Colloids and Surfaces B: Biointerfaces

PURPOSEBenzalkonium chloride (BAK), a common preservative in eye drops, has a major side effect of dry eye. The mechanisms are typically attributed to BAK cytotoxicity. However, due to its surfactant properties, BAK can disrupt the tear film lipid layer (TFLL), leading to dry eye. This study examined the stability and compressibility of the TFLL and the impact of the presence of BAK. METHODSMeibomian gland secretion (meibum, source of the TFLL) was collected from sacrificed cows eyelids. Lipids were extracted by dissolving meibum in chloroform to a final concentration of 1 mg/mL, with one solution additionally containing 0.1 mg/mL BAK. Each solution was overlaid on a water subphase in a Langmuir Trough-Blodgett trough. The changes of surface pressure ({pi}) with area (A) for the lipid film upon compression were monitored, and the corresponding compression modulus (Cs-1) at each data point was determined. RESULTSThe {pi}-A isotherms for meibum lipid monolayers exhibited near-reversible behavior with a smooth profile with a maximum {pi} of approximately 32 mN/m. The Cs-1-{pi} isotherms of the meibum lipid monolayer show that the films are gel-like with a constant compressive modulus of 24-32 mN/m within the surface pressure range of 8-30 mN/m. In contrast, adding BAK dramatically decreased the maximum surface pressure to only 10 mN/m and the compressive modulus to only 2-10 mN/m. CONCLUSIONSThis study demonstrated that BAK disrupts the meibum lipid layer by forming a monolayer with decreased stability and reduced compressive resistance, a mechanism that may underlie its dry-eye side effect yet has largely been neglected.

This study presents the first investigation of the adsorption behaviour of Afatinib on gold nanoparticle (AuNP) monolayers, employing a combination of AFM-SEIRA and SERS techniques. Two types of AuNPs with distinct sizes, synthesized using different reagents, were employed to elucidate the influence of surface type on drug adsorption. The first type of AuNPs was synthesized using sodium borohydride (SB), whereas the second type was obtained using hydroxylamine hydrochloride (HH) as the reducing agent. AFM-SEIRA revealed that Afatinib interacts to the AuNPs primarily through the quinazoline ring, amide group, and amino moiety, with adsorption geometry strongly dependent on nanoparticle type. Contributions from CH3 and CH2 moieties were also identified, indicating their role in stabilizing the molecule/metal interface. Time-resolved SERS studies demonstrated that the adsorption process is dynamic and involves molecular reorientation, followed by gradual desorption, which is accelerated at physiological temperature (37 {degrees}C). Competitive adsorption experiments with phenylboronic acid (PBA) showed that Afatinib exhibits higher affinity toward AuNPs, however, co-adsorption leads to reduced stability of both species on the surface. The results reveal molecular insights into drug/nanoparticle interactions and emphasize the role of surface functionalization in efficient nanocarrier design. This work deepens understanding of adsorption at plasmonic interfaces for biomedical use.

Many biochemical processes are dependent on the presence or absence of molecular oxygen (O2). Palladium-tetrapyrrol derivatives can be used to measure O2-concentrations and O2-turnover during biochemical reactions and microbial growth in standard microtiter plates (MTPs). Palladium(II)-5,10,15,20-(tetrapentafluorophenyl)-porphyrin (1; CAS 72076-09-6) and Palladium(II)-5,10,15,20-(tetraphenyl)tetrabenzoporphyrin (2; CAS 119654-64-7) are introduced with this study. Spectral analyses of both compounds revealed that fluorescence quenching by O2 is not evenly distributed throughout all wavelengths and can therefore be used ratiometrically. Experimentally determined fluorescence lifetimes are around 500 {micro}s and 300 {micro}s for 1 and 2, respectively. A simple protocol is disclosed, how to immobilize the indicators on the bottom of MTP wells to give clear transparent dye doped polymer layers. We propose a straightforward procedure of how fluorescence data can be processed and calibrated in terms of O2 concentrations. Diverse applications are demonstrated and discussed, which include oxygen consumption and production by microorganisms as well as by enzymatically catalysed biochemical reactions. Various aspects are critically considered, as there are e.g. the dependence of O2 solubility on temperature and salinity, the diffusion of O2 across diverse phase boundaries, the unwanted O2 ingress into the reaction volume, the oxygen binding capacity of the MTP plastic material and the pH-dependence of the sensor layer. The findings and methods presented here open up a broad variety of high throughput assays involving changes of dissolved O2 as measurands for biochemical and biological activity. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=98 SRC="FIGDIR/small/718663v1_ufig1.gif" ALT="Figure 1"> View larger version (30K): org.highwire.dtl.DTLVardef@4daa4dorg.highwire.dtl.DTLVardef@e7ab8aorg.highwire.dtl.DTLVardef@1af1149org.highwire.dtl.DTLVardef@97fea5_HPS_FORMAT_FIGEXP M_FIG C_FIG

Significant cellular processes, including protein sorting, signal transduction, and pathogen entry, amongst others, are associated with membrane microdomains, also known as lipid rafts. Lipid rafts, due to their unique biophysical properties compared to their surrounding environment, which stem from their distinct lipid and protein profiles, have garnered interest in methods and techniques that tune their coexisting liquid-ordered/liquid-disordered state, aiming to disrupt or destabilize them. Since cholesterol stabilizes the membrane domain, cholesterol-depleting compounds like cyclodextrin can be used to destabilize and disrupt the membrane rafts. Overall, given the membrane rafts importance in biological processes, it is crucial to understand the biophysical factors that influence its stability. In this study, we present a new method for disrupting and dissolving lipid rafts in a model system of phase-separated supported lipid bilayer (SLB) patches composed of DOPC, DPPC, and cholesterol. Using fluorescence microscopy to monitor the liquid ordered (Lo) and liquid disordered (Ld) phases of the SLB patches, we observed that adding DOPC liposomes causes a transformation of the co-existing Ld and Lo phases into a single-phase bilayer. On the other hand, adding liposomes that match the lipid content of the phase-separated SLB patch increase the areas of the existing Ld and Lo phases. This work also offers a new method for redistributing raft-localized molecules, confirmed by tracking the redistribution of cholera toxin bound to GM1 after domain dissolution with DOPC liposomes. The work describes an alternative method for dynamically altering membrane composition and dissolving domains via liposome addition, rather than lipid depletion or exchange.

Nowadays N95 facial mask has gain huge attention due to COVID19 pandemic situation and it serves as the prime PPE. Though the microbes can be restricted to get inside the human body due to the presence of mask temporarily, but over the time, bacteria and other microbes may get entrapped into the threads of the mask itself and thus acting as a storage chamber of microbes. It is necessary to eliminate them from the mask surface. To do so different floral structured Nano-ZnO with variable oriented arrangement of petals were fabricated on the surface of the N95 mask and further characterized through instrumentations including XRD, FTIR,UV-Vis, Fluorescence-Spectroscopy, SEM, DLS. The average crystallite size calculated for synthesized four different ZnO nanoflower were 25.19 nm, 23.46 nm, 27.27 nm and 31.78 nm (for glycerol, PEG, EDTA, Chitosan assisted) respectively. The antimicrobial activity was investigated by standard microbial broth dilution assay and Kirby-Bauer test which assured the inhibition of the bacterial growth. The MIC-MBC value of ZnO nanoflowers for E.coli and B. subtilis were found to be effective at dilution of 250 {micro}g/ml and 100 {micro}g/ml. Additionally a modified Kirby-Bauer assay has been designed to investigate the killing efficiency of the bacteria (E.coli). O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=145 SRC="FIGDIR/small/719592v1_ufig1.gif" ALT="Figure 1"> View larger version (35K): org.highwire.dtl.DTLVardef@a76030org.highwire.dtl.DTLVardef@9bf1b3org.highwire.dtl.DTLVardef@19232forg.highwire.dtl.DTLVardef@54fe68_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOFig. - Graphical AbstractC_FLOATNO C_FIG

This study presents the development and spectroscopic characterization of an erlotinib-functionalized gold nanoparticle (erlotinib:AuNP) nanosystem designed for targeted delivery to metastatic non-small cell lung cancer H1299 cells. Initial MTS assays demonstrated that free erlotinib induced a concentration-dependent reduction in cell viability, while 0.1 {micro}M erlotinib exhibited negligible cytotoxicity and was therefore selected for nanosystem fabrication. AuNPs alone showed minimal toxicity toward H1299 cells over the investigated concentration range. Following conjugation of erlotinib with AuNPs, the resulting nanosystems reduced cell viability to approximately 60%, indicating enhanced biological activity of the drug after nanoparticle-assisted delivery. Fluorescence microscopy confirmed the intracellular internalization of the nanosystems in H1299 cells, with nanoparticle aggregates predominantly localized in the perinuclear and perimitochondrial regions. Three-dimensional Raman spectroscopy (3D RS) mapping further verified the intracellular localization of the conjugates through characteristic Raman signatures of erlotinib:AuNPs. Importantly, 3D RS enabled detection of nanosystems at concentrations below the sensitivity limit of fluorescence imaging, demonstrating superior analytical performance for intracellular nanosystem tracking. Atomic force microscopy-infrared (AFM-IR) spectroscopy coupled with principal component analysis (PCA) demonstrated substantial biochemical modifications induced by the erlotinib:AuNP nanosystems, including enhanced lipid-related spectral features and significant alterations in protein secondary structure, particularly the increased contribution of unordered and antiparallel {beta}-turn conformations. The obtained results demonstrate that combining plasmonic nanocarriers with advanced vibrational spectroscopy enables highly sensitive monitoring of intracellular drug delivery and nanosystem-induced biochemical responses in cancer cells.

Dendrimers are nanosized molecules with potential antimicrobial applications. This study evaluates the antibacterial and anti-biofilm properties of two cationic dendrimers, NVX-G6 (G6) and NVX-G9 (G9), against clinically relevant bacterial pathogens. Minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC99) were determined for Escherichia coli, Pseudomonas aeruginosa, methicillin-sensitive Staphylococcus aureus (MSSA), and methicillin-resistant Staphylococcus aureus (MRSA). The synergy of dendrimers with ceftazidime and vancomycin was evaluated using checkerboard assays. Furthermore, biofilm formation inhibition assays and fluorescent microscopy were performed to assess dendrimer interactions with bacterial biofilms. The results indicate that G6 and G9 exhibit limited direct antibacterial activity at high concentrations (MIC > 1024 {micro}g/mL) but demonstrate synergistic effects when combined with ceftazidime against E. coli and P. aeruginosa (FIC < 0.5). Notably, both dendrimers penetrated and colocalized within established biofilms, with time-dependent reductions in biomass observed after extended incubation, suggesting a role in progressive biofilm disruption rather than acute inhibition of formation, although significant biomass reduction was not observed under standard assay conditions. These findings contribute to the understanding of dendrimer-antibiotic interactions and their implications in antimicrobial and nanomedicine therapy.

Biofilm extracellular matrix (ECM) varies with environmental conditions and substrate properties. Understanding the surface-biofilm relationship helps to perfect antibacterial strategies and to design new engineered living materials (ELMs). In this work, we studied how cationic and anionic polyelectrolyte coatings affect macroscopic features of Escherichia coli curli-producing biofilms, as well as the properties of their curli amyloid fibers. Cationic coatings limited biofilm spreading, increased their surface density and water absorption, which correlated with a higher yield of curli amyloid fibers with looser structure. In contrast, anionic surfaces allowed for standard biofilm spreading, with a lower fiber yield but a more compact and chemically stable fiber structure. Higher biofilm rigidity and adhesion were measured on both types of charged surfaces. Thus, we propose that the differences in biofilm macroscopic properties result from a trade-off between curli quantity and quality in the ECM, namely fiber density and molecular packing, as well as their interaction with water. Our findings provide insights on how the biophysical properties of the ECM can be controlled by tuning the substrate physico-chemical characteristics with charged coatings. This work opens up new avenues for developing antimicrobial strategies, as well as tailoring the properties of amyloid-based ELMs. TOC figure O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=82 SRC="FIGDIR/small/721109v1_ufig1.gif" ALT="Figure 1"> View larger version (22K): org.highwire.dtl.DTLVardef@191cd79org.highwire.dtl.DTLVardef@148f914org.highwire.dtl.DTLVardef@1d8c2f8org.highwire.dtl.DTLVardef@1e84eaf_HPS_FORMAT_FIGEXP M_FIG C_FIG

The rapid emergence of antimicrobial resistance, particularly among multidrug-resistant (MDR) and extended-spectrum {beta}-lactamase (ESBL)-producing Escherichia coli, necessitates the development of novel therapeutic strategies. In this study, we report the green synthesis and functionalization of silver nanoparticles (AgNPs) using Azadirachta indica leaf extract conjugated with amoxicillin (Amoxicillin-AI-AgNPs) to enhance antibacterial efficacy. The synthesized nanoparticles were characterized using UV-Vis spectroscopy, FTIR, XRD, DLS, SEM, EDAX, and TEM analyses, confirming the formation of stable, spherical, crystalline nanoparticles with an average size of [~]87 nm and a zeta potential of -28.73 mV. High conjugation efficiency ([~]94%) of amoxicillin with AgNPs was achieved after 96 hours of incubation. Antimicrobial activity assessed against 88 clinical MDR and ESBL-producing E. coli isolates demonstrated significantly enhanced efficacy of Amoxicillin-AI-AgNPs compared to amoxicillin alone, with minimum inhibitory concentrations (MIC) ranging from 1.56 to 6.25 {micro}g/mL and minimum bactericidal concentrations (MBC) between 25-100 {micro}g/mL. Cytotoxicity evaluation on HEK-293 cells revealed a relatively high IC50 value (382.14 {+/-} 6.59 {micro}g/mL), indicating low toxicity at antibacterial doses. The synergistic interaction between AgNPs and amoxicillin likely contributes to improved bacterial inhibition and overcoming resistance mechanisms. Overall, this study highlights the potential of plant-mediated antibiotic- nanoparticle conjugates as an effective and biocompatible approach to combat antibiotic-resistant bacterial infections.

In this paper, we aim to present a new intravital cells visualization method, which is based on use of a dye called ABDS ("A Beautiful dye for staining"), which can be prepared using a marker pen and is useful for eukaryotic cell research. Using a wide range of instruments, including optical measurements, microscopy studies and wet biology techniques, we have shown that ABDS is close by properties to Rhodamine 6G dye (R6G), which is well known as endoplasmic reticulum stainer. However, by the careful examination of the ABDS and R6G images (ABDS/R6G), we have proved for the first time that these dyes also stain the cytoplasmic membranes. The significant contrast between ABDS/R6G signal from cell membrane and endoplasmic reticulum allows them to be distinguished in the fluorescence photographs. Other important properties of ABDS are its availability, simplicity in manufacturing, safety for living cells in vitro, and bright stable fluorescence, which in contrast to commercial dye like DiBAC allows us to study cells in space and time with high detalization. The paper includes a method for preparing ABDS, a data set with its characteristics, comparison with other commercial dyes, as well as examples of ABDS usage in cells research. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=198 SRC="FIGDIR/small/717455v1_ufig1.gif" ALT="Figure 1"> View larger version (65K): org.highwire.dtl.DTLVardef@f1ceacorg.highwire.dtl.DTLVardef@137abd2org.highwire.dtl.DTLVardef@1f19efcorg.highwire.dtl.DTLVardef@1fcbc9e_HPS_FORMAT_FIGEXP M_FIG C_FIG HighlightsO_LIA protocol for high-resolution vital staining of the cells using an inexpensive dye based on permanent marker ink is proposed. C_LIO_LIThe absorption, emission and Raman spectra of the proposed dye are presented, and a direct comparison with commercial dyes Rhodamine 6G, DiBAC and Deep Red Cell Mask dye is made. C_LIO_LIThe main characteristics of the proposed dye are low toxicity, long-term fluorescence, and the ability to separately stain the endoplasmic reticulum and cytoplasmic membrane. C_LIO_LIThe ability of the Rhodamine 6G dye to stain cell membranes also has been proved. C_LI

Collagen type II (Col-II) and collagen type I (Col-I) are major components of articular cartilage present at different ratios at its surface. Understanding how each of these components mediates the assembly of molecular films derived from synovial fluid (SF), the native lubricant of synovial joints, is critical to explain the loss of mechanical performance in pathological conditions, guide the design of biomaterial implants meant to be in contact with SF, and develop molecular therapies to restore SF properties. This work demonstrates that Col-II articular surface model assists in scaffolding of full SF-derived films, while Col-I model lacks SF film scaffolding capabilities. However, when Col-II and Col-I are exposed to recombinant lubricin (rLub) alone, the major boundary lubricant in SF, both adsorbed and retained similar amounts. These insights, deduced from quartz crystal microbalance with dissipation, diffuse reflectance circular dichroism, and atomic force microscopy, reveal possible mechanisms underlying the loss of mechanical performance of synovial joints in pathology, where Col-I becomes the major collagenous component of the articular cartilage surface, as well as considerations for designing functional biomaterial implants. Furthermore, this work reinforces the idea of rLub as an intra-articular osteoarthritis therapy with the ability to bind to Col-II and Col-I, irrespectively. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=170 SRC="FIGDIR/small/726594v1_ufig1.gif" ALT="Figure 1"> View larger version (69K): org.highwire.dtl.DTLVardef@138a77borg.highwire.dtl.DTLVardef@7b8512org.highwire.dtl.DTLVardef@15d7060org.highwire.dtl.DTLVardef@17ccf20_HPS_FORMAT_FIGEXP M_FIG C_FIG

The emergence of multidrug-resistant (MDR) and extended-spectrum {beta}-lactamase (ESBL)-producing Escherichia coli poses a significant threat to global public health, necessitating the development of alternative antimicrobial strategies. In this study, biogenic silver nanoparticles (AgNPs) were synthesized using aqueous seed extract of Azadirachta indica as a green, eco-friendly reducing and stabilizing agent. Successful synthesis of nanoparticles was confirmed by a visible color change and characterized using UV-Visible spectroscopy, FTIR, XRD, DLS, SEM, EDAX, and TEM analyses. The synthesized AgNPs exhibited a strong surface plasmon resonance peak at 420 nm and were predominantly spherical with an average size of [~]38 nm and a zeta potential of -24.26 mV, indicating moderate stability. The antimicrobial efficacy of the synthesized AgNPs was evaluated against 88 clinical isolates of MDR and ESBL-producing E. coli. The nanoparticles demonstrated potent antibacterial activity with a minimum inhibitory concentration (MIC) ranging from 1.5625 to 3.125 {micro}g/mL and bactericidal effects at low concentrations, significantly outperforming the neem seed extract alone. Cytotoxicity assessment using HEK-293 cell lines revealed a relatively high IC50 value (297.01 {+/-} 10.04 {micro}g/mL), suggesting low toxicity at effective antimicrobial doses. Overall, the study highlights the potential of A. indica seed-mediated AgNPs as an effective and biocompatible antimicrobial agent against resistant bacterial strains, warranting further in vivo investigations for clinical applications.

Underwater adhesion research increasingly draws on bioinspired systems to uncover the molecular mechanisms that enable strong interfacial binding in aqueous environments. The biofilm adhesin Bap1 from Vibrio cholerae contains a short peptide motif, SYWFFGWHTK (CP), which exhibits exceptional adhesive performance, surpassing mussel foot protein mfp5 under comparable conditions. Despite its promise, the roles of ionic environments and aggregation behavior in governing CP adhesion remain unclear. In this study, we investigate how ion identity influences CP aggregation, film formation, and interfacial properties. Using dynamic light scattering, we identify the formation of micron-scale assemblies of aggregated molecular clusters (AAMCs), with size distributions modulated by salt type. Quartz crystal microbalance with dissipation and liquid atomic force microscopy reveal that CP film formation is both surface- and ion-dependent. On gold substrates, AAMCs preferentially adsorb and collapse into rigid, smooth nanofilms, consistent with hydrophobic-driven compaction. In contrast, silicate surfaces inhibit such collapse, yielding distinct morphologies and interfacial energetics. These findings demonstrate that surface chemistry and ionic conditions jointly regulate peptide aggregation and adhesion. This work provides mechanistic insight into hydrophobic-rich peptide systems and informs the rational design of next-generation wet adhesives, with broader implications for biomaterials and peptide-based formulations. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=130 SRC="FIGDIR/small/733527v1_ufig1.gif" ALT="Figure 1"> View larger version (40K): org.highwire.dtl.DTLVardef@1bd012aorg.highwire.dtl.DTLVardef@1977892org.highwire.dtl.DTLVardef@16cf79borg.highwire.dtl.DTLVardef@f405bf_HPS_FORMAT_FIGEXP M_FIG C_FIG

Culture conditions were optimized for the production of melanin nanoparticle by the bacterial strain Pseudomonas stutzeri BTCZ 305. Response surface methodology was employed for determining the most significant fermentation conditions using variables including, pH, temperature and L-tyrosine concentration identified through one-factor-at-a time approach. Box-behnken design consisting of 17 different combinations of all these factors were performed. Using this methodology, a quadratic regression model was built and the optimal combinations of media constituents for maximum melanin production 1192.27 microg/mL were determined as temperature (32.5 degreeC), pH (8.5) and L-tyrosine concentration (7 g/L). Melanin production was obtained experimentally coincident with the predicted value and the model was proven to be adequate. The nanostructural distribution, its stability in colloidal suspension and particle size were also characterized with the help of TEM, particle size analysis and Zeta potential. The potent applicability of this molecule in anti-inflammation and wound healing was also elucidated.

Environmental pollution from leather industries have become a menace. The microbial remediation of industrial waste and its reuse for agriculture could be a beneficial outcome. In present study, the bioremediated Cr III in the effluents are further converted to value product - Chromium oxide NP. This ensures double edged benefit as effluent is bioremediated and Chromium oxide NP with several applications is derived. A noteworthy advancement of the research involved the green synthesis of chromium oxide nanoparticles using Tridax procumbens. The effluent bioremediated can be used for agricultural purposes. By effectively characterizing tannery effluent and isolating chromium-tolerant bacteria, the study not only demonstrate a practical bioremediation solution but also showcase the potential of green synthesis in producing chromium oxide nanoparticles. In conclusion, this research marks a significant advancement in environmental science, leveraging both biological and nanotechnological innovations to address pressing challenges in pollution control. The present study focuses on a novel process of obtaining chromium oxide nanoparticle from tannery effluent with several applications derived from bioremediated tannery effluent using a cost-effective and eco-friendly process. The nanoparticle has a stable particle size and exhibit antioxidant, anti-diabetic properties. This product offers a breakthrough solution for the leather industry and healthcare sector. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=132 SRC="FIGDIR/small/720289v1_ufig1.gif" ALT="Figure 1"> View larger version (52K): org.highwire.dtl.DTLVardef@192e96borg.highwire.dtl.DTLVardef@1aae28org.highwire.dtl.DTLVardef@19fd282org.highwire.dtl.DTLVardef@1b562f9_HPS_FORMAT_FIGEXP M_FIG C_FIG

Liposomes are self-assembled lipid vesicles capable of encapsulating both hydrophilic and hydrophobic therapeutics, making them versatile platforms in drug delivery and biomedical technology. In this study, the limitations of the classical thin-film hydration method were critically evaluated, and a sustainable, systematically optimized strategy was established for generating defined liposomal lamellar phases. Hydration conditions were optimized, and 4 mL of buffer per 10 mg of lipid was determined to be optimal for effective rehydration and improved statistical reliability of vesicle measurements. A refined probe-sonication protocol (20% amplitude, 5 s ON/55 s OFF pulse) enabled controlled transformation of multivesicular vesicles into stable multilamellar and unilamellar vesicles at net ON-times of 90 s and 185 s, respectively, without overheating or contamination. In addition, a Python-based machine-learning tool was developed for vesicle size characterization. Collectively, these optimizations provided a reproducible and sustainable framework for preparing liposomes across different lamellar phases.

This study investigates a novel light-activated drug delivery system designed to produce on-demand drug release. The light-activated system was developed by incorporating a photostable photothermal agent, croconium dye, into liposomes to enable thermally triggered drug release. The drug release from the liposomes was determined at three powers of 210, 295, and 380 mW under 0-, 1-, and 2-minute light irradiation. A continuous wave 808 nm laser was used as the light source. Dexamethasone sodium phosphate (DSP) released from the liposomes was tunable depending on the power and irradiation time with a range of 1 -19 g released depending on irradiation power and time. For local temperature measurement during the photothermal activation, polymerized 10, 12 - Pentacosadiynoic acid (PCDA) was incorporated in the lipid bilayer. Under heating polymerized PCDA undergoes a transition into a red phase from a blue phase. Utilizing the spectrum changes under known temperatures a regression model was developed to calculate the local temperature of the liposomes under irradiation. The ability of the liposomes to release DSP under irradiation in the presence of a phantom tissue was tested under different attenuation coefficients to match various common biological tissues. The liposomes were still able to release DSP in the presence of tissue phantoms for a certain thickness of the tissue. Finally, the cytotoxicity of the liposomes with the croconium dye for chemical and thermal toxicity was determined. The liposomes displayed good biocompatibility with Human Microvascular Endothelial Cell line-1 (HMEC-1). The results support the use of croconium dye as a potential alternative to commonly photothermal agents used in drug delivery such as metal nanoparticles. Future work will focus on optimization of absorbance spectrum for drug release, and in vivo studies for efficacy and safety.

Manufacturing and storage processes can expose microbes to oxidative stress, reducing viability and limiting their use in biotechnological applications. Here, we evaluate graphene quantum dots (GQDs) containing hydroxyl and carboxyl groups as protective additives that mitigate peroxide-induced oxidative stress in Escherichia coli. GQDs did not adversely affect bacterial growth under basal conditions and restored growth in the presence of hydrogen peroxide. Using the membrane-partitioning fluorescent probe C11-BODIPY, we found that GQDs reduced peroxide-induced oxidation in bacterial membranes. We further used redox-sensitive roGFP2 probes to monitor intracellular oxidative stress and found that GQDs suppressed intracellular hydrogen peroxide accumulation and attenuated disruption of glutathione redox homeostasis. Together, these results show that GQDs protect bacteria by limiting peroxide-driven oxidative damage at both membrane and intracellular levels. This work supports the potential use of GQDs as protective additives for microbial formulations that are susceptible to oxidative stress.

Staphylococcus aureus (S. aureus) is an opportunistic pathogen that is a global health concern for its ability to cause a wide spectrum of clinical infections. Due to the emergence of resistance to commonly used antibiotics, there has been interest in exploring the use of antimicrobial peptides to treat S. aureus infections. However, changes in the lipid composition of the lipid bilayer membrane can alter the activity of peptides, and S. aureus is able to induce variations in lipid composition in response to environmental stress. Here, we explore how the main lipid components in S. aureus are altered when exposed to LL-37, a human cathelicidin involved in primary immune response, and ATRA-1, a short antimicrobial peptide derived from the snake Naja atra venom. A lipidomic study is conducted through HPLC-MS-MS (LC-ESI-MS/MS) to quantify phosphatidylglycerol, cardiolipin, lysyl-phosphatidylglycerol, monogalacto- and digalacto-diacylglycerol, and carotenoids. In addition, menaquinones, responsible for electron transport during oxidative phosphorylation, were also quantified. Biophysical properties such as membrane electric surface potential and lipid packing were assessed. We find that lipid adaptation is specific to the type of antimicrobial peptide, where ATRA-1 mainly induces changes in the electric surface potential through variations in Lysyl-PG, while exposure to LL-37 changes carotenoid levels, inducing an increase in membrane rigidity as measured by FTIR. In addition, both peptides induce a reduction in menaquinone and DGDG levels. These findings highlight the role of membrane lipid remodeling as a peptide-specific response mechanism in S. aureus, with implications for the development of AMP-based therapies. HighlightsO_LIStaphylococcus aureus responds through shifts in lipid composition and membrane biophysical properties to exposure to the antimicrobial peptides LL-37 and ATRA-1. C_LIO_LIBoth LL-37 and ATRA-1 lead to shifts in the glycolipids MGDG and DGDG; two lipids involved in regulating negative membrane curvature stress and responsible for shifting resistance to antimicrobial peptide activity in Staphylococcus aureus. C_LIO_LILL-37 treatment leads to an overall reduction in carotenoid content in Staphylococcus aureus, including the carotenoid end-product staphyloxanthin and the precursor 4,4-diaponeurosporenoic acid. Both lipids regulate membrane biophysical properties and protect Staphylococcus aureus from oxidative stress. C_LIO_LIBoth LL-37 and ATRA-1 lead to a reduction in menaquinone levels, which are involved in the electron transport chain during oxidative phosphorylation. Reduction in these menaquinones have been associated to the formation of small colony variants that are often observed in chronic Staphylococcus aureus infections. C_LI

The protein corona (PC) formed on the surface of nanoparticles (NPs) upon exposure to human biofluids is a dynamic interface that reflects the physiological and pathological status of the host. In this study, we investigated how maternal body mass index (BMI) influences the composition of the NPs PC during late pregnancy. Polystyrene NPs were incubated with plasma samples collected from third-trimester pregnant individuals across normal weight, overweight, and obese BMI categories. Comprehensive characterization using dynamic light scattering (DLS), zeta potential measurements, and transmission electron microscopy (TEM) confirmed BMI-dependent differences in PC thickness and colloidal stability. SDS-PAGE and label-free quantitative proteomics revealed distinct molecular compositions: PCs from obese individuals were enriched in inflammatory and lipid metabolism-associated proteins (e.g., APOE, CRP), while normal weight-derived PCs showed higher levels of complement regulators and extracellular matrix proteins. Principal component analysis (PCA) demonstrated clear clustering of proteomic profiles by BMI group, suggesting BMI-specific PC fingerprints. These findings indicate that maternal metabolic phenotype shapes nano-bio interactions at the proteomic level and highlight the potential of PC profiling as a non-invasive approach for assessing maternal health and metabolic status. This work lays the foundation for integrating NP-based proteomics into precision nanomedicine for maternal-fetal health monitoring.