The investigated water-insoluble polysaccharide was derived from the fruiting bodies of Pleurotus citrinopileatus, a golden oyster mushroom. The structural studies included compositional and linkages analyses, size-exclusion chromatography, and 2D NMR, FT-IR as well as Raman spectroscopy. The presence of (1 → 3)-α-d-glucan in the cell wall of P. citrinopileatus mycelium was confirmed using fluorophore-labelled antibodies. The isolated polymer predominantly comprised d-glucose (94.3%). Small amounts of xylose and mannose were also detected. Hexoses are linked primarily by (1 → 3)-glycosidic bonds (82.1%). Terminal glucose and →3)-linked mannose residues were also observed. Additionally, small amounts of doubly substituted hexoses [namely →3,4)-Glcp-(1→ and →3,6)-Glcp-(1→] were also present. The molecular weight of the polymer was estimated to be approximately 552 kDa. FT-IR and Raman spectroscopy, along with NMR studies indicated that this polymer has structure typical for a (1 → 3)-α-d-glucan. Acid hydrolysis yields a mixture of glucose and glucooligosaccharides (GOS), with degree of polymerization (Dp) ranging from 2 to 10. The in vitro fermentability of GOS by potential probiotic strains was assessed and compared with reference prebiotics, fructooligosaccharides and inulin. GOS supported the growth of all the tested strains and exhibited a strong bifidogenic effect.

Creating perfusable vascular networks that replicate physiological flow remains a major challenge in tissue engineering. We present a rapid (<1 min) volumetric additive manufacturing (VAM) method for fabricating tunable, biologically relevant vascular structures that support controlled perfusion. A recyclable GelMA/PEGDA resin was optimized to produce high-fidelity hydrogels with embedded channels. To study how flow, structure, and function interact, we designed modular perfusion platforms providing precise control over physiological shear stress (3-50 dyne/cm(2)), flow rates (1-15 mL/min), and pulsatile or continuous flow. These systems enable endothelial attachment, stable perfusion, and permeability measurements, supporting physiologically relevant flow and mass-transport studies for vascularized grafts and drug-delivery assays. In addition to controlled perfusion, the VAM process allows fast prototyping (<45 s) and improved biomimicry by generating vascular architectures within soft tissue-like hydrogels (similar to 10-12 kPa). The platform also permits independent pressure modulation inside the vessel and surrounding matrix, with simulations closely reflecting experimental flow and permeability data and confirming diffusion-dominated transport. Together, this framework provides a versatile resource for translational vascular modeling and drug-delivery research.

Objective To develop and evaluate a soft capsule formulation of Artemisia annua L. extract, with the aim of improving formulation performance of artemisinin, particularly in relation to its poor aqueous solubility and limited stability. Methods The extract was prepared using supercritical CO₂ extraction followed by ethyl acetate purification. Chemical constituents were identified by UPLC-MS/MS. The soft capsule formulation was optimized through single-factor experiments. Artemisinin content and dissolution behavior were determined by HPLC. Quality evaluation (fill weight variation, disintegration time, microbial limits, and stability) was conducted according to the Chinese Pharmacopoeia (2025 Edition). Safety was assessed through acute oral toxicity and 28-day repeated-dose toxicity studies in rats. Results The optimal extraction conditions were 25 MPa, 50°C, with 50% ethanol as co-solvent, yielding 2.5% extract after purification. A total of 78 components were identified, with artemisinin and representative sesquiterpene lactones as major constituents. The prepared soft capsules met pharmacopoeial quality standards, showing consistent artemisinin content (~ 6.4 mg/capsule), favorable dissolution (> 85% within 45 min), and good stability. The acute oral LD₅₀ was greater than 2000 mg/kg, and the NOAEL in the 28-day study was 250 mg/kg, with only reversible liver changes observed at high doses. Conclusion This study establishes a reproducible preparation process and provides initial evidence regarding the quality attributes and short-term safety of Artemisia annua soft capsules. The findings suggest improved formulation performance in terms of dissolution and stability; however, further studies, including long-term toxicity, pharmacokinetics, and clinical evaluation, are required.

Berberine, curcumin, biochanin A, cucurbitacin E, and caffeic acid phenethyl ester (CAPE) are plant-derived compounds with long histories of use in traditional medicine for inflammatory and proliferative conditions. Their known capacity to modulate NF-κB signaling makes them candidates for anticancer investigation, particularly in mesenchymal malignancies such as fibrosarcoma, which arise in muscle-rich environments shared with normal myogenic tissue. To evaluate the selective anticancer potential of these compounds in fibrosarcoma (WEHI-164) and normal muscle (L6) cells, with focus on mitochondrial function, mitophagy, cellular senescence, and NF-κB-related metabolic pathways, alongside preliminary in vivo toxicity assessment. IC50 values were determined using MTT and PrestoBlue® assays. Mitochondrial membrane potential was assessed using JC-1 and normalized to the matched untreated control for each cell line, and mitophagy by PINK1/PARKIN immunofluorescence colocalisation together with a mitophagy dye assay. Cellular senescence was measured using a β-galactosidase assay, and ATP levels by a luminescence-based method. Gene expression of NF-κB pathway components and PFKFB3 was analyzed by RT-qPCR. In vivo-like toxicity was assessed using the Galleria mellonella model, including PBS handling, DMSO vehicle, and 70% ethanol utility controls, with survival data analyzed by Kaplan-Meier curves and the log-rank test. The compounds differentially affected normal and cancer cells, indicating selectivity toward malignant phenotypes. Decreased ATP and mitochondrial depolarization suggest disruption of bioenergetic homeostasis, supported by modulation of mitophagy. Stronger effects in WEHI-164 cells indicate higher susceptibility to mitochondrial dysfunction. Increased cellular senescence suggests inhibition of tumor proliferation. These findings indicate that natural NF-κB modulators may exert anticancer effects by targeting mitochondrial and metabolic homeostasis. Differential sensitivity between normal and tumor cells highlights therapeutic potential. In the G. mellonella model, berberine and curcumin did not differ significantly from the PBS or DMSO controls, whereas CAPE, CurE, and particularly biochanin A produced significantly greater larval mortality. The G. mellonella assay should be regarded only as a preliminary acute toxicity screen, and further in vivo studies in mammalian models are required to clarify mechanisms and clinical relevance.

Powder Bed Fusion with an Electron Beam (PBF-EB) enables the fabrication of patient-specific titanium implants with complex geometries; however, the inherent roughness and unique surface features of PBF-EB-produced alloys complicate the optimization of both corrosion resistance and biological performance. This study investigates the combined effects of mechanical surface pre-treatment and anodic oxidation on PBF-EB-manufactured Ti-6Al-4V ELI. Raw, sandblasted, and machined substrates were anodized in orthophosphoric acid at 15 V and 30 V, and their surface topography, surface chemistry, electrochemical behaviour, early-stage bacterial adhesion, and osteoblast colonization were evaluated.Mechanical pre-treatment was found to define the primary surface topography, while anodization modified surface morphology and chemistry in a voltage- and surface-condition-dependent manner. X-ray photoelectron spectroscopy showed that titanium was present exclusively as Ti4+, consistent with fully oxidized TiO₂ at the investigated surfaces, while phosphate- and calcium-containing species were detected after anodization. Electrochemical testing indicated that corrosion behaviour depended on both the initial surface condition and anodizing voltage. Anodization at 30 V was associated with favourable electrochemical responses on machined and sandblasted surfaces but with reduced corrosion resistance on the highly rough as-built substrate. Qualitative and semi-quantitative analysis showed that bacterial fluorescence intensity varied between the investigated surface conditions and may be associated with both surface morphology and chemistry. The highest osteoblast-associated Neutral Red signals were observed for the anodized sandblasted surfaces.Overall, the results indicate that the response of PBF-EB-produced Ti-6Al-4V ELI to anodization depends on the initial surface condition. The combined analysis provides useful guidance for selecting surface preparation and anodization conditions for biomedical applications.

Introduction:There is an evident need for novel ophthalmic therapeutics that operate outside the conventional antibiotic and chemically synthetized antiseptic paradigms. Medicinal plants represent a chemically rich and cost-effective source of bioactive compounds, though their use requires careful toxicological assessment. Methods:In this study, extracts from four Papaveraceae species - Chelidonium majus (roots), Corydalis cheilanthifolia (aerial parts and roots), Glaucium flavum (aerial parts and roots), and Fumaria officinalis (herb) - were investigated across three experimental levels: in silico, in vitro, and in vivo, using the Galleria mellonella larval model. Results:Chemical profiling identified several isoquinoline alkaloids, including berberine, coptisine, protopine, chelidonine, allocryptopine, glaucine, and tetrahydropalmatine. In silico assessment suggested low systemic toxicity but possible blood-brain barrier permeability for some compounds. In vitro antimicrobial testing against Staphylococcus aureus ATCC 6538 and Pseudomonas aeruginosa ATCC 15442, performed using broth microdilution and bacterial-cellulose-based diffusion assays, demonstrated moderate but reproducible activity of extracts, particularly in Ch. majus roots (MIC equal 1.56 mg/mL for S. aureus and 6.25 mg/mL for P. aeruginosa) and in C. cheilanthifolia, where depending on the type of extract (herb_2, herb_1, root), range of MIC was 0,78-1.56 mg/mL for S. aureus and 3,12-6,25 mg/mL for P. aeruginosa. Cytotoxicity testing on fibroblast cell lines showed no significant toxicity in applied concentrations. In vivo testing showed that all extracts, except C. cheilanthifolia (herb_1) and Ch. majus (root) were virtually non-lethal to G. mellonella larvae, indicating a favorable safety profile in 0.05 mg/mL to 0.002 mg/mL concentrations. To evaluate their suitability for ocular application, wettability tests were performed using commercial tear substitute formulations. The incorporation of the C. cheilanthifolia herb_2 extract at bactericidal concentrations did not alter these physicochemical properties. Discussion:Overall, the Papaveraceae extracts demonstrated moderate antibacterial activity and low cytotoxicity in applied concentrations, supporting their potential use as adjuvant components in eye drop formulations - enhancing therapeutic efficacy, reducing required antiseptic dosage, and potentially lowering the risk of resistance development.

(1→3)-α-d-Glucan is an important component of the cell wall of most fungi. The polymer has many applications, including as a therapeutic agent in the prevention or treatment of various diseases, as well as a heavy metal sorbent and a component of new materials used in the plastics industry. The presence of (1→3)-α-d-glucan (water-insoluble, alkali-soluble polysaccharide) in the cell wall of Pleurotus djamor (pink oyster mushroom) was confirmed using specific fluorophore-labeled antibodies. Therefore, the water-insoluble fraction (WI-ASF) of P. djamor B123 fruiting bodies was isolated by alkaline extraction and used for further analyses. The structural features of the WI-ASF were determined by composition analysis, linkage analysis, Fourier transform infrared and Raman spectroscopy, 1H and 13C nuclear magnetic resonance spectroscopy, scanning electron microscopy, as well as viscosity, specific rotation, and gel permeation chromatography. These studies revealed the presence of glucose units linked by α-glycosidic bonds and scanty amounts of mannose and xylose. Furthermore, methylation analysis of WI-ASF demonstrated that the (1→3)-linked glucopyranose (Glcp) is the primary moiety (86.4%) of the polymer, while the 3,4- and 3,6-substituted hexoses are the branching residues of the glucan. The results of chemical and spectroscopic investigations indicated that the analyzed WI-ASF is a (1→3)-linked α-d-glucan type with a molecular weight of 552 kDa.

Pharyngitis is a leading cause of outpatient antibiotic use, despite its typically viral or self-limiting nature. Such unnecessary antibiotic therapies are not only the cause of increasing antibiotic resistance, but also significant changes in the human microbiota in the intestines and other locations, which translate into immune disorders and an increased risk of developing several chronic diseases. Orally administered octenidine-containing lozenges provide a topical alternative; however, their effects on the host microbiota of the oral cavity, throat, and intestine remain unclear. In this study, we evaluated the antimicrobial and antibiofilm in vitro activity of octenidine lozenges against 106 microbial strains, including pathogens and commensals from the oral cavity, pharynx, and large intestine. Minimal biocidal concentrations (MBCs) and minimal biofilm eradication concentrations (MBECs) were determined under physiologically relevant exposure times: 23 min for oral contact and 24 h for intestinal transit. ADME in silico analysis confirmed the lack of absorption of octenidine through the blood-brain barrier and the gastric intestinal mucosa. At concentrations achievable in saliva and the intestinal lumen, octenidine effectively eradicated in vitro all oropharyngeal pathogens while leaving intestinal commensals unaffected. Its impact on oral commensals resembled that of routine mechanical cleaning. These in vitro findings are of high translative value because they support the use of octenidine lozenges as a safe topical treatment for pharyngeal infections, "sore throat", without adverse effects on the gut microbiota.

Acne vulgaris is one of the most common dermatological diseases and has a complex etiology. Despite the wide range of available therapeutic options, modern and effective solutions are still being sought, particularly in the area of topical therapy. The aim of this study was to develop hydrogel formulations that provide stability for the antibiotics they contain—tetracycline or chlortetracycline enriched with azeloglycine—the latter an ingredient supporting acne-prone skin care. The physicochemical parameters, stability, and antimicrobial activity of the obtained formulations were analyzed. HPLC analysis showed that tetracycline exhibited greater stability than chlortetracycline, especially in mildly acidic and neutral environments. The addition of azeloglycine improved the rheological properties of the hydrogels, reduced tetracycline degradation under alkaline conditions, and enhanced the penetration of active ingredients into the model sebum. All tested formulations demonstrated antimicrobial activity against Staphylococcus aureus. In the artificial sebum biofilm model, hydrogels containing azeloglycine more effectively reduced staphylococcal biofilm mass. No formulations showed toxicity towards Galleria mellonella larvae. The results indicate the potential usefulness of the developed hydrogels as modern multifunctional formulations for topical acne treatment. Hydrogel formulations containing tetracycline and azeloglycine may represent a promising future anti-acne preparation exhibiting synergistic antibacterial, anti-inflammatory, and sebum-cleansing effects.

Essential oils (EOs) have long been studied for their antimicrobial properties, yet most investigations rely on simplistic models, limited strain panels, and anecdotal interpretations—failing to meet the standards expected of modern anti-infective agents. Advancing beyond this tradition, we implement a framework developed over several years of systematic investigation. Using this approach, we assessed the antibiofilm activity of Rosmarinus officinalis L. and Thymus vulgaris L. EOs against a panel of clinical Staphylococcus aureus isolates obtained from non-healing wounds. By applying infection-relevant conditions, such as wound-mimicking media and surfaces, strain-level resolution, and both contact and volatile exposure, we revealed substantial inter-strain variability in susceptibility, challenging the notion of EOs as uniformly effective agents. This variability was quantified using robust statistics, lending confidence to the reproducibility and translational relevance of the findings. These results underscore the need for essential oil stewardship: a reproducible, interdisciplinary framework for EO testing, interpretation, and clinical translation. Our work demonstrates that such approach is feasible and sets the foundation for its broader adoption. The key message from this study is that EOs cannot meaningfully support or complement antibiotics and antiseptic agents in combating infections unless they are evaluated with the same methodological rigor.

Pharyngitis is a leading cause of outpatient antibiotic use, despite its typically viral or self-limiting nature. Such unnecessary antibiotic therapies are not only the cause of increasing antibiotic resistance, but also significant changes in the human microbiota in the intestines and other locations, which translate into immune disorders and an increased risk of developing several chronic diseases. Orally administered octenidine-containing lozenges offer a topical alternative, but their effects on the host microbiota of the oral cavity, throat, and intestine remain unclear. In this study, we evaluated the antimicrobial and antibiofilm in vitro activity of octenidine lozenges against 106 microbial strains, including pathogens and commensals from the oral cavity, pharynx, and large intestine. Minimal biocidal concentrations (MBCs) and minimal biofilm eradication concentrations (MBECs) were determined under physiologically relevant exposure times: 23 minutes for oral contact and 24 hours for intestinal transit. At concentrations achievable in saliva and the intestinal lumen, octenidine effectively eradicated all oropharyngeal pathogens while leaving intestinal commensals unaffected. Its impact on oral commensals resembled that of routine mechanical cleaning. These in vitro findings are of high translative value because they support the use of octenidine lozenges as a safe topical treatment for pharyngeal infections - “sore throat”, without adverse effects on the gut microbiota. ### Competing Interest Statement The authors have declared no competing interest. Wroclaw Medical University, D230.25.047

The rising resistance of bacterial and fungal strains, particularly in biofilm form, is diminishing the efficacy of available therapies and poses a major threat to human health. This highlights the need for new antimicrobial agents. A review of biological studies has shown that 2,7-naphthyridine derivatives exhibit a wide spectrum of pharmacological properties, including antimicrobial activity, which has contributed to the development of new compounds containing this scaffold. In this work, the obtained compounds were tested to assess their ability to eradicate biofilm formed by selected reference strains of opportunistic pathogens: Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans as well as towards normal microbiota representative, referred to as the Lactobacillus crispatus. The tested 2,7-naphthyridine derivatives showed selective antimicrobial activity, exclusively against S. aureus. 10j demonstrated the highest, among tested compounds, activity on this pathogen (MIC = 8 mg/L), while compound 10f exhibited ~100-fold stronger activity (MIC = 31 mg/L) than the majority of the library compounds. The in vitro assessment on fibroblast cell lines demonstrated low cytotoxicity of both compounds 10f and 10j, which was subsequently confirmed in vivo using the Galleria mellonella larval model, where no signs of systemic toxicity were observed during the 5-day observation period. Due to the structural similarity of the compounds 10f and 10j to typical gyrase/topoisomerase IV inhibitors, molecular dynamics simulations were performed on a ternary complex containing protein, DNAds, and a 1,5-naphthyridine inhibitor (PDB ID: 6Z1A). Molecular dynamics of the gyrase–DNA ternary complex supported stable binding of both hydrazone derivatives, with 10j showing slightly more favorable MM/GBSA energetics driven by electrostatics and halogen bonding, consistent with its ~4-fold lower MIC versus 10f. Taken together, our data highlight compound 10j as a promising microbiota-sparing antibacterial candidate, particularly suitable for selective interventions against S. aureus, for instance in vaginal infections, where targeted eradication of the pathogen without disturbing protective commensals is highly desirable.

This study evaluates the in vitro antimicrobial efficacy and cytotoxicity of acidified sodium chlorite (ASC), a source of chlorine dioxide. Despite its controversial promotion in alternative medicine as a cure-all solution, known as "Miracle Mineral Solution" (MMS), the data on its factual medicinal activity is very limited. Therefore, we aimed to elucidate the activity of ASC against biofilms of Staphylococcus aureus, Pseudomonas aeruginosa, Enterococcus faecalis, Streptococcus mutans, Pseudomonas aeruginosa, Escherichia coli, and Lactobacillus sp. or an organic acid (ASC1, ASC2, respectively). The lowest antimicrobial concentration of ASC registered was 0.002992% (29.92 ppm) but did not exhibit stronger antimicrobial activity than polyhexamethylene biguanide. Biofilms of S. mutans and E. coli were the most susceptible to tested formulations. Biofilm formed by L. rhamnosus displayed susceptibility to concentrations lower than the minimum biofilm eradication concentration (0.09575%, 957.5 ppm). In the in vitro cytotoxic assay towards eukaryotic fibroblasts and in vivo model of Galleria mellonella larvae concentration-related increase of cytotoxic effects was observed. Our findings demonstrate that these concentrations of ASC which can effectively eradicate biofilms, also pose potential health risks due to their in vitro and in vivo cytotoxicity. It implies that ASC applied in humans can lead to damage to the mucous membrane of the gastrointestinal tract. This research contributes to the ongoing debate on the safety and efficacy of chlorine dioxide in clinical applications, highlighting the need for precise dosing to avoid mucosal damage in therapeutic contexts.

1,8-cineole, renowned for its versatile therapeutic properties, has long been utilized in the treatment of respiratory system disorders. Its potential for oral administration offers a new dimension as an effective systemic therapy with anti-inflammatory and antibacterial effects. Maintaining stable levels of the compound in the body enhances treatment efficacy and reduces the risk of recurrence. Additionally, 1,8-cineole exhibits anti-biofilm activity, addressing biofilms often associated with chronic respiratory infections, thereby making it an effective tool in managing hard-to-treat infections. This article reviews the latest research and potential applications of 1,8-cineole in the treatment of respiratory infections, emphasizing its multifaceted mechanism of action.

Helicobacter pylori is a common Gram-negative bacterium that inhabits the human stomach and causes a variety of gastric pathologies. One of the growing concerns is its dynamic spread of antibiotic resistance, a process in which biofilm formation is involved. Therefore, it is necessary to find an appropriate, high-throughput research model for the in vivo biofilm development by H. pylori. The aim of the current research report was to determine the usefulness of G. mellonella larvae in assessing the survival of a multidrug-resistant, strong biofilm producing H. pylori strain during its exposure to stress caused by clarithromycin. Using infection models lasting for 3 or 6 days, we confirmed the ability of the tested H. pylori strain to survive in the larvae. We noticed that exposure to clarithromycin significantly reduced the number of cultured bacteria relative to the control, although we did not observe any differences in the number of bacteria using time-lapse, live analysis of fluorescently stained larval hemolymph samples. In conclusion, we confirmed that the examined H. pylori strain can produce biofilm in G. mellonella larvae organism and is able to survive exposure to minimal inhibitory concentrations of clarithromycin (established in vitro) in in vivo conditions. Further refinement of methodologies for monitoring the viability of clinical H. pylori strains in the greater wax moth larvae will enhance the accuracy and reliability of this promising research model.

It is the first comprehensive study showing the effect of amber particles in a biodegradable polymer matrix. The effect of amber powdered waste (AbW) on poly(ε-caprolactone) (PCL) processing was evaluated, along with the influence of different processing methods on the structure and properties of the PCL/AbW systems. AbW was melt-mixed with PCL in proportions of 1 %, 2.5 %, or 5 % wt, without the use of any compatibilizer. Following the twin-screw extrusion mixing, the composites were further processed using injection molding or cast-film extrusion. The materials obtained were tested for processing properties (melt flow rates, density), morphology (scanning electron microscopy), chemical composition (infrared spectroscopy), thermal properties (differential scanning calorimetry, thermogravimetry, Vicat softening temperature, and heat deflection temperature), strength (static tensile test, flexural test, hardness, and impact strength) and surface properties (contact angle). The addition of amber to PCL did not require changes to the parameters of subsequent processing. The interactions between the low-molecular weight compounds from AbW and the matrix appear more significant than any potential degradation of the polymeric chain during processing. Regardless of the processing method, the PCL/AbW system exhibited strong interfacial bonding while maintaining the ductile nature of the PCL matrix. The addition of 1 % and 2.5 % AbW enhanced the reinforcement and strengthening of the composites. Nevertheless, the addition of 5 % AbW resulted in lower mechanical performance and increased heterogeneity in thermal events. Amber powdered waste from the jewelry industry appears to be a promising sustainable filler and compatibilizer for PCL, while also potentially adding biological functionalities.

Post-processing of DLP-printed polyethylene glycol diacrylate (PEGDA)/polyvinyl alcohol (PVA) resins with tannic acid (TA) enhanced their mechanical properties and introduced bioactive functionalities.

The rising interest in essential oils (EOs) as antimicrobial agents demands evaluation frameworks that provide structured, reproducible assessments. In this study, we examined the strain-dependent response of Pseudomonas aeruginosa to pharmacopoeia-grade Thyme Essential Oil (obtained from Thymus vulgaris L., TEO) or polyhexamethylene biguanide antiseptic (PHMB) using a panel of ten genetically diversified strains in planktonic and biofilm forms, and by complementary in vitro models. Chemical composition of TEO was assessed using Gas Chromatography-Mass Spectrometry (GC–MS), and the main components were thymol, p-cymene, and γ-terpinene. Despite uniform test conditions, we observed striking inter-strain variability: TEO Minimal Inhibitory Concentrations (MICs) differed by up to 1,000-fold, and biofilm susceptibility profiles ranged from full tolerance to near-complete eradication. Notably, strains with low metabolic activity and sparse cell populations—but high matrix biomass—exhibited reduced responsiveness to TEO, while susceptibility to PHMB was more consistent, though not absolute. These findings highlight the critical influence of both microbial phenotype and agent formulation on antimicrobial outcomes. Rather than framing EOs as superior or inferior alternatives, our results advocate for their integration into a stewardship paradigm—one that values standardization, model-based evaluation, and informed formulation. In this context, we position essential oil stewardship not as a constraint but as a necessary evolution for their credible inclusion in antimicrobial strategies.

The extracts from fruits of Chaenomeles japonica (Thunb.) Lindl. ex Spach (CJE), Cornus mas L. (CME), and Hippophaё rhamnoides L. (HRE) are known inhibitors of a variety of eukaryotic hydrolases, engaged in the digestion of fats and polysaccharides. However, there are no data on their potential interaction with the bacterial hydrolases participating in the replication of microbial nucleic acids. This analysis predicted the interaction of the most abundant constituents of HRE, CJE, and CME with the bacterial nucleases. The analysis covered the molecular docking of isorhamnetin glycosides, procyanidins C1 and B2, epicatechin, loganic acid, and cornuside with bacterial enzymes (Escherichia coli endonuclease 1, colicin E9, and ribonuclease H; or Staphylococcus aureus thermonuclease and nuclease SbcCD). The suggested complexes have been subjected to molecular mechanics with generalized Born and surface area solvation (MM/GBSA) calculations. The second aim was the in vitro evaluation of the influence of the CJE, HRE, and CME on the metabolic activity of bacterial biofilm of selected microbial strains, as well as fibroblasts (L929) and adenocarcinoma intestinal cells (Caco-2) toxicity. Among all extracts, CME showed the most relevant effect on the survival of planktonic cells and biofilm of E. coli and Pseudomonas aeruginosa. As a result of in silico studies, most virtual hits were predicted to inhibit the proteins under investigation, except for procyanidin C1. Further research on the direct interaction of phytochemicals and selected enzymes in vitro is required and challenged.