Human Dermal Fibroblast Cells Research Articles

Development of alginate and alginate sulfate/polycaprolactone nanoparticles for growth factor delivery in wound healing therapy

Connective tissue growth factor (CTGF) holds great promise for enhancing the wound healing process; however, its clinical application is hindered by its low stability and the challenge of maintaining its effective concentration at the wound site. Herein, we developed novel double-emulsion alginate (Alg) and heparin-mimetic alginate sulfate (AlgSulf)/polycaprolactone (PCL) nanoparticles (NPs) for controlled CTGF delivery to promote accelerated wound healing. The NPs’ physicochemical properties, cytocompatibility, and wound healing activity were assessed on immortalized human keratinocytes (HaCaT), primary human dermal fibroblasts (HDF), and a murine cutaneous wound model. The synthesized NPs had a minimum hydrodynamic size of 200.25 nm. Treatment of HaCaT and HDF cells with Alg and AlgSulf2.0/PCL NPs did not show any toxicity when used at concentrations <50 µg/mL for up to 72 h. Moreover, the NPs’ size was not affected by elevated temperatures, acidic pH, or the presence of a protein-rich medium. The NPs have slow lysozyme-mediated degradation implying that they have an extended tissue retention time. Furthermore, we found that treatment of HaCaT and HDF cells with CTGF-loaded Alg and AlgSulf2.0/PCL NPs, respectively, induced rapid cell migration (76.12% and 79.49%, P<0.05). Finally, in vivo studies showed that CTGF-loaded Alg and AlgSulf2.0/PCL NPs result in the fastest and highest wound closure at the early and late stages of wound healing, respectively (36.49%, P<0.001 on day 1; 90.45%, P<0.05 on day 10), outperforming free CTGF. Double-emulsion NPs based on Alg or AlgSulf represent a viable strategy for delivering heparin-binding GF and other therapeutics, potentially aiding various disease treatments.

Development and functional testing of a novel in vitro delayed scratch closure assay.

As the development of chronic wound therapeutics continues to expand, the demand for advanced assay systems mimicking the inflammatory wound microenvironment in vivo increases. Currently, this is performed in animal models or in in vitro cell-based models such as cell culture scratch assays that more closely resemble acute wounds. Here, we describe for the first time a delayed scratch closure model that mimics some features of a chronic wound in vitro. Chronic wounds such as those suffered by later stage diabetic patients are characterised by degrees of slowness to heal caused by a combination of continued localised physical trauma and pro-inflammatory signalling at the wound. To recreate this in a cell-based assay, a defined physical scratch was created and stimulated by combinations of pro-inflammatory factors, namely interferon, the phorbol ester PMA, and lipopolysaccharide, to delay scratch closure. The concentrations of these factors were characterised for commonly used human keratinocyte (HaCaT) and dermal fibroblast (HDF) cell lines. These models were then tested for scratch closure responsiveness to a proprietary healing secretome derived from human Wharton's jelly mesenchymal stem cells (MSCs) previously validated and shown to be highly effective on closure of acute wound models both in vitro and in vivo. The chronically open scratches from HaCaT cells showed closure after exposure to the MSC secretome product. We propose this delayed scratch closure model for academic and industrial researchers studying chronic wounds looking for responsiveness to drugs or biological treatments prior to testing on explanted patient material or in vivo.

Biologically active sodium pentaborate pentahydrate and Hypericum perforatum oil loaded poly vinyl alcohol: chitosan membranes

In this study, sodium pentaborate pentahydrate (NaB) and Hypericum perforatum (HP) oil were incorporated into polyvinyl alcohol (PVA) and chitosan (CH) polymer blend to obtain membranes by solution casting method. In order to see the synergistic effects of NaB and HP oil on the biological and physical properties of the membranes NaB and HP oil were incorporated into membrane matrix in different ratios. Fourier-transform infrared spectroscopy (FTIR) results showed that no significant bond formation between the bioactive components and the PVA:CH matrix. According to mechanical test results, Young's Modulus and elongation at break decreased from 426 MPa to 346 MPa and 52.23 % to 15.11 % for neat PVA:CH membranes and NaB and HP oil incorporated PVA:CH (PVA:CH@35NaB:HP) membranes, respectively. Antimicrobial activity tests have shown the membranes were over 99 % effective against Escherichia coli, Staphylococcus aureus, and Candida albicans, underlining their potential for infection control. Cytocompatibility assay performed with Human Dermal Fibroblast (HDFa) cells highlight the biocompatibility of the membranes, revealing 74.84 % cell viability after 72 h. The properties of NaB and HP oil doped PVA:CH based membranes obtained from these experiments reveal the promise of a versatile membrane for applications in wound healing, tissue engineering and other biomedical fields.

Burn Wound Healing Abilities of a Uronic Acid Containing Exopolysaccharide Produced by the Marine Bacterium Halomonas malpeensis YU-PRIM-29T.

Bacterial exopolysaccharides (EPS) are an emerging class of biopolymers with extensive applications in different fields due to their versatile physico-chemical and biological properties. The role of EPS in healing of different wound types is gaining interest in the tissue engineering sector. Burn is one of the devitalizing injuries that causes greater physical harm and can be fatal. Appropriate treatment modalities have to be followed for faster healing outcomes and to minimize the risk. In this study, a bacterial EPS (EPS-H29) from the marine bacterium Halomonas malpeensis YU-PRIM-29T was used to treat the burn wound in vivo. The biochemical and structural characterizations of EPS-H29 were carried out using standard methods. In addition, FE-SEM, conformational, rheological, and HP-GPC analyses were carried out. In vitro biocompatibility of EPS-H29 was studied in human dermal fibroblasts (HDFs) and keratinocytes (HaCaT). Scratch assay was used to study the wound healing in vitro. For in vivo evaluation, burn wound (second-degree) was created on Wistar albino rats and treated with EPS-H29 along with appropriate control groups. The total sugar and protein contents of EPS-H29 were 72.0 ± 1.4% and 4.0 ± 0.5%, respectively, with a molecular weight of 5.2 × 105Da. The lyophilized samples exhibited porous surface features, and in solution, it showed triple helical conformation and shear thickening behavior. In vitro cell-based assays showed biocompatibility of EPS-H29 up to 200μg/mL concentration. At a concentration up to 50μg/mL, EPS-H29 promoted cell proliferation. Significant increase in the HDF cell migration was evident with EPS-H29 (15μg/mL) treatment in vitro and induced significantly higher (p ≤ 0.0001) closure of the scratch area (90.3 ± 1.1%), compared to the control (84.3 ± 1.3%) at 24h. Enhanced expression of Ki-67 was associated with the cell proliferative activities of EPS-H29. The animals treated with EPS-H29 showed improved healing of burn wounds with significantly higher wound contraction rate (80.6 ± 9.4%) compared to the positive control (54.6 ± 8.0%) and untreated group (49.2 ± 3.7%) with histopathological evidence of epidermal tissue formation at 15days of treatment. These results demonstrate the biocompatibility and burn wound healing capability of EPS-H29 and its potential as an effective topical agent for the burn wound care.

Green synthesis of stable and biocompatible silver nanoparticles with natural flavonoid apigenin

The synthesis of silver nanoparticles has been carried out, controlling the size and using only apigenin as a reducing and coating agent. Unlike citrate-stabilized silver nanoparticles, the apigenin-coated nanoparticles are very stable in physiological environments. This unique stability is a result of a thick polyphenolic coating on the surface of the particles, which has been observed by electron microscopy. Based on Raman scattering, the phenolic coating originates from apigenin. Hemolysis induced by nanoparticles is less than 1% up to a concentration of 64 μg/ml and less than 5% up to a concentration of 128 μg/ml. These particles do not cause significant toxicity in human dermal fibroblast cells up to a concentration of 256 μg/ml. In the evaluation of the antioxidant properties of particles, it was found that they have antioxidant behavior up to a concentration of 32 μg/ml and pro-oxidant behavior at higher concentrations. Considering the biocompatibility of these nanoparticles, many biomedical applications can be considered for them.

Bead-Free Electrospun Nanofibrous Scaffold Made of PVOH/Keratin/Chitosan Using a Box–Behnken Experimental Design and In Vitro Studies

Bead-free nanofibrous scaffolds composed of PVOH/keratin/chitosan were prepared using electrospinning after optimising the process parameters using a Box–Behnken experimental design. Two quadratic models were developed to optimise the fibre diameter and the diameter of fibre beads considering the voltage, flow rate, spinning distance, and amount of biopolymer as independent variables. All independent variables were found to be significant in determining responses, although not all interactions among these were significant. The models were highly effective in describing responses, with an R2 of 98.58 and 99.67%. The optimum conditions were determined to be 15.82 kV voltage, 0.25 mL/h flow rate, 105 mm spinning distance, and 30% biopolymers. The accuracy of the models was verified and found to be within an acceptable range. The bead-free nanofibrous scaffold exhibited no cytotoxicity to Human Aneuploid Immortal Keratinocyte (HaCaT) and Normal Human Dermal Fibroblast (NHDF) cell lines, enabling cell adhesion and proliferation. Both cell lines remained attached with perfect cell morphology when co-cultured on the scaffold for 30 days, indicating the scaffold’s potential for biomedical applications.

Investigation of the effects of thiazole compounds on thioredoxin reductase 1 (TrxR1), glutathione S-transferase (GST), and glutathione reductase (GR) targeted human brain glioblastoma cancer (U-87 MG).

Cancer is a fatal disease that kills thousands of people worldwide. Despite the information produced by research on cancer treatment, applications in cancer treatment are limited. Therefore, scientists' efforts to develop more effective treatment approaches continue. In the study, we aimed to determine the anticancer potential of amino thiazole compounds on human glioblastoma (U-87 MG) and human dermal fibroblast (HDFa) cells and their inhibition effects on enzymes that cause multidrug resistance in cancer cells. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide cell viability test was performed to understand the cytotoxic properties of thiazole derivatives. The cellular death mechanisms behind thiazole application were investigated using flow cytometry analysis. According to cell viability analysis, thiazole derivatives exhibited a greater effect on U-87 MG than the HDFa cell line in terms of cytotoxicity. Flow cytometry showed higher apoptotic cell death in U-87 MG cells than in the HDFa cell line. It can be concluded that thiazole compounds exert anticancer effects on U-87 MG and HDFa as well as show apoptotic properties. Their effects on thioredoxin reductase 1 (TrxR1), glutathione S-transferase (GST), and glutathione reductase (GR) activities, which are important in the development of chemotherapeutic methods, were also examined. From the results obtained, it was determined that the 2-amino-4-(p-tolyl)thiazole (T7) compound significantly suppressed both TrxR1 and GST activities, and the 2-amino-6-methylbenzothiazole (T8) compound significantly suppressed both TrxR1 and GST activities. Compound T7 was determined to be a selective inhibitor for TrxR1 and GST targeting, and compound T8 was determined to be a selective inhibitor for TrxR1 and GR targeting glioblastoma treatment.

Dissolvable microarray patches of levodopa and carbidopa for Parkinson’s disease management

Carbidopa and levodopa remain the established therapeutic standard for managing Parkinson’s disease. Nevertheless, their oral administration is hindered by rapid enzymatic degradation and gastrointestinal issues, limiting their efficacy, and necessitating alternative delivery methods. This work presents a novel strategy employing dissolving microarray patches (MAPs) loaded with carbidopa and levodopa, formulated with Tween® 80 to improve their transdermal delivery. The fabricated MAPs demonstrated an acceptable mechanical strength, resisting pressures equivalent to manual human thumb application (32 N) onto the skin. Additionally, these MAPs exhibited an insertion depth of up to 650 µm into excised neonatal porcine skin. Ex vivo dermatokinetic studies could achieve delivery efficiencies of approximately 53.35 % for levodopa and 40.14 % for carbidopa over 24 h, demonstrating their significant potential in drug delivery. Biocompatibility assessments conducted on human dermal fibroblast cells corroborated acceptable cytocompatibility, confirming the suitability of these MAPs for dermal application. In conclusion, dissolving MAPs incorporating carbidopa and levodopa represent a promising alternative for improving the therapeutic management of Parkinson’s disease.

Statistical modeling, optimization and characterization of andrographolide loaded emulgel for its therapeutic application on skin cancer through enhancing its skin permeability

Andrographolide is a natural diterpene lactone with multiple biological effects. In the present study, a total of 11 andrographolide-loaded emulgels (ANG 1- ANG 11) were prepared by emulsification and solvent evaporation method using flaxseed oil and xanthan gum in different ratios, as suggested by the Design-Expert software. A 2-factor-5-level design was employed with different responses including spreadability, extrudability, viscosity, and drug release after 1 h (h) and 24 h. Based on the Design-Expert software response, the optimized emulgel ANG 12 was formulated and evaluated. The 24 h In-vitro drug release was found to be 95.7 % following Higuchi kinetics. Ex-vivo skin retention of 784.78 ug/cm2 was observed during the study. MTT assay performed on Human epidermoid carcinoma (A-431) cells demonstrated cell growth arrest at G0/G1 and G2/M phase after 24 h of ANG 12 treatment (IC50: 11.5 µg/ml). The cellular permeability of ANG-12 was assessed by Fluorescein isothiocyanate (FITC) assay. Compared to untreated cells (0.54 % uptake) the ANG-12 treated cells had shown 87.17 % FITC permeation. The biocompatibility study performed on non-cancerous human dermal fibroblast cells (HDF cells) shows 91.54 % viability after 24 h of the treatment showing the non-toxic nature of ANG-12. Confocal imaging had shown a significant time-dependent increase in in-vivo cellular uptake with enhanced, progressive penetration of the emulgel into the skin. An in-vivo skin irritation study conducted on Swiss albino mice confirmed the safety aspects of the ANG 12. Hence, it can be concluded that nanoemulgel of andrographolide (ANG 12) could be a novel approach to treating skin cancer.

Rapid eradication of vancomycin and methicillin-resistant Staphylococcus aureus by MDP1 antimicrobial peptide coated on photocrosslinkable chitosan hydrogel: in vitro antibacterial and in silico molecular docking studies.

Antibiotic resistance and weak bioavailability of antibiotics in the skin due to systemic administration leads to failure in eradication of vancomycin- and methicillin-resistant Staphylococcus aureus (VRSA and MRSA)-associated wound infections and subsequent septicemia and even death. Accordingly, this study aimed at designing a photocrosslinkable methacrylated chitosan (MECs) hydrogel coated by melittin-derived peptide 1 (MDP1) that integrated the antibacterial activity with the promising skin regenerative capacity of the hydrogel to eradicate bacteria by burst release strategy. The MECs was coated with MDP1 (MECs-MDP1), characterized, and the hydrogel-peptide interaction was evaluated by molecular docking. Antibacterial activities of MECs-MDP1 were evaluated against VRSA and MRSA bacteria and compared to MECs-vancomycin (MECs-vanco). Antibiofilm activity of MECs-MDP1 was studied by our novel 'in situ biofilm inhibition zone (IBIZ)' assay, and SEM. Biocompatibility with human dermal fibroblast cells (HDFs) was also evaluated. Molecular docking showed hydrogen bonds as the most interactions between MDP1 and MECs at a reasonable affinity. MECs-MDP1 eradicated the bacteria rapidly by burst release strategy whereas MECs-vanco failed to eradicate them at the same time intervals. Antibiofilm activity of MECs-MDP1 were also proved successfully. As a novel report, molecular docking analysis has demonstrated that MDP1 covers the structure of MECs and also binds to lysozyme with a reasonable affinity, which may explain the inhibition of lysozyme. MECs-MDP1 was also biocompatible with human dermal fibroblast skin cells, which indicates its safe future application. The antibacterial properties of a photocrosslinkable methacrylated chitosan-based hydrogel coated with MDP1 antimicrobial peptide were successfully proved against the most challenging antibiotic-resistant bacteria causing nosocomial wound infections; VRSA and MRSA. Molecular docking analysis revealed that MDP1 interacts with MECs mainly through hydrogen bonds with reasonable binding affinity. MECs-MDP1 hydrogels eradicated the planktonic state of bacteria by burst release of MDP1 in just a few hours whereas MECs-vanco failed to eradicate them. inhibition zone assay showed the anti-biofilm activity of the MECs-MDP1 hydrogel too. These findings emphasize that MECs-MDP1 hydrogel would be suggested as a biocompatible wound-dressing candidate with considerable and rapid antibacterial activities to prevent/eradicate VRSA/MRSA bacterial wound infections.

Copper(I) complexes of acylthiourea ligands: structural insights and cytotoxic potential

The reaction of substituted acylthiourea derivatives (L1–L4) with [(PPh3)2Cu(μ‐Cl)2Cu(PPh3)] in a proper stoichiometric proportion gave the complexes (1–4) in good yields. The structures of the compounds (ligands and complexes) were advocated through spectroscopic analyses and x‐ray diffraction (XRD) studies; the latter confirmed their molecular structures. The complexes (1–4) were assessed for their cytotoxic potential through MTT assay against A549, T24, and HepG2 cancer cells, and their toxicity was evaluated using Vero and MCF‐10a normal cells. The complexes (1–4) displayed better cytotoxic action toward HepG2 cancer cells than cisplatin. Complexes 3 and 4 significantly destroyed A549 cancer cells with the IC50 values of 16.25 and 4.90 μM, respectively, which were lower than that of cisplatin (IC50 = 17.73 μM). Furthermore, the higher IC50 values in normal cells showed that the complexes were less hazardous to the normal cells except complex 3. The biocompatibility study showed that the complexes had minimal impact on normal human dermal fibroblast (NHDF) cells. The mode of cell death of complexes 3 and 4 against cancer cells was evaluated using staining assays. Both compounds demonstrated a significant reduction in live cells, an increase in early apoptotic cells, and pronounced proapoptotic effects, as evidenced by fragmented nuclei. Flow cytometry revealed a distinctive subpopulation in the subG0 phase, highlighting the compounds' ability to induce apoptosis and inhibit cell cycle progression.

Bacterial nanocellulose loaded with bromelain and nisin as a promising bioactive material for wound debridement

The bacterial nanocellulose (BnC) membranes were produced extracellularly by a novel aerobic acetic acid bacterium Komagataeibacter melomenusus. The BnC was modified in situ by adding carboxymethyl cellulose (CMC) into the culture media, obtaining a BnC-CMC product with denser fibril arrangement, improved rehydration ratio and elasticity in comparison to BnC. The proteolytic enzyme bromelain (Br) and antimicrobial peptide nisin (N) were immobilized to BnC matrix by ex situ covalent binding and/or adsorption. The optimal Br immobilization conditions towards the maximized specific proteolytic activity were investigated by response surface methodology as factor variables. At optimal conditions, i.e., 8.8 mg/mL CMC and 10 mg/mL Br, hyperactivation of the enzyme was achieved, leading to the specific proteolytic activity of 2.3 U/mg and immobilization efficiency of 39.1 %. The antimicrobial activity was observed against Gram-positive bacteria (S. epidermidis, S. aureus and E. faecalis) for membranes with immobilized N and was superior when in situ modified BnC membranes were used. N immobilized on the BnC or BnC-CMC membranes was cytocompatible and did not cause changes in normal human dermal fibroblast cell morphology. BnC membranes perform as an efficient carrier for Br or N immobilization, holding promise in wound debridement and providing antimicrobial action against Gram-positive bacteria, respectively.

AESIS-1, a Rheumatoid Arthritis Therapeutic Peptide, Accelerates Wound Healing by Promoting Fibroblast Migration in a CXCR2-Dependent Manner.

In patients with autoimmune disorders such as rheumatoid arthritis (RA), delayed wound healing is often observed. Timely and effective wound healing is a crucial determinant of a patient's quality of life, and novel materials for skin wound repair, such as bioactive peptides, are continuously being studied and developed. One such bioactive peptide, AESIS-1, has been studied for its well-established anti-rheumatoid arthritis properties. In this study, we attempted to use the anti-RA material AESIS-1 as a therapeutic wound-healing agent based on disease-modifying antirheumatic drugs (DMARDs), which can help restore prompt wound healing. The efficacy of AESIS-1 in wound healing was assessed using a full-thickness excision model in diabetic mice; this is a well-established model for studying chronic wound repair. Initial observations revealed that mice treated with AESIS-1 exhibited significantly advanced wound repair compared with the control group. In vitro studies revealed that AESIS-1 increased the migration activity of human dermal fibroblasts (HDFs) without affecting proliferative activity. Moreover, increased HDF cell migration is mediated by upregulating chemokine receptor expression, such as that of CXC chemokine receptor 2 (CXCR2). The upregulation of CXCR2 through AESIS-1 treatment enhanced the chemotactic reactivity to CXCR2 ligands, including CXC motif ligand 8 (CXCL8). AESIS-1 directly activates the ERK and p38 mitogen-activated protein kinase (MAPK) signaling cascades, which regulate the migration and expression of CXCR2 in fibroblasts. Our results suggest that the AESIS-1 peptide is a strong wound-healing substance that increases the movement of fibroblasts and the expression of CXCR2 by turning on the ERK and p38 MAPK signaling cascades.

Abstract 5232: Placental circulating T cells expressing CD16 in combination with trastuzumab demonstrate robust anti-tumor antibody-dependent cellular cytotoxicity (ADCC) against gastric cancer

Abstract Introduction: Celularity is developing PT-CD16VS as a novel platform for use in combination with approved monoclonal antibodies for the potential treatment of multiple cancers. PT-CD16VS is an allogeneic cell therapy derived from human postpartum placental circulating T (P-T) cells that are genetically modified to express a proprietary CD16 variant and endogenous T cell receptor (TCR) knockout. Here we report the characterization and preclinical evaluation of PT-CD16VS against HER2+ gastric cancer cells in combination with Trastuzumab. In vitro safety studies evaluating the potential for on-target/off-tumor activity of PT-CD16VS against HER2 low expressing lung epithelial cells (LEC) and human dermal fibroblasts (HDF) are also highlighted. Methods: P-T cells were activated and transduced with a lentiviral vector containing a CD16 construct expressing a high affinity CD16 variant, CD16VS. Following transduction, P-T cells were transfected to knock out the TCR. In vitro, the functional activity of PT-CD16VS cells in combination with Trastuzumab was assessed against the HER2+ gastric cancer NCI-N87 cell line in cytotoxicity, cytokine release, and proliferation assays. For on-target/off-tumor studies, similar assays were utilized to assess activity against the HER2 low expressing LEC and HDF cell lines. In vivo, PT-CD16VS with Trastuzumab was evaluated in a subcutaneous NCI-N87 xenograft model in NSG mice. Tumor volume was measured and tumor samples were evaluated for PT-CD16VS infiltration. Results: High CD16 transduction efficiency (&amp;gt;60%) was achieved across donors. In vitro, PT-CD16VS plus Trastuzumab demonstrated potent cytotoxicity against NCI-N87 cells at low E:T ratios (0.625:1) and at 12 hours of co-culture with moderate cytokine secretion at 24-hour of co-culture. PT-CD16VS proliferated following 48-hour incubation with NCI-N87 cells and Trastuzumab, as compared to an IgG1 control. PT-CD16VS in combination with Trastuzumab did not elicit on-target/off-tumor cytotoxicity or cytokine release in response to low HER2 expression on normal LEC or HDF cells. In vivo, PT-CD16VS with Trastuzumab demonstrated significant reduction in tumor volume compared to vehicle, Trastuzumab alone, and the positive control Enhertu groups. In addition, PT-CD16VS infiltration into the tumor was shown to be Trastuzumab dependent, with infiltrating cells expressing high levels of CD16 and Ki67. Conclusions: Our results show that PT-CD16VS has robust in vitro and in vivo ADCC activity against gastric cancer tumor cells when combined with Trastuzumab. Furthermore, PT-CD16VS activity is specific to tumor HER2 antigen expression and does not target low HER2 expression on normal healthy cells. PT-CD16VS is being developed in combination with various monoclonal antibodies for the treatment of multiple cancers. Citation Format: Kathy Karasiewicz-Mendez, Joseph Gleason, Kristina Tess, Mansour Djedaini, Shengchen Lin, Chenfei Huang, Shuyang He, Robert Hariri, Adrian Kilcoyne. Placental circulating T cells expressing CD16 in combination with trastuzumab demonstrate robust anti-tumor antibody-dependent cellular cytotoxicity (ADCC) against gastric cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5232.

Development and in vitro characterization nanoemulsion containing the methanol extract of Hypericum linarioides for wound healing: In vitro scratch assay

It was aimed to develop and in vitro characterize the methanol extract of Hypericum linarioides (MHL)-containing nanoemulsion formulation (MHL-NEF) to heal the wound effectively. MHL-NEF was prepared and its droplet size, polydispersity index (PDI), viscosity, pH and zeta potential (ZP) values were determined. The effect of MHL and MHL-NEF on human dermal fibroblast cell viability was examined by MTT assay. Also, their wound healing potential was evaluated using the scratch assay. The droplet size, PDI, ZP, pH and viscosity values for the MHL-NEF were found to be 157.67±7.61 nm, 0.268±0.036, (-)40.20±6.79 mV, 4.27±0.01, and 1.82±0.01 cP, respectively. In the cell viability assay, the highest percentage of living cells was obtained at concentration of 0.4 µg/mL for the MHL and MHL-NEF. In the scratch assay, MHL-NEF showed significantly ameliorated wound closure (%) rates at 24 and 30 h than the groups of control, MHL and blank-NEF (NEF without MHL) (p < 0.001). Immunofluorescence staining was carried out. TGF-β1 and FGFR-2 expressions in MHL-NEF-treated fibroblast cells increased significantly at 24 h compared to the groups of control, MHL, and B-NEF (p < 0.001). Our findings show that MHL-NEF may be promising for better wound healing.

Modulating the Wnt Signaling Pathway in Wound Healing: The Potential of L-Carnosine, Zinc Sulfate, and B-Complex Vitamins Cocktail

Background: The process of wound healing is a dynamic event that starts with inflammation, proliferation, and migration of various types of fibroblast cells and aims to restore the integrity and functionality of damaged tissue. One crucial signaling pathway involved in wound healing is the Wnt signaling pathway. This study aimed to investigate the effects of a cocktail consisting of L-carnosine, zinc sulfate, and B-complex vitamins on the modulation of the Wnt signaling pathway during wound healing. Methods: The optimal dosages of L-carnosine, zinc sulfate, and B-complex were used to prepare the cocktail. Then, the cocktail-mediated wound healing was measured through scratch assay on human dermal fibroblast (HDF) cells. Finally, the effects of the cocktail on the expression of SFRP1, SFRP2, SFRP4, MMP7, and RSPO2 genes involved in the Wnt signaling pathway were measured during the wound closure phenomenon by real-time quantitative polymerase chain reaction (RT-qPCR). Results: The scratch assay demonstrated the wound-healing effects of the prepared cocktail on HDF cells. Additionally, the RT-qPCR results showed that the cocktail can activate the Wnt signaling pathway by down-regulating the expression of SFRP1, SFRP2, SFRP4, and MMP7 and up-regulating the expression of RSPO2. Conclusions: The data suggest that the L-carnosine, zinc sulfate, and B-complex vitamin cocktail exhibits wound healing properties by increasing fibroblast proliferation via the activation of the Wnt pathway.

Effects of Blood-Derived Products on Cellular Senescence and Inflammatory Response: A Study on Skin Rejuvenation.

Blood-derived products, such as citrate platelet-rich plasma (CPRP) and hyperacute serum (HAS), are recognized for their rich growth factor content. When human dermal fibroblast (HDF) cells are exposed to combined mitogenic and DNA-damaging stimuli, it can lead to an increased burden of senescent cells and a modified senescence-associated secretory phenotype. In this study, the senescent state was comprehensively assessed through various methods, including phosphorylated histone H2AX (γH2AX) staining, p21 and p16 q-PCR, p21-western blot, growth curves, and senescence-associated ß-galactosidase staining. Two primary treatments with blood products were administered, one early (immediately after etoposide) and the other late (11 days after etoposide treatment). The effects of the blood product treatment were evaluated by measuring interleukin 6 and 8 (IL-6 and IL-8) levels, as well as collagen 1 (COL1) and p21 mRNA expression. Additionally, 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) assays, cell size measurements, viability assays, and cell number calculations were conducted. The results revealed that cells treated with hyperacute serum in the early treatment phase exhibited the lowest observed IL-6 and IL-8 levels. In contrast, a clear inflammatory response for IL-8 was observed in cells treated with hyperacute serum and citrate platelet-rich plasma during the late treatment. Furthermore, an upregulation of COL1 expression was observed in the early treatment, while cells in the late treatment group remained unaffected. Notably, citrate platelet-rich plasma-treated cells showed a decrease in COL1 expression. Overall, the treatment with blood products appears to have slightly positive effects on skin rejuvenation.

Three-dimensional macro/micro-porous curcumin releasing polycaprolactone/chitosan nanofiber scaffolds as wound dressing

In this study, three-dimensional (3D) polycaprolactone (PCL)-chitosan (Chi)-curcumin (Cur) nanofibrous wound dressings were fabricated with a dispersion shaping method. After conventional electrospinning of PCL-Chi and PCL-Chi-Cur mixture, bead-free and smooth nanofibers with the mean diameters between 90 – 160 nm were successfully obtained. Then, 3D nanofibrous PCL-Chi and PCL-Chi-Cur wound dressings with mean diameters of 235 ± 50 nm and 226 ± 47 nm, respectively, were fabricated in macro/micro-porous structure. According to the micro-computerized tomography analysis, 3D nanofibrous PCL-Chi and PCL-Chi-Cur materials had the mean pore sizes of 97 µm and 208 µm with the total porosity of 90%. When chitosan was added to the structure, the hydrophilicity and water uptake capacity of the developed materials were significantly improved. Furthermore, curcumin was successfully loaded into 3D nanofibrous structures with 61% encapsulation efficiency and 3% drug loading capacity. When the curcumin release was analysed, it was observed that 55% of the loaded curcumin was released within the first 3 h, and the release was completed at the end of 10 h with the released amount of 64%. Kinetic analyzes revealed that the release profile was diffusion controlled in accordance with the Higuchi model. Cell culture studies were performed with human dermal fibroblasts (HDFs) and cell metabolic activity and morphology were monitored by MTT and SEM analyses, respectively. The presence of curcumin in 3D nanofibrous wound dressings had a positive effect on the viability and proliferation of HDF cells, albeit decreasing, especially in the first 7 days of culture, but this effect almost disappeared on day 14. In SEM analyses, it was observed that HDFs in all groups migrated to the depths of the material through macropores, but it can be said that cell-cell and cell-matrix interactions were much better in the PCL-Chi-Cur group. Hence, curcumin-incorporated 3D nanofibrous material containing macro/micro pores within the structure have potential to be a good alternative wound dressing in the future when curcumin release is further extended.

Photoluminescent Self-Healable Waterborne Polyurethane/Mo and S Codoped Graphitic Carbon Nitride Nanocomposite with Bioimaging and Encryption Capability.

Creating polymers that combine various functions within a single system expands the potential applications of such polymeric materials. However, achieving polymer materials that possess simultaneously elevated strength, toughness, and self-healing capabilities, along with special properties, remains a significant challenge. The present study demonstrates the preparation of S and Mo codoped graphitic carbon nitride (g-C3N4) (Mo@S-CN) nanohybrid and the fabrication of self-healing waterborne polyurethane (SHWPU)/Mo@S-CN (SHWPU/NS) nanocomposites for advanced applications. Mo@S-CN is an intriguing combination of g-C3N4 nanosheets and molybdenum oxide (MoOx) nanorods, forming a complex lamellar structure. This unique arrangement significantly improves the inborn properties of SHWPU to an impressive degree, especially mechanical strength (28.37-34.11 MPa), fracture toughness (73.65-140.98 MJ m-2), and thermal stability (340.17-348.01 °C), and introduces fluorescence activity into the matrix. Interestingly, a representative SHWPU/NS0.5 film is so tough that a dumbbell of 15 kg, which is 53,003 times heavier than the weight of the film, can be successfully lifted without any significant crack. Remarkably, fluorescence activity is developed because of electronic excitations occurring within the repeating polymeric tris-triazine units of the Mo@S-CN nanohybrid. This fascinating feature was effectively harnessed by assessing the usability of aqueous dispersions of the Mo@S-CN nanohybrid and photoluminescent SHWPU/NS nanocomposites as sustainable stains for bioimaging of human dermal fibroblast cells and anticounterfeiting materials, respectively. The in vitro fluorescence tagging test showed blue emission from 365 nm excitation, green emission from 470 nm excitation, and red emission from 545 nm excitation. Most importantly, in vitro hemocompatibility assessment, in vitro cytocompatibility, cell proliferation assessment, and cellular morphology assessment supported the biocompatibility nature of the Mo@S-CN nanohybrid and SHWPU/NS nanocomposites. Thus, these materials can be used for advanced applications including bioimaging.

Assessment of the biocompatibility and bioimaging potential of fluorescent carbon dots derived from waste biomass

The fluorescent carbon dots (C-dots) are attractive platforms in bioimaging, biosensing, and disease detection applications due to their attractive physicochemical characteristics. Herein, we synthesized fluorescent and biocompatible C-dots from banana biomass through heat treatment. The C-dots were soluble in water and showed a fluorescent behavior under UV light (365 nm). The synthesized C-dots demonstrated an excitation-dependent emission behavior with a satisfying quantum yield of 15 %. The C-dots exhibited different functional groups in their structure. The C-dots have no adverse effects on human dermal fibroblast cells (HDFs), indicating their biocompatibility. The C-dots have successfully delivered inside cells and showed fluorescent behavior. These results showed that the prepared materials could potentially explore high-performance bioimaging applications.