3T3 Fibroblasts Research Articles

FFAR-mediated signaling drives migration of pancreatic cancer cells in hypoxic fibroblast co-cultures

The tumor microenvironment (TME) comprises cancer and non-cancerous stromal cells, including fibroblasts. Free fatty acids (FFAs) regulate various biological responses by binding to G protein-coupled FFA receptors (FFARs). In this study, we examined the impact of FFAR1 and FFAR4 on the cell migration of pancreatic cancer PANC-1 cells co-cultured with 3T3 fibroblast cells under hypoxic conditions. PANC-1 cells cultured at 1 % O2 exhibited elevated FFAR1 expression and decreased FFAR4 expression compared to those at 21 % O2. Cell migration of PANC-1 cells was reduced under 1 % O2 conditions. FFAR1 knockdown enhanced PANC-1 cell migration, whereas FFAR4 knockdown inhibited it. Co-culture of PANC-1 cells with 3T3 cells at 1 % O2 significantly increased FFAR4 expression, while FFAR1 expression remained unchanged. To evaluate the effects of FFAR1 and FFAR4 on PANC-1 cell migration in co-culture with 3T3 cells, we conducted a wound healing assay using the Culture-Insert 2 Well. PANC-1 and 3T3 cells were individually seeded into the two wells and incubated at both 21 % and 1 % O2 for 13 h. The cell migration of PANC-1 cells co-cultured with 3T3 cells at 1 % O2 was notably higher compared to 21 % O2. TUG-770 reduced and TUG-891 enhanced the cell migration of PANC-1 cells co-cultured with 3T3 cells under both 21 % and 1 % O2 conditions. These findings suggest that FFAR1 and FFAR4 play important roles in regulating the cell migration of PANC-1 cells co-cultured with 3T3 cells under hypoxic conditions.

The role of TGF-β in the electrotactic reaction of mouse 3T3 fibroblasts in vitro.

Endogenous electric fields (EFs) serve as a crucial signal to guide cell movement in processes such as wound healing, embryonic development, and cancer metastasis. However, the mechanism underlying cell electrotaxis remains poorly understood. A plausible hypothesis suggests that electrophoretic or electroosmotic forces may rearrange charged components of the cell membrane, including receptors for chemoattractants which induce asymmetric signaling and directional motility. This study aimed to explore the role of Transforming Growth Factor Beta (TGFβ) signaling in the electrotactic reaction of 3T3 fibroblasts. Our findings indicate that inhibiting canonical and several non-canonical signaling pathways originating from the activated TGF-β receptor does not hinder the directed migration of 3T3 cells to the cathode. Furthermore, suppression of TGF-β receptor expression does not eliminate the directional migration effect of 3T3 cells in the electric field. Additionally, there is no observed redistribution of the TGF-β receptor in the electric field. However, our studies affirm the significant involvement of Phosphoinositide 3-Kinase (PI3K) in electrotaxis, suggesting that in our model, its activation is likely associated with factors independent of TGFβ action.

The Potential Of Superoxide Dismutase Enzyme From Tomato Fruit (Solanum Lycopersicum) To Repair Collagen Damage In 3t3 Fibroblast Cells Exposed To Ultraviolet A Radiation

UV A radiation can cause photoaging to the skin which is show by a decrease in the amount of collagen and death of fibroblast cell. This study aimed to determine the SOD enzyme activity of tomatoes (Solanum lycopersycum Mill.) influences the increase of viability of fibroblast cell and deposition of collagen. The study design used a completely randomized design (CRD) with 11 treatments and 3 repetitions. The procedures of this study include: (1) extracting SOD enzymes (2) making bradford reagents (3) making standard protein solutions (4) measuring total protein content (5) testing the antioxidant activity of SOD enzymes (6) preparing 3T3 fibroblast cells (7) Cell exposure 3T3 fibroblasts with UV A rays (8) preparation of tomato SOD enzyme stock solutions (9) determination of cell viability and collagen deposition. Data analized by normality test, homogeneity test, one way ANOVA test, tuckey test and dunnet test. The results of SOD enzyme activity showed the highest percent inhibition was 77.83%. Highest value of percent viability of fibroblast cells that were exposed to UV A rays after given 90% SOD enzyme treatment was 233.00% and collagen deposition was 157.67%. The results showed that variations in the SOD enzyme concentration of tomatoes had a significant effect on cell viability and collagen deposition in fibroblast cells. with a value of sig = 0,000 (p <α). Based on the result, we can concluded that SOD enzyme from tomatoe fruit show significant activity as anti-photoaging.

The synergistic impact of graphene and copper on microstructure, corrosion behaviors and biocompatibility of a magnesium metal matrix composite in Hank's solution

BackgroundThis research aimed to explore the combined effects of graphene (Gr) and copper (Cu) in a magnesium metal composite (MMC) on its microstructure, biocompatibility, electrochemical, and immersion corrosion behaviors. MethodsThe MMCs were prepared using the mechanical stir-casting method. Alloys with a preset weight percentage of Gr (0.15 wt.%) and various weight percentages of Cu (0.5 and 1 wt.%) were produced with pure Mg as the matrix. Significant findingsThis study demonstrates that Gr and Cu enhance crystallinity and increase grain size in composite materials. The introduction of Gr into the magnesium matrix alone elevates the corrosion rate. However, there was a further addition of 0.5 wt.% Cu (0.5Cu) showed a slight reduction in grain size growth and corrosion rate. The addition of more copper (1 wt.% Cu (1Cu)) increases both grain size and the corrosion rate again. Cytotoxicity test results indicate that Gr and Cu enhance the cell viability of 3T3 fibroblasts in vitro. These findings shed light on the intricate interplay between Gr and Cu in the microstructure and corrosion resistance of Mg-based composites and inspire confidence in developing magnesium metal composites for clinical applications.

Chitosan films synthesized via thiol-ene click chemistry: Toward safe and versatile platforms for packaging, cosmetics and biomedical applications

The first synthesis of chitosan-based films using bio-based reticulating agents via Thiol-Michael addition click reaction was described, along with their successful application in packaging, cosmetics, and biomedical fields. The impact of chitosan (CS) concentration and the type of synthesized bio-sourced biacids (GD) and triacids (GT and TT) on the physicochemical properties of these films were assessed. Films prepared with dibasic and tribasic acids (4%-CS) demonstrated superior mechanical properties and lower solubility compared to those containing 2% CS. The incorporation of the bio-sourced GD, GT, and TT acids led to a reduction of 53%, 60% and 60.4%, respectively, in the water vapor permeability (WVP) of 4% CS-based films compared to the control film. Remarkably, CS-TT-4% films exhibited the highest mechanical strength (TS), elongation at break (%E), elasticity, water vapor, and light barrier properties. Cytotoxicity tests performed on the NIH 3T3 fibroblast cell line showed that CS-GD-4% films exhibited the highest compatibility. The efficiency of these films as drug carriers was tested with challenging anticancer and cosmetic molecules, particularly etoposide (ETP) caffeine (Caf), showing very high drug loading with a progressive release of less than 20% after 72h of incubation. Moreover, ex-vivo permeation studies in rat skin revealed that the percutaneous absorption of Caf is significantly increased by a factor of 5.5 compared to that of CS-GD-4%-Caf films with no significant histological alterations of skin layers. Altogether, this work evidences the most suitable casting solvent for developing safe chitosan films, promising their potential applications in the cosmetic and biomedical fields.

Biocompatibility and Antibacterial Properties of NiTiAg Porous Alloys for Bone Implants.

In order to reduce infections, porous NiTi alloys with 62% porosity were obtained by self-propagating high-temperature synthesis with the addition of 0.2 and 0.5 at. % silver nanoparticles. Silver significantly improved the alloys' antibacterial activity without compromising cytocompatibility. An alloy with 0.5 at. % Ag showed the best antibacterial ability against Staphylococcus epidermidis. All alloys exhibited good biocompatibility with no cellular toxicity against embryonic fibroblast 3T3 cells. Clinical evaluation of the results after implantation showed a complete absence of purulent-inflammatory complications in all animals. Even distribution of silver nanoparticles in the surface layer of the porous NiTi alloy provides a uniform antibacterial effect.

Fabrication of a self-healing hydrogel with antibacterial activity using host-guest interactions between dopamine-modified alginate and β-cyclodextrin dimer

Self-healing hydrogels possess an ability to recover their functionality after experiencing damage by regenerating cross-links. The main challenge in making self-healing hydrogels based on host-guest (HG) interactions is their limited mechanical strength, which can be solved using beta-cyclodextrin dimers (β-CDsD). Here, β-CDsD as a host cross-linker was used to increase the mechanical property of the HG interactions. Alginate with acceptable biocompatibility was modified by dopamine (ALG-DOP) and employed as a guest polymer. Self-healing hydrogel was developed between them, and Ag nanoparticles were added to create an antibacterial activity. Dopamine with appropriate size and suitable adhesiveness established HG interactions with β-CDsD, and cells were able to grow well on hydrogel. This hydrogel showed an impressive self-healing capability <5 min. These hydrogels revealed a respectable porosity from 15 to 55 μm essential for exchanging the substances required for cell growth and cell waste elimination. Biocompatibility was investigated against NIH 3 T3 fibroblasts cells, and the results showed that the cells grew well. The in vitro release of curcumin from the hydrogel was examined in PBS at pH of 7.4. The hydrogel can be a perfect candidate for controlled drug release, and wound-dressing due to self-healing property, antibacterial activity, adhesion, and biocompatibility.

A promising eco-sustainable wound dressing based on cellulose extracted from Spartium junceum L. and impregnated with Glycyrrhiza glabra L extract: Design, production and biological properties

Glycyrrhiza glabra extract is widely known for its antioxidant and anti-inflammatory properties and can improve the wound healing process. The aim of this work was to shorten the time of the healing process by using an eco-sustainable wound dressing based on Spanish broom flexible cellulosic fabric by impregnation with G. glabra extract-loaded ethosomes. Chemical analysis of G. glabra extract was performed by LC-DAD-MS/MS and its encapsulation into ethosomes was obtained using the ethanol injection method. Lipid vesicles were characterized in terms of size, polydispersity index, entrapment efficiency, zeta potential, and stability. In vitro release studies, biocompatibility, and scratch test on 3T3 fibroblasts were performed. Moreover, the structure of Spanish broom dressing and its ability to absorb wound exudate was characterized by Synchrotron X-ray phase contrast microtomography (SR-PCmicroCT). Ethosomes showed a good entrapment efficiency, nanometric size, good stability over time and a slow release of polyphenols compared to the free extract, and were not cytotoxic. Lastly, the results revealed that Spanish broom wound dressing loaded with G. glabra ethosomes is able to accelerate wound closure by reducing wound healing time. To sum up, Spanish broom wound dressing could be a potential new green tool for biomedical applications.

Fabrication of pH-stimuli hydrogel as bioactive materials for wound healing applications

Hydrogels exhibit exceptional suitability as wound dressing due to their remarkable three-dimensional (3D) characteristics. Here, we have reported the fabrication of hydrogels from biopolymers carboxymethyl cellulose (CMC), polyvinyl alcohol (PVA), and gelatin via a simple blending method to mimic the natural extracellular matrix. Scanning electron microscopy (SEM), water contact meters (WCM), and Fourier-transform infrared spectroscopy (FTIR) were used to evaluate the chemical structural, morphological, and wettability behavior. The wetting and degradation behavior were also found to be different for different formulations (Min. (51.60o) and Max. (113.60o)) and (Min. (38.82 mg) and Max. (3.72 mg)), respectively. Swelling was investigated in different media, including phosphate buffer saline solution (PBS) and aqueous media. It was observed that the hydrogel displayed the highest degree of swelling in an aqueous medium (Min. (597.32–1121.49 %) and Max. (1089.51–2139.73 %)) compared to PBS media (Min. (567.01–1021.85 %) and Max. (899.13–1639.17 %)). The release of Neomycin was studied in a PBS medium via the Franz diffusion method at 37 °C. The maximal release in various media demonstrated pH-responsive behavior. The viability and proliferation of fibroblast (3T3) cell lines were examined in vitro to evaluate cytocompatibility. Human Embryonic Kidney (HEK) 293 cells were used to evaluate the hydrogels' ability to promote vascularization and angiogenesis. Therefore, the data demonstrate that hydrogels that have been manufactured have qualities that make them promising for use as wound dressings in wound healing applications.

Facile preparation of zwitterionic core-shell nanogels for dual-functional coatings: Combining antifouling and antibacterial features in silicon implant model surface modification

To develop surface-coating technology for fabricating functionalized biomedical implant and overcome implant-associated bacterial infections, this study presented an innovative antifouling and antibacterial dual-functional nanogel coating methodology for engineering silicon-based biomedical implants/biochips. Firstly, zwitterionic core-shell PAA@(GMA/SBMA) nanogels were designed and prepared via reflux-precipitation polymerization and were characterized through NMR, FTIR, DLS, AFM and TEM. The nanogels exhibited uniform and spherical morphology, salt-ion responsiveness, and high lysozyme loading capability. Subsequently, the PAA@(GMA/SBMA) nanogels were covalently grafted onto the surface of silicon wafers, via an epoxy-amine ring-opening reaction, to prepare nanogel-coated silicon wafers (Si@nanogels). A comprehensive array of characterization techniques, including SEM-EDS, contact angle measurements, and XPS, validated the effective nanogel-coating. These coatings demonstrated notable resistance to protein adsorption, highlighting their robust antifouling properties. Upon loading with the antibacterial agent lysozyme, the nanogel coating displayed remarkable antibacterial efficacy against both Gram-negative (E. coli) and Grampositive (S. aureus) bacteria and effectively inhibited biofilm formation. Furthermore, the nanogel coatings exhibited good biocompatibility and dynamic stability, thus facilitating the surface-mediated adhesion and growth of fibroblast 3T3 cells. Significantly, this work provided a facile, economic and efficient surface modification approach to fabricate nanogel-coated antifouling/antibacterial dual-functional biomedical implant model towards potential clinical applications.

Effect of Peptide-Polymer Host-Guest Electrostatic Interactions on Self-Assembling Peptide Hydrogels Structural and Mechanical Properties and Polymer Diffusivity.

Peptide-based supramolecular hydrogels are an attractive class of soft materials for biomedical applications when biocompatibility is a key requirement as they exploit the physical self-assembly of short self-assembling peptides avoiding the need for chemical cross-linking. Based on the knowledge developed through our previous work, we designed two novel peptides, E(FKFE)2 and K(FEFK)2, that form transparent hydrogels at pH 7. We characterized the phase behavior of these peptides and showed the clear link that exists between the charge carried by the peptides and the physical state of the samples. We subsequently demonstrate the cytocompatibility of the hydrogel and its suitability for 3D cell culture using 3T3 fibroblasts and human mesenchymal stem cells. We then loaded the hydrogels with two polymers, poly-l-lysine and dextran. When polymer and peptide fibers carry opposite charges, the size of the elemental fibril formed decreases, while the overall level of fiber aggregation and fiber bundle formation increases. This overall network topology change, and increase in cross-link stability and density, leads to an overall increase in the hydrogel mechanical properties and stability, i.e., resistance to swelling when placed in excess media. Finally, we investigate the diffusion of the polymers out of the hydrogels and show how electrostatic interactions can be used to control the release of large molecules. The work clearly shows how polymers can be used to tailor the properties of peptide hydrogels through guided intermolecular interactions and demonstrates the potential of these new soft hydrogels for use in the biomedical field in particular for delivery or large molecular payloads and cells as well as scaffolds for 3D cell culture.

Synthesis of Rhodamine-Conjugated Lupane Type Triterpenes of Enhanced Cytotoxicity.

Various conjugates with rhodamines were prepared by starting with betulinic acid (BA) and platanic acid (PA). The molecules homopiperazine and piperazine, which were identified in earlier research, served as linkers between the rhodamine and the triterpene. The pentacyclic triterpene's ring A was modified with two acetyloxy groups in order to possibly boost its cytotoxic activity. The SRB assays' cytotoxicity data showed that conjugates 13-22, derived from betulinic acid, had a significantly higher cytotoxicity. Of these hybrids, derivatives 19 (containing rhodamine B) and 22 (containing rhodamine 101) showed the best values with EC50 = 0.016 and 0.019 μM for A2780 ovarian carcinoma cells. Additionally, based on the ratio of EC50 values, these two compounds demonstrated the strongest selectivity between malignant A2780 cells and non-malignant NIH 3T3 fibroblasts. A375 melanoma cells were used in cell cycle investigations, which showed that the cells were halted in the G1/G0 phase. Annexin V/FITC/PI staining demonstrated that the tumor cells were affected by both necrosis and apoptosis.

Bacteriophage-cocktail hydrogel dressing to prevent multiple bacterial infections and heal diabetic ulcers in mice.

Bacteriophage (phage) has been reported to reduce the bacterial infection in delayed-healing wounds and, as a result, aiding in the healing of said wounds. In this study we investigated whether the presence of phage itself could help repair delayed-healing wounds in diabetic mice. Three strains of phage that target Salmonella enterica, Escherichia coli, and Pseudomonas aeruginosa were used. To prevent the phage liquid from running off the wound, the mixture of phage (phage-cocktail) was encapsulated in a porous hydrogel dressing made with three-dimensional printing. The phage-cocktail dressing was tested for its phage preservation and release efficacy, bacterial reduction, cytotoxicity with 3T3 fibroblast, and performance in repairing a sterile full-thickness skin wound in diabetic mice. The phage-cocktail dressing released 1.7%-5.7% of the phages embedded in 24 h, and reduced between 37%-79% of the surface bacteria compared with the blank dressing (p <.05). The phage-cocktail dressing exhibited no sign of cytotoxicity after 3 days (p <.05). In vivo studies showed that 14 days after incision, the full-thickness wound treated with a phage-cocktail dressing had a higher wound healing ratio compared with the blank dressing and control (p <.01). Histological analysis showed that the structure of the skin layers in the group treated with phage-cocktail dressing was restored in an orderly fashion. Compared with the blank dressing and control, the repaired tissue in the phage-cocktail dressing group had new capillary vessels and no sign of inflammation in its dermis, and its epidermis had a higher degree of re-epithelialization (p <.05). The slow-released phage has demonstrated positive effects in repairing diabetic skin wounds.

Colchicine modulates TIMP1, MMP1, and pMAP4 expression to enhance extracellular matrix degradation in 3T3 fibroblast-mediated myocardial infarction

Context: Cardiovascular disease is a leading cause of mortality worldwide. Following acute myocardial infarction (AMI), cardiac cells release many cytokines and proteolytic enzymes that regulate the extracellular matrix (ECM), including matrix metalloproteinases (MMPs), tissue inhibitor of metalloproteinases (TIMPs), and phosphorylated microtubule-associated protein 4 (Phospho/pMAP4). Aims: To analyze the role of colchicine in ECM regulation post-ischemic in cell line culture towards MMPs, TIMPs, and pMAP4. Methods: This in vitro experimental study was conducted on the 3T3 fibroblast cell line subjected to hypoxic conditions (CoCl2 treatment) and compared with 3T3 cells experiencing hypoxia/ischemic and colchicine treatment. MMP-1, TIMP-1, and pMAP4 expression were measured using flow cytometry. Data analysis was performed using One-Way ANOVA with a 95% confidence level. Results: Treatment with colchicine in the 3T3 fibroblast cell culture regulated the extracellular matrix by increasing TIMP-1 expression and reducing MMP-1 and pMAP4 expression (p&lt;0.0001) following ischemic conditions induced by CoCl2. Conclusions: Colchicine effectively mitigates ECM degradation by enhancing TIMP-1 expression and inhibiting MMP-1 activity and pMAP4 expression post-ischemic conditions.

Amine functionalised graphene embedded polyvinyl alcohol (PVA) and PVA-chitosan hydrogel composites

A novel amine-functionalized graphene oxide (AFG) doped polyvinyl alcohol (PVA)/chitosan (PVA-Ch) composite film was developed using an eco-synthesis approach, eliminating the need for halogenated compounds. The resulting AFG-doped PVA/Chitosan (PVA-Ch/AFG) polymer film exhibited promising properties for controlled delivery and biosensing applications. The investigation included assessing the swelling behaviour, dissolution percent, gel fraction, and mechanical properties of the polymer film. The swelling characteristics of PVA-Ch and PVA-Ch/AFG were found to be pH and temperature-dependent across various pH ranges (3, 5, 7, and 9). Interestingly, PVA-Ch/AFG demonstrated a stable swelling pattern at pH 5 and 7, unaffected by changes in chitosan concentration, indicating enhanced stability compared to PVA-Ch. The study also explored the use of PVA-Ch/AFG in a drug delivery system, revealing controlled release of the model antibiotic amphicillin, emphasizing its potential in medical applications. Furthermore, the eco-friendly synthesis route underscored the safety of PVA-Ch/AFG for use in food and medical applications. Biocompatibility assessments, including biodegradability studies and cytotoxicity tests on fibroblasts (3T3 cells), confirmed the safety profile of PVA-Ch/AFG. In conclusion, the study suggests that PVA-Ch/AFG holds promise for bio-sensing applications, offering a flexible and colorimetric platform capable of encapsulating, adsorbing, and desorbing biomolecules such as drugs and sensing compounds.

Arabinoxylan-based psyllium seed hydrocolloid: Single-step aqueous extraction and use in tissue engineering

Biomacromolecules derived from natural sources offer superior biocompatibility, biodegradability, and water-holding capacity, which make them promising scaffolds for tissue engineering. Psyllium seed has gained attention in biomedical applications recently due to its gel-forming ability, which is provided by its polysaccharide-rich content consisting mostly of arabinoxylan. This study focuses on the extraction and gelation of Psyllium seed hydrocolloid (PSH) in a single-step water-based protocol, and scaffold fabrication using freeze-drying method. After characterization of the scaffold, including morphological, mechanical, swelling, and protein adsorption analyses, 3D cell culture studies were done using NIH-3 T3 fibroblast cells on PSH scaffold, and cell viability was assessed using Live/Dead and Alamar Blue assays. Starting from day 1, high cell viability was obtained, and it reached 90 % at the end of 15-day culture period. Cellular morphology on PSH scaffold was monitored via SEM analysis; cellular aggregates then spheroid formation were observed throughout the study. Collagen Type-I and F-actin expressions were followed by immunostaining revealing a 9- and 10-fold increase during long-term culture. Overall, a single-step and non-toxic protocol was developed for extraction and gelation of PSH. Obtained results unveiled that PSH scaffold provided a favorable 3D microenvironment for cells, holding promise for further tissue engineering applications.

Microwave-Assisted Atom Transfer Radical Cyclization in the Synthesis of 3,3-Dichloro-γ- and δ-Lactams from N-Alkenyl-Tethered Trichloroacetamides Catalyzed by RuCl2(PPh3)3 and Their Cytotoxic Evaluation.

An expeditious synthesis of γ- and δ-lactams from tethered alkenyl trichloroacetamides in the presence of 5% of RuCl2(PPh3)3 is reported. In this investigation we have demonstrated that microwave activation significantly enhances reaction rates, leading to the formation of the corresponding lactams in yields ranging from good to excellent. Thus, we have been able to prepare a wide range of lactams, including indole and morphan bicyclic scaffolds, where the corresponding reactions were completely diastereoselective. This process was successfully extended to α,α-dichloroamides without affecting either their yield or their diastereoselectivity. Some of the lactams prepared in this work were evaluated for their hemolytic and cytotoxic responses. All compounds were found to be non-hemolytic at the tested concentration, indicating their safety profile in terms of blood cell integrity. Meanwhile, they exhibited interesting cytotoxicity responses that depend on both their lactam structure and cell line. Among the molecules tested, γ-lactam 2a exhibited the lowest IC50 values (100-250 µg/mL) as a function of its cell line, with promising selectivity against squamous carcinoma cells (A431) in comparison with fibroblasts (3T3 cell line).

Understanding the antibacterial mechanism of a phytochemical derived from Urginea indica against Methicillin-Resistant Staphylococcus aureus: A phytochemical perspective to impede antibiotics resistance

Bacterial resistance to conventional antibiotics is a pressing concern in the realm of infectious disease treatment, given its rapid evolution. This underscores the urgency of identifying novel therapeutic compounds. Recent efforts have been concentrated on exploring natural sources of antibacterial compounds, with a particular focus on plant-based derivatives due to their enhanced biocompatibility. In this context, our research has led to the isolation and purification of a groundbreaking plant-based phytochemical derivative known as N-ethylacetamide. We meticulously tested its antibacterial activity against bacterial strains, Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus. Remarkably, N-ethylacetamide exhibited substantial antibacterial and anti-biofilm effects within the concentration range of 5–15 µg/mL, offering promise for combating these pathogens. Our investigations revealed N-ethylacetamide's compatibility with mammalian cells, as evidenced by tests conducted on RAW 264.7 and 3 T3 fibroblast cell lines. The potential antibacterial efficacy of this purified compound was validated through in-vitro infection studies, and a positive immune response was observed in an in-vivo mice model (Balb/c). The accumulated experimental evidence underscores the potential of N-ethylacetamide as a therapeutic agent against bacterial infections. It presents the exciting possibility of addressing these challenges with minimal side effects, offering hope for a more effective and safer approach to combat bacterial diseases.

Dysregulation of miR-146a is associated with exacerbated inflammation, oxidative and endoplasmic reticulum stress in the progression of diabetic foot ulcer.

Recent evidence has implicated the role of microRNA-146a (miR-146a) in regulating inflammatory responses. In the present study, we investigated the role of miRNA-146a in the progression of diabetic foot ulcer (DFU) in type 2 diabetes mellitus patients (T2DM) and studied its correlation with stress mediators such as Endoplasmic Reticulum (ER) and oxidative stress. Ninety subjects were enrolled and evenly distributed among three groups: Controls (n = 30), T2DM without complications (n = 30) and T2DM with foot ulcers (n = 30). Subsequently, each group was further subdivided based on the University of Texas classification. Peripheral blood was collected from all the study subjects, while tissue biopsies were taken only from DFU patients. Total RNA from both PBMCs and wound tissues were isolated using miRNA isolation kit and qPCR was performed to check the expression of miR-146a, ER stress and oxidative stress markers. Our findings revealed a significant decrease in miR-146a expression among T2DM patients with Grade 2 and Grade 3 DFUs compared with those with Grade 0 and Grade 1 DFUs. Notably, inflammatory genes regulated by miR-146a, including TRAF6, IRAK-1 and ADAM, were all upregulated in T2DM patients with Grade 2 and Grade 3 DFUs. Moreover, reduced miR-146a levels were correlated with increased markers of ER stress and oxidative stress in Grade 2 and Grade 3 DFU patients. Furthermore, our in vitro experiment using mouse 3T3 fibroblasts demonstrated a downregulation of miR-146a following induction of hyperglycaemia, ER stress and oxidative stress in these cells. These findings suggest a potential link between diminished miR-146a expression and heightened oxidative and ER stress in T2DM patients with more severe grades of DFUs. Our results imply that targeting miR-146a may hold therapeutic promise for managing disease progression in DFU patients, as it could help alleviate oxidative and ER stress associated with diabetic complications.

Incorporating silver nanoparticles into electrospun nanofibers of casein/polyvinyl alcohol to develop scaffolds for tissue engineering

Developing novel antimicrobial wound dressings that have the potential to address the challenges associated with chronic wounds is highly imperative in providing effective infection control and wound healing support. Biocompatible electrospun nanofibers with their high porosity and surface area enabling efficient drug loading and delivery have been investigated in this regard as viable candidates for chronic wound care. Here, we design Casein/Polyvinyl alcohol (CAN/PVA) nanofibers reinforced with silver nanoparticles (Ag NPs) by the electrospinning technique to develop diabetic wound healing scaffolds. The prepared samples were characterized using spectroscopic and electron microscopic techniques. The biocompatibility of the polymer samples were assessed using 3 T3 fibroblast cell lines and the maximum cell viability was found to 95 % at a concentration of 50 μg/mL for the prepared nanofibers. Scratch assay tests were also performed to analyze the wound healing activity of the nanofibers wherein they demonstrated increased migration and proliferation of fibroblast 3 T3 cells. Moreover, these nanofibers also exhibit antibacterial efficiency against Gram-negative bacteria, Escherichia coli (E.coli). Therefore, the antimicrobial nature of the electrospun nanofibers coupled with their moisture absorption properties and wound healing ability render them as effective materials for wound dressing applications.