their composition. A change in the ratio between the relative intensities of the characteristic bands of compounds in the IR spectrum is obvious only with a significant change in the content of these compounds in cartilage. This change occurs after trypsinization, when IR spectrometric express analysis of cartilage tissue employing the principal component regression method allows a correct determination of the ratio of the main components in the cartilage matrix, those of collagen and glycosaminoglycans. The proposed technique includes one measurement, does not require prolonged and laborious sample preparation, does not require long, multi-stage and laborious chemical manipulations to determine each of the components, and makes it possible to determine the features and changes in the composition for a large set of samples of cartilage tissue of different types. In future, this approach can be used for non-invasive diagnostics of cartilage tissue. The main components of the matrix of cartilage tissues are collagen  and sulfated polysaccharides — glycosaminoglycans (GAGs), which form complex compounds with protein (proteoglycans and their aggregates) , 2. The Classical chemical analysis of biological tissues usually requires multi-stage sample preparation, which includes obtaining soluble forms (the first stage) and the analysis of solutions for each of the components (the second stage). At the same time, the transfer of tissue into a soluble form cannot be performed using the same method for both collagen and GAGs, and the analysis of each component requires special analytical equipment and highly skilled personnel.Multistage sample preparation and analysis, and, consequently, the use of expensive consumable reagents, can be excluded by measuring a set of physical values of an integral structure. In this case, the establishment of a relationship between the obtained set of physical values and chemical characteristics will remain challenging. These methods include IR spectroscopy. The theoretical basis for the use of IR spectroscopy for the analysis, interpretation, and mathematical processing of IR spectra of biological fluids and tissues is presented in the paper . The pap–1 changes considerably [The usefulness of IR spectra of cartilage tissue to assess the content of the main components in it was discussed already more than 20 years ago . The eariderably , 17. Chaiderably , 18, 19.iderably , althougThe situation has changed with the use of formal methods of multivariate analysis of large complex arrays of experimental data (IR spectra of multicomponent systems) in IR spectroscopy. Thus, researchers, creating their own processing algorithms with the use of multivariate analysis of the IR spectra of cartilage materials, have reached a good accord on the amounts of GAGs determined by spectral and chemical methods of analysis in joint cartilage , nasal sAttenuated total reflectance (ATR) spectroscopy is of particular interest. It provides performing express measurements of cartilage spectra using a prism tip, which is only in contact with the sample, which significantly simplifies sample preparation. However, only the near-surface layer of the sample, where the evanescent field penetrates during total reflection, participates in the spectrum measurements. It is clear that the IR spectrum of the surface layer of the sample may differ from the IR spectrum averaged over the volume. The question of the adequacy of determining the composition of cartilage by ATR IR spectroscopy is still open.The aim of the study was to develop a method for diagnosing cartilage tissue of various types with a quantitative determination of the main components based on multivariate analysis of IR spectra and to verify the data using classical chemical analysis.Porcine nasal septum and rib cartilage, as well as a cow knee joint and fragments of rabbit and sheep spines, were acquired from the farm and were pre-treated no later than 12 h after removal.2 were cut out from cartilage tissue. Such plates were derived from the middle part of the transverse sections of the nasal septum and costal cartilage. Samples of articular cartilage were taken separately from visually smooth areas and from rough areas with pronounced fibrousness. The nuclei pulposi were removed from the intervertebral discs. After isolation, the specimens were dried in a thermostat at 37°C in the air for 18 h, and their IR spectra were collected. Thereafter, each preparation was divided into equal parts, the GAG content was determined in one half of the sample, and the collagen content was determined in the other one.The samples were washed in a NaCl solution, and the fragments intended for the study were derived. For spectral and subsequent biochemical analyses, 0.5–0.8-mm thick plates with an area of approximately 0.5×0.5 cmOne of the components of the calibration mixture was chondroitin-4-sulfate (ChS) , one of the most common GAGs in cartilage tissue. Type I collagen  was used as the second component. A calibration solution with a mass ratio of collagen and ChS from 0.11 to 49.0 was placed on a glass slide and dried in a thermostat at 37°C for 18 to 24 h. As a result, a film was formed on the glass slide, which was used as a standard for obtaining the IR spectrum.Cartilage tissue specimens weighing 1.5–2.0 mg were placed in a solution containing trypsin at a concentration of 1 mg/ml and sodium azide at a concentration of 0.2 mg/ml, kept in a thermostat at 37°C for 24 h. Part of the sample material passed into the supernatant (C1), another part turned out to be undissolved (O1). In a separate experiment with cartilage of the nasal septum, the supernatant was decanted, dried in a glass Petri dish, and the IR spectrum of the obtained film was measured. The undissolved material was washed, dried and its IR spectrum was measured. In other cases, the entire system was subjected to heating and repeated trypsinization.The supernatant and undissolved residue remaining after the first trypsinization were heated in a water bath at 85°C for 15–20 min, then trypsin was added to a final concentration of 2 mg/ml and left in a thermostat at 37°C for another 24 h. The resulting colloidal mixture was centrifuged, the supernatant (C2) was decanted. In a separate experiment, centrifugate (O2) and supernatant C2 were dried, and their IR spectra were collected. In all other cases, C2 from the second trypsinization was analyzed for GAG content using a spectrophotometric reaction.The amount of GAGs was determined in the supernatant after the second trypsinization of the dried samples. The assay is based on the complexation reaction of anionic GAGs with dimethylmethylene blue (DMMB) which is accompanied by metachromasia. The analysis was carried out according to the method proposed by Farndale et al.  with minKchem.The ratio of mass fractions of collagen and ChS in cartilage samples was designated as An acid hydrolyzate of dried cartilage samples weighing 2–5 mg was prepared in a mixture of hydrochloric and trifluoroacetic acids in sealed glass ampoules at a temperature of 166°C. The amino acid analysis was performed on a Hitachi-835 analyzer  in the standard mode for the analysis of protein hydrolysates. For each hydrolyzate, measurements were performed 3 times. The content of collagen was estimated assuming that hydroxyproline is 13.5% (mass) in the amino acid pool of collagen.–1 with a resolution of 0.5 cm–1 and averaged over 32 scans, converted to log (1/R) units using the OMNIC software algorithm. The value of log (1/R) is similar to absorbance, since R is the ratio of the intensity of infrared energy reflected from the sample to that from a diamond prism without a sample.The sample spectra were recorded using a Nicolet iS10 infrared Fourier spectrometer  by the ATR method using a special accessory with a diamond prism. The IR spectra were collected in the reflection mode in the range of 400–4000 cm–1 were processed using the TQ Analyst software .IR spectra in the range of 750–1800 cmKsp values for articular cartilage samples taken from visually smooth areas and from rough areas with pronounced fibrousness was assessed by a two-sample Student’s t-test. The a priori significance level was set as p=0.05.The experimental data of chemical analysis and spectral analysis of several samples of the same type were processed using the OriginPro 2015 software package . The data were presented as М±SD, where М is the mean value and SD is the standard deviation. The difference between the mean The experimental data of spectral analysis using the calibration method by principal component regression were processed using the TQ Analyst software, which provided the root-mean-square errors of calibration, prediction , and corKchem ratio of 2.33 is also shown there.The IR spectra of pure components (collagen and ChS) are presented in The following bands are most characteristic for the collagen IR spectrum:–1 and less intense bands with a maximum of ~1236 cm–1 associated with vibrations in the peptide group (Amide I and Amide II bands);intense bands with maxima ~1628 and ~1530 cm–1 associated with bending vibrations of СН2 and СН3 groups;bands with maxima ~1447 and ~1394 cm–1, presumably associated with the bending vibration of the СН2 group in the proline residue in the polypeptide chain.the band representative for collagen with a maximum of ~1338 cmThe following bands appear in the IR spectrum of ChS:–1, associated with vibrations in the N-acetyl group  and C=O stretching vibrations in the COO– group;a wide band, in the region of 1650–1500 cm–1, associated with bending vibrations of CH2 and CH3 or COO– groups;bands centered at ~1410 and ~1375 cm–1, associated with symmetric and antisymmetric stretching vibrations in the SO3– group and, possibly, with C-O-C skeletal vibrations (~1025 cm–1);intense bands centered at ~1223 and ~1025 cm–1, associated with the pyranose ring;a band centered at ~1125 cm–1, associated with skeletal vibrations of С-О-С and С-О-S groups.bands centered at ~924 and ~855 cmRс2=0.997) and predicting (Rt2=0.9978). The calibration and test lines practically merge. Based on calibration dependence, Ksp was determined as the ratio of the mass fractions of collagen and ChS.Kchem and Ksp values in a sample of cartilage tissue of the nasal septum. The IR spectrum of the original tissue is shown in curve 2).Ksp=0.91, the spectral analysis showed that there was a sharp increase in the proportion of collagen in the insoluble O1 residue (Ksp=4.55) and a decrease in this value in the C1 supernatant film (Ksp=0.32). Nevertheless, after the first trypsinization, a noticeable part of the initial amount of ChS remained in the insoluble O1 residue. After trypsinization, denaturation, and repeated trypsinization of the sample with Ksp=0.89, the vast majority of ChS was found in the C2 supernatant solution (Ksp=0.25), while the insoluble O2 residue contained predominantly collagen (Ksp=13.5). The IR spectra of the O2 centrifugate (curve 1) and the C2 film (curve 3) differ markedly and approach the IR spectra of collagen and ChS, respectively.After the primary trypsinization of the sample with rifugate , curve 1 C2 film , curve 3Ksp value averaged over 11 nasal septal cartilage samples.Kchem and Ksp values for two samples of costal cartilage.It should be noted that the IR spectra of specimens of hyaline cartilage  did not fundamentally differ from those for nasal septum cartilage. Kchem and Ksp values for articular cartilage samples with different textures (two samples of each texture). The Table also shows the Ksp value averaged for 12 samples from a smooth surface and 8 samples from a rough fibrous surface of the joint.Articular cartilage samples were taken from both the smooth and rough fibrous surfaces of the joint. Kchem and Ksp values for different samples of the nucleus pulposus of rabbit and sheep.curve 3) and cartilage tissues  and sheep nucleus pulposus (Ksp~1.2) shown in Changes in the ratios between the characteristic bands of compounds in the IR spectrum are evident with a significant change in the content of these compounds in cartilage. For example, when comparing the IR spectra of rabbit nucleus pulposus  was obtained on the whole sample, and the other one — as the amount of the dyed complex of GAGs with DMMB in trypsinate, requires explanation. Let us assume that the error in estimating the content of collagen determined by the most accurate of the analytical methods used is minimal. At the same time, the constants of binding GAG and DMMB dye into a complex for different GAGs differ and the calibration curves for them do not coincide [An important result is the coincidence of the coincide , while tcoincide .K value is the difference in the composition of calibration mixtures and tissue specimens. However, the similarity of their IR spectra suggests that other components (other than proteins and GAGs) have little effect on the type of the IR spectrum of cartilage tissues. Once again, we underline that we estimate not the absolute amount of collagen and GAGs, but only their ratio, which coincides with the value of the coefficient determined on the basis of chemical analysis.There are other factors that stipulate the inaccuracy of the method for determining GAGs using DMMB , but thiKsp) obtained by employing ATR does not require multi-stage sample preparation, which is costly, time-consuming, and requires highly skilled analysis performers. In addition, the chemical determination of collagen and GAG cannot be done in one and the same sample, and the analyzed samples may not be identical. Knowing the spectral ratio in the sample, it is possible to select and conduct an analysis of one of the components that is more convenient for the researcher. The amount of the second component is easily calculated. Determination of the Kchem value is often the first one when studying cartilage tissue changes, and the number of samples can amount to several tens. In this case, the express characterization of the variability of the chemical composition is extremely important.Experimental determination of a collagen/GAG ratio based on the IR spectrum (Infrared spectrometric analysis in the mode of attenuated total reflection employing the method of principal component regression makes it possible to determine correctly the ratio of collagen and glycosaminoglycans, the main components of the cartilage matrix, and to conduct a preliminary non-invasive diagnosis of cartilage tissue. We have proposed an express method to determine the features and changes in the composition for a large set of cartilage tissue samples of several types. An additional advantage is an extremely simple sample preparation, which does not require multi-stage and time-consuming chemical manipulations and the reagent consumption.


Background: Osteoarthritis is a degenerative joint disorder of articular cartilage and is the most common type of arthritis in the elderly. There are only a few reports regarding the use of Hydroxychloroquine in the treatment of osteoarthritis. Methods: To investigate the effects of Hydroxychloroquine on the symptoms of mild to moderate knee osteoarthritis (Kellgren and Lawrence grade II and III), we performed a double-blind, placebo-controlled study in 44 patients. The patients were randomly assigned to two groups: one group received Hydroxychloroquine pills (200 mg twice daily) and the other group received placebo pills. Symptoms were assessed by the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) at baseline and at the end of weeks 4, 8, 12, 16, 20, and 24.Results: Approximately, 98% of the patients were women at an average age of 47 years. There was no significant difference in the baseline characteristics between the two groups. In the placebo group, maximum improvement occurred at the 4th week; and during the remaining time, there was no significant improvement. In the Hydroxychloroquine group, maximum improvement occurred at the 8th week and persisted over the entire remaining follow-up period. There were significant differences between the two groups regarding the degree of reduction in the WOMAC total score and the WOMAC subscales scores of pain, stiffness, and function at the end of weeks 4, 8, 12, 16, 20, and 24.Conclusion: Hydroxychloroquine conferred significant improvement in the symptoms of mild to moderate knee osteoarthritis in our patients and may, accordingly, be recommended for knee osteoarthritis treatment. Currently, the pharmacological treatment of osteoarthritis is primarily aimed at controlling symptoms and analgesics and non-steroidal anti-inflammatory drugs (NSAIDs) are commonly prescribed. There are at present no specific pharmacologic therapies that can slow the progression of this condition.2Treatment of osteoarthritis can be frustrating for patients and physicians.5,6 Lipid peroxidation is thought to play a role in apoptosis. Over the last two decades, there has been increasing evidence showing the importance of classic apoptosis in the creation of osteoarthritis.7 Antimalarial agents also have antioxidant properties and may provide protection against tissue damage by free radicals.5,6Antimalarial agents have immunomodulatory and anti-inflammatory properties, although their precise mechanism of action in rheumatic diseases is unknown. The anti-inflammatory properties of the antimalarials include effects on the arachidonic acid cascade, by downregulation of phospholipase A2 and C, which contribute to the production of proinflammatory prostaglandins and lipid peroxidation.The purpose of the present study was to investigate the potential effect of Hydroxychloroquine (HCQ) on the symptoms of knee osteoarthritis.8 Eligible patients were those who met all of the following criteria: 1) primary knee osteoarthritis; 2) knee osteoarthritis Kellgren and Lawrence grade II or III;9 3) knee pain for at least the preceding 6 months; 4) minimum age of 30 years; and 5) literacy. Patients were excluded if they had any of the following: 1) secondary osteoarthritis; 2) knee arthroscopy during the preceding 6 months; 3) intra-articular injection of corticosteroids during the last 6 months; 4) presence of other inflammatory diseases; 5) history of hypersensitivity to antimalarial drugs; and 6) any kind of eye disease.This 24-week, randomized, double-blind, parallel-group study was conducted on knee osteoarthritis patients. All the patients fulfilled the American College of Rheumatology classification criteria for knee osteoarthritis.The trial was registered in the Iranian Registry of Clinical Trials database, accessible at www.rct.ir (IRCT138709121479N1). The study protocol received approval from the Ethics Committee of Mashhad University of Medical Sciences, and all the patients provided written informed consent prior to study participation.Patients were enrolled from outpatient clinics. All the patients underwent a baseline eye examination by an ophthalmologist. The patients were thereafter divided into two groups via a random number table: the first group received 200 mg HCQ  with dosage of 200 mg twice daily for 6 months and the second group received placebo tablets with the same schedule. The HCQ and placebo tablets were identical in shape and appearance. The placebo pills were produced by the Faculty of Pharmacy, Mashhad University of Medical Sciences. All the drugs were labeled with the randomization numbers of the participants. The patients and the staff members involved in data collection were unaware of the group assignment. The patients were allowed to use their usual painkillers . However, they were told to discontinue their analgesics and NSAIDs 48 hours before the physician’s visit to avoid confounding the test results. The patients were also asked to record the name and number of the analgesics and NSAIDs that they consumed during each day. All the patients were requested at each visit to report any eventual side effect. All the patients were trained how to answer the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC).The WOMAC index questionnaires were completed by each patient at the baseline visit and at the end of weeks 4, 8, 12, 16, 20, and 24. The WOMAC measures five items for pain (score range 0–50), two for stiffness (score range 0–20), and 17 for functional limitation (score range 0–170). t test or the Mann–Whitney U test for the quantitative variables (chosen according to the nature or distribution of these variables). A repeated measure ANOVA was performed to assess the change over time in the WOMAC total score and the WOMAC subscales scores of pain, stiffness, and function. The level of significance was a P value smaller than 0.05.The data were analyzed by SPSS statistical software (version 16) . The numerical variables are described by mean and standard deviation. The baseline characteristics were compared between the groups using the Fisher exact text for the qualitative variables and Student’s Sixty-four patients were introduced to the study from our Rheumatology Clinics. Of them, six patients were not eligible and seven refused to give consent. Fifty-one patients were, therefore, randomized into two groups ; however, forty-four patients completed the study : four patients were lost to follow-up  and 3 patients  discontinued the treatment due to drug side effects . Both grth week and there was no significant improvement during the remaining time. In the HCQ group, maximum improvement occurred at the 8th week and lasted over the entire remaining follow-up period. There were significant differences between the two groups as regards the degree of reduction in the WOMAC total score and the WOMAC subscales scores of pain, stiffness, and function at weeks 4, 8, 12, 16, 20, and 24 . Three patients in the HCQ group discontinued the treatment due to drug side effects (skin rash in 2 and vertigo in one), whereas there were no drug side effects in the placebo group. The difference between the two groups with respect to the frequency of drug side effects was statistically significant (P=0.001).10,11 Pharmacologic treatment categories for osteoarthritis are typically set up to designate whether drugs are symptom-relieving or disease-modifying. Nonetheless, the evidence has thus far proved insufficient as to which drugs have optimal disease-modifying properties in osteoarthritis.2The knee joint is commonly afflicted by osteoarthritis. As much as knee osteoarthritis is a disease of high prevalence in world populations and is associated with high morbidity, the treatment has shown little progress in the last decades.5,6 Be that as it may, their precise mechanism of action in rheumatic diseases is still unknown.Antimalarial agents have immunomodulatory and anti-inflammatory properties, and the effectiveness of these drugs in the therapy of some rheumatic diseases has been well known in the medical literature for many years.12 in a controlled, randomized, double-blind study assessed the effectiveness of HCQ on knee osteoarthritis and concluded that although their two groups of HCQ and placebo exhibited improvement, HCQ had no superiority over the placebo in the treatment of knee osteoarthritis. Bryant, Desrosier, and Carpenter13 treated 8 patients with erosive hand osteoarthritis and obtained satisfactory results in 6 patients, all of whom showed improvement in a period ranging from 7 weeks to 7 months. Gianantonio Saviola et al.14 compared the efficacy of Clodronate (a bisphosphonate) with HCQ for treating active erosive osteoarthritis of the hand and demonstrated that while the former probably played an efficacious role in the treatment of active erosive osteoarthritis, the latter seemed to be ineffective. Vuolteenaho et al.15 found that HCQ suppressed nitric oxide production induced by IL-1 β in osteoarthritic cartilage and concluded that this drug could be useful in treating this disease. There are only a few reports regarding the use of HCQ in osteoarthritis. Indeed, there is only one report on the use of HCQ in knee osteoarthritis. Herval de Lacerda Bonfante et al.12 study (48 years vs. 60 years). The lower age of our patients may be due to the affliction of younger people by knee osteoarthritis in Iran compared with some other countries.16,17 In contrast to the Herval de Lacerda Bonfante study, the results of our study showed statistically significant improvement in the total WOMAC score, WOMAC pain score, WOMAC stiffness score, and WOMAC function score in the patients using HCQ. Although drug side effects statistically were more frequent in the HCQ group, the symptoms were mild to moderate and were improved immediately after the discontinuation of HCQ. The present study was performed to investigate the effect of HCQ on mild to moderate knee osteoarthritis symptoms. The average age of our patients was less than that in the Herval de Lacerda BonfanteThe results of the present study showed that HCQ improved the symptoms of knee osteoarthritis in our patients. Nevertheless, no conclusion can be drawn apropos the disease-modifying action of HCQ because the focus of the study was merely on clinical changes. Larger new clinical trials are required to evaluate not only the symptoms but also the joint space if one is to demonstrate the validity of the use of HCQ as a disease-modifying drug. HCQ conferred significant improvement in the symptoms of mild to moderate knee osteoarthritis in our patients and can, therefore, be deemed useful in the treatment of knee osteoarthritis.


Chemotherapy-induced nausea and vomiting are the most important complications for cancer patients as its prevalence has been reported to be about 54-96 percent. ginger has been used for medicinal purposes including nausea and vomiting in traditional Persian, Chinese and Indian pharmacopoeia.The objective of this study was to evaluate the efficacy of complimentary ginger among cancer patients experiencing nausea and vomiting.A randomized cross-over clinical trial was carried out on patients under chemotherapy treatment for at least 2 episodes of chemotherapy and at least 2 episodes of previous experience of nausea and vomiting. Subjects of this study received 2 different complementary regimes with 250mg ginger capsule in regime A and placebo capsule in regime B. subjects of the study were crossed over to receive the other regime during the two cycles of chemotherapy.Findings of the study indicated that subjects receiving ginger showed significant reduction in frequency and intensity of nausea and vomiting compared to placebo receiving subjects.According to finding of this study, in accordance to most of other researches, ginger is an effective agent to reduce chemotherapy-induced nausea and vomiting. However, there are some researches supporting ginger as a moderate antiemetic agent among cancerous patients under chemotherapy. Chemotherapy - induced nausea and vomiting are the most common intensive side effects and an important source of anxiety among cancer patients as its prevalence has been reported about 54 - 96 percent . ChemothSince, the substitute and complementary therapies may be used as therapy or supportive methods. These methods can be applied as independent therapy methods or along with other standard methods for cancer patients . HistoriThe major pharmacological activity of ginger, Zingiber official, is related to its active components named of gingered and shogalos. The effects of these combinations are anti inflammation, anti vomiting, anti fever, anti cough, anti blood pressure, anti cancer, decreasing of prostaglandin, and the sedation of digestive problems. The effect of ginger products as an antiemetic is implemented by several mechanisms. For example, ginger and shogaols decrease the stomach contractions, but increase the activity of gastro intestinal trace (GIT). These combinations have anti-cancer effect and exert the effects of garbage against of the free radicals . SontakiThere are some scientific studies in which contradictory findings have been reported. The effects of ginger on nausea and vomiting after surgery by laparoscopy genital system on 180 under surgery women indicates that the ginger had no effect on the intensity of nausea and vomiting in comparison with control group . FurtherWith consideration to contradictory results of antiemetic effects in different studies, this study was aimed to determine the effect of ginger on chemotherapy induced nausea and vomiting in cancer patients.This study is a randomized, prospective, cross-over double - blinded clinical trial, conducted on patients under chemotherapy treatment. Subjects of this study were selected from cancer patients attending in hematology ward in university hospital. In this research information was gathered by using questions and check list after the following steps:1)Supplying and editing the check list 2)Determining the validity and reliability 3)Presenting the letter of introduction from university.4)Acquiring the permission from behavior committee. Hospital authority Oncology specialist, center of blood and patient research (acquiring the form of conscious satisfaction) During the study the researcher attended in hematology ward during the week from Saturday to Thursday from 8 am. For about 6.5 month (from September 1st until March 20th 2007). After interviewing with available patients and filling up the information form, she was making certain that the chosen person has the appropriate conditions to be a sample for the research. This study was doing on the basis of the block randomization with the four block method. The researcher was choosing A and B regimes and evaluating the effects of these two regimes on the patients who were under chemotherapy on the basis of the selected group of the plan. The A regime was a routine antiemetic drugs along with 4 ginger capsules. Two of them should be taken 30 minutes before prescription of chemotherapy in the edible form. The other capsules were received 6 hours after chemotherapy.The B regime was a routine antiemetic along with 4 placebo capsules. Two of them 30 minutes before the chemotherapy prescription and two others were received 6 hours after chemotherapy. Routine antiemetic regimen consists of one Ampoule of Grainestrone (keitril) 3 milligrams and one Ampoule of Degzametazone of 8 mg which patient was received 30 minutes before chemotherapy as basic drug for controlling chemotherapy nausea and vomiting. This drug may be used with 10 mg Metocolopramid which was prescribed within 24 hours, if it was requested by the patient.It is necessary to explain that ginger capsule, with Zintoma trade mark, consists of 250 mg ginger powder which was produced by Isfahan Gol daru and placebo consists of 250 mg ineffective powder (Chickpea powder). The shape, color, and fragrance of this powder were similar to ginger and both of them were provided by the same company. Patients were chosen on the basis of inclusion criteria containing the following items:1)Age over 18 2)Having the experience of chemotherapy with nausea and vomiting, 3)Having at least two chemotherapy episodes. Consists of 50-100 mg Cisplatin, with or without other chemotherapy agents with similar amount and prescription in two chemotherapy cycles without having any plans for radiotherapy among cycles 4)Presence in ward for 24 hours ( in order to investigate the PRN) 5)Having no nausea and vomiting experiences for some reasons except for chemotherapy, 6)Lack of receiving PRN in 24 past hours, 7)Lack of treatment with corticosteroid drugs during considered cycles, 8)Lack of afflictions to hepatitis, digestion system blockage, brain malignancy and cerebral metastasis and clotting disorder on the basis of recorded information in files 9)Lack of using the anti-clot drug.The style of assigning scores to strain tool of intensity of vomiting and the way of investigation, by kortila tools was explained to the patient. If a patient was under radiotherapy or the process of investigation was stopped for any reason  the trend was, he or she left out of the plan. The process of patient's inspection was as follows. The researcher gave two edible 250mg capsules (ginger or placebo) to the patient half an hour before chemotherapy and simultaneously with Prescription Ampoule ketril 3 mg and Dexamethazon 8mg. afterwards gave two similar edible capsules 6 hours after chemotherapy. As mentioned, the capsules were chosen so that the patient received one kind of regimen during each cycle. Thus finally, all of the patients who were under investigation received both regimes A and B randomly, at least for 28 days, during two episodes.The nausea's frequencies and score were measured 5 times during 24 hours. These measurements were done 1,2,3,4 hours after prescription of the second dose of capsules and 24 hours after chemotherapy in the next morning. In each time the severity of the measurement was determined by using the strain tools of severity of nausea. In this tool the flat surface or zero equals to the lack of nausea existence, steps 1 to 3 equal to mild nausea, steps 4 to 6 equal to average nausea, steps 7to 9 equals to sever nausea and step 10 equals to the severest possible nausea. The period of vomiting and retching was also observed during 24 hours, simultaneously with observation of nausea severity. The severity was also determined by Kortilla tools. The measurement of the severity of vomiting by using this tool was as follows:•no retching equals to the lack of vomiting , •less than 3 times of retching with or without eliminating of stomach content equals to mild vomiting ,•3 to 5 times of retching with or without the eliminating of stomach content equals to average vomiting •More than 5 times of retching with or without eliminating of stomach content equals to sever vomiting It is necessary to mention that within the hours of measurement of severity of the nausea and vomiting and at the end of 24 hours, Methocolopramid does which was used by the patient as PRN, was investigated and recorded in check list. In this research the effect of period, group and treatment was measured, in a way that the difference, sum of the average and the standard deviation of nausea score (the frequency of vomiting) the severity of nausea ( the intensity of vomiting ) was measured as   in periFinally, the information was analyzed statistically by using SPSS version 16, in a way that the description was applied the data descriptive statistic descriptive and categorize and comprehensive statistic was used to test the hypothesis. Kolmogorov - Smirnov Test was applied to prove adaption to normal distribution. The independent t-test and data was used to analyze the effect of treatment, period and group in data with normal distribution and for data without normal distribution non-parametric test (Mann- Whitney U test) was implemented.2 of Cisplatin. Most of the units in the first cycle (%81.8) and the second cycle (%46.6) had 2 - 4 times chemotherapy period and %52.3 received Cisplatin in combination with Fluorouracil 5. The intensity of acute phase nausea (24 hours) in different cycles for patient who received regime A (routine anti – emesis regime and ginger) and regime B (routine anti emesis regime and placebo) was observed. These measurements showed that the severity of the nausea in the first, second, third, fourth and the end of 24 hours in people who received regime A rather than regime B reduced %9, %18.2, %13.7, %22.7, %13.7, %27.3 respectively. In the second cycle these values were %7.2, %8 (in regime B) %5.3 , %14.1 (in regime B) and %24.1. About acute phases of vomiting, in the considered cycles, for patients who received regime A (Routine anti-emesis and ginger) and regime B (Routine anti-emesis and placebo) were studied. The results showed that the severity of the vomiting in the first, second, third, fourth hours and the end of 24 hours, in the first cycle of measurement for people who received ginger were respectively %9.1, %9.1, %9.1, %4.6,%4.7 less than (placebo). The results of the second cycle were showed that in the first hour there wasn’t any significant difference between the intensity of vomiting in two groups. In second, third, fourth hours and at the end of 24 hours respectively %4, %6.3, %6.7 and % 8.3 decreases have been seen in regime B in comparison the regime A. The frequencies of nausea in the first, second, third, fourth hours and at the end of 24 hours were measured. Then the measurement of the lack of group and period effect, and the effect of treatment were measured with statistical Mann- Whitney U test. There wasn’t any significant statistical difference between two groups in two cycles of treatments. Nausea scores in the first, second, third and fourth hours were measured. After elimination of the group and period effects, the effect of treatment was measured. In these hours the nausea score in ginger group also decreased, but there wasn’t any statistical significant difference between two groups in the mentioned hours. Nausea score in 3rd and from 4th hour to the end of 24 hours was measured. The effect of group and period was measured in the mentioned hours. After making certain of the lack of their effects, the effect of treatment was measured. These measurements became meaningful after using independent t test Mann – Whitney U test with P < 0.01 and P < 0.05 , vomiting and capsule elimination and lack of capsule swallowing (Two person), and relinquishment from continuing the treatment by a person without any special description. Five people of participants were under radiotherapy between the cycles of chemotherapy, which this situation was also considered as exclusion criteria, so these samples were left out of the study. Findings showed that most of the researched units (56.8%) were over 50 years (50.3 ± 13.1), (%59.1) male and many of units afflicted with the esophagus cancer and most of them (%47.7) were under therapy with 60 - 69 mg /mP < 0.05  and 3. TThe findings of this research showed that the severity of the nausea in acute phase (the first 24 hours) in the first and second cycles in ginger consignee group was less that placebo. About the effectiveness of ginger in different hours of measured cycles can refer to the study, done by Nanthakoma et al. The results of this study showed that in nausea was appeared in %48.3 of ginger group members and %66.7 of placebo group members, so the ginger was 18.4% more effective than placebo .Another approving study is the study which is done by Vutyavanich et al, The results of this study showed that , the decrease of nausea average score for all participants in ginger group was more than place. In this research after receiving the considered regime, the improvement of nausea was 28 out of 32 people in ginger group (%87.5)and 10 out of 35 people in placebo (%28.2) .With regThe result in second cycle showed that in the first hour there wasn’t any difference between the intensity of vomiting in two regimes. There was respectively %4, %6.3, %6.7,%8.3 reduction in regimen A compared to regimen B for second , third , fourth hours and at the end of 24 hours. The study done by Visalyputra was from researches in which the effect of placebo was considerable. In this study there was no difference in the amount of nausea and vomiting incidence between placebo and ginger users. The results of the mentioned study showed that nausea incidence for placebo was %32, for ginger users was %22, and for dipridamol and ginger users was %33. For vomiting these values were %32, %25 and %25 respectively. Thus the amount of nausea and vomiting incidence in placebo and ginger group were 2 gram and for dipridamol was similar to each other and there wasn’t any difference .The of the frequencies of nausea in the first, second, third, fourth hours and at the end of 24 hours, after omitting of the effect of group and period, by using Mann - Whitney U test in measuring the effect of therapy presented the lack of significant statistical difference. These results were similar to the results gained by Leopold et al. In the mentioned study the nausea incidence after operation were %49, 58%, and 53% respectively for placebo group, 300 and 600 mg ginger group, in which ginger group the effectiveness of therapies wasn’t significant P = 0.69 . In thisNausea score in the third hour and from the fourth hour to the end of 24 hours was also measured. In the mentioned hours and after measuring the effect of period and group and after certainty of the lack of their effect, the effect of therapy measured. By using the Mann - Whitney U test were respectively significant with P < 0.01 and P < 0.05. Here we can refer to the research done by Sontaki et al. This research presents the reduction of nausea score for %62 of under therapy with ginger that was significant from the statistical point of view . The freOn the other hand, Pangrojpaw et al. showed that ginger was more effective than Diphenhydramin, in addition it has less side effects . Ozgoli Conclusion: although there wasn’t any significant difference in the present research in all statistical tests between ginger and placebo, in most hours of measurement the reduction of intensity score and the frequencies of nausea were observed. This change was less in the frequencies of vomiting. Therefore, on the basis of present research and the other studies, done in all over the world, ginger can be used as a simple method, inexpensive and secure and as complimentary of antiemetic in controlling of nausea and vomiting. In this study, the lack of attainment to expected results can be assigned to factors such as the lack of sufficient ginger does’ sufficiency, the little amount of available sample, the lack of exclusiveness of the cancer, and the hours of giving ginger. Thus by implementation of widespread studies and with solving the mentioned problems, it is possible to gain the better results.