Ex-vivo Study Research Articles

Formulation and evaluation of salicylic acid hydrogel based on hydrotropic solubility enhancement technique

Background and objective: Salicylic acid (SA) has keratolytic activities and it is used topically to treat dandruff and seborrheic dermatitis. The hydrotropic phenomenon in pharmaceutical preparations is utilized to enhance the solubility of water-insoluble drug molecules that promoted to prepare the salicylic acid as a hair gel for local effect and it can extend its keratolytic action for a longer period compared to the salicylic acid solution. Methods: Preformulation study was performed to exclude any unwanted chemical interaction between SA and excipients using Fourier-transform infrared spectroscopy (FTIR). The solubility of SA was determined separately in sodium acetate and sodium citrate solutions at a concentration of 1, 3, and 5 %w/v, using distilled water as a solvent. An optimum gel formulation was developed and it was used to prepare the SA gel formulation. Characterizations were performed in terms of physical appearance, viscosity, pH, and spreadability, in-vitro studies were performed in physiological pH, and ex-vivo diffusion studies were performed utilizing rats’ skin. Results: FTIR did not show any chemical interaction between the drug and sodium citrate. The hydrotropic solution of sodium citrate with a concentration of 5% w/v increased the solubility of SA by 28 folds, while the sodium acetate solutions with a concentration of 5% w/v increased the solubility of SA by 19 folds. The optimum gel formula (F1) with a drug content of 97% showed a slow dissolution rate and minimum diffusion through the skin. Conclusion: The hydrotropic solubilization technique significantly influenced the solubilization of salicylic acid in the water and the highest solubility rate was achieved from 5% w/v sodium citrate solution. The formulated hydrogels using carbopol 971P as gelling agent decreased the diffusion rate of SA.

Exploration of Potential Anti-inflammatory cum Anti-Rheumatoid-arthritis Phyto-molecule through Integrated Green Approach: Network Pharmacology, Molecular Docking, Molecular dynamics, In-vitro and Ex-vivo study.

Rheumatoid arthritis (RA) and associated inflammatory complications are the most prevalent illnesses and can turn into fatal conditions if left untreated. Allopathic medicine is not satisfactory for curing RA. Scientific literature reports reveal that several phyto-compounds viz. flavonoids, saponins, and terpenoids, can heal joints and organs from auto-inflammatory rheumatoid arthritis and pain. Gene ontology, gene network analysis, molecular clustering, and literature review were used to optimise RA-specific highly expressed genes. In-silico molecular docking was performed to short-out potential phytomolecules (Neohesperidin dihydrochalcone (NHDC)) from 1000 datasets-library against RA and validate using MD simulation running at 100 ns. In-vitro anti-inflammatory assays of NHDC inhibited egg-albumin denaturation, IC50 of 47.739 ± 0.51 µg/ml. The ex-vivo MTT assay with NHDC rendered 67.209% inhibition at 100 µM against fd-FLS-cells. NHDC downregulated pro-inflammatory cytokine IL-17A production by 61.11% and 50% at 300 and 200 μM, respectively. Thus, this Studies recommend that NHDC may be highlighted as a novel multi-target PADI4 and JAK3 inhibitor with better efficacy and minimal toxicity in RA warranted to In-Vivo and clinical investigation. The current findings have uncovered remarkable genes and signalling pathways linked to RA, which could enhance our existing comprehension of the molecular mechanisms that drive its development and progression.

Revitalizing antimicrobial strategies: paromomycin and dicoumarol repurposed as potent inhibitors of M.tb's replication machinery via targeting the vital protein DnaN

Despite the WHO's recommended treatment regimen, challenges such as patient non-adherence and the emergence of drug-resistant strains persist with TB claiming 1.5 million lives annually. In this study, we propose a novel approach by targeting the DNA replication-machinery of M.tb through drug-repurposing. The β2-Sliding clamp (DnaN), a key component of this complex, emerges as a potentially vulnerable target due to its distinct structure and lack of human homology. Leveraging TBVS, we screened ∼2600 FDA-approved drugs, identifying five potential DnaN inhibitors, by employing computational studies, including molecular-docking and molecular-dynamics simulations. The shortlisted compounds were subjected to in-vitro and ex-vivo studies, evaluating their anti-mycobacterial potential. Notably, Dicoumarol, Paromomycin, and Posaconazole exhibited anti-TB properties with a MIC value of 6.25, 3.12 and 50 μg/ml respectively, with Dicoumarol and Paromomycin, demonstrating efficacy in reducing live M.tb within macrophages. Biophysical analyses confirmed the strong binding-affinity of DnaNdrug complexes, validating our in-silico predictions. Moreover, RNA-Seq data revealed the upregulation of proteins associated with DNA repair and replication mechanisms upon Paromomycin treatment.This study explores repurposing FDA-approved drugs to target TB via the mycobacterial DNA replication-machinery, showing promising inhibitory effects. It sets the stage for further clinical research, demonstrating the potential of drug repurposing in TB treatment.

Tailoring of spanlastics for promoting transdermal delivery of irbesartan: In vitro characterization, ex vivo permeation and in vivo assessment

Irbesartan (IRB), a selective AT1 subtype angiotensin II receptor blocker used for management of hypertension, exhibiting low and variable oral bioavailability because of its poor aqueous solubility. Transdermal delivery is increasingly being used to deliver drugs since it avoids the drawbacks associated with oral delivery. The purpose of the current study was to develop a transdermal gel of IRB through encapsulation of the drug into spanlastics vesicles containing penetration enhancers to enhance transdermal permeation and sustain IRB release. By ethanol injection method, ten IRB loaded spanlastics (IRB-SPs) were prepared; Span 60 was used as the main vesicle component and the used edge activator (EA) was Tween 80. The prepared formulations were studied in regards to entrapment efficiency (EE%), particles size (PS), zeta potential (ZP), polydispersity index (PDI), and in vitro release study. The selected formula (SF) which comprised of Span 60 and Tween 80 at weight ratio of 1:1 and Labrafil as a penetration enhancer, exhibited EE% of 90.06 ± 1.44 %, PS of 311.6 ± 2.45 nm, PDI of 0.536 ± 0.044, and the % drug released after 8 h is 58.2 ± 1.87 %. Transmission electron microscopy confirmed the spherical shape of the prepared spanlastics. Differential scanning calorimetry (DSC) confirmed the drug encapsulation within the spanlastics. The SF and pure drug were incorporated into HPMC K4M based hydrogel (2.5 % w/v). Histopathological and ex-vivo skin permeation studies of hydrogel were assessed as well as its in vitro characteristics. Skin permeation was enhanced by 3.5 folds with the SF loaded gel compared to the control gel. The pharmacokinetic study revealed better bioavailability of the SF-gel by 1.72 and 2.53 fold as compared to the oral suspension and the control gel, respectively. Furthermore, the pharmacodynamic assessment was carried out on systolic blood pressure (SBP) of various IRB formulations in dexamethasone induced hypertensive rat; the antihypertensive effect of SF loaded transdermal gel showed significantly (p < 0.001) much greater blood pressure reduction than the market product's oral suspension. Finally, spanlastics-based gels could be an excellent way to improve IRB transdermal delivery.

Revolutionizing treatment for topical fungal infections: evaluating penetration-enhancer-containing vesicles as a fluconazole delivery system: Ex-vivo and in-vivo dermal testing

Fungal infections pose a significant challenge in numerous developing nations and worldwide, necessitating urgent solutions. Oral administration of antifungal medications often leads to severe adverse reactions. Hence, employing topical delivery systems is preferred to ensure efficient dermal delivery of antifungal agents while minimizing side effects. Furthermore, the incorporation of penetration enhancers into nanocarriers loaded with antifungal agents has demonstrated enhanced efficacy in combating mycotic infections. Consequently, ultra-deformable penetration enhancer-containing vesicles (PEVs) were developed to explore this promising approach. In this study, Labrasol® and Transcutol® were used as penetration enhancers in formulating ultra-deformable PEVs containing the antifungal agent Fluconazole (FCZ). The PEVs underwent comprehensive characterization, including measurements of particle size (PS), charge, and entrapment efficiency (EE%). The results revealed that the size of tested PEVs ranged from 100 to 762 nm. All particles exhibited a negative charge, with a minimum zeta potential (ZP) of −38.26 mV, and an intermediate entrapment efficiency (EE%) that reached approximately 40%w/w. Ex-vivo studies demonstrated the ability of PEVs to deliver FCZ to the dermis while minimizing transdermal delivery. The selected formula was tested in-vivo using candidiasis-induced rat model and showed a superiority in its antifungal effect against Candida Albicans compared to the drug control. Stability studies were executed for the selected formula, and revealed good stability shown by the insignificant change in the PS, ZP& EE% over a six-month period.

Antipsoriatic Effect of Silymarin NLCs Based Gel: In Vitro and In Vivo Activity.

Psoriasis is a chronic inflammatory disorder affecting over 100 million people, requires long-term therapy. Current treatments offer only symptomatic relief. However, phytoconstituents-based therapies like Silymarin (SLM) have shown promising effects. The study aims to develop, optimize, and evaluate a novel stable SLM NLC gel to improve anti-psoriatic activity by enhancing its permeability and retention into the dermal layer. SLM NLC formulation was prepared and optimized using 32 full factorial designs. The formulation was evaluated for the particle size, PDI, zeta potential, and % entrapment efficiency, evaluated by Transmission electron microscopy and thermal analysis. The freeze dried and prepared NLC-loaded gel was evaluated for physicochemical parameters, ex-vivo, and in-vivo studies. SLM-loaded NLC shows 624nm particle size, 0.41 PDI, 92.95% entrapment efficiency, and -31.6mV zeta potential. The sphere form of NLCs was confirmed using TEM. Controlled drug release was observed in ex vivo studies, low PASI score compared to disease control. Further, the levels of IL-6, TNF-α, and NF-κB were also reduced. The results are supported by histopathology showing minimal parakeratosis indicated in the SLM NLC-treated group. Prepared NLC-based shows enhance topical penetration and decrease the thickness of psoriatic plaques in the in vivo study.

In vitro drug release profiling of Sirolimus polymeric microparticles coated long-acting stents

In the realm of arterial disease interventions, drug-eluting stents (DES) have become a vital therapeutic choice in preventing atherosclerotic plaque formation and restenosis and facilitating vessel healing. Sirolimus-encapsulated poly Lactic-co-Glycolic acid (PLGA) Microparticles (MPs) were developed using solvent evaporation. MPs were freeze-dried with a cryoprotectant and coated on the stent surface using an efficient and reproducible nitrogen-assisted spray coating technique. The MPs displayed a uniform distribution particle size of 4.38 ± 1.1 μm, span value of 0.88 ± 0.02, coating mass transfer efficiency of 13.45 ± 1.1 % on the stent, and a coating time of ≤ 2 min per stent. Post sterilization, the particle size and morphology of the coated stents remained unchanged. Accelerated in vitro drug release profiles were evaluated under different conditions, indicating significant influences based on dissolution methods ranging from 28.2 %±4.3 %, 42.5 %±5.3 %, 76.6 %±4.7 %, and 84.25 %±3.1 % for dialysis bag (DB), vessel simulating flow-through cell (vFTC), flow-through cell (FTC), and sample and separate (SS) technique respectively for 48 h. The drug release mechanism from the coated stents is governed by the combination of the Korsmeyer Peppas and Higuchi models. The developed dissolution method exhibited discriminative effectiveness when evaluated with critical formulation attributes and process parameter variations. The 48 h accelerated drug release studies correlated well with the 6-month real-time release rate with an R2 value of 0.9142 and Pearson’s R2 of 0.9561. Ex-vivo studies demonstrated the permeation of MPs into artery tissues. Stability studies confirmed that MPs coated stents maintained desired properties at 4 °C and 30 °C/65 % RH for 6 months. Overall, these findings contribute to advancing stent technology, suggesting the potential for improvement of arterial interventions and enhanced patient outcomes.

Mitochondrial bioenergetic dysfunction linked to myxomatous mitral valve degeneration explored by PBMCs metabolism analysis

Impaired mitochondria cause an impressive decrease in ATP production becoming a common condition of cardiovascular diseases. Myxomatous mitral valve disease (MMVD) is characterized by mitochondrial dysfunction. By a non-invasive procedure of metabolism analysis on peripheral blood mononuclear cells, we exploit ex-vivo studies that directly constitute a translational approach to evaluate the cell bioenergetics. Cell ATP production decreased in the presence of MMVD, whereas glycolysis was unaffected. In MMVD, the mitochondrial activity underwent a significant reduction of basal respiration, maximal respiration, and ATP production. Our results depicted a pathological condition of MMVD characterized by cell metabolism deprived of mitochondrial energy support.

Antibiofilm effect of different concentrations of silver nanoparticles combined with calcium hydroxide against Enterococcus faecalis biofilm: An exvivo study.

This study aimed to evaluate the antibiofilm activity of different concentrations of silver nanoparticles (AgNPs) in combination with calcium hydroxide [Ca(OH)2] against Enterococcus faecalis biofilm. On an E. faecalis biofilm on dentin discs, the following medicaments were applied for 7 days (n = 13/group): 0.005% AgNPs+Ca(OH)2, 0.01% AgNPs+Ca(OH)2, 0.02% AgNPs + Ca(OH)2, Ca(OH)2 and saline/control. Specimens were stained with LIVE/DEAD® BacLight™ dye and analysed with confocal laser scanning microscopy. Proportion of dead bacteria was calculated and analysed. There was a significant reduction in E. faecalis biofilm in all medicament groups (43.5%, 49.1%, 69.1%, 48.7%) respectively, compared with control group (2.54%) (p < 0.001). The 0.02% AgNPs + Ca(OH)2 group demonstrated the most significantly superior antibiofilm effect, with no significant difference between remaining groups. In conclusion, combining 0.02% AgNPs enhanced the antibiofilm effect of Ca(OH)2 on E. faecalis biofilm compared with lower AgNPs concentrations.

Development a novel nano-platform for Thrombolysis acceleration by Thrombin sensitive polymer-peptide hybrid nancapsules

According to the importance of time in treatment of thrombosis disorders, faster than current treatments are required. For the first time, this research discloses a novel strategy for rapid dissolution of blood clots by encapsulation of a fibrinolytic (Reteplase) into a Thrombin sensitive shell formed by polymerization of acrylamide monomers and bisacryloylated peptide as crosslinker. Degradability of the peptide units in exposure to Thrombin, creates the Thrombin-sensitive Reteplase nanocapsules (TSRNPs) as a triggered release system. Accelerated thrombolysis was achieved by combining three approaches including: deep penetration of TSRNPs into the blood clots, changing the clot dissolution mechanism by altering the distribution pattern of TSRNPs to 3D intra-clot distribution (based on the distributed intra-clot thrombolysis (DIT) model) instead of peripheral and unidirectional distribution of unencapsulated fibrinolytics and, enzyme-stimulated release of the fibrinolytic. Ex-vivo study was carried out by an occluded tube model that mimics in-vivo brain stroke as an emergency situation where faster treatment in short time is a golden key. In in vivo, efficacy of the developed formulation was confirmed by PET scan and laser Doppler flowmetry (LDF). As the most important achievements, 40.0 ± 0.7 (n = 3) % and 37.0 ± 0.4 (n = 3) % reduction in the thrombolysis time (faster reperfusion) were observed by ex-vivo and in-vivo experiments, respectively. Higher blood flow and larger digestion mass of clot at similar times in comparison to non-encapsulated Reteplase were observed that means more effective thrombolysis by the developed strategy.

A Comparison Study Using Microneedle, Transdermal Gel, and Tape Strip-Based Delivery of Caffeine Infused with Chemical Enhancers: Characterizations and Ex-vivo Study

A Comparison Study Using Microneedle, Transdermal Gel, and Tape Strip-Based Delivery of Caffeine Infused with Chemical Enhancers: Characterizations and Ex-vivo Study

Sustained release microneedle patch for pronounced systemic delivery of doxazosin mesylate

Introduction: Microneedle patch is one of the fascinating drug delivery approaches that offers low invasiveness and a painless physical application to enhance the delivery of micro and macro-molecules into the skin. Methods: Variable contents of chitosan and polyvinyl alcohol were used for the development of doxazosin mesylate containing sustained release microneedle patches via solvent casting technique. The prepared patches were evaluated for microscopic evaluation, mechanical strength, drug loading (%) and Fourier transform infrared spectroscopy (FTIR) etc. The skin penetration study was performed by using pig ear skin and results were captured through confocal microscopy. Ex-vivo release study and pharmacokinetic evaluation were also performed. Results: Sharp needle tips with a height of 600µm and a base of 200µm were confirmed through microscopic examination. Optimized formulation (SRF-6) exhibited loading of 92.11% doxazosin mesylate with appreciable strength up to 1.94N force. Ex-vivo release studies revealed 87.24% release within 48 hours. Moreover, the pharmacokinetic parameters in case of optimized patch formulation (SRF-6) were markedly improved i.e. MRT (19.46 h), AUC (57.12 μg.h /mL), Cmax (2.16 µg /mL), tmax (10.10 h) and t1/2 (6.32 h) as compared to commercially available tablet. Biocompatibility of the developed patches was validated from skin irritation studies. Conclusion: Results confirmed the successful fabrication of microneedle patch having sufficient strength and effective penetration ability into the skin to ensure controlled release of incorporated drug for the intended duration. It can be employed as an efficient carrier system for other therapeutics those are prone to bioavailability issues due to first pass effect after their oral administration.

A pharmacokinetic framework describing antibiotic adsorption to cardiopulmonary bypass devices.

Cardiopulmonary bypass (CPB) can alter pharmacokinetic (PK) parameters and the drug may adsorb to the CPB device, altering exposure. Cefazolin is a beta-lactam antibiotic used for antimicrobial prophylaxis during cardiac surgery supported by CPB. Adsorption of cefazolin could result in therapeutic failure. An exvivo study was undertaken using CPB devices primed and then dosed with cefazolin and samples were obtained over 1 hour of recirculation. Twelve experimental runs were conducted using different CPB device sizes (neonate, infant, child, and adult), device coatings (Xcoating™, Rheoparin®, PH.I.S.I.O), and priming solutions. The time course of saturable binding, using Bmax (binding capacity), Kd (dissociation constant), and T2off (half-time of dissociation), described cefazolin adsorption. Bmax estimates for the device sizes were neonate 40.0 mg (95% CI 24.3, 67.4), infant 48.6 mg (95% CI 5.97, 80.2), child 77.8 mg (95% CI 54.9, 103), and adult 196 mg (95% CI 191, 199). The Xcoating™ Kd estimate was 139 mg/L (95% CI 27.0, 283) and the T2off estimate was 98.4 min (95% CI 66.8, 129). The Rheoparin® and PH.I.S.I.O coatings had similar binding parameters with Kd and T2off estimates of 0.169 mg/L (95% CI 0.01, 1.99) and 4.94 min (95% CI 0.17, 59.4). The Bmax was small (< 10%) relative to a typical total patient dose during cardiac surgery supported by CPB. A dose adjustment for cefazolin based solely on drug adsorption is not required. This framework could be extended to other PK studies involving CPB.

Quantitative analysis of loxoprofen sodium loaded microspheres comprising pectin and its thiolated conjugates: In-vivo evaluation of their sustained release behavior

Continuous use of oral NSAIDs can damage mucosal membrane, which results in decreased bioavailability and non-compliance with the therapy. But the use of sustained release drug delivery systems might offer a solution. Objective was to synthesize mucoadhesive SR microspheres by using different combinations of pectin (PEC) and its thiolated derivative (T-PEC3100) for improved loxoprofen (LS) permeation. Thiolated pectin (T-PEC) was synthesized by the esterification method using thioglycolic acid. Thiolation was confirmed by thiol group quantification and charring point determination. Further characterization was done by Fourier Transform Infrared spectroscopy (FTIR), and Scanning electron microscopy (SEM). Ex-vivo mucoadhesion study was performed to confirm the improved characteristics. Microspheres (MS) were prepared using different ratios of PEC/T-PEC by solvent evaporation method and their particle size and surface morphology were evaluated. Mucus permeation study was carried out using the trans-well plate method. Sustained release behavior of prepared microspheres was investigated through the edema inhibition method in albino rats. T-PEC3100 was considered the optimum formulation for further evaluation and contained maximum thiol group content. FTIR spectra showed a characteristic peak of –SH and charring point was also changed considerably confirming the successful thiolation of PEC. SEM results showed spherical microspheres in the size range of 2–10 μm. Thiol-rich formulation of MS exhibited more than 80 % release after 12 h and maximum absorbable dose (MAD) was calculated as 400 μg % inhibition of edema in MS treated group was slowly attained initially but the reduction in inflammation was detected even after 24 h as compared to control group. Promising results from In-vivo edema inhibition study suggest the possible use of these thiolated MS in formulating sustained release formulation for arthritis.

Boosting of retinol activity using novel lecithin: Retinol acyltransferase inhibitors.

Lecithin:retinol acyltransferase (LRAT) is the main enzyme catalysing the esterification of retinol to retinyl esters and, hence, is of central importance for retinol homeostasis. As retinol, by its metabolite retinoic acid, stimulates fibroblasts to synthesize collagen fibres and inhibits collagen-degrading enzymes, the inhibition of LRAT presents an intriguing strategy for anti-ageing ingredients by increasing the available retinol in the skin. Here, we synthesized several derivatives mimicking natural lecithin substrates as potential LRAT inhibitors. By exploring various chemical modifications of the core scaffold consisting of a central amino acid and an N-terminal acylsulfone, we explored 10 different compounds in a biochemical assay, resulting in two compounds with IC50 values of 21.1 and 32.7 μM (compounds 1 and 2), along with a simpler arginine derivative with comparative inhibitory potency. Supported by computational methods, we investigated their structure-activity relationship, resulting in the identification of several structural features associated with high inhibition of LRAT. Ultimately, we conducted an exvivo study with human skin, demonstrating an increase of collagen III associated with a reduction of the skin ageing process. In conclusion, the reported compounds offer a promising approach to boost retinol abundance in human skin and might present a new generation of anti-ageing ingredients for cosmetic application.

Do the number of zirconia implants and the thickness of CBCT image reconstruction affect the detection of peri-implant bone defect? A diagnostic accuracy exvivo study.

To evaluate the influence of multiplanar reconstruction thickness on the detection of peri-implant bone defects with a standalone zirconia implant and compare it to when another implant is in the vicinity using cone-beam computed tomography (CBCT). Five dry human mandibles were used to create twenty implant sites in the second premolar and first molar regions. The OP300 Maxio was used to acquire CBCT images (90 kVp, 6.3 mA, 5 × 5 cm FOV, and 0.125 mm3 voxel size) before and after creating 3 mm peri-implant bone defects in the buccal aspect of the premolar region. Half of the scans featured a single zirconia implant in the premolar region, while the others had two implants in the premolar and molar regions. Three reconstruction thicknesses (0.125 mm, 1 mm, and 2 mm) were considered for the multiplanar reconstruction analyses. Five oral and maxillofacial radiologists assessed the detection of peri-implant bone defects using a 5-point scale. Diagnostic parameters were calculated and compared using Two-way ANOVA (α = .05). The studied factors showed no significant influence on the diagnosis of peri-implant bone defects (p > .05). Diagnostic performance was notably higher with a single implant, especially with a 2-mm reconstruction thickness (AUC = 0.88, sensitivity = 0.68, specificity = 0.94). Although the differences were not statistically significant, the results were more modest when two implants were present (AUC = 0.80, sensitivity = 0.58, specificity = 0.82). The presence of an adjacent zirconia implant and variations in reconstruction thickness did not influence the detection of 3 mm buccal peri-implant bone defects on CBCT images.

A novel inhibitory pathway of synovial inflammation exerted by glucocorticoids and tumor necrosis factor inhibitors via lymphocyte activation gene-3 up-regulation: an ex-vivo study.

To investigate the impact of glucocorticoids (GCs) and anti-rheumatic drugs on the lymphocyte activation gene-3 (LAG-3) and on programmed cell death-1 (PD-1) expression on synovial and peripheral cells ex-vivo. Synovial fluid mononuclear cells (SFMCs) from psoriatic arthritis (PsA, n = 26) and rheumatoid arthritis (RA, n = 13) patients, SFCs from osteoarthritis (OA, n = 5) patients and peripheral blood mononuclear cells (PBMCs) of healthy donors (n = 14) were co-cultured with GCs, glucocorticoid receptor antagonist RU486, methotrexate (MTX) and biologics. LAG-3 and PD-1 expressions on immune subsets were analyzed by flow cytometry. GCs in PsA inhibited SFMCs growth vs medium (2.3 ± 0.4X105 vs 5.3 ± 0.7X105, respectively, p < 0.01) and markedly upregulated CD14+LAG-3+ cells (11.7 ± 2.4% vs 0.8 ± 0.3%, p < 0.0001, respectively), but not CD3+LAG-3+ and CD14+PD-1+ cells. MTX had no effect on CD14+LAG-3+ cells (0.7 ± 0.3%). The TNFi inhibitors, infliximab (IFX) and etanercept, but not IL-12/23i, upregulated CD14+LAG-3+ cells vs medium (2.0 ± 0.6% and 1.6 ± 0.4% vs 0.5 ± 0.1%, p < 0.03, respectively). SFMCs growth inhibition in both PsA and RA correlated with CD14+LAG-3+ cell upregulation (r = 0.53, p = 0.03). RU486 inhibited GC-induced CD14+LAG-3+ cell up-regulation in a dose-dependent manner compared with GC alone (5µM 5.3 ± 1.2% and 50µM 1.3 ± 0.5% vs 7.0 ± 1.4%, p < 0.003), but had no significant effect on CD14+LAG-3+ cells co-cultured with IFX. GCs in healthy donors' PBMCs upregulated the immune subsets CD3+LAG-3+, CD14+LAG-3+ and CD14+PD-1+ cells. This study proposes a novel regulatory mechanism of GCs and of TNFi mediated by LAG-3 upregulation in synovial monocytes and PBMCs. LAG-3 modulation may be a promising target for development of novel therapies for inflammatory arthritis.

Novel anti-reflux ureteral skirt: proof of concept in a Yorkshire-Landrace pig model.

Stents with anti-reflux properties have been previously described in various studies but not widely adopted. We designed, constructed and tested a novel umbrella-looking anti-reflux skirt on a double J stent at the level of the ureteral orifice in a previous ex-vivo study. This study serves as a proof of concept of testing this prototype in a porcine model. 3 Yorkshire-Landrace pigs were put under general anaesthesia. Conventional bilateral retrograde stenting with white-colored double-J stents were performed. Deployment of anti-reflux skirt was performed unilaterally. Capsaicin mixed with methylene blue and saline were instilled in the bladder. Both stents were removed and methylene blue stain were inspected to assess the level of reflux. Bilateral stenting was performed again and stents/skirt prototype were left in-situ for 1 week. The procedure was repeated on Day 7 to re-test the anti-reflux property of the prototype after ureteral dilatation. Intraluminal reflux was evident, with methylene blue stain seen on the inner surface of the stents. Extraluminal reflux was proved with blue stain seen on the outer surface of the stents, both in undilated and dilated ureters. Study also proved the anti-reflux property of the skirt as the blue stain didn't go beyond the skirt, whereas the contralateral stent was stained up to proximal ureter. In this porcine in-vivo study, we elucidated that reflux occurred were both intraluminal and extraluminally. As a proof of concept, it also showed the anti-reflux property of a novel ureteral skirt.

A physiologically-based pharmacokinetic modeling approach for dosing amiodarone in children on ECMO.

Extracorporeal membrane oxygenation (ECMO) is a cardiopulmonary bypass device commonly used to treat cardiac arrest in children. The American Heart Association guidelines for cardiopulmonary resuscitation (CPR) and emergency cardiovascular care recommend using amiodarone as a first-line agent to treat ventricular arrhythmias in children with cardiac arrest. However, there are no dosing recommendations for amiodarone to treat ventricular arrhythmias in pediatric patients on ECMO. Amiodarone has a high propensity for adsorption to the ECMO components due to its physicochemical properties leading to altered pharmacokinetics (PK) in ECMO patients. The change in amiodarone PK due to interaction with ECMO components may result in a difference in optimal dosing in patients on ECMO when compared with non-ECMO patients. To address this clinical knowledge gap, a physiologically-based pharmacokinetic model of amiodarone was developed in adults and scaled to children, followed by the addition of an ECMO compartment. The pediatric model included ontogeny functions of cytochrome P450 (CYP450) enzyme maturation across various age groups. The ECMO compartment was parameterized using the adsorption data of amiodarone obtained from exvivo studies. Model predictions captured observed concentrations of amiodarone in pediatric patients with ECMO well with an average fold error between 0.5 and 2. Model simulations support an amiodarone intravenous (i.v) bolus dose of 22 mg/kg (neonates), 13 mg/kg (infants), 8 mg/kg (children), and 6 mg/kg (adolescents). This PBPK modeling approach can be applied to explore the dosing of other drugs used in children on ECMO.

Hydrophobic binary mixtures containing amphotericin B as lipophilic solutions for the treatment of cutaneous leishmaniasis

Cutaneous leishmaniasis, caused by Leishmania parasites, requires treatments with fewer side effects than those currently available. The development of a topical solution based on amphotericin B (AmB) was pursued. The considerable interest in deep eutectic solvents (DESs) and their remarkable advantages inspired the search for a suitable hydrophobic excipient. Various mixtures based on commonly used hydrogen bond donors (HBDs) and acceptors (HBAs) for DES preparations were explored. Initial physical and in-vitro screenings showed the potential of quaternary phosphonium salt-based mixtures. Through thermal analysis, it was determined that most of these mixtures did not exhibit eutectic behavior. X-ray scattering studies revealed a sponge-like nanoscale structure. The most promising formulation, based on a combination of trihexyl(tetradecyl)phosphonium chloride and 1-oleoyl-rac-glycerol, showed no deleterious effects through histological evaluation. AmB was fully solubilized at concentrations between 0.5 and 0.8 mg·mL−1, depending on the formulation. The monomeric state of AmB was observed by circular dichroism. In-vitro irritation tests demonstrated acceptable viability for AmB-based formulations up to 0.5 mg·mL−1. Additionally, an ex-vivo penetration study on pig ear skin revealed no transcutaneous passage, confirming AmB retention in healthy, unaffected skin.