Concentrations Of Indomethacin Research Articles

Neuroendocrine modulation by metamizole and indomethacin: investigating the impact on neuronal markers and GnRH release.

Some evidence that non-steroidal anti-inflammatory drugs have neuroprotective effects indicates their potential for use in a new field. However, their effects on hormone secretion have yet to be adequately discovered. Therefore, we aimed to evaluate the effects of metamizole and indomethacin on neuronal markers as well as the GnRH expression in the GT1-7 cell line. The effects of these drugs on proliferation were evaluated by MTT analysis. The effect of 10-50-250 µM concentrations of the drugs also on the expression of neuronal factors and markers, including NGF, nestin and βIII Tubulin, and additionally GnRH, was determined by the RT-qPCR method. NGF and nestin mRNA expressions were increased in all concentrations of both metamizole and indomethacin. No changes were detected in βIII Tubulin. While metamizole showed an increase in GnRH mRNA expression, there was no change at 10 and 50 µM concentrations of indomethacin, but a remarkable decrease was observed at 250 µM concentrations. The results of our study showing an increase in the expression of neuronal factors reveal that metamizole and indomethacin may have possible neuroprotective effects. Moreover, the effects on the GnRH expression appear to be different. Animal models are required to confirm these effects of NSAIDs on neurons.

Anti-Adhesive Resorbable Indomethacin/Bupivacaine-Eluting Nanofibers for Tendon Rupture Repair: In Vitro and In Vivo Studies.

The treatment and surgical repair of torn Achilles tendons seldom return the wounded tendon to its original elasticity and stiffness. This study explored the in vitro and in vivo simultaneous release of indomethacin and bupivacaine from electrospun polylactide-polyglycolide composite membranes for their capacity to repair torn Achilles tendons. These membranes were fabricated by mixing polylactide-polyglycolide/indomethacin, polylactide-polyglycolide/collagen, and polylactide-polyglycolide/bupivacaine with 1,1,1,3,3,3-hexafluoro-2-propanol into sandwich-structured composites. Subsequently, the in vitro pharmaceutic release rates over 30 days were determined, and the in vivo release behavior and effectiveness of the loaded drugs were assessed using an animal surgical model. High concentrations of indomethacin and bupivacaine were released for over four weeks. The released pharmaceutics resulted in complete recovery of rat tendons, and the nanofibrous composite membranes exhibited exceptional mechanical strength. Additionally, the anti-adhesion capacity of the developed membrane was confirmed. Using the electrospinning technique developed in this study, we plan on manufacturing degradable composite membranes for tendon healing, which can deliver sustained pharmaceutical release and provide a collagenous habitat.

Wastewater surveillance of pharmaceuticals during the COVID-19 pandemic in Mexico City and the Mezquital Valley: A comprehensive environmental risk assessment

This study tracked five pharmaceutically active compounds (PhACs) in Mexico City's sewage, namely, famotidine, indomethacin, dexamethasone, azithromycin, and ivermectin, which were used to treat COVID-19. The monitoring campaign was carried out over 30 months (May 2020 to November 2022), covering the five COVID-19 waves in Mexico. In the Central Emitter, the main sewage outflow, famotidine displayed levels of 132.57 ± 28.16 ng L−1 (range from < LOQ to 189.1 ng L−1), followed by indomethacin (average 672.46 ± 116.4 ng L−1, range from 516.7 to 945.2 ng L−1), dexamethasone (average 610.4 ± 225.7 ng L−1, range from 233.4 to 1044.5 ng L−1), azithromycin (average 4436.2 ± 903.6 ng L−1, range from 2873.7 to 5819.6 ng L−1), and ivermectin (average 3413.3 ± 1244.6 ng L−1, range from 1219.8 to 4622.4 ng L−1). The concentrations of dexamethasone, azithromycin and ivermectin were higher in sewage from a temporary COVID-19 care unit, by a factor of 3.48, 3.52 and 2.55, respectively, compared with those found in municipal wastewater. In the effluent of the Atotonilco Wastewater Treatment Plant (AWWTP), which treats near 60 % of the Mexico City's sewage, famotidine was absent, while concentrations of indomethacin, dexamethasone, azithromycin and ivermectin were 78.2 %, 76.7 %, 74.4 %, and 88.1 % lower than those in the influent, respectively. The occurrence of PhACs in treated and untreated wastewater resulted in medium to high environmental risk since Mexico City's wastewater is reused for irrigation in the Mezquital Valley. There, PhACs were found in irrigation canals at lower levels than those observed in Mexico City throughout the monitoring. On the other hand, famotidine, indomethacin, and dexamethasone were not found in surface water resulting from the infiltration of wastewater through soil in Mezquital Valley, while azithromycin and ivermectin sporadically appeared in surface water samples collected through 2021. Using an optimized risk assessment based on a semi-probabilistic approach, the PhACs were prioritized as ivermectin > azithromycin > dexamethasone > famotidine > indomethacin.

Detailed Experimental and In Silico Investigation of Indomethacin Binding with Human Serum Albumin Considering Primary and Secondary Binding Sites

The interaction of indomethacin with human serum albumin (HSA) has been studied here considering the primary and secondary binding sites. The Stern-Volmer plots were linear in the lower concentration range of indomethacin while a downward curvature was observed in the higher concentration range, suggesting the presence of more than one binding site for indomethacin inside HSA due to which the microenvironment of the fluorophore changed slightly and some of its fraction was not accessible to the quencher. The Stern-Volmer quenching constants (KSV) for the primary and secondary sites were calculated from the two linear portions of the Stern-Volmer plots. There was around a two-fold decrease in the quenching constants for the low-affinity site as compared to the primary binding site. The interaction takes place via a static quenching mechanism and the KSV decreases at both primary and secondary sites upon increasing the temperature. The binding constants were also evaluated, which show strong binding at the primary site and fair binding at the secondary site. The binding was thermodynamically favorable with the liberation of heat and the ordering of the system. In principle, hydrogen bonding and Van der Waals forces were involved in the binding at the primary site while the low-affinity site interacted through hydrophobic forces only. The competitive binding was also evaluated using warfarin, ibuprofen, hemin, and a warfarin + hemin combination as site markers. The binding profile remained unchanged in the presence of ibuprofen, whereas it decreased in the presence of both warfarin and hemin with a straight line in the Stern-Volmer plots. The reduction in the binding was at a maximum when both warfarin and hemin were present simultaneously with the downward curvature in the Stern-Volmer plots at higher concentrations of indomethacin. The secondary structure of HSA also changes slightly in the presence of higher concentrations of indomethacin. Molecular dynamics simulations were performed at the primary and secondary binding sites of HSA which are drug site 1 (located in the subdomain IIA of the protein) and the hemin binding site (located in subdomain IB), respectively. From the results obtained from molecular docking and MD simulation, the indomethacin molecule showed more binding affinity towards drug site 1 followed by the other two sites.

Development of an Intrauterine Device Releasing Both Indomethacin and Levonorgestrel During the First Months of Use: Pharmacokinetic Characterization in Healthy Women.

Post-placement menstrual bleeding pattern changes with intrauterine contraceptives (IUCs), including levonorgestrel-releasing intrauterine systems (LNG-IUS), can be a reason for avoidance or early discontinuation. Prostaglandins play an important role in menstrual bleeding and pain. The key drivers of prostaglandin synthesis are cyclooxygenase (COX) enzymes, which are inhibited by non-steroidal anti-inflammatory drugs. In this study, we report the findings from pharmacokinetic (PK) analyses undertaken with an LNG-IUS (LNG-IUS8) modified with an additional reservoir containing indomethacin (IND). The IND/LNG-IUS8 is a proof-of-concept device studied in a phase II proof-of-concept/dose-finding study. IND/LNG-IUS8 contains the same LNG content as the unmodified LNG-IUS8 (13.5 mg) but was prepared with three different IND doses (low, 6.5 mg; middle, 12.5 mg; and high, 15.4 mg), resulting in different daily release rates. Overall, 174 healthy, premenopausal women were randomized to one of the four treatment arms (low-, middle-, high-dose IND/LNG-IUS8 or LNG-IUS 8). Initial and residual IND and LNG content were collected and the amount of IND and LNG released in vivo over the period of use was calculated. A subset of 62 participants underwent dense blood sampling for PK analysis. Concentrations of IND and LNG in plasma were determined by validated liquid chromatography-tandem mass spectrometry methods and plotted over time. Descriptive statistics were calculated for plasma drug concentrations and PK parameters. High-dose IND/LNG-IUS8 initially released much higher levels of IND than expected based on in vitro release data, followed by a steep decline, with the reservoir emptied by 4.5 months. Middle- and low-dose IND/LNG-IUS8 demonstrated steady sustained release of IND over time, emptying after 7.4 and 8.4 months, respectively. Peak plasma concentrations of IND for low- and middle-dose IND/LNG-IUS8 remained below the 20% maximal inhibitory concentration (IC20) values for COX enzymes. The average daily IND release rate in vivo was 49 µg/day for low-dose and 112 µg/day for middle-dose IND/LNG-IUS8. The IND release rate profile and IND plasma concentrations in vitro both decreased steadily over time with low- and middle-dose IND/LNG-IUS8. The LNG release rate profile was comparable for all IND/LNG-IUS8 dose groups and LNG-IUS8. This PK study demonstrates that two different drugs can be released at different rates from an IUS designed with two drug reservoirs. Inclusion of IND does not impact the LNG PK profile. Low- and middle-dose IND/LNG-IUS8 were associated with a systemic IND exposure that should preclude the occurrence of adverse events typically observed after oral IND dosing. ClinicalTrials.gov identifier number: NCT03562624.

Ketoprofen, piroxicam and indomethacin-suppressed quorum sensing and virulence factors in Acinetobacter baumannii.

Quorum sensing (QS) inhibition is a promising strategy to suppress bacterial virulence and control infection caused by Gram-negative and Gram-positive bacteria. This study explores the QS inhibiting activity of the non-steroidal anti-inflammatory drugs (NSAIDs) in Acinetobacter baumannii. Ketoprofen, piroxicam and indomethacin revealed QS inhibition via elimination of violacein production of the reporter strain Chromobacterium violaceum ATCC 12472 without affecting bacterial growth. The minimal inhibitory concentration (MIC) of ketoprofen, piroxicam and indomethacin was determined against A. baumannii strains ATCC 17978, ATCC 19606, A1, A11 and A27 by the microbroth dilution method. The MICs of ketoprofen against tested isolates were 0.7-6.25 mg ml-1 , piroxicam MICs were 1.25-2.5mg ml-1 , and indomethacin MICs were 3.12-12.5mg ml-1 . Those compounds significantly inhibited QS-associated virulence factors such as biofilm formation, and surface motility, as well as, significantly increased bacterial tolerance to oxidative stress without affecting bacterial growth. On the molecular level, the three compounds significantly inhibited the transcription of QS regulatory genes abaI/abaR and biofilm-regulated genes cusD and pgaB. Molecular docking analysis revealed the potent binding affinity of the three compounds with AbaI via hydrogen and/or hydrophobic bonds. These results indicate that NSAIDs, ketoprofen, piroxicam and indomethacin, could be potential inhibitors of the QS and could suppress the QS-related virulence factors of A. baumannii. Ketoprofen, piroxicam and indomethacin could provide promising implications and strategies for combating the virulence and pathogenesis of A. baumannii.

Experimental and in silico investigation on the interaction of indomethacin with bovine serum albumin: Effect of sodium dodecyl sulfate surfactant monomers on the binding

This paper describes the interaction of indomethacin with bovine serum albumin (BSA) using experimental and computational approaches. Inner filter effect (IFE) had significantly impacted the fluorescence results as uncorrected data shown the considerable involvement of tyrosine residues which became very low after correcting the data with the former, thus, only the tryptophan residues were found to be associated with fluorescence quenching. Analyses of the corrected data suggested that there was 1:1 strong binding between BSA and indomethacin that taken place via static quenching mechanism. The binding was exothermic in nature and energetically favorable with ordering of the system. The binding of indomethacin remained almost unchanged in presence of low amount of sodium dodecyl sulfate (SDS) surfactants monomers while it significantly diminished when the concentration of SDS monomers increased. Low concentrations of indomethacin didn’t affect the secondary structure of BSA, however, high concentrations led to the partial unfolding of the protein. Molecular docking along with molecular dynamics simulation results explained that this drug molecule occupied new binding site with strong binding affinity of −8.35 kcal/mol, which is oriented in the interface between domain 1 and domain 2. The interaction was facilitated by both hydrophobic forces as well as hydrogen bonding.

Effect of indomethacin on the oxidative stress of Cyprinus carpio using of biomarkers

In recent years, drugs have been identified as newly emerging and threatening the aquatic and terrestrial environments. For this purpose, in this study, the effect of indomethacin on oxidative stress in Cyprinus carpio was investigated. In order to determine the LC50, the fishes were exposed to various concentrations of indomethacin and daily losses were recorded during hours. Then the lethal toxicity (LC50) of indomethacin was obtained for carp (328.49 mg/l). In the next step, the fishes were exposed to various concentrations of indometacin in five treatments including T1 (control), T2 (0.01 mg), T3 (0.02 mg), T4 (0.05 mg), T5 (0.1 mg) for 28 days. At the end of the experiment, antioxidant enzymes such as catalase (CAT), glutathione peroxidase(GPX) and malondialdehyde(MDA) were measured in the liver and gill tissues treated fishes. The results showed that GPX enzyme activity in gill and liver tissue in all treatments increased significantly in compared to the control treatment (p<0.05). MDA levels in liver tissue in T4 and T5 groups and gill tissue in T3 and T4 groups showed a significant increase in compared to the control group (p<0.05). Also, CAT activity in liver and gill tissue in T4 and T5 groups was increased significantly in compared to the control group (p<0.05). The results of this study demonstrated that keeping indomethacin in an aqueous environment can influenc the antioxidant status and health of the fishes.

The Protective Effect of 5-Aminosalicylic Acid Against Non-Steroidal Anti-Inflammatory Drug-Induced Injury Through Free Radical Scavenging in Small Intestinal Epithelial Cells.

Background and objectives: Non-steroidal anti-inflammatory drugs (NSAIDs) have been among the major causes of small intestinal injury in clinical practice. As such, the current study investigated the protective effect of 5-aminosalicylic acid (5-ASA) against an NSAID-induced small intestinal injury. Materials and Methods: IEC-6 cells were treated with various concentrations of indomethacin with or without 5-ASA in a serum-free medium, after which an 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Dromide (MTT) assay, a cell apoptosis assay, a caspase-3 activity assay, a reactive oxygen species (ROS) content and Superoxide dismutase 2 (SOD2) activity measurement, a Western blotting for occludin and zonula occludens-1 (ZO-1) and a wound healing assay were conducted. Results: 5-ASA ameliorated indomethacin-induced cell apoptosis and an increase in the intracellular ROS content while augmenting the indomethacin-induced suppression of SOD2 activity in IEC-6 cells. Moreover, 5-ASA reversed the indomethacin-induced attenuation of occludin and ZO-1 expression and promoted faster wound healing effects in IEC-6 cells following an indomethacin-induced injury. Conclusions: Our results suggested that 5-ASA protects small intestinal cells against an NSAID-induced small intestinal injury by scavenging free radicals. Therefore, 5-ASA could be a potential treatment for an NSAID-induced small intestinal injury.

Fmoc-diphenylalanine-based hydrogels as a potential carrier for drug delivery

Abstract Self-assembled hydrogels from 9-fluorenylmethoxycarbonyl-modified diphenylalanine (Fmoc-FF) peptides were evaluated as potential vehicles for drug delivery. During self-assembly of Fmoc-FF, high concentrations of indomethacin (IDM) drugs were shown to be incorporated into the hydrogels. The β-sheet arrangement of peptides was found to be predominant in Fmoc-FF–IDM hydrogels regardless of the IDM content. The release mechanism for IDM displayed a biphasic profile comprising an initial hydrogel erosion-dominated stage followed by the diffusion-controlled stage. Small amounts of polyamidoamine dendrimer (PAMAM) added to the hydrogel (Fmoc-FF 0.5%–IDM 0.5%–PAMAM 0.03%) resulted in a more prolonged IDM release compared with Fmoc-FF 0.5%–IDM 0.5% hydrogel. Furthermore, these IDM-loaded hydrogels demonstrated excellent thixotropic response and injectability, which make them suitable candidates for use as injectable self-healing matrices for drug delivery.

Противовоспалительное и анальгетическое действие нового сложноэфирного производного индометацина

Сложный эфир индометацина и ментола (ИМЛ) при введении внутрь в дозе 12,5 мг/кг оказывает профилактическое противовоспалительное действие на моделях острого экссудативного воспаления, вызванного каррагенином, гистамином и серотонином, и хронического пролиферативного воспаления у крыс. При введении за 12 ч до каррагенина антиэкссудативное действие ИМЛ превосходит эффект индометацина на 36 % (p &lt; 0,05), при введении ИМЛ за 8 ч до применения гистамина и серотонина — на 32 % (p &lt; 0,05). Максимальная концентрация индометацина, освобождаемого при гидролизе ИМЛ, создается в плазме крови крыс через (8 ± 0,3) ч, а максимальная концентрация индометацина — через (4 ± 0,5) ч. ИМЛ тормозит развитие грануляционной ткани слабее индометацина на 25,6 % (p &lt; 0,05), подавляет гипералгезию у мышей больше, чем индометацин на 16 % (p &lt; 0,05), не повреждает слизистую оболочку желудка крыс и создает в ней концентрацию индометацина в 3,2 – 5,3 раза более низкую, чем та, которая возникает при применении индометацина (p &lt; 0,05).

Effect of supersaturation on absorption of indomethacin and tadalafil in a single pass intestinal perfusion rat model, in the absence and presence of a precipitation inhibitor

The effect of the degree of supersaturation (DS) on absorption of the model drugs indomethacin and tadalafil was elucidated in a single-pass intestinal perfusion (SPIP) model in rats. In addition, the performance of the precipitation inhibitor (PI) hydroxypropylmethylcellulose (HPMC) was evaluated when added at a concentration of 0.1% (w/v) to fasted state simulated intestinal fluid (FaSSIF and FaSSIFHPMC) used as perfusion medium.A supersaturated state was created by a solvent shift method where indomethacin or tadalafil dissolved in dimethyl sulfoxide (DMSO) were administered to a segment of the small intestine, which subsequently was perfused with FaSSIF or FaSSIFHPMC. The perfusate was collected for 60 min, and for one group of rats dosed with 30 mg tadalafil, for 120 min. Blood samples were drawn every 15 min. The solubility of indomethacin and tadalafil in the perfusate was determined. The DS of each drug in the perfusate was calculated by dividing the concentration in the perfusate at selected time points with the solubility. The DS was above one for all timepoints for both drugs, thus showing supersaturation during the time of perfusion. For indomethacin, no improvement of the DS was seen when perfusing with FaSSIFHPMC, compared to FaSSIF. For tadalafil, a higher DS was achieved when perfusing with FaSSIFHPMC compared to FaSSIF. Perfusing the drugs with FaSSIFHPMC resulted in a significantly lower area under the curve (AUC0-60 min) for plasma concentrations of indomethacin, and no increase in the AUC0-60 min of plasma concentrations of tadalafil compared to perfusion with FaSSIF.The importance of simultaneously estimating the intraluminal DS and absorption of a drug was demonstrated by the SPIP model in the present study. Further, the study highlights the discrepancy between optimal in vitro supersaturation, intraluminal supersaturation and in vivo performance of two poorly soluble drugs, and further emphasizes the importance of optimization of in vitro methods in order to predict in vivo supersaturation and precipitation of drugs.

Induction of cystic ovarian follicles (COFs) in cattle by using an intrafollicular injection of indomethacin.

The aim of this study was to establish a model to induce cystic ovarian follicles (COFs) in cattle using the cyclooxygenase inhibitor, indomethacin. Eighteen Holstein-Frisian cattle were synchronized with prostaglandin F2alpha (PGF2α) and gonadotropin-releasing hormone (GnRH). Ultrasound-guided transvaginal intrafollicular injections were performed in 23 preovulatory follicles with different concentrations of indomethacin 16 h after GnRH administration. An injection of 0.2 ml 35 µM indomethacin solution (resulting in a final concentration of 8 µg/ml in the follicular fluid) was the minimal dosage leading to COF formation. The induced COFs reached a maximum mean diameter of 36.9 ± 4.5 mm eleven days after injection. The estrous cycle was extended to 25–39 days. Luteinization was first observed 4 days after injection, accompanied by a slight increase in plasma progesterone concentration. The bioactivity of indomethacin was demonstrated by the decrease of prostaglandin E2 in the follicular fluid of three animals. The method presented here is minimally invasive and allows for the generation of defined COFs for further investigations.

TOXICOLOGICAL EVALUATION OF INDOMETHACIN AS A RISK FACTOR FOR WORKERS’ HEALTH

Introduction. The toxicity of indomethacin was studied for its hygienic regulation. Material and methods. The toxic properties of indomethacin in the experiments on out-bred and linear mice, rats, Guinea pigs and rabbits contained in standard vivarium conditions and quarantined have been studied. In the experiments, various modes (single, repeated, chronic) and ways of exposure (intragastric, inhalation, epicutaneous) were modeled. The average lethal dose (LD50) of Indomethacin and the threshold of a single acute action (Limac) were determined; irritant effect on the skin and mucous membranes, cumulative and allergenic activity were revealed. In subacute and chronic intake to the body, the main target organs were determined on the based of the results of biochemical and hematological studies. Results. DL50 for male rats, females and male mice, when introduced into the stomach, were have been established to be 20, 15 and 25.6 mg/kg respectively. It refers to the substances of hazard class 2. DL50 in the intraperitoneal introduction for the rats accounted for 13.8 mg/kg, for Guinea pigs - 500 mg/kg. The clinical picture of acute poisoning in mice and rats was characterized by low mobility, decreasing breathing, ataxia, muscle relaxation, anorexia, diarrhea, ulceration with the perforation of the intestines, and the death on the 2-4th days after the poisoning. In the experiments on Guinea pigs, the ulcerogenic effect was not detected. Local irritant effect on the skin and mucous membranes of the eyes was not revealed. It has a marked skin-resorptive action causing ulcerogenic effect and the death of the animals after 6 applications. The introduction of verospiron to the rats in a dose of 25 mg/kg simultaneously with the application of indomethacin ointment on the skin prevented the ulcer development in the gastrointestinal tract and the death of the animals. No sensitizing effect was detected. It has an average cumulative ability: the cumulation coefficient amounted to 2.6. In a subacute experiment, there was a decrease in the body temperature and summation-threshold index, an increase in the vertical motor activity and a threshold of pain sensitivity. During the study of blood serum an increase in AcAt activity, a rise of chlorides in the blood serum and their decrease in the urine, and an increase in the number of erythrocytes and hemoglobin in peripheral blood were revealed. In the pathomorphological study, there was an increase in the coefficients of liver mass and ulceration of the stomach and intestines. The threshold of acute inhalation action accounted for 0.52 mg/m3 (by the reduction of the summation-threshold index and the content of sodium and chlorides in the urine). Conclusion. The maximum permissible concentration of indomethacin in the air of the working area was of 0.05 mg/m3 with the mark “special protection of the skin and eyes”, hazard class 1, aerosol.

Differential effects of dexamethasone and indomethacin on Tenon's capsule fibroblasts: Implications for glaucoma surgery

Dysregulated wound healing and subsequent fibrosis represents the most common cause of failure in glaucoma filtration surgery. Primary means to prevent this outcome are the anti-metabolite surgical adjuvants, however, topical corticosteroids are commonly used postoperatively to permit further control of wound healing and development of the filtration bleb. Unfortunately, they carry important side effects such as raised intraocular pressure, cataract and increased infection risk. Non-steroidal anti-inflammatory drugs (NSAIDs) show promising results in clinical trials as an alternative wound modulatory drug. NSAIDs exhibit non-inferiority to steroids in terms of post-operative intraocular pressure control and secondary IOP lowering interventions, however there is little known about the differing effects these drugs exert on human Tenon's capsule fibroblast (HTCF) mediated wound healing. The purpose of this study was to assess the individual effects of dexamethasone and indomethacin on the extracellular matrix modifying actions of HTCFs in vitro. To this end, HTCFs were cultured in 3D collagen matrices as well as in 2D monolayers and exposed to clinically relevant concentrations of dexamethasone or indomethacin for up to seven days. HTCF-mediated wound healing functions were assayed through collagen matrix contraction, extracellular matrix morphology, estimation of HCTF proliferation and differentiation into myofibroblasts within the collagen matrices, as well as western blot. Both drugs significantly reduced HTCF-mediated collagen contraction relative to control however there was a significant trend towards greater inhibition with indomethacin exposure compared to dexamethasone. Indomethacin exposure significantly reduced HTCF-mediated collagen remodelling activity compared vehicle control, whereas dexamethasone was unable to reduce remodelling activity at any of the studied exposures. Both drugs reduced myofibroblast differentiation, however indomethacin alone demonstrated an inhibitory effect on final cell number relative to control whereas dexamethasone had no significant effect at any studied exposure. These findings demonstrate that both steroidal and NSAID treatment can mitigate HTCF-mediated collagen contraction and αSMA expression. However, NSAIDs may function to better impede HTCF proliferation and remodelling activity. Taken in the context of previous glaucoma surgical trials, NSAIDs appear to be a viable alternative to steroids for post-operative wound modulation.

The effect of non-steroidal anti-inflammatory drugs on osteosarcoma cells.

Osteosarcoma (OS), an aggressive malignancy, is the most common primary bone tumor in children. Nonsteroidal anti-inflammatory drugs (NSAIDs) are widely used to reduce pain and inflammation. NSAIDs have shown to be toxic to certain malignancies such as colorectal, breast, and pancreatic cancers, but are not well-studied in OS. The purpose of this study is to assess whether ketorolac induces apoptosis in OS cells, compare this to indomethacin, which has been shown to inhibit OS proliferation, and explore the underlying mechanism. A rat OS cell line (UMR-108) was exposed to various concentrations of ketorolac and indomethacin. Cell viability, cytotoxicity, apoptosis induction, DNA fragmentation and the expression of apoptosis-related markers were examined by MTT assay, colony formation assay, flow cytometry, agarose gel electrophoresis, and Western blot respectively. The results indicated that ketorolac and indomethacin could induce apoptosis of rat OS cells in a dose- and time-dependent manner. Apoptosis was confirmed by cell morphology and annexin positivity. The molecular data showed that NSAIDs affected expression of Bcl-2, survivin, and Poly (ADP-ribose) polymerase-1 (PARP). These findings demonstrated that NSAIDs induced apoptosis in rat OS cells in vitro. Further research focusing on the potential cytotoxicity of NSAIDs in vivo is needed.

An investigation into the possibility of molecular inclusion complexation of indomethacin with starch by the alkaline method

Starch systems containing indomethacin were prepared by the alkaline method and characterized in order to investigate the potential of this method to produce amylose molecular inclusion complexes as targeted and controlled drug delivery carriers. Three types of starches with different amylose/amylopectin composition were used to prepare the systems. Their structure was probed by X-ray diffraction (XRD) and Raman spectroscopy, whereas their morphology was investigated by Optical microscopy and Confocal laser scanning microscopy (CLSM). Raman spectroscopy revealed the presence of p-chlorobenzoic acid, the degradation product of the alkaline hydrolysis of indomethacin in the matrix of the starch systems. The results were confirmed by XRD measurements and the degree of crystallinity of the starch systems ranged from 35 to 90%. In vitro release testing of indomethacin from the starch systems in the simulated gastric and small intestine fluid showed very low concentrations of indomethacin. CLSM micrographs and optical microscopic examination revealed the presence mostly of the p-chlorobenzoic acid, in the matrix of the starch systems, rather than of indomethacin. The results, in general, indicated that the p-chlorobenzoic acid molecules were either included in the amylose helices or they have been entrapped between the helices of the complexed amylose whereas the few indomethacin molecules present in the starch matrix probably were also mechanically entrapped.

Controlled Release Binary and Ternary HPMC-HPC-Gelucire® 50/13 Hybrids Based Solid Dispersions of Indomethacin: In vitro Evaluation and In vivo Anti-inflammatory Studies

Aim: The aim of this study is to formulate and evaluate solid dispersions of indomethacin based on Gelucire 50/13, Hydroxylpropyl methylcellulose and/or Hydroxylpropyl cellulose hybrid binary or ternary system, with a view of enhancing oral bioavailability, sustaining drug release with optimum anti-inflammatory activity and minimal side effects. Methodology: Solid dispersions were prepared by solvent-evaporation method at varying polymer ratios (with HPMC and Gelucire 50/13 at 1:1, 2:1, 3:1, 4:1, 1:2, 1:3, 1:4 and HPMC, HPC and Gelucire 50/13 at 1:1:1) with a constant concentration of indomethacin (100 mg). The formulations were characterized in terms of morphology, stability and drug content. The release profiles of indomethacin from the solid dispersions were examined in vitro using two different media i.e. SGF and SIF, without enzymes. The anti-inflammatory properties were evaluated. Results: The formulated solid dispersions were stable and almost spherical to irregularly-shaped. The drug content of the formulations was accepted according to the Pharmacopoeial limit. In vitro dissolution studies showed an increased dissolution rate at pH 7.4 compared to pH 1.2 for the various batches with solid dispersions showing a faster and sustained dissolution rate than the pure crystalline drug. The percentage inhibitions of edema produced by the formulations in the in vivo studies were significantly higher and sustained than the pure drug. Conclusion: Solid dispersions prepared with these biodegradable polymer hybrids, HPMC/HPC/Gelucire® 50/13, showed promising potential in clinical practice for enhancing the delivery of indomethacin.

Effect of CYP2C9 Polymorphisms on the Pharmacokinetics of Indomethacin During Pregnancy.

Cytochrome P450 (CYP)2C9 catalyzes the biotransformation of indomethacin to its inactive metabolite O-desmethylindomethacin (DMI). The aim of this work was to determine the effect of CYP2C9 polymorphisms on indomethacin metabolism in pregnant women. Plasma concentrations of indomethacin and DMI at steady state were analyzed with a validated LC-MS/MS method. DNA was isolated from subject blood and buccal smear samples. Subjects were grouped by genotype for comparisons of pharmacokinetic parameters. For subjects with the *1/*2 genotype, the meansteady-state apparent oral clearance (CL/Fss)of indomethacin was 13.5 ± 7.7 L/h (n = 4) and the mean metabolic ratio (AUCDMI/AUCindomethacin) was 0.291 ± 0.133. For subjects with the *1/*1 genotype, these values were 12.4 ± 2.7 L/h and 0.221 ± 0.078, respectively (n = 14). Of note, we identified one subject who was a carrier of both the *3 and *4 alleles, resulting in an amino acid change (I359P) which has not been reported previously. This subject had a metabolic ratio of 0.390 and a CL/Fss of indomethacin (24.3 L/h) that was nearly double the wild-type clearance. Although our results are limited by sample size and are not statistically significant, these data suggest that certain genetic polymorphisms of CYP2C9 may lead to an increased metabolic ratio and an increase in the clearance of indomethacin. More data are needed to assess the impact of CYP2C9 genotype on the effectiveness of indomethacin as a tocolytic agent.

Development of micellar electrokinetic chromatography method for the determination of three defined impurities in indomethacin.

A micellar electrokinetic chromatography method for the determination of indomethacin impurities (4-chlorobenzoic acid, 5-methoxy-2-methyl-3-indoleacetic acid, and 3,4-dichloroindomethacin) has been developed. A 64.5/56-cm fused silica 50μmid capillary with extended light path (150μmid) detection window was used. Internal standard was 1-naphthylacetic acid. The analytes were separated at 30kV with DAD detection at 224nm. A central composite face-centered design was applied for the optimization of the separation conditions. The effect of SDS concentration, content of methanol, concentration of phosphate buffer, and pHof the buffer were studied at three levels. The optimized background electrolyte was 20mmol/L phosphate buffer (pH7.57) containing 58mmol/L SDS and 0% MeOH. Sufficient resolution of all compounds with Rs≥3.5 was achieved within 10min. The method was validated for a range of 1.25-80μg/mL of each impurity corresponding to 0.05-3.2% relative to the concentration of indomethacin (2.5mg/mL). The calibration curves were rectilinear with correlation coefficients r2 exceeding 0.9994. The lower limit of quantification was 0.05% or 1.25μg/mL that complies with the reporting limits regarding the ICH Q3A guideline. The method was applied to purity assay of indomethacin in both bulk drug and gel.