Search Results (15,168)

Sustainable revalorization of agave bagasse, a lignocellulosic residue from mezcal production, is essential for environmental management. This study evaluated its potential as a substrate for the in vitro cultivation of the wild edible mushroom Pleurotus agaves. Characterization revealed a robust lignocellulosic matrix (70.9–75.87% NDF, 42.05–51.18% ADF and 10% lignin) and significant antioxidant potential, particularly in A. marmorata, which also exhibited higher total reducing sugars (11.94 mg/mL). This provides an energetic advantage for initial mycelial growth. Substrate microstructure was analyzed via microscopy (CLSM/SEM) before and after thermal pretreatment (55 °C). The IE-2038 strain was tested in five formulations: straw (P-55), bagasse (B-55), and straw–bagasse mixtures at 50–50%, 25–75%, and 75–25%. Mycelial growth rates indicated that PB-55 and pB-55 exhibited the fastest fungal colonization (8.2 mm/day and 8.3 mm/day). Microstructural analysis revealed significant damage to the polymeric organization of the bagasse, caused by mezcal production techniques and thermal treatment. This damage made lignin and cellulose more accessible for P. agaves. This synergy is supported by the adaptation of P. agaves to agave stalks. These findings confirm the capacity of bagasse as a sustainably bioprocessed substrate for edible mushroom cultivation, providing an effective alternative for the revalorization of agro-industrial residues that contribute to the circular economy. Full article

Background: Cancer has emerged as the primary cause of death worldwide in recent years. Current cancer treatment strategies require improvement, creating a pressing need for the development of novel therapeutic agents. This study investigated the anticancer effects of a series of newly synthesized tri- and difluoromethylated spiro[5.5]trienone compounds and evaluated the antitumor efficacy of a lead compound, 3s. Methods: The methyl thiazolyl tetrazolium (MTT) assay was used to assess the effect of the trienone compounds on the growth of cancer cells. Cell cycle distribution and intracellular reactive oxygen species (ROS) levels were analyzed by flow cytometry. Protein expression was examined by Western blot. A mouse xenograft model was utilized to test the anticancer effects and toxicity of 3s in vivo. Results: All 21 tri- and difluoromethylated spiro[5.5]trienones exhibited inhibitory effects on the growth of cancer cells. Among them, compound 3s showed the strongest inhibitory effect. It induced cell cycle arrest at the G2/M phase and promoted apoptosis. Mechanistically, 3s activated JNK and ERK signaling and elevated intracellular ROS levels. Furthermore, in a mouse xenograft model, 3s significantly inhibited tumor growth with minimal toxicity. Conclusions: Compound 3s exhibits potent anticancer efficacy both in vitro and in vivo. The discovery of 3s offers new potential for cancer therapy. Full article

Background: To evaluate the potential of cefiderocol as a topical ophthalmic antibiotic by determining the susceptibility of keratitis isolates from an extensive panel of Gram-negative bacterial species to this siderophore-cephalosporin class antibiotic. Methods: Minimum Inhibitory Concentrations (MICs) of cefiderocol were determined by the broth dilution method using iron-depleted, cation-adjusted Mueller–Hinton broth. The following Gram-negative bacteria were included: Acinetobacter baumannii (n = 13), Achromobacter xylosoxidans (n = 14), Escherichia coli (n = 15), Klebsiella aerogenes (n = 14), Klebsiella pneumoniae (n = 13), Klebsiella oxytoca (n = 14), Moraxella spp. (n = 15), Proteus mirabilis (n = 13), Pseudomonas aeruginosa (n = 17), Serratia marcescens (n = 14) and Stenotrophomonas maltophilia (n = 12). MIC₉₀ values were calculated for each of the species. Results: MIC₉₀ values (µg/mL): A. baumannii (0.5), A. xylosoxidans (0.25), E. coli (0.5), K. aerogenes (1.0), K. oxytoca (0.5), K. pneumoniae (0.5), Moraxella spp. (0.5), P. mirabilis (0.25), P. aeruginosa (0.5), S. marcescens (0.5), and S. maltophilia (0.25). In total, 100% of the isolates were determined to be susceptible to cefiderocol in vitro except for A. xylosoxidans and Moraxella spp., for which there are no established breakpoints for cefiderocol. Conclusions: Cefiderocol demonstrated in vitro activity against the tested panel of Gram-negative keratitis isolates. The results of this study suggest cefiderocol may be useful for the treatment of keratitis caused by numerous Gram-negative pathogens. Further development of cefiderocol for the topical treatment of Gram-negative keratitis is indicated. Full article

M. crystallinum is an edible halophytic succulent plant rich in phenolic compounds with potential pharmaceutical applications. However, it is known that these phytocompounds generally present low absorption, which hinders their direct use in formulations. Therefore, delivery systems, such as phytosomes, can be regarded as a potential strategy to overcome this disadvantage. This study aimed, for the first time, to prepare extracts from the ice plant using different solvents and to incorporate them into phytosomes. Physicochemical characterization of these phytosomes, their antioxidant activity, as well as the quantification and in vitro release profile of their phenolic and flavonoid compounds were studied. Different extraction solvents were assayed, and Ethanol:Acetone (80:20) achieved a strong antioxidant activity (reaching ca. 71.16%), extracting 3200.3 mg of GAE/100 g and 761.7 mg of QE/100 g of phenolic and flavonoid compounds, respectively. The phytosomal formulation exhibited a mean particle size of 233.80 nm, a polydispersity index of 0.23, and a zeta potential of −27.27 mV. Furthermore, a high encapsulation efficiency (96.63%) of the extracts in the phytosomes was obtained. The in vitro release test indicated that the antioxidant activity was retained, reaching a maximum of 42%, accompanied by a release of 51% of the flavonoid content at the end of the 3 h assay, under the experimental conditions. These findings highlight the potential of phytosomes formulated with Mesembryanthemum crystallinum extract as a delivery system for antioxidant phytochemicals. Full article

The Epi2SensA method is a method similar to the validated EpiSensA assay for assessing the skin sensitization potential of chemicals. The Epi2SensA protocol includes adaptation (changes to exposure conditions and the controls) for using an alternative reconstructed human epidermis (RhE) model, the EpiDerm™ model. The interlaboratory validation study evaluated the reliability and predictive capacity of Epi2SensA according to OECD Performance Standards. Four laboratories (Mattek, Now Part of Sartorius, Eurofins Munich, Burleson Research Technologies, Inc., and Food and Drug Safety Center) conducted blinded testing of 20 coded reference substances representing various chemical categories and sensitization potencies. Statistical analysis using modified acceptance criteria (a 60% cell viability threshold) and a modified prediction model (requiring at least two positive gene markers) demonstrated substantially improved performance compared to the original EpiSensA criteria. The between-laboratory reproducibility (BLR) was 85%, the average within-laboratory reproducibility (WLR) was 83.3%, and the average predictivity parameters were 88.1% for sensitivity, 88.9% for specificity, and 88.3% for accuracy. Epi2SensA achieved performance metrics comparable to the validated reference method (EpiSensA), supporting regulatory acceptance of the Epi2SensA assay using the EpiDerm™ model (Mattek Corporation, Now Part of Sartorius, Ashland, MA, USA) as an alternative RhE source for OECD TG 442D skin sensitization testing. Full article

Anticancer peptides (ACPs) offer a promising alternative to conventional chemotherapy but face challenges, including poor selectivity, limited tumor penetration, low cellular uptake, and rapid degradation in serum. To address these barriers, we developed synthetic mRNAs encoding chimeric ACPs designed for enhanced intracellular delivery and activity. mRNAs for constructs SAK6L9AS(1X), SAK6L9AS(4X), and WTAS-K6L9(4X) were transcribed in vitro and tested against 4T1 breast cancer cells. Cytotoxicity was assessed by cell confluence and MTT assays, while apoptosis was evaluated using caspase 3/7 activation, PI staining, and Annexin V flow cytometry. Our results demonstrate that all SAK6L9AS variants induced robust apoptosis and cellular toxicity in 4T1 cells. Importantly, this work provides the first demonstration of intracellular expression of an mRNA-encoded ACP fused to a cell-penetrating peptide, thereby validating a modular platform for RNA-based delivery of anticancer agents. This study highlights the feasibility of mRNA-encoded peptide therapeutics as a scalable and customizable strategy for cancer treatment. By combining the advantages of mRNA delivery with rational peptide design, ACP chimeras can be expressed directly inside tumor cells, overcoming the limitations of exogenous peptide administration. Our findings support further development of synthetic mRNA therapeutics to generate potent, selective anticancer peptides with reduced systemic toxicity and improved translational potential. Full article

Persistent endodontic infections are frequently associated with Enterococcus faecalis, a microorganism capable of penetrating dentinal tubules and surviving conventional disinfection procedures. This in vitro study evaluated the antimicrobial activity of Chihuahuan propolis against E. faecalis using planktonic and intratubular infection models. Propolis extract was tested at concentrations of 15, 35, and 70 mg/mL and compared with triple antibiotic paste (TAP) as a clinically relevant intracanal medicament. Antimicrobial efficacy was assessed by disk diffusion, minimum inhibitory concentration (MIC), colony-forming unit (CFU) reduction in infected dentinal tubules, and scanning electron microscopy (SEM). Chihuahuan propolis exhibited concentration-dependent antimicrobial activity, with a MIC of 17.5 mg/mL. In the intratubular model, propolis at 70 mg/mL achieved a CFU reduction comparable to TAP after seven days of application. SEM analysis confirmed a marked reduction of bacterial colonization within dentinal tubules. Within the limitations of this in vitro, monoespecies model, Chihuahuan propolis demonstrated antimicrobial efficacy against E. faecalis comparable to TAP, supporting its further investigation as a potential non-antibiotic intracanal medicament. Full article

Dental tissue regeneration, particularly alveolar bone and gingival repair, remains a major challenge in regenerative medicine. 3D bioprinting offers patient-specific and anatomically precise constructs, representing an advanced alternative to conventional grafting. In this study, nanohydroxyapatite (nHA), chitosan (CS), and collagen (CoL) were combined to fabricate and characterize 3D bioprinted dental grafts. SEM revealed a highly porous, interconnected architecture favorable for cell infiltration and nutrient exchange. EDS confirmed Ca/P ratios of 2.06 for nHA/CoL and 1.83 for nHA/CS/CoL, both of which are above the stoichiometric 1.67, indicating the presence of additional mineral phases and ion substitutions. FTIR and XRD verified characteristic functional groups and crystalline phases, including B-type HA with carbonate substitution. Mechanical testing showed that pure nHA exhibited the lowest compressive strength, whereas CoL incorporation improved stiffness. The nHA/CS/CoL composite achieved the highest compressive strength, elastic modulus, and toughness, demonstrating superior mechanical resilience. DSC analysis indicated endothermic peaks at 106.49 °C and 351.91 °C, with enthalpy values (264.91 J/g and 15.09 J/g) surpassing those of nHA alone. TGA revealed ~28.8% weight loss across three degradation stages, confirming enhanced thermal stability. In vitro cytocompatibility testing using L929 fibroblasts validated the biocompatibility of the composites. Collectively, the synergy between bioceramics and biopolymers markedly improved both mechanical and thermal performance. These findings position the nHA/CS/CoL scaffold as a promising candidate for clinical applications in dental tissue regeneration. Unlike conventional grafting materials, this study introduces a synergistically optimized nHA/CS/CoL bio-ink formulation specifically designed for extrusion-based 3D bioprinting of patient-specific dental constructs. The core innovation lies in the precise integration of nHA within a dual-polymer matrix (CS/CoL), which bridges the gap between mechanical resilience and biological signaling, achieving a compressive strength that mimics native alveolar bone while maintaining high cytocompatibility. Full article

This study investigates the antioxidant, enzyme inhibitory, and antimicrobial activities of water (WEHL) and ethanol (EEHL) extracts of hop (Humulus lupulus) cones. Phytochemical analyses revealed higher total phenolic content in EEHL (271.52 ± 0.13 mg GAE/g) than in WEHL (251.84 ± 0.06 mg GAE/g), as well as higher total flavonoid content (182.56 ± 0.45 mg QE/g for EEHL versus 179.39 ± 0.46 mg QE/g for WEHL). Antioxidant activity, determined by DPPH and ABTS assays, showed that EEHL had stronger radical scavenging capacity with IC₅₀ values of 19.13 ± 4.66 μg/mL (DPPH) and 12.66 ± 1.94 μg/mL (ABTS), compared to WEHL (DPPH: 20.90 ± 2.39 μg/mL; ABTS: 32.41 ± 4.29 μg/mL). In reducing assays, EEHL also showed better absorbance values in FRAP (0.77 ± 0.01), CUPRAC (2.09 ± 0.05), and Fe³⁺ reducing (1.95 ± 0.01) tests. EEHL likely outperformed WEHL due to solvent polarity and extraction efficiency. Moderately polar ethanol extracts a broader range of phenolics and flavonoids, including fewer polar bioactive compounds that contribute to antioxidant capacity and enzyme inhibition. This matches higher TPC/TFC in EEHL and explains stronger radical scavenging, reducing power, and multi-enzyme inhibition. Enzyme inhibition studies revealed that EEHL inhibited acetylcholinesterase (IC₅₀: 26.06 μg/mL), butyrylcholinesterase (IC₅₀: 44.00 μg/mL), α-glycosidase (IC₅₀: 119.31 μg/mL), and carbonic anhydrase isoenzymes hCA I (IC₅₀: 59.78 μg/mL) and hCA II (IC₅₀: 21.19 μg/mL). LC–MS/MS analysis identified major phenolic compounds such as isoquercitrin (3.14 ng/mL), rutin (0.60 ng/mL), and hesperidin (0.43 ng/mL) in EEHL. Antimicrobial screening showed selective activity against Staphylococcus aureus with an inhibition zone of 18.50 ± 0.58 mm, while no inhibition was observed against Escherichia coli and Candida albicans. These findings provide a solvent-dependent in vitro profile that can guide extraction strategies, support antioxidant and multi-enzyme screening (including hCA I and II), and identify candidates for selective antimicrobial evaluation and further preclinical investigation. Despite extensive use of hop extracts, comparative solvent-dependent profiling that links LC–MS/MS phenolic composition with a broad multi-enzyme inhibition panel, including the less frequently evaluated hCA I/II isoenzymes, remains limited. Therefore, the objective of this study was to systematically compare WEHL and EEHL in terms of phytochemical content and in vitro antioxidant, enzyme inhibitory, and antimicrobial activities. Overall, these results provide a solvent-dependent, comparative in vitro profile of WEHL vs. EEHL that can support antioxidant, multi-enzyme screening (including hCA I and II), and selective antimicrobial assays. Full article

Non-albicans Candida (NAC) species have emerged as significant opportunistic pathogens due to their reduced susceptibility to antifungal agents combined with their strong ability to form biofilms. The severity of systemic candidiasis caused by NAC species highlights the need for novel antifungal strategies. Retinoids, a group of compounds derived from vitamin A, have been demonstrated to possess significant antifungal activity against the reference strain C. albicans ATCC 2091. This study aimed to assess the antifungal potential of three retinoids, all-trans retinoic acid (ATRA), trifarotene, and tazarotene, against NAC clinical isolates. Various concentrations of the retinoids (from 1 mM to 0.06 mM) were tested in vitro against the growth, adhesion, and biofilm development of NAC species, including Candida glabrata, Candida krusei, and Candida tropicalis, as well as a reference strain of C. auris (CDC B11903). At 1 mM, all three compounds maximally inhibited the growth, adhesion, and biofilm formation of all tested NAC species. At lower concentrations (0.5–0.06 mM), C. krusei remained the most susceptible, especially to tazarotene. Tazarotene also showed a strong inhibitory effect on C. auris and C. glabrata at 0.5–0.25 mM; however, this effect was weaker than that observed against C. krusei. At low concentrations (0.12–0.06 mM), only trifarotene induced a mild but statistically significant inhibition of C. tropicalis growth. Trifarotene at 0.5 mM was also the most effective retinoid in inhibiting C. glabrata and C. tropicalis adherence and biofilm formation, with inhibitory activity maintained even at sub-0.5 mM concentrations (0.25–0.12 mM). Overall, the results suggest that all three retinoids exhibited dose-dependent and species-specific antifungal activity against NAC species, supporting their potential as novel, tailored antifungal agents against drug-resistant Candida strains. Full article

Degradable polymers, such as poly(L-lactide) (PLLA), are widely investigated for biomedical applications, including drug delivery systems and temporary implants. Their functionality can be expanded by incorporating degradable metal microparticles that may influence degradation behaviour and enable additional surface modification strategies. In this study, the feasibility of composites consisting of PLLA and biodegradable iron microparticles was investigated. Composites were fabricated by solvent casting, providing a gentle alternative to thermal processing methods, which often compromise polymer integrity. Composites were evaluated by thermogravimetric analysis, differential scanning calorimetry, scanning electron microscopy (SEM), tensile testing, dynamic mechanical analysis, and X-ray photoelectron spectroscopy (XPS). Incorporation of iron altered thermal behaviour and crystallinity of PLLA, indicating interactions between polymer matrix and dispersed metal phase that may affect degradation kinetics and material stability. While iron addition reduced Young’s modulus, tensile strength, and elongation at break, composites maintained sufficient structural integrity for potential biomedical applications. XPS and SEM confirmed the embedding of particles within the polymer matrix, enabling potential post-processing approaches. In vitro direct contact and eluate tests demonstrated good cell viability, whereas exposure to free iron particles resulted in dose- and time-dependent cytotoxic effects. Overall, the results demonstrate the feasibility of solvent-cast PLLA–iron composites for resorbable biomedical applications. Full article

Background/Objectives: Spotty liver disease (SLD), caused by Campylobacter hepaticus, is an emerging disease that leads to substantial production losses in the egg industry. The shift toward antibiotic-free and cage-free production systems has further intensified the impact of SLD. The current control measures largely rely on autogenous killed vaccines; however, their use is constrained by the slow and fastidious growth of C. hepaticus and inconsistent efficacy. To overcome these limitations, this study aimed to identify immunogenic mimotopes as vaccine candidates and express them on the surface of an avian pathogenic Escherichia coli (APEC) vector. Methods: To identify immunogenic mimotopes, Ph.D.-12 phage display peptide library was screened using the hyperimmune serum raised against killed whole-cell C. hepaticus in specific pathogen-free chickens. Subsequently, the outer membrane protein C (OmpC) of E. coli was used as a scaffold for constructing a surface display library. A single restriction site, PstI, located in the seventh external loop of OmpC, was strategically utilized to insert each 12-amino-acid mimotope with a six-histidine (6xHis) tag sequence at its N-terminus, generating ompC + mimotope fusion constructs. These constructs were cloned into the inducible expression vector pTrc and electroporated into an E. coli DH5α ∆ompC strain, which lacked ompC. The surface expression of the mimotopes was confirmed in vitro. The verified ompC + mimotope constructs were subsequently subcloned into the pYA3422 constitutive expression vector and electroporated into the APEC PSUO78 ∆aroA ∆asd vaccine vector strain. A chicken vaccination–challenge trial was conducted using nine groups of chickens, including an unvaccinated challenged control and an unvaccinated–unchallenged negative control. Each experimental group received a mixture of two recombinant E. coli strains carrying different mimotopes at a dose of 1 × 10⁹ CFU, which were administered orally twice at 16 and 18 weeks of age. Results: Fourteen immunogenic mimotopes corresponding to 13 different C. hepaticus proteins were identified as potential vaccine candidates. The expression of these mimotopes on the surface of the E. coli was successfully demonstrated using the OmpC-mediated surface display system. Of the 14 mimotopes tested, two flagellar-related peptides and one major outer membrane protein (MOMP)-derived peptide elicited significant immune responses and conferred protection against the C. hepaticus challenge. Conclusions: We successfully developed a functional E. coli surface display system that was capable of expressing 12-amino-acid mimotopes of C. hepaticus, providing a robust platform for evaluating vaccine candidates against SLD. Immunogenicity and efficacy studies in chickens demonstrated that three identified mimotopes conferred protection against C. hepaticus colonization of the bile and liver. Future in vivo investigations are necessary to develop and evaluate the immunogenicity and protective efficacy of a multivalent mimotope vaccine consisting of three identified mimotopes against both C. hepaticus and APEC, utilizing the ΔaroA Δasd APEC PSU078 strain as the vaccine vector. Full article

Background/Objectives: Worldwide, diabetes is a 21st century disease with continuously increasing prevalence. Current medications often have long-term adverse effects, which is why new substances are needed to help combat these disadvantages. Methods: In this respect, the present study develops a series of compounds with potential antidiabetic activity, including synthesis, physicochemical–spectral characterization and in vitro–in silico evaluation. Results: The sulfonamide derivatives were obtained by condensation reactions of para-toluenesulfonamide (p-TSA) with two different isocyanates, directly or after the condensation reaction with urea. The spectroscopic methods, IR, ¹H-NMR, ¹³C-NMR, were used for the structural elucidation of the compounds to confirm the presence of the functional groups responsible for the antihyperglycemic action, namely amide, azomethine and sulfonyl groups. Cytotoxicity screening on NCTC fibroblasts confirmed the excellent safety profile of the most synthesized derivatives across the tested range (100–1500 μg/mL). In contrast, the p-TSA-c-d derivative showed a clear transition from a biocompatible profile at 100 μg/mL to a more cytotoxic phenotype at concentrations exceeding 750–1500 μg/mL. The synthesized derivatives, particularly p-TSA-c-d, exhibited remarkable antidiabetic potential by effectively inhibiting α-amylase and α-glucosidase, with IC₅₀ values as low as 46.54 μM, outperforming the standard reference acarbose. The molecular docking tests revealed different mechanisms for the inhibitory activity exerted by the p-TSA derivatives on the two targeted enzymes. Conclusions: Although these developed compounds can be considered promising antidiabetic agents, studies can be further deepened in the future by performing in vivo tests. Full article

The 2025 approval of the selective NaV1.8 blocker suzetrigine for acute pain marked a pivotal advance in analgesic drug development. Yet the subsequent failure of Vertex’s next-generation NaV1.8 inhibitor VX993 to demonstrate clinical analgesia underscores enduring challenges in translating mechanistic promise into patient benefit. This review examines why promising targets and compounds, spanning NaV and TRP channels, often falter and outlines a path toward more reliable target selection and validation. I first summarize the pain pathway, from nociceptor transduction through spinal processing to cortical perception, emphasizing how inflammation and peripheral sensitization reshape excitability. Historically serendipitous, pain drug discovery now prioritizes molecular precision. Most approved chronic pain therapies act in the CNS and are limited by modest efficacy and adverse effects. Nociceptor-enriched targets (NaV1.7/1.8/1.9; TRP channels) remain attractive, yet redundancy among NaV subtypes and the necessity of blocking targets at the correct anatomical sites complicate translation. Human genetics and multi-omics provide a powerful, unbiased engine for target discovery. Rare high-impact variants offer strong causal hypotheses, while common polygenic contributions illuminate broader susceptibility. Large biobanks increasingly reveal a mismatch between legacy pain targets and genetically supported candidates across neuronal and non-neuronal cells. Human DRG transcriptomics highlight NaV channel redundancy. Human in vitro electrophysiology and PK/PD analyses show suzetrigine achieves ~90–95% NaV1.8 engagement, yet neurons can still fire unless additional channels are blocked. Species differences and drug distribution (including BBB/PNS penetration and P-gp efflux) critically influence efficacy; centrally accessible blockade (e.g., for NaV1.7 or TRPA1) may be necessary to achieve robust analgesia, challenging peripherally restricted strategies. Osteoarthritis illustrates how obesity-driven metabolic inflammation, synovial immune activation, subchondral bone remodeling, and specific nociceptor subtypes converge to drive mechanical pain. Multi-omic integration across diseased human tissues can pinpoint causal processes and cell types, enabling more selective and safer target choices. I propose a practical framework for target validation that integrates: (i) rigorous human genetic support; (ii) cell-type and site-of-action mapping; (iii) human-relevant electrophysiology and PK/PD with verified target engagement; (iv) species-appropriate models; (v) consideration of modality (small molecule, biologic, RNA, targeted protein degradation). Advancing genetically and anatomically aligned targets, tested at the right sites and exposures, offers the best path to genuinely effective, better-tolerated pain therapeutics. Full article

Background/Objectives: Staphylococcus epidermidis (RP62A) and Staphylococcus aureus (UAMS-1) are clinically relevant pathogens frequently implicated in implant-associated infections due to their ability to form biofilms. RP62A is typically linked to persistent, chronic, low-grade infections, whereas UAMS-1 is associated with acute, invasive disease. Both strains serve as representative models for chronic and acute periprosthetic joint infections (PJIs). The objective of this study was to examine and compare in vitro biofilm formation by RP62A and UAMS-1 on orthopaedic materials/disc surfaces of defined composition. Methods: In vitro biofilm formation assays were performed using orthopaedic disc surfaces composed of cobalt–chromium alloy (CoCr), titanium alloy (Ti), and polyethylene (PE) after 72 h of incubation. Biofilm biomass was quantified using crystal violet staining, with absorbance measured at OD₅₇₀. A polystyrene (PS) surface served as a control. Additionally, retrieved orthopaedic explant components were used as substrates for in vitro biofilm assays, in which RP62A was incubated for 72 h on the explanted surfaces. Supporting assays on glass slides were conducted to examine strain-specific biofilm-related architecture. Results: In vitro biofilm mass quantification assays showed strong biofilm formation by RP62A across all tested surfaces, with the highest absorbance on CoCr (OD₅₇₀ = 5.80 ± 0.19). Notably, biofilm formation on CoCr was 76% higher compared to PS (p < 0.0001). No significant differences were observed among all three surface discs (p > 0.1). Biofilm formation was highest on PE for UAMS-1 (OD₅₇₀ = 1.29 ± 0.09) and was significantly greater than on Ti (178%, p < 0.001) and CoCr (196%, p < 0.0001). In the in vitro assays performed on retrieved explant components, RP62A showed pronounced biofilm accumulation on polyethylene tibial inserts, particularly in regions of mechanical wear and friction. Supporting assays on glass slides were performed to examine strain-specific surface microstructural, revealing dense network-like structures for RP62A and thinner, discontinuous layers for UAMS-1. Conclusions: RP62A formed dense biofilms in vitro on multiple orthopaedic implant materials and retrieved explant components, consistent with its association with chronic periprosthetic joint infections. Increased biofilm accumulation was observed on mechanically worn polyethylene surfaces. In contrast, UAMS-1 showed lower biofilm formation on metallic disc surfaces, indicating strain- and material-dependent differences. These findings highlight the relevance of implant material selection and surface integrity for strategies targeting biofilm-associated implant infections. Full article