
Deep eutectic solvents (DESs) have garnered significant attention in pharmaceutical research in recent years due to their high efficiency, facile separability and biocompatibility. Characterised by hydrogen-bond donors (HBDs) and hydrogen-bond acceptors (HBAs), DESs exhibit low toxicity, favourable biodegradability, and excellent solubilising properties, rendering them promising alternatives to conventional organic solvents in drug synthesis, natural active ingredient extraction and drug delivery. This review systematically evaluates the broad pharmaceutical applications of DESs, with particular emphasis on enhanced drug solubility and bioavailability. In plant extraction, DESs outperform traditional solvents (e.g., diethyl ether) by improving the recovery efficiency of bioactive compounds. In drug synthesis, certain DESs function simultaneously as reaction media and catalysts, enhancing reaction efficiency and selectivity while minimising environmental impact. In drug delivery, DESs facilitate transdermal and oral absorption through interactions with biological membranes, making improved delivery efficiency. Despite these advantages, challenges remain, including high viscosity, formulation complexity, and unresolved regulatory considerations. Future research must focus on physico-chemical optimisation, safety evaluation, and scalable production to fully realise the potential in pharmaceutical applications.
Daratumumab is a fully human anti-CD38 monoclonal antibody with strong potential as a targeting vector for therapeutic radionuclides. This study aimed to develop a freeze-dried daratumumab immunoconjugate kit by selecting a suitable chelator (DOTA-NHS, p-SCN-Bn-DOTA, or p-SCN-Bn-1B4M-DTPA) for the 177Lu-labelling, optimising the freeze-drying formulation, and evaluating the physicochemical properties and purity profiles. Conjugation performed in carbonate buffer at elevated temperature enhanced chelator incorporation and supported the selection of daratumumab-p-SCN-Bn-DOTA as the most suitable candidate, achieving radiolabelling yield up to 99.8 % without additional purification. Among the evaluated freeze-dried formulations, a saline-based, buffer-free sucrose-mannitol formulation containing polysorbate 20 (S.F5) provided the most favourable characteristics, including minimal residual moisture and the highest monomer purity with non-detectable HMWS species under the applied SE-HPLC conditions. ATR-FTIR and Raman spectroscopy confirmed preservation of the antibody structural integrity after conjugation and freeze-drying. In an in vitro study using human serum, [177Lu]Lu-daratumumab-p-SCNBn-DOTA maintained higher radiochemical purity for 168 h than [177Lu]Lu-daratumumab-p-SCN-Bn-1B4MDTPA, indicating greater stability. These results support the feasibility of a ready-to-use freeze-dried daratumumab-p-SCN-Bn-DOTA kit for the 177Lu-labelling.
Diabetic retinopathy (DR) is one of the primary causes of vision impairment, affecting individuals with diabetes, and is marked by the neurodegeneration of the retina along with increased intraocular pressure (IOP). This study sought to determine the effects of pelargonidin on extracellular matrix (ECM) modulation and the inhibition of transforming growth factor-β (TGF-β) and Janus Kinase 2/Signal Transducer and Activator of Transcription 3 (JAK2/STAT3) pathway in retinal ganglion cells of streptozotocin-induced diabetic rats. Male Sprague-Dawley rats (180-200 g) were rendered diabetic by intraperitoneal administration of streptozotocin (STZ). The rats were divided into 5 groups: control, diabetic model (STZ), STZ + low dose pelargonidin (12.5 mg kg-1 per day), STZ + medium dose pelargonidin (25 mg kg-1 per day) and STZ + high dose pelargonidin (50 mg kg-1 per day). IOP was monitored using a tonometer. Whole-mount retinal immunofluorescence staining using RNA-binding protein with multiple splicing (RBPMS) was performed to assess retinal ganglion cell (RGC) density. Protein expression levels of apoptotic markers, ECM components, and TGF-β and JAK2/STAT3 signalling pathways were evaluated by Western blotting. Pelargonidin treatment dose-dependently reduced the elevated IOP. Importantly, immunofluorescence analysis revealed a marked dose-dependent preservation of retinal ganglion cell (RGC) density: STZ-induced RGC loss was significantly reversed by pelargonidin, with the highest dose restoring RGC density to near-control or higher levels in both the central and peripheral retina. This was achieved via modulation of apoptosis-related proteins through the upregulation of Bcl-xL, Bcl-2, and downregulation of Bad, Bax and cleaved caspase-3. Furthermore, pelargonidin modulated ECM remodelling protein expression in the RGC layer. In particular, TGF-β2/Smad2/3 signalling was downregulated, and the JAK2/STAT3 pathway was upregulated. By reducing IOP, preserving RGC density, modulating ECM deposition, inhibiting TGF-β and upregulating the JAK2/STAT3 pathway, pelargonidin exerts protective effects against diabetic retinal injury. The results of this study further confirm the pharmacological potential of pelargonidin as a therapeutic agent for diabetic retinopathy.
The effects of vilazodone (VZD) on the acquisition of active avoidance behavior were examined in C57BL/6J mice. Both female and male mice were assigned to three groups (n = 8 per group per sex): the vehicle control group (VEH), the 0.5 mg kg-1 vilazodone lower dose group (VZD0.5) and the 1 mg kg-1 vilazodone higher dose group (VZD1.0). Spontaneous locomotion and anxiety-like behaviour were assessed after drug administration intraperitoneally in an open field test (OFT). Another cohort of mice was trained in a three-day shuttle box active avoidance test (AAT) after drug administrations with the aim of evaluating the effects of VZD on the acquisition of active avoidance behaviour. In the OFT, VZD decreased freezing time in the corner area in both female and male mice, indicating reduced anxiety-like behaviours. In the AAT, the active avoidance rate was significantly improved on day 1 in female mice and day 2 in male mice, suggesting that VZD facilitated active avoidance learning with sexual dimorphism. Furthermore, the increased active avoidance rates were negatively correlated with freezing time during training. Interestingly, these group differences and correlations diminished on day 3, implying that the facilitation was restricted to early training phases. Collectively, VZD facilitates the acquisition of active avoidance behaviour in mice with distinct sexual dimorphism and temporal dynamics.
Alzheimer's disease (AD) is characterised by β-amyloid (Aβ) plaque accumulation and tau hyperphosphorylation. H3 relaxin, a neuro-peptide, is known to exert neuroprotective effects. In this study, we investigated how H3 relaxin confers neuroprotection in a streptozotocin (STZ)-induced mouse model and modulates PI3K/Akt-mTOR signalling. Mice were divided into four groups (n = 6 per group): control (saline), STZ, STZ + H3 relaxin, and STZ + donepezil. Following STZ induction, H3 relaxin (1 µg per day) was administered intracerebro ventricularly (ICV) for 14 consecutive days, whereas donepezil (2.5 mg kg-1 per day) was administered orally for the same duration. Cognitive performance was assessed using the Morris water maze (MWM) test. Aβ deposition in the cortex was evaluated through immunohistochemistry. Western blotting was conducted for tau phospho rylation, PI3K/Akt/mTOR signalling, and autophagy markers in the hippocampus. Oxidative stress and inflammation markers were measured using ELISA. H3 relaxin markedly improved memory by decreasing escape latency and duration while spending more time in the target quadrant in the MWM test. Additionally, H3 relaxin reduced Aβ plaque burden and tau phosphorylation (Ser396/404) while enhancing PI3K/Akt-mTOR signalling. Oxidative stress was attenuated, as evidenced by increased GSH and HO-1 levels and reduced MDA and H2O2 concentrations. Moreover, markers of inflammation, NF-κB and TNF-α were suppressed. Overall, H3 relaxin ameliorated cognitive deficits in STZ-induced AD mice through modulation of impaired PI3K/Akt-mTOR signalling, reduction of Aβ and tau pathology, and promotion of autophagy.
During in-use stability testing of cefixime granules for oral suspension, an impurity with a relative retention time of 0.19 was consistently detected and increased during storage. To ensure regulatory compliance and patient safety, the impurity was structurally identified and toxicologically qualified. A laboratory-scale formulation and commercial products were studied under refrigerated and ambient conditions. The impurity was isolated by automated fraction collection and characterised by liquid chromatography-mass spectrometry and tandem mass spectrometry. Kinetic evaluation showed pseudo-first-order formation, with faster accumulation at ambient temperature. The impurity was identified as a γ-lactone degradation product of cefixime, present as multiple stereoisomers stabilised under acidic conditions. In silico toxicological assessment using complementary platforms indicated no additional structural alerts, no mutagenic potential, and negligible acute toxicity. The impurity forms only after prolonged storage of reconstituted suspensions and is classified as an ICH M7 Class 5 impurity, requiring no further genotoxicity testing. The applied analytical-computational workflow provides an efficient approach for impurity qualification in β-lactam antibiotics.
The objective of this study was to explore the protective effect of astragaloside IV on a model of isoproterenol-induced (ISO) hyper-trophic injury in rat cardiomyocytes H9c2 (cell line derived from embryonic BD1X rat heart tissue). A cell hypertrophy injury model was established (H9c2 cells treated with 100 μmol L-1 ISO). The cells were divided into normal control, a model group, and an astragalo-side IV group at several concentrations. Astragaloside IV was pre-administered for 2 hours, followed by ISO treatment for 24 hours. Cell viability, cell surface area, apoptosis rate, lactate dehydrogenase (LDH) activity, reactive oxygen species (ROS), superoxide dismutase (SOD), the mRNA levels of Bcl-2, Bax, p62, and LC3, the protein expressions of Sirt1, p62, caspase-3, beclin, and p53 and the LC3II/LC3I ratio were detected. Astragaloside IV significantly alleviated ISO-induced hypertrophy injury in H9c2 cells, reduced cell surface area and LDH release, decreased apoptosis rate and intracellular ROS levels, increased SOD levels, upregulated the expressions of autophagy-related mRNA and proteins, and downregulated the expressions of apoptosis-related mRNA and proteins. Astragaloside IV can effectively inhibit ISO-induced hypertrophy and apoptosis in H9c2 cells, and its mechanism may be related to promoting auto-phagy and reducing oxidative stress.
This study examined the effects of omega-3 fatty acids supplementation on gut barrier integrity, systemic inflammation, neurotrans-mission and oxidative stress, in an aged rat model of propionic acid (PPA)-induced neurotoxicity. Twenty-four aged male rats were divided into four groups: control, omega-3, PPA and PPA + omega-3. Serum cytokines, tight-junction proteins (TJP1), dopamine, serotonin, short-chain fatty acids (SCFAs), oxidative stress markers, and histopathology of the brain and small intestine were evaluated. PPA exposure significantly increased tumour necrosis factor-α (TNF-α) and interleukin-6 (IL-6) and reduced TJP1 expression, confirming gut barrier disruption and systemic inflammation. Omega-3 fatty acids supplementation selectively reduced IL-6 but did not reverse PPA-induced TNF-α elevation or oxidative stress. CLDN2 expression increased in PPA + omega-3 rats, suggesting a compensatory but incomplete barrier response. Dopamine, serotonin, and SCFA levels showed upward trends with supplementation but were not statistically significant. Histological analysis demonstrated partial preservation of neuronal and intestinal structure in the PPA + omega-3 group. Overall, omega-3 fatty acids exerted modest anti-inflammatory effects but failed to fully restore oxidative balance or barrier integrity in aged rats, suggesting that omega-3 fatty acids may be more effective as a preventive rather than restorative intervention in ageing-related gut-brain axis disruption.
The main protease 3CL pro of the SARS-CoV-2 virus is a well--established therapeutic target for the treatment of COVID-19. In this study, we screened an in-house compound library and identified a series of α-heteroarylthiomethyl ketones as inhibitors of 3CL pro . Among these, analogues 31 and 33 emerged as the most interesting candidates with IC 50 values of 95.4 ± 3.1 and 95.0 ± 6.9 µmol L – 1 , respectively. Preliminary in vitro studies suggest a potential covalent mode of inhibition, although further studies are required to confirm this mechanism. These findings provide a new chemical scaffold for the development of 3CL pro -targeting inhibitors.
The voltage-gated potassium channel Kv1.3 is a key regulator of T-cell activation and a validated therapeutic target for autoimmune and inflammatory diseases. In this study, a ligand-based design strategy was employed to expand a library of benzamide-derived K v 1.3 inhibitors. Starting from a previously optimised thiophene-based inhibitor, structu ral modifications were introduced to the 2-methoxybenzamide moiety and the central tetrahydropyran or cyclohexane scaffold. A series of ketone, hydroxy, and carbamate derivatives was synthesised and evaluated for K v 1.3 inhibition using whole-cell patch-clamp electrophysiology. Structure-activity relationship analysis revealed that cis -isomers in the hydroxy series exhibited stronger activity than their trans counterparts, with some analogues displaying submicro-molar IC 50 values. In the carbamate series, trans -isomers were generally more potent, with trans - 18 and trans - 16 achieving IC 50 values of 122 and 166 nmol L–1, respectively. These results provide valuable insights into the design of K v 1.3 inhibitors and support further development of these compounds for immunomodulatory applications.
This study aimed to examine the motility, biofilm production, endotoxin release, and antibiotic resistance of 81 Ralstonia pickettii isolates collected from different pharmaceutical water systems in Croatia. Swimming and twitching motility were detected in all isolates, while swarming was not observed. Biofilm production was detected in approximately 40 % of the isolates under the tested conditions. Notably, extracellular polymeric substance (EPS) production was a common trait among all isolates. Endotoxin production was detected with the Limulus Amoebocyte Lysate test. Antibiotic susceptibility testing revealed consistent resistance to colistin, as well as significant resistance rates to β-lactam antibiotics, ertapenem, amoxicillin/clavulanic acid, ticarcillin and ampicillin. High susceptibility to first-generation cephalosporins, cephalexin, cefoxitin and chloramphenicol was observed. All isolates were susceptible to tigecycline and tetracycline. The isolates were grouped into three genetically closely related clusters, yet notable phenotypic diversity in biofilm production and antibiotic susceptibility persisted within these groups. The study highlights R. pickettii's adaptability in pharmaceutical water systems, marked by its motility, biofilm-forming capabilities, and multidrug resistance. These results emphasise the importance of rigorous monitoring of water systems to reduce transmission risks and prevent the emergence of resistant strains in clinical environments.
Antibiotic resistance is a growing global health threat, with patient non-adherence to prescribed antibiotic regimens representing an important contributing factor. This prospective interventional study investigated the impact of clinical pharmacist counselling at hospital discharge on adherence to oral antibiotic therapy. The study was conducted at the Department of Nephrology and Endocrinology, General Hospital "Dr. Tomislav Bardek", Koprivnica, between March 2022 and July 2025. A total of 98 participants aged ≥18 years who were prescribed oral antibiotics at discharge were randomised into intervention (counselling) and non-intervention groups. Baseline socio-demographic and clinical data were collected, and adherence was assessed following treatment completion using a structured questionnaire developed by the authors. Of the 98 participants, 94 were included in the final analysis. Non-adherence was significantly lower in the intervention group, compared to the non-intervention group (2.1 % vs. 36.2 %, p < 0.001). Urinary tract infections represented the most common indication for antibiotic therapy, with ciprofloxacin and amoxicillin-clavulanic acid being the most frequently prescribed medicines. This study demonstrates that pharmacist-led discharge counselling markedly improves adherence to short-term antibiotic therapy. These findings provide preliminary evidence supporting integration of clinical pharmacists into hospital discharge processes to promote rational antibiotic use and combat antimicrobial resistance.
Glucocorticoids are a group of drugs increasingly used in modern medical practice due to their pronounced anti--inflammatory and immunosuppressive properties. In this study, prednisolone disodium phosphate and prednisolone acetate were analysed, with the aim of developing and validating an HPLC method in accordance with ICH Q2(R2) guidelines and quantifying their content in the active pharmaceutical ingredient powder and a model in--house sample. By applying the HPLC-DAD method with gradient elution, effective separation of the analytes was achieved. The method met all validation parameter requirements. The obtained results showed that the content of both analytes in the tested samples (bulk API powders and in-house prepared model formulation) was within the prescribed limits according to current pharmacopoeial standards. The proposed HPLC-DAD method was assessed for its applicability and environmental profile utilizing a range of green and blue metric tools. This comprehensive evaluation confirms that the method adheres to green analytical principles, making it suitable for sustainable pharmaceutical analysis.
l-Met-enkephalin is a neuropeptide known to exert protective effects in various experimental models of autoimmune and inflammatory diseases. These effects are mediated through opioid receptors and can be abolished by the opioid receptor antagonist naltrexone. Investigation of peptide enantiomerism and the incorporation of d-amino acids are crucial for designing novel peptides with altered structural and biological properties compared with their native l-forms. Since no data are currently available on the properties or biological activity of the d-Met-enkephalin enantiomer, we evaluated its hepatoprotective potential in a mouse model of acetaminophen-induced hepatotoxicity. Male CBA mice were treated with d-Met-enkephalin, and hepatoprotection was assessed by measuring plasma alanine aminotransferase (ALT) and aspartate amino-transferase (AST) activities, along with histological liver necrosis scores. The peptide's secondary structure and antisense peptide binding were analysed using circular dichroism and fluorescence spectroscopy, respectively. d-Met-enkephalin demonstrated dose-dependent hepatoprotective effects within the range of 0.5-20 mg kg-1, with maximal protection observed at 5 mg kg-1, a dose comparable to that of the l-enantiomer (7.5 mg kg-1). This preservation of biological activity may be attributed to the presence of the achiral amino acid glycine at positions 2 and 3, which maintains the functional conformation of the d-enantiomer. The role of opioid receptor involvement was further examined through direct receptor blockade using naltrexone and indirect inhibition with the antisense peptide IPPKY.
The COVID-19 pandemic introduced substantial changes to clinical practice, including widespread antibiotic use. These changes raised concerns about a potential rise in healthcare-associated infections, particularly Clostri dioi des difficile infection (CDI). This study aimed to investigate the hidden impact of COVID-19 treatment strategies on the incidence of CDI, with a specific focus on antibiotic use, advanced age, comorbidities, and the administration of proton pump inhibitors (PPIs) and corticosteroids. A retrospective observational study was conducted using anonymised patient data from the University Clinical Hospital Mostar. The number of CDI cases significantly increased during the COVID-19 peak in 2021, showing a perfect positive correlation with COVID-19 incidence (ρ = 1.0, p < 0.05). Antibiotic use was strongly associated with CDI (69 % vs. 12 %; p < 0.05), as was advanced age (≥ 65 years; 71 %; p < 0.05). The combined use of proton pump inhibitors and corticosteroids was significantly more frequent in the CDI group (54 % vs. 24 %; p < 0.05). The findings highlight how COVID-19 treatment strategies can unintentionally raise CDI risk, stressing the need for prudent antibiotic use, careful drug management and targeted prevention for elderly and high-risk patients.
The aim of this study was to evaluate the antibacterial and antibio-film potential of two naturally occurring naphthoquinones, 2-hydroxy-1,4-naphthoquinone (2-HNQ) and 2-methoxy-1,4-naphthoquinone (2-MNQ), against the Gram-negative bacterium Escherichia coli strain ATCC 25922. In the first step of the study, the minimum inhibitory concentrations (MICs) of 2-HNQ and 2-MNQ were determined using the microdilution method. Subsequently, possible mechanisms underlying the antibacterial activity of 2-HNQ and 2-MNQ against E. coli were investigated by assessing intracellular reactive oxygen species (ROS) production and membrane permeability. Finally, the impact of 2-HNQ and 2-MNQ on swarming motility and on pre- and post-biofilm formation of E. coli was evaluated. The MIC of 2-HNQ against E. coli was 500 µg mL–1, while that of 2-MNQ was 100 µg mL–1. Both compounds increased intracellular ROS production and altered the membrane permeability of E. coli. Moreover, 2-HNQ and 2-MNQ reduced swarming motility and inhibited both pre- and post-biofilm formation. The results of this study indicate that both naphthoquinones possess antibacterial potential, with 2-MNQ showing greater potency against E. coli. Their antibacterial activity appears to involve ROS generation and disruption of membrane permeability. Importantly, both tested naphthoquinones also impaired E. coli motility and bio-film formation, suggesting potential applications in the treatment of infections caused by E. coli.
Methyl caffeate (MC), a naturally occurring methyl ester of caffeic acid (CA), exhibits potent antioxidant activity and a broad spectrum of biological effects. This study investigates the antioxidant mechanism of MC through its reaction with the stable radical DPPH•, employing both experimental and computational approaches. Kinetic measurements were conducted in a predo minantly nonpolar medium (1,4-dioxane with phosphate buffer), revealing concerted proton and electron transfer. This experimental evidence was supported by values of kinetic isotope effects (KIEs) and thermodynamic activation parameters. Analysis of the intrinsic bond orbitals (IBOs) along the calculated intrinsic reaction coordinate (IRC) trajectories supported the proposed proton- coupled electron transfer (PCET) reaction mechanism. Additionally, the Fe(II) complexation ability of MC was evaluated spectrophotometrically, demonstrating stable complex formation at pH 7.0, suggesting potential for mitigating hydroxyl radical generation in physiological conditions. These findings offer new insights into the antioxidant behaviour of MC and its potential applications in pharmaceutical and nutraceutical formulations.
Preeclampsia (PE) is a complex pregnancy disorder that may cause adverse outcomes for mother and baby. Combining risk factors with clinical, laboratory, and ultrasonographic data can help identify women at risk. This study investigated the relationship between maternal risk factors, soluble fms-like tyro-sine kinase 1 (sFlt-1), placental growth factor (PlGF), their ratio, and pregnancy outcomes, involving 68 women with PE risk factors and 21 controls. There were no significant differences in the frequency of adverse outcomes (PE, foetal death, and infants with abnormal birth weight), sFlt-1, PlGF, the sFlt-1/PlGF ratio, or birth weight centiles between the PE-risk and control groups. The most frequently recorded high-risk factor was gestational diabetes mellitus, whereas moderate risk was a pre-pregnancy body mass index of over 30 kg m-2. The most prominent difference was observed in the subgroups with gestational hypertension and first-time pregnant women as risk factors, with significantly higher sFlt-1/PlGF ratios compared to the control group. Combining multiple risk factors increased the sFlt-1/PlGF ratio compared to both the control group and the group with only one risk factor. The study documented PE risk factors and outcomes at a Croatian hospital where angiogenic markers are not routinely used in screening. Findings highlighted the importance of integrating PE screening into standard practice.
Optimising medication adherence is essential for effective heart failure (HF) management, yet nonadherence remains common, particularly among hospitalised and advanced-stage patients. This study evaluated the internal consistency reliability and score association of the Medication Adherence Report Scales, MARS-5 and MARS-10, self-report questionnaires in clinical pharmacist-led adherence assessments in hospitalised HF patients. Tools were administered during structured pharmacist-led interviews. To complement quantitative findings, four clinical cases were presented to illustrate the clinical relevance of adherence assessment in real--world HF management. Results showed a strong association between MARS-5 and MARS-10 (n = 70) responses (unweighted Cohen’s kappa 0.820 (95 % CI 0.683–0.957; p < 0.001) for categorizing patients as nonadherent or adherent and Pearson’s r coefficient of 0.899 (95 % CI 0.847–1.000; p < 0.001) for continuous score correlation), supporting score association and flexible use in clinical settings, with MARS-5 reliably identifying nonadherence (defined as score < 20 for MARS-5 and ≤ 8 for MARS-10), with potentially reduced respondent burden due to fewer items. Serious clinical complications were documented in nonadherent patients (41.43 % by MARS-5 and 35.71 % by MARS-10), illustrated through selected cases including stent thrombosis, embolic stroke, graft dysfunction, and deterioration in glycaemic control. These findings indicate the potential of MARS-5 as a practical, time-efficient tool for routine adherence assessment in acute settings. Case analyses underscore the critical role of the clinical pharmacist in proactively identifying nonadherence and enabling timely, targeted interventions to mitigate risk and improve patient outcomes in HF care.
This double-blind randomised controlled trial aimed to evaluate the efficacy of an emollient cream with and without 1 % supercritical CO2 extract of Calendula officinalis in contact dermatitis. Twenty healthy volunteers without pre-existing dermatological conditions participated in this single-centre study. Each participant's forearm was divided into three test sites: control (no treatment), placebo (base emollient), and intervention (calendula cream). The study employed random allocation of test sites using Microsoft Excel software. Both investigators and participants were blinded to the treatment assignments. The main outcomes were changes in skin hydration and transepidermal water loss (TEWL), measured with non-invasive probes (Corneometer CM 825, Tewameter TM 300), and erythema measured by Mexameter MX 18. Following irritant exposure, all sites showed increased TEWL and reduced hydration, confirming skin barrier impairment. The intervention site demonstrated significantly greater hydration compared with both the control (p = 0.017) and placebo sites (p = 0.035) on day 4, with this improvement persisting on day 8 (p = 0.043). TEWL values at the intervention site were significantly lower than control on day 3 (p = 0.022), indicating faster barrier recovery. No significant differences in erythema were observed between groups, and no adverse events occurred. Results of this study indicate that the addition of 1 % Calendula officinalis extract to an emollient cream enhanced skin hydration and accelerated recovery after irritant exposure, suggesting potential benefit in managing contact dermatitis with non-pharmacological skincare formulations.