NIH3T3 Cells Research Articles

Rational design of bioengineered recombinant collagen-like protein enhances GelMA hydrogel for diabetic wound healing

Gelatin methacryloyl (GelMA) holds significant potential in tissue engineering; however, its clinical applications are often constrained by its lack of functional groups. To overcome this limitation, recombinant proteins with multiple biofunctional domains present a promising strategy for GelMA functionalization, enhancing its biological properties. In this study, we developed a rationally designed recombinant collagen-like protein (RC) engineered with multiple biofunctional domains, which demonstrated the ability to upregulate collagen 1α (COL-1α) expression in NIH-3 T3 cells. By utilizing EDC/NHS chemistry, the purified RC was conjugated to GelMA, resulting in GelMA-RC hydrogels that significantly improved cell viability and migration compared to unmodified GelMA. Subsequent in vivo studies showed that RC-modified GelMA exhibited superior wound healing efficacy, largely attributed to enhanced expression of cytokeratin-14 (CK-14) and COL-1α. These findings underscore the potential of RC-functionalized GelMA in promoting diabetic wound repair and suggest broader applicability for functionalizing other biomaterials.

Aβ-targeting synNotch Receptor for Alzheimer's Disease: Expanding Applications to Extracellular Protein Aggregates.

The synthetic Notch receptor (synNotch) system is a versatile platform that induces gene transcription in response to extracellular signals. However, its application has been largely confined to membrane-bound targets due to specific activation requirements. Whether synNotch can also target extracellular protein aggregates, such as amyloid beta (Aβ) in Alzheimer's disease (AD), is unclear. To address this, we engineered an Aβ-targeting synNotch receptor controlling the production of chimeric human-mouse versions of Lecanemab (Leqembi®) or Aducanumab (Aduhelm®), both FDA-approved antibodies for AD. We demonstrate that NIH 3T3 cells expressing this synNotch system detect and respond to extracellular Aβ aggregates by synthesizing and secreting Aducanumab or Lecanemab. These findings broaden the potential applications of synNotch, extending its targets beyond membrane-bound proteins to extracellular protein aggregates, providing obvious benefits to research in this scientific arena.

Preparation and Characterization of Photo-Cross-Linkable Methacrylated Silk Fibroin and Methacrylated Hyaluronic Acid Composite Hydrogels.

Composite biomaterials with excellent biocompatibility and biodegradability are crucial in tissue engineering. In this work, a composite protein and polysaccharide photo-cross-linkable hydrogel was prepared using silk fibroin methacrylate (SFMA) and hyaluronic acid methacrylate (HAMA). SFMA was obtained by the methacrylation of degummed SF with glycidyl methacrylate (GMA), while HA was methacrylated by 2-aminoethyl methacrylate hydrochloride (AEMA). We investigated the effect of the addition of 1 wt % HAMA to 5, 10, and 20 wt % SFMA, which resulted in an increase in both static and cycling mechanical strengths. All composite hydrogels gelled under UV light in <30 s, allowing for rapid stabilization and stiffness increases. The biocompatibility of the hydrogels was confirmed by direct and indirect contact methods and by evaluation against the NIH3T3 and MC3T3 cell lines with a live-dead assay by confocal imaging. The range of obtained mechanical properties from developed composite and UV-cross-linkable hydrogels sets the basis as possible future biomaterials for various biomedical applications.

Atypical Exon 2/3 Mutants G48C, Q43K, and E37K Present Oncogenic Phenotypes Distinct from Characterized NRAS Variants.

NRAS belongs to the RAS family of GTPases. In colorectal cancer (CRC), NRAS mutations are rare compared to KRAS, but may lead to worse outcomes. We report the functional characterization of the novel NRAS mutants-G48C, Q43K, and E37K-identified in Filipino young-onset CRC patients. Unlike previously characterized NRAS mutants with no apparent effects on cell proliferation, these mutants enhanced proliferation of both HCT116 and NIH3T3 cells. This was confirmed in 3D spheroid assays to mimic the spatial organization of cells. G48C and E37K showed apoptosis resistance in both cell lines, and Q43K showed resistance in HCT116 cells. All three showed no effect on cellular migration in NIH3T3, but G48C enhanced the migration rate of HCT116 cells. Actin staining of NIH3T3 cells expressing the mutants showed a shrunken cytoplasm and transient structures associated with motility and invasiveness. Docking simulations show that GDP is only able to bind fully within the binding pocket of wild-type NRAS, but not in the mutants. Further, G48C, Q43K, and E37K all have less negative ΔG values, indicating a weaker GDP-binding affinity compared to wild-type NRAS. Taken together, the results suggest that oncogenic readouts of NRAS mutants are codon- and mutation-specific, with potential repercussions on the aggressiveness, resistance, and therapeutic response.

Enhancing antifungal and antibacterial properties of denture resins with nystatin-coated silver nanoparticles

Long-term oral health issues caused by fungi and bacteria are a primary concern for individuals who wear dentures. Denture stomatitis, primarily caused by Candida albicans (C. albicans), is a prevalent condition among denture users. Metal nanoparticles exhibit improved antimicrobial effectiveness and fewer adverse effects. This study aimed to evaluate the antifungal and antibacterial effects of nystatin-coated silver nanoparticles (Nys-coated AgNPs) embedded in acrylic resin as a more biocompatible material for denture resins. AgNPs and Nys-coated AgNPs were synthesized and characterized using UV-Vis, SEM, EDX, and DLS. Specimens of polymethyl methacrylate (PMMA) with three different concentrations of Nys, AgNPs, and Nys-coated AgNPs (0.1%, 1%, 10% w/w) were prepared. The water absorption properties of the disks and drug release were investigated for 14 days and 120 h, respectively. The hydrophilic and hydrophobic properties of the samples and their contact angles were evaluated using the sessile drop technique. The antifungal and antimicrobial activity of the prepared discs was studied against C. albicans and Streptococcus mutans, respectively. Adding Nys-coated AgNPs decreased the contact angle of discs from 67° to 49°. Furthermore, the water absorption rates of the different discs were not significantly different from those of the control groups. Results showed that Nys-coated AgNPs (10% w/w) in PMMA effectively inhibited C. albicans growth better than Nys composites (10% w/w). Additionally, Nys-coated AgNPs composites, as well as AgNPs-containing composites, showed considerable antibacterial activity against S. mutans. Nys-coated AgNPs (10% w/w) had no toxic effect on NIH3T3 cells. In conclusion, Nys-coated AgNPs could be considered a good candidate for incorporation into denture resins to address chronic oral diseases.

Preparation of Disulfide/Trisulfide Core-Cross-Linked Polycarbonate Nanocarriers for Intracellular Reduction-Triggered Drug Release.

Polymeric nanocarriers have attracted significant attention in the field of anticancer drug delivery due to their unique advantages. However, designing nanocarriers that can maintain stability in the bloodstream while achieving specific drug release within tumor cells remains a major challenge. To address this issue, constructing reversible cross-linked polymeric nanocarriers that are sensitive to the intracellular reducible glutathione (GSH) characteristic of the tumor microenvironment is a promising strategy. Based on this, we designed and synthesized two novel six-membered bicyclic carbonate monomers containing disulfide (DSBC) and trisulfide (TSBC) bonds. Through a one-step ring-opening polymerization, a series of reduction-sensitive polycarbonate copolymers (i.e., PEG-PDSBC and PEG-PTSBC) were prepared, and doxorubicin (DOX)-loaded nanoparticles were fabricated using a nanoprecipitation method. The in vitro drug release behaviors of these nanoparticles were systematically investigated. The results showed that these polymers, due to the cross-linked structure formed by the ring-opening polymerization of their bicyclic monomers, could self-assemble into stable nanoparticles. Under different concentrations of glutathione, DOX-loaded PEG-PTSBC nanoparticles demonstrated faster drug release, indicating more optimized intracellular drug release properties. Further cytotoxicity experiments revealed that both types of blank nanoparticles exhibited good biocompatibility with the 4T1 and NIH-3T3 cells. Fluorescence microscopy and flow cytometry results further indicated that DOX-loaded PEG-PTSBC nanoparticles released more drugs in 4T1 cells, significantly inhibiting tumor cell growth compared with DOX-loaded PEG-PDSBC nanoparticles, with no noticeable difference in NIH-3T3 normal cells. In conclusion, this study suggests that trisulfide cross-linked polycarbonate-based nanocarriers hold promise as an anticancer drug delivery system that combines stability in the bloodstream with specific intracellular drug release, offering new insights for the development of novel, efficient, and safe anticancer nanomedicines.

TRAF7 determines circadian period through ubiquitination and degradation of DBP.

D-site binding protein, DBP, is a clock-controlled transcription factor and drives daily rhythms of physiological processes through the regulation of an array of genes harboring a DNA binding motif, D-box. DBP protein levels show a circadian oscillation with an extremely robust peak/trough ratio, but it is elusive how the temporal pattern is regulated by post-translational regulation. In this study, we show that DBP protein levels are down-regulated by the ubiquitin-proteasome pathway. Analysis using 19 dominant-negative forms of E2 enzymes have revealed that UBE2G1 and UBE2T mediate the degradation of DBP. A proteomic analysis of DBP-interacting proteins and database screening have identified Tumor necrosis factor Receptor-Associated Factor 7 (TRAF7), a RING-type E3 ligase, that forms a complex with UBE2G1 and/or UBE2T. Ubiquitination analysis have revealed that TRAF7 enhances K48-linked polyubiquitination of DBP in cultured cells. Overexpression of TRAF7 down-regulates DBP protein level, while knockdown of TRAF7 up-regulates DBP in cultured cells. Knockout of TRAF7 in NIH3T3 cells have revealed that TRAF7 mediates the time-of-the-day-dependent regulation of DBP levels. Furthermore, TRAF7 has a period-shortening effect on the cellular clock. Together, TRAF7 plays an important role in circadian clock oscillation through destabilization of DBP.

Preparation of a chitosan/polyvinyl alcohol-based dual-network hydrogel for use as a potential wound-healing material for the sustainable release of drugs

Treating chronic wounds poses significant challenges in clinical medicine due to bacterial infection, reactive oxygen species (ROS) accumulation, and excessive inflammation. This study aimed to address these issues by developing a wound dressing with antibacterial, antioxidant, and anti-inflammatory properties. Chitosan was functionally modified with acrolein to covalently bind to epigallocatechin gallate (EGCG), enabling a high EGCG load. Subsequently, polyvinyl alcohol (PVA) and EGCG-modified chitosan were crosslinked to prepare a new double-network hydrogel with added cysteine (CSAEC/P50). CSAEC/P50 demonstrated optimal mechanical properties (low swelling rate, high water retention, and optimal flexibility), low hemolysis, high coagulation properties, and antibacterial and antioxidant activities. Cell scratch tests indicated that CSAEC/P50 can promote NIH3T3 cell migration. Immunofluorescence results showed that CSAEC/P50 promoted the transformation of proinflammatory M1 macrophages to anti-inflammatory M2 macrophages. These findings suggest that CSAEC/P50 has significant potential for use in wound dressing applications.

Synthesis of cytocompatible gum Arabic-encapsulated silver nitroprusside nanocomposites for inhibition of bacterial pathogens and food safety applications.

Silver nitroprusside (AgN) exhibits significant antibacterial activity; however, its inherent toxicity poses a major concern. This study synthesized AgN with enhanced antibacterial properties while minimizing toxicity. Gum Arabic (GA), a natural polysaccharide widely utilized in food and biomedical applications owing to its exceptional cytocompatibility, was selected for encapsulating AgN to mitigate toxicity while preserving or enhancing its biological activity. The resulting composite material, GA-AgN nanocomposites (NCs), was systematically characterized using various analytical techniques. Transmission electron microscopy analysis revealed that GA-AgN NCs exhibited a rectangular morphology, with an average size of 230.13±62.8nm. The zeta potential of GA-AgN NCs was measured at -29.3±0.70mV. Furthermore, GA-AgN NCs demonstrated stability over diverse storage durations, incubation periods, and pH conditions by maintaining its size and surface charge. X-ray diffraction results indicated a reduction in the crystallinity of AgN when incorporated into the amorphous GA matrix, while Fourier-transform infrared spectroscopy analysis confirmed that the functional properties of both AgN and GA were preserved in the NCs. The release of Ag and Fe ions from the NCs was observed to be time- and pH-dependent. Importantly, the incorporation of GA did not compromise the antibacterial or antibiofilm efficacy of AgN against bacterial pathogens. Additionally, GA significantly mitigated the cytotoxic effects of AgN on NIH3T3 cells and red blood cells. Furthermore, GA-AgN NCs effectively extended the shelf-life of Salmonella enterica-infected green grapes. Thus, this study illustrates that GA-fabricated AgN NCs exhibit potential as an antibacterial agent in food preservation applications.

A silver lining in MRSA treatment: The synergistic action of poloxamer-stabilized silver nanoparticles and methicillin against antimicrobial resistance.

Increasing antibiotic resistance in bacterial infections, including drug-resistant strains like methicillin-resistant Staphylococcus aureus (MRSA), necessitates innovative therapeutic solutions. Silver nanoparticles are promising for combating infections, but toxicity concerns emphasize the importance of factors like dosage, size, shape, and surface chemistry. Hence, exploring poloxamer as a stabilizing agent to reduce its toxicity and enhance the antibacterial effect on MRSA is investigated. Silver nanoparticles stabilized with poloxamer (AgNPs@Pol) were synthesized through the chemical reduction method and characterized using UV-visible spectrophotometer, HR-TEM, DLS, and Zeta potential measurements. Subsequently, the antibacterial activity of AgNPs@Pol alone and in combination with methicillin against MRSA and methicillin-susceptible S. aureus (MSSA) was evaluated using the broth microdilution method. AgNPs@Pol showed significant efficacy against MRSA and MSSA, achieving a 100% reduction in colony-forming units (CFU) at 9.7 μg/ml. The minimum inhibitory concentration (MIC) against MRSA and MSSA was 8.6 μg/ml and 4.3 μg/ml, respectively. A synergistic effect was observed when AgNPs@Pol was combined with methicillin. Treatment with AgNPs@Pol increased reactive oxygen species (ROS) production in both strains, contributing to its antibacterial activity. Real-time qPCR analysis indicated the downregulation of genes involved in antimicrobial resistance and cell adhesion in both strains. Further, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay demonstrated low cytotoxicity for AgNPs@Pol against MCF-7, MG-63, and NIH-3T3 cell lines. The developed AgNPs@Pol demonstrated extensive colloidal stability, potent antibacterial activity and synergistic effect with methicillin against MRSA and MSSA. Further studies in primary cells and in vivo models may validate its potential for clinical applications.

PH-sensitive and redox-responsive poly(tetraethylene glycol) nanoparticle-based platform for cancer treatment

Effective drug delivery with precise tumour targeting is crucial for cancer treatment. To address the challenges posed by the specificity and complexity of the tumour microenvironment, we developed a poly(tetraethylene glycol)-based disulfide nanoparticle (NP) platform and explored its potential in cancer treatment, focusing on drug loading and controlled release performance. Poly(tetraethylene glycol) NPs were characterised using nuclear magnetic resonance spectroscopy, mass spectrometry, and ultraviolet-visible spectroscopy. Additionally, we evaluated physicochemical properties, including dynamic light scattering, zeta potential analysis, drug loading capacity (DLC), and drug loading efficiency (DLE). The impact of NPs on the mouse colorectal cancer cell line (CT26) and NIH3T3 cells was assessed using a cytotoxicity assay, live/dead staining assay, flow cytometry, and confocal fluorescence microscopy. The experimental results align with the expected chemical structure and physicochemical properties of poly(tetraethylene glycol) NPs. These NPs exhibit high DLE (78.7%) and DLC (12%), with minimal changes in particle size over time in different media.In vitroexperiments revealed that the NPs can induce significant cytotoxicity and apoptosis in CT26 cells. Cellular uptake notably increases with increasing concentration and exposure time. The confocal microscopic analysis confirmed the effective distribution and accumulation of NPs within cells. In conclusion, poly(tetraethylene glycol) NPs hold promise for improving drug-delivery efficiency, offering potential advancements in cancer treatment.

Regulation of alternative polyadenylation isoforms of Timp2 is an effector event of RAS signaling in cell transformation.

Alternative polyadenylation (APA) generates mRNA isoforms with different lengths of the 3' untranslated region (3' UTR). The tissue inhibitor of metalloproteinase 2 (TIMP2) plays a key role in extracellular matrix remodeling under various developmental and disease conditions. Both human and mouse genes encoding TIMP2 contain two highly conserved 3'UTR APA sites, leading to mRNA isoforms that differ substantially in 3'UTR size. APA of Timp2 is one of the most significantly regulated events in multiple cell differentiation lineages. Here we show that Timp2 APA is highly regulated in transformation of NIH3T3 cells by the oncogene HRAS G12V . Perturbations of isoform expression with long 3'UTR isoform-specific knockdown or genomic removal of the alternative UTR (aUTR) region indicate that the long 3'UTR isoform contributes to the secreted Timp2 protein much more than the short 3'UTR isoform. The short and long 3'UTR isoforms differ in subcellular localization to endoplasmic reticulum (ER). Strikingly, Timp2 aUTR enhances secreted protein expression but no effect on intracellular proteins in reporter assays. Furthermore, downregulation of Timp2 long isoform mitigates gene expression changes elicited by HRAS G12V . Together, our data indicate that regulation of Timp2 protein expression through APA isoform changes is an integral part of RAS-mediated cell transformation and 3'UTR isoforms of Timp2 can have distinct impacts on expression of secreted vs. intracellular proteins.

Enhanced Anticancer Activity of 7MeERT over Ertredin: A Comparative Study on Cancer Cell Proliferation and NDUFA12 Binding.

We have previously identified Ertredin (3-(2-amino-5-bromophenyl) quinoxalin-2(1H)-one) as a compound that suppresses 3D spheroid formation and tumorigenesis in NIH3T3 cells induced by Epidermal Growth Factor Receptor variant III (EGFRvIII) transduction. One of its targets has been shown to be NDUFA12 (NADH Dehydrogenase (Ubiquinone) 1 Alpha Subcomplex Subunit 12), a component protein of oxidative phosphorylation complex I. In this report, we compared the growth inhibitory activity of Ertredin with its methylated analogue 7MeERT (3-(2-amino-5-bromophenyl)-7-methylquinoxalin-2(1H)-one) on human cancer cells. 7MeERT induced the inhibition of the proliferation of various cancer cells similarly to Ertredin and showed higher activity in glioblastoma cells, A431 cells overexpressing EGFR (wild type), and multiple myeloma cells. Molecular docking analysis and a Cellular Thermal Shift Assay (CETSA) suggested that 7MeERT binds to NDUFA12 similarly to Ertredin. The binding of 7MeERT and Ertredin to NDUFA12 in glioblastoma was further supported by the inhibition of the oxygen consumption rate. These results suggest that 7MeERT also binds to NDUFA12, inhibits oxidative phosphorylation, and has a higher anti-cancer cell growth inhibitory activity than Ertredin.

In vitro anticancer efficacy of Calendula Officinalis extract-loaded chitosan nanoparticles against gastric and colon cancer cells

Objective This study assessed the anticancer activities of Calendula officinalis-loaded chitosan nanoparticles in gastric and colon cancer cells compared with fibroblast cells and examined the balance between ROS and antioxidants. Methods Considering this information, we synthesized Calendula officinalis-loaded chitosan nanoparticles (CO-CSNPs) via the ionic gelation method. Their characterizations were carried out with ZetaSizer, UV-Vis, FTIR and SEM devices including size, morphology and surface zeta potential analysis, loading capacity, encapsulation efficiency, in vitro drug release, and chemical interactions. The anticancer activities of CO, CSNPs, and CO-CSNPs were tested against AGS, Caco-2, and normal NIH-3T3 cells using an XTT assay. The anticancer effects were evaluated using DAPI staining, scratch assay, reactive oxygen species (ROS) detection and the CUPRAC method on cellular and non-cellular processes that promote anticancer mechanisms. Results Results showed that CO and CO-CNPs exhibited anticancer activity against AGS and Caco-2. Further, the formulation of CO with CSNPs enhanced the anticancer activity of CO while having no cytotoxicity on NIH-3T3. DAPI staining, scratch assay, ROS, and CUPRAC method confirmed the anticancer activity of CO and CO-CSNPs, which resulted in a reduction in the number of apoptotic cells, inhibited migration, triggered apoptotic pathway via ROS, and higher antioxidant activity. Conclusions The results of the study indicate that CO-CSNPs are a promising therapeutic formulation for gastric and colon cancer treatment. We consider that this study will lead to the investigation of molecular mechanisms of CO-CSNPs in cancer treatment and their investigation in clinical studies.

Exploring the Wound Healing Potential of Hispidin.

Hispidin, a polyphenol component mainly derived from the medicinal mushroom species Phellinus and Inonotus, shows promise for biomedical applications, yet its potential in wound healing remains largely unexplored. This research investigates the wound healing effects of hispidin through in vitro and in vivo experiments, while also evaluating its antimicrobial properties and safety profile. In vitro scratch assays were conducted to evaluate the impact of hispidin on the migration of NIH-3T3 cells. The wound healing potential of hispidin was assessed in rats using excision wounds, dead space wounds, and linear incisions, treated with various topical ointments including a simple ointment, 2.5% (w/w) and a 5% (w/w) hispidin ointment, and a 0.2% (w/w) nitrofurazone ointment, administered at 0.2 g daily for 14 days. Hispidin demonstrated antimicrobial properties and was particularly effective against Staphylococcus epidermidis. Hispidin enhanced NIH-3T3 cell viability, and promoted wound closure in scratch assays, correlating with increased levels of FGF21, TGF-β1, EGF, and VEGF. In excision wound models, the 5% (w/w) hispidin ointment improved wound contraction, epithelialization, tissue regeneration, fibroblast activity, and angiogenesis. In the granulation tissue from dead space wound models, hispidin reduced pro-inflammatory cytokines (TNF-α, IL-6, IL-1β) and lipid peroxidation, while increasing anti-inflammatory cytokines (IL-10) and antioxidant activities (SOD, GPx, CAT), along with connective tissue markers like hydroxyproline, hexosamine, and hexuronic acid. Hispidin also enhanced wound breaking strength in incision models. Acute dermal toxicity studies indicated no adverse effects at 2000 mg/kg. These findings highlight hispidin's potential in wound care, demonstrating its antimicrobial, regenerative, and safety properties.

Gadolinium doped carbon dots for anti-gram-negative bacteria and visible light photodynamic enhancement of antibacterial effect

Infection with gram-negative bacteria is the main source of the most serious infectious pathogens. Developing new antibacterial materials that break through their external membranes and stay in the bacterial body to result in an antibacterial effect is the key to achieving high efficiency against Gram-negative bacteria. A Gd-doped carbon dot (GRCD) was prepared using the approved therapeutic diagnostic agents Rose Bengal (RB) and gadolinium ions (Gd3+), which was used to resist Gram-negative bacteria (e.g. E. coli, Escherichia coli). GRCD not only showed strong antibacterial activity by destroying the external membranes of E. coli (inhibition rate against E. coli was 92.0 % at 20 μg/mL) but also bound to E. coli DNA and generated single oxygen (1O2) (quantum yield was 0.50) through visible light-driven catalysis, thus decomposing the DNA of E. coli and further enhancing the antibacterial performance of GRCD. Under visible light conditions, the inhibition rate against E. coli reached 95.8 % at a low concentration of 2.5 μg/mL, without obvious cytotoxicity to NIH3T3 cells. The use of GRCD in treating wound infections in mice caused by E. coli was quite good, without side reactions on the mice’s essential organs. In this study, a new approach has been provided to the design and synthesis of carbon dot nanocomposites for use against Gram-negative bacteria.

Tanshinone IIA Loaded Inhaled Polymer Nanoparticles Alleviate Established Pulmonary Fibrosis.

Idiopathic pulmonary fibrosis (IPF) is a fatal respiratory disease characterized by chronic, progressive scarring of the lung parenchyma, leading to an irreversible decline in lung function. Apart from supportive care, there is currently no specific treatment available to reverse the disease. Based on the fact that tanshinone IIA (TAN) had an effect on protecting against TGF-β1-induced fibrosis through the inhibition of Smad and non-Smad signal pathways to avoid myofibroblasts activation, this study reported the development of the inhalable tanshinone IIA-loaded chitosan-oligosaccharides-coated poly(lactic-co-glycolic acid) (PLGA) nanoparticles (CPN@TAN) for enhancing the pulmonary delivery of tanshinone IIA to treat pulmonary fibrosis. The CPN@TAN with a size of 206.5 nm exhibited excellent in vitro aerosol delivery characteristics, featuring a mass median aerodynamic diameter (MMAD) of 3.967 ± 0.025 μm and a fine particle fraction (FPF) of 70.516 ± 0.929%. Moreover, the nanoparticles showed good stability during atomization and enhanced the mucosal penetration capabilities. The results of confocal spectroscopy confirmed the potential of the nanoparticles as carriers that facilitated the uptake of drugs by NIH3T3, A549, and MH-S cells. Additionally, the nanoparticles demonstrated good in vitro biocompatibility. In a mouse model of bleomycin-induced pulmonary fibrosis, noninvasive inhalation of aerosol CPN@TAN greatly suppressed collagen formation and facilitated re-epithelialization of the destroyed alveolar epithelium without causing systemic toxicity compared with intravenous administration. Consequently, our noninvasive inhalation drug delivery technology based on polymers may represent a promising paradigm and open the door to overcoming the difficulties associated with managing pulmonary fibrosis.

Preparation of injectable self-healing hydrogels using carboxymethyl chitosan and oxidized dextran with incorporating quercetin-loaded PF127 micelles for wound healing

Rapid achievement of the irregularly shaped post-wound closure is closely related to the effectiveness of clinical hydrogel-based wound dressings to inhibit microbial invasion and promote wound healing. Herein, an injectable and self-healing hydrogel with rapid gelation was engineered based on carboxymethyl chitosan (CMCS) and oxidized dextran (Odex). The hydrophobic drug, quercetin, with antioxidant and antibacterial effects, was loaded in the PF127 micelles with an encapsulation efficiency of 59.06 %, and then quercetin-loaded PF127 micelles were ingeniously incorporated into the hydrogels. The prepared composite hydrogels not only showed a stable and sustained release behavior of quercetin, but also possessed excellent antioxidant activities and antibacterial properties against Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli. Moreover, the composite hydrogels with microstructure (150–250 μm) also exhibited self-healing and injectable properties. In addition, the survival rates of NIH 3T3 cells when cultured with hydrogel extract were all over 80 %, illustrating the excellent biocompatibility of hydrogels. Therefore, our prepared multifunctional composite hydrogels showed a broad prospect as wound dressings for treating acute skin injuries.

Use microfluidics to study cell migration in response to fluid shear stress gradients

Cells respond not only to biological regulatory factors but also to physical stimuli such as electric fields, light, and shear stress in their environment. These stimuli can lead to cell migration and morphological changes. In the human body, cells encounter fluid shear stress induced by interstitial flow, lymphatic flow, blood flow, or organ-specific conditions within their micro-environments. Therefore, fluid shear stress, a classic mechanical force, has gained significant attention in wound healing and cancer metastasis. In this study, a microfluidic chip was developed to both culture cells and generate a shear stress gradient to direct cell migration. The design of this device’s geometry allows the generation of a shear stress gradient perpendicular to the direction of medium flow. This greatly eliminates the influence of flow-induced cell responses. Using mouse fibroblast cells (NIH3T3) and human lung cancer cells (CL1-5) as models, their migration directionality, migration rates, and alignment in response to the shear stress gradient were investigated. Within the stress range of 0.095–0.155 Pa and a gradient of 0.015 Pa/mm, NIH3T3 cells did not exhibit significant directional migration, differences in migration rates, or specific alignment patterns. In contrast, CL1-5 cells preferred higher shear stress environments and alignment parallel to the medium flow, suggesting that these conditions could induce higher mobility in these cancer cells.

Cepharanthine attenuates pulmonary fibrosis via modulating macrophage M2 polarization

BackgroundIdiopathic pulmonary fibrosis (IPF) is a group of chronic interstitial pulmonary diseases characterized by myofibroblast proliferation and extracellular matrix (ECM) deposition. However, current treatments are not satisfactory. Therefore, more effective therapies need to be explored. Cepharanthine (CEP) is a naturally occurring alkaloid that has recently been reported to have multiple pharmacological effects, particularly in chronic inflammation.MethodsFor in vivo experiments, first, a pulmonary fibrosis murine model was generated via tracheal injection of bleomycin (BLM). Second, the clinical manifestations and histopathological changes of the mice were used to verify that treatment with CEP might significantly reduce BLM-induced fibrosis. Furthermore, flow cytometric analysis was used to analyze the changes in the number of M2 macrophages in the lung tissues before and after treatment with CEP to explore the relationship between macrophage M2 polarization and pulmonary fibrosis. In vitro, we constructed two co-culture systems (THP-1 and MRC5 cells, RAW264.7 and NIH 3T3 cells), and measured the expression of fibrosis-related proteins to explore whether CEP could reduce pulmonary fibrosis by regulating macrophage M2 polarization and fibroblast activation.ResultsThe results showed that the intranasal treatment of CEP significantly attenuated the symptoms of pulmonary fibrosis induced by BLM in a murine model. Our findings also indicated that CEP treatment markedly reduced the expression of fibrosis markers, including TGF-β1, collagen I, fibronectin and α-SMA, in the mouse lung. Furthermore, in vitro studies demonstrated that CEP attenuated pulmonary fibrosis by inhibiting fibroblast activation through modulating macrophage M2 polarization and reducing TGF-β1 expression.ConclusionsThis study demonstrated the potential and efficacy of CEP in the treatment of pulmonary fibrosis. In particular, this study revealed a novel mechanism of CEP in inhibiting fibroblast activation by regulating macrophage M2 polarization and reducing the expression of fibrosis-associated factors. Our findings open a new direction for future research into the treatment of pulmonary fibrosis.