Wound Neutrophils Research Articles

Application of silver nanoparticles containing Gundelia tournefortii L. leaf aqueous extract in the treatment of microbial diseases and cutaneous wound healing

AbstractDuring the past few years, silver nanoparticles (AgNPs) have become some of the most investigated and explored nanotechnology‐derived nanostructures, given the fact that nanosilver‐based materials have interesting, challenging, and promising characteristics suitable for various biomedical applications. AgNPs have excellent medical and nonmedical effects and applications compared with other metallic nanoparticles. As regards modern biomedical applications of AgNPs, tremendous interest is oriented toward therapeutically enhanced personalized healthcare practice. AgNPs have genuine features and impressive potential for the development of novel performance‐enhanced therapeutic alternatives, complex healthcare condition strategies, tissue restoration and regeneration materials, biomaterial and medical device coatings, detection and diagnosis platforms, drug‐delivery formulations, and antimicrobial agents. In this study, fresh leaves of Gundelia tournefortii L. were used for the synthesis of AgNPs. Also, we assessed the cytotoxic, antioxidant, antifungal, antibacterial, and cutaneous wound healing properties of AgNO3, G. tournefortii, and the synthesized nanoparticles (AgNPs@GT). These nanoparticles were characterized using various techniques. In X‐ray diffraction analysis, 16.7 nm was measured for the crystal size of the nanoparticles. Also, scanning and transmission electron microscopy images revealed an average diameter of 16.5 nm. Fourier transform infrared spectroscopic results indicated that antioxidant compounds in the plant were the sources of reducing power, reducing silver ions to AgNPs@GT. In vitro biological experiments indicated that AgNPs@GT has excellent antioxidant potential against DPPH, antifungal effects against Candida krusei, C. guilliermondii, C. glabrata, and C. albicans, and antibacterial activities against Pseudomonas aeruginosa, Staphylococcus aureus, Salmonella typhimurium, Streptococcus pneumoniae, Escherichia coli O157:H7, and Bacillus subtilis. Also, these nanoparticles showed no cytotoxic effect against human umbilical vein endothelial cells. In vivo biological tests revealed that AgNPs@GT ointment significantly (p ≤ 0.01) increased the levels of wound contracture, blood vessels, hydroxyproline, hexuronic acid, hexosamine, fibrocytes, fibroblasts, and fibrocyte‐to‐fibroblast ratio and significantly (p ≤ 0.01) decreased the levels of wound area, total cells, neutrophils, and lymphocytes in rats. These results indicate that the inclusion of G. tournefortii extract ameliorates the solubility of AgNPs, which led to a marked enhancement in fungicidal, bactericidal, and cutaneous wound healing effects under in vitro and in vivo conditions.

Synthesis, characterization, and evaluation of cytotoxicity, antioxidant, antifungal, antibacterial, and cutaneous wound healing effects of copper nanoparticles using the aqueous extract of Strawberry fruit and l-Ascorbic acid

Fragaria ananassa, also known as “Strawberry” is a common species in Iran and widely used for its anti-inflammatory, anti-ulcer, astringent, anti-allergic, antibacterial, antifungal, and antidiarrheal activities and also in the treatment of skin wounds. The purpose of the study was chemical characterization and assessment of cytotoxicity, antioxidant, antifungal, antibacterial, and cutaneous wound healing properties of copper nanoparticles (CuNPs) using the aqueous extract of Strawberry fruit and l-Ascorbic acid as reducing and stabilizing agents. These nanoparticles were characterized by FT-IR, UV–visible spectroscopy, EDS, FE-SEM, and TEM analysis. TEM images exhibited a uniform spherical morphology and diameters of 10–30 nm for the biosynthesized nanoparticles. DPPH free radical scavenging test revealed similar antioxidant properties for Strawberry, CuNPs, and butylated hydroxytoluene. The Strawberry and synthesized CuNPs had great cell viability dose-dependently against HUVEC cell line. In the microbiological part of this study, CuNPs showed higher antibacterial and antifungal properties than all standard antibiotics (p ≤ 0.01). Also, CuNPs prevented the growth of all bacteria at 2–8 mg/mL concentrations and destroyed them at 2–16 mg/mL concentrations (p ≤ 0.01). In the case of antifungal property of CuNPs, they inhibited the growth of all fungi at 2–4 mg/mL concentrations and destroyed them at 2–8 mg/mL concentrations (p ≤ 0.01). In vivo design, the use of CuNPs ointment in the treatment groups substantially remarkably raised (p ≤ 0.01) the wound contracture, hydroxyl proline, hexosamine, hexuronic acid, fibrocyte, and fibrocytes/fibroblast rate and reduced (p ≤ 0.01) the wound area, total cells, neutrophil, macrophage, and lymphocyte compared to Strawberry, CuSO4, tetracycline, Eucerin basal, and untreated control groups. In conclusion, the results of chemical characterization confirm that the Strawberry fruit can be consumed to produce copper nanoparticles with a remarkable amount of remedial effects without any cytotoxicity against HUVECs.

Application of titanium nanoparticles containing natural compounds in cutaneous wound healing

The aim of the study was the rapid green synthesis of titanium nanoparticles using the aqueous extract of Falcaria vulgaris leaves (TiNPs@FV) and exploring their antioxidant, cytotoxicity, antifungal, antibacterial, and cutaneous wound healing activities under in vitro and in vivo condition. These nanoparticles were characterized by UV‐Vis, Fourier transform‐infrared(FT‐IR), X‐ray diffraction XRD), field emission‐scanning electron microscopy FE‐SEM), and transmission electron microscopy TEM) analyses. The synthesized TiNPs@FV had great cell viability on human umbilical vein endothelial cells and indicted this method was nontoxic. DPPH (2,2‐diphenyl‐1‐picrylhydrazyl) test revealed similar antioxidant potentials for F. vulgaris, TiNPs@FV, and butylated hydroxytoluene. All data of antibacterial, antifungal, and cutaneous wound healing tests were analyzed by SPSS 22 software. In the antimicrobial part of this study, TiNPs@FV indicated higher antifungal and antibacterial effects than all standard antibiotics (p ≤ 0.01). Minimal inhibitory concentration (MIC) and minimal fungicidal concentration of TiNPs@FV against all fungi were at 2–4 mg/mL and 2‐8 mg/mL ranges, respectively. But, MIC and minimal bactericidal concentration of TiNPs@FV against all bacteria were at 2‐8 mg/mL and 2‐16 mg/mL ranges, respectively. In the part of cutaneous wound healing, use of TiNPs@FV ointment significantly (p ≤ 0.01) raised the wound contracture, vessel, hydroxyl proline, hexuronic acid, hexosamine, fibrocyte, and fibrocytes/fibroblast rate and significantly (p ≤ 0.01) decreased the wound area, total cells, neutrophil, and lymphocyte compared to other groups in rats. The results of FT‐IR, UV‐Vis, XRD, TEM, and FE‐SEM confirm that the aqueous extract of F. vulgaris leaves can be used to yield titanium nanoparticles with a notable amount of remedial effects.

The Sinergy of Isoquercetin and D-Galacturonic acid of Cyclea barbata Miers extract on the Migration and Apoptosis of Neutrophil in Rat Burn Wound

Second-degree burn is one of acute traumas that has complexities in its healing process; one of which is the disruption of neutrophils activity and function as the initial cellular defense during inflammatory phase. Appropriate wound treatment can modulate the quantity of neutrophils in burn wounds. Cyclea barbata Miers (CBM) is a local plant that has a potential to influence inflammatory process. This study was aimed to determine the polyphenol and monosaccharide contents of CBM and the effect of CBM on neutrophils migration and apoptosis. We used 30 adult male Wistar rats as experimental animals, wich were randomly separated in two groups according the eutanized time. Each group divided in 3 groups (n=5 per group). Cyclea barbata Miers’ content was analyzed using liquid chromatography mass spectrometry (LCMS/MS) method, while neutrophils migration and apoptosis were observed using immunofluorescence assay method. The result of LCMS/MS assay showed that CBM contains isoquercetin (93.83ng/mL) and d-galacturonic acid (0.114ng/mL). Further result showed that CBM extract can decrease neutrophil migration and improve neutrophil apoptosis during the inflammatory phase of the burn wound. This ability may be enabled by the anti-inflammatory and antiradical properties of isoquercetin and DGA in Cyclea barbata Miers. Further study is required to confirm this hypothesis, specially addition of comparasion groups that treated by single compound such as isoquercetin and DGA.

Biosynthesis of iron nanoparticles using Allium eriophyllum Boiss extract: Chemical characterization, antioxidant, cytotoxicity, antibacterial, antifungal, and cutaneous wound healing effects

Recently, metal nanoparticles have been used for the treatment of several disorders, such as infectious diseases. Indeed, finding the antibiotics of nanoparticles is in researching priority of many countries. The aim of this experiment was the green synthesis of iron nanoparticles from aqueous extract of Allium eriophyllum Boiss (FeNPs@AE) and evaluation of their antioxidant, cytotoxicity, antibacterial, antifungal, and cutaneous wound healing effects under in vitro and in vivo conditions. These nanoparticles were characterized by FT‐IR, UV, XRD, EDS, TEM, and FE‐SEM analysis. According to the XRD analysis, 40 nm was measured for the crystal size of nanoparticles. SEM and TEM images exhibited a uniform spherical morphology and average diameters of 45 nm for the nanoparticles. FTIR findings suggested antioxidant compounds in the nanoparticles were the sources of reducing power, reducing iron ions to FeNPs@AE. DPPH test indicated similar antioxidant potentials for FeNPs@AE and butylated hydroxytoluene. The synthesized FeNPs@AE had great cell viability dose‐dependently and revealed this method was nontoxic. In the antimicrobial part of this study, FeNPs@AE prevented the growth of all bacteria and removed them at 2‐4 mg/ml concentrations (p ≤ 0.01). In the case of antifungal effects of FeNPs@AE, they inhibited the growth of all fungi at 1‐2 mg/ml concentrations and destroyed them at 2‐4 mg/ml concentrations (p ≤ 0.01). In part of cutaneous wound healing effect of FeNPs@AE, after creating the cutaneous wound, the rats were randomly divided into six groups: treatment with 0.2% FeNPs@AE ointment, treatment with 0.2% A. eriophyllum ointment, treatment with 0.2% FeCl3.6H2O ointment, treatment with 3% tetracycline ointment, treatment with Eucerin basal ointment, and untreated control. These groups were treated for 10 days. Use of FeNPs@AE ointment in the treatment groups substantially raised (p ≤ 0.01) the wound contracture, vessel, hydroxyl proline, hexuronic acid, fibrocyte, and fibrocytes/fibroblast rate and remarkably decreased (p ≤ 0.01) the wound area, total cells, neutrophil, and lymphocyte compared to other groups. The results of FT‐IR, UV, XRD, TEM, and FE‐SEM confirm that the aqueous extract of A. eriophyllum can be used to produce iron nanoparticles with a significant amount of antioxidant, antibacterial, antifungal, and cutaneous wound healing properties without any cytotoxicity.

Ethnomedicinal plant‐extract‐assisted green synthesis of iron nanoparticles usingAllium saralicumextract, and their antioxidant, cytotoxicity, antibacterial, antifungal and cutaneous wound‐healing activities

The aim of the experiment was the evaluation of antioxidant, cytotoxicity, antibacterial, antifungal and cutaneous wound‐healing activities of green synthesized iron nanoparticles usingAllium saralicumR.M. Fritsch leaves (FeNPs@AS). These nanoparticles were spherical with a size range of 40–45 nm, and were characterized using various analysis techniques including ultraviolet–visible spectroscopy to determine the presence of FeNPs@AS in the solution. We studied functional groups ofA. saralicumextract in the reduction and capping process of FeNPs@AS by Fourier transform‐infrared spectroscopy; crystallinity and FCC planes by X‐ray diffraction pattern; and surface morphology, shapes and size of FeNPs@AS by scanning electron microscopy and transmission electron microscopy. Agar diffusion tests were done to determine the antibacterial and antifungal characteristics. FeNPs@AS prevented the growth of all bacteria and removed them at 2–8 mg/ml concentrations (P ≤ 0.01). In the case of antifungal potentials of FeNPs@AS, they inhibited the growth of all fungi and destroyed them at 2–4 mg/ml concentrations (P ≤ 0.01). The 2,2‐diphenyl‐1‐picrylhydrazyl test revealed similar antioxidant potentials for FeNPs@AS and butylated hydroxytoluene. The synthesized FeNPs@AS had great cell viability dose‐dependently and indicated this method was non‐toxic. For thein vivoexperiment, after creating the cutaneous wound, the rats were randomly divided into six groups: treatment with 0.2% FeNPs@AS ointment; treatment with 0.2%A. saralicumointment; treatment with 0.2% FeCl3·6H2O ointment; treatment with 3% tetracycline ointment; treatment with Eucerin basal ointment; and untreated control. These groups were treated for 10 days. Use of FeNPs@AS ointment in the treatment groups significantly decreased (P ≤ 0.01) the wound area, total cells, neutrophils and lymphocytes, and significantly raised (P ≤ 0.01) the wound contracture, hydroxyl proline, hexosamine, hexuronic acid, fibrocyte and fibrocytes/fibroblast rate compared with other groups. These results show that the inclusion ofA. saralicumextracts improves the therapeutical properties of FeNPs, which led to a significant enhancement in the antioxidant, non‐cytotoxicity, antibacterial, antifungal and cutaneous wound‐healing activities of the nanoparticles.

A Frog-Derived Immunomodulatory Peptide Promotes Cutaneous Wound Healing by Regulating Cellular Response.

Wound healing-promoting peptides exhibit excellent therapeutic potential in regenerative medicine. However, amphibian-derived wound healing-promoting peptides and their mechanism of action remain to be further elucidated. We hereby characterized a wound healing-promoting peptide, Ot-WHP, derived from Chinese concave-eared frog Odorrana tormota. It efficiently promoted wound healing in a mouse model of full-thickness wounds. Ot-WHP significantly increased the number of neutrophils in wounds, and modestly promoted neutrophil phagocytosis and phorbol myristate acetate (PMA)-induced neutrophil extracellular trap formation. Ot-WHP also significantly increased the number of macrophages in wound sites, and directly induced chemokine, cytokine and growth factor production in macrophages by activating mitogen-activated protein kinases (MAPKs) and nuclear factor-κB (NF-κB) signaling pathways. Of note, Ot-WHP did not act as a chemoattractant for neutrophils and macrophages, suggesting its chemotactic activity depends on inducing chemoattractant production in macrophages. Besides, Ot-WHP directly promoted keratinocyte migration by enhancing integrin expression and cell adhesion. In addition, Ot-WHP significantly enhanced the cross-talk between macrophages and keratinocytes/fibroblasts by promoting keratinocyte/fibroblast proliferation, and fibroblast-to-myofibroblast transition despite having no direct effects on keratinocyte/fibroblast proliferation, and fibroblast differentiation. Collectively, Ot-WHP directly elicited the production of regulatory factors in macrophages, consequently initiated and accelerated the inflammatory phase by recruiting neutrophils and macrophages to wounds, and in turn enhanced the cross-talk between macrophages and keratinocytes/fibroblasts, additionally promoted keratinocyte migration, and finally promoted cutaneous wound healing. Our findings provide a promising immunomodulator for acute wound management and new clues for understanding the mechanism of action of amphibian-derived wound healing-promoting peptides.

Synthesis of titanium nanoparticles using Allium eriophyllum Boiss aqueous extract by green synthesis method and evaluation of their remedial properties

The application of nanotechnologies to medicine, or nanomedicine, which has already demonstrated its tremendous impact on the pharmaceutical and biotechnology industries, is rapidly becoming a major driving force behind ongoing developments in the antimicrobial and wound healing fields. This study confirms the potential of Allium eriophyllum Boiss aqueous extract for the green synthesis of titanium nanoparticles (TiNPs). Also, we reveal the antioxidant, cytotoxicity, cutaneous wound healing, antifungal and antibacterial properties of TiNPs. These nanoparticles were characterized using Fourier transform infrared (FT‐IR) and UV–visible spectroscopies, X‐ray diffraction (XRD), transmission electron microscopy (TEM) and field emission scanning electron microscopy (SEM). FT‐IR findings suggested molecular interactions of compounds in the plant were the sources of reducing power, reducing titanium ions to TiNPs. In XRD analysis, 22.8 nm was measured for the crystal size of the nanoparticles. SEM and TEM images indicated a uniform spherical morphology and average diameters of 22 nm. 2,2‐Diphenyl‐1‐picrylhydrazyl experiments were conducted to assess the antioxidant activities, which indicated similar antioxidant potentials for TiNPs and butylated hydroxytoluene. In the antimicrobial part of this study, agar diffusion experiments were done to determine the antibacterial and antifungal characteristics. TiNPs had antifungal activities against Candida guilliermondii, C. krusei, C. albicans and C. glabrata and antibacterial potentials against Gram‐negative bacteria (Pseudomonas aeruginosa, Salmonella typhimurium and Escherichia coli O157:H7) and Gram‐positive bacteria (Staphylococcus aureus, Streptococcus pneumoniae and Bacillus subtilis). Also, these nanoparticles did not have any cytotoxicity against human umbilical vein endothelial cells. In in vivo experiments, TiNP ointment significantly decreased (p ≤ 0.01) wound area, total cells, macrophages, neutrophils and lymphocytes and notably increased (p ≤ 0.01) wound contracture, vessels, hydroxyproline, hexosamine, hexuronic acid, fibrocytes and fibrocyte/fibroblast ratio in rats. The results of FT‐IR, UV, XRD, TEM and SEM analyses confirm that aqueous extract of A. eriophyllum leaves can be used to yield TiNPs with notable antioxidant, antibacterial, antifungal and cutaneous wound healing potentials without any cytotoxicity. Further clinical trials are necessary for confirmation of these remedial properties of TiNPs in humans.

Novel synthesis of Falcaria vulgaris leaf extract conjugated copper nanoparticles with potent cytotoxicity, antioxidant, antifungal, antibacterial, and cutaneous wound healing activities under in vitro and in vivo condition

Facile green synthesis of copper nanoparticles from different biological procedures has been indicated, but among all, biosynthesis of copper nanoparticles from medicinal plants is considered as the most suitable method. The use of medicinal plant material increases the therapeutical effects of copper nanoparticles. The aim of this study was green synthesis of copper nanoparticles from aqueous extract of Falcaria vulgaris leaf (CuNPs) and assessment of their cytotoxicity, antioxidant, antifungal, antibacterial, and cutaneous wound healing properties. These nanoparticles were characterized by X-ray diffraction (XRD), fourier-transform infrared spectroscopy (FT-IR), ultraviolet–visible spectroscopy (UV), transmission electron microscopy (TEM), and field emission scanning electron microscopy (FE-SEM) analysis. The synthesized CuNPs had great cell viability dose-dependently (Investigating the effect of the CuNPs on human umbilical vein endothelial cell (HUVEC) line) and indicated this method was nontoxic. Also, 2,2-diphenyl-1-picrylhydrazyl (DPPH) test was done to assess the antioxidant activities, which indicated similar antioxidant potentials for CuNPs and butylated hydroxytoluene. In part of cutaneous wound healing property of CuNPs, after creating the cutaneous wound, the rats were randomly divided into six groups: treatment with 0.2% CuNPs ointment, treatment with 0.2% CuSO4 ointment, treatment with 0.2% F. vulgaris ointment, treatment with 3% tetracycline ointment, treatment with Eucerin basal ointment, and untreated control. These groups were treated for 10 days. Treatment with CuNPs ointment remarkably increased (p ≤ .01) the wound contracture, vessel, hexosamine, hydroxyl proline, hexuronic acid, fibrocyte, and fibrocytes/fibroblast rate and substantially reduced (p ≤ .01) the wound area, total cells, neutrophil, and lymphocyte compared to other groups. In antibacterial and antifungal parts of this research, the concentration of CuNPs with minimum dilution and no turbidity was considered minimum inhibitory concentration (MIC). To determine minimum fungicidal concentration (MFC) and minimum bactericidal concentration (MBC), 60 μL MIC and three preceding chambers were cultured on Sabouraud Dextrose Agar and Muller Hinton Agar, respectively. The minimum concentration with no fungal and bacterial growth were considered MFC and MBC, respectively. CuNPs inhibited the growth of all fungi at 2–4 mg/mL concentrations and removed them at 4–8 mg/mL concentrations (p ≤ .01). In case of antibacterial effects of CuNPs, they inhibited the growth of all bacteria at 2–8 mg/mL concentrations and removed them at 4–16 mg/mL concentrations (p ≤ .01). The results of XRD, FT-IR, UV, TEM, and FE-SEM confirm that the aqueous extract of F. vulgaris leaf can be used to yield copper nanoparticles with notable amount of antioxidant, antifungal, antibacterial, and cutaneous wound healing potentials without any cytotoxicity. Further clinical trials are necessary for confirmation these therapeutical effects of CuNPs in human.

Green synthesis and chemical characterization of silver nanoparticles from aqueous extract of Falcaria vulgaris leaves and assessment of their cytotoxicity and antioxidant, antibacterial, antifungal and cutaneous wound healing properties

Nanotechnology encompasses the understanding of the fundamental physics, biology, chemistry and technology of nanometre‐scale objects. In recent decades, nanotechnology has grown rapidly owing to its widespread application in science and industry. The aim of the work reported was the green synthesis and chemical characterization of silver nanoparticles from aqueous extract of Falcaria vulgaris leaves (AgNPs@Falcaria) and evaluation of their cytotoxicity and antioxidant, antibacterial, antifungal and cutaneous wound healing effects under in vitro and in vivo conditions. These nanoparticles were characterized using Fourier transform infrared and UV–visible spectroscopies, scanning and transmission electron microscopies and atomic force microscopy. 2,2‐Diphenyl‐1‐picrylhydrazyl free radical scavenging experiments were conducted to evaluate the antioxidant potential, which indicated similar antioxidant potentials for AgNPs@Falcaria and butylated hydroxytoluene. The synthesized AgNPs@Falcaria had great cell viability dose‐dependently, indicating their non‐toxicity. Minimum inhibitory, minimum bactericidal and minimum fungicidal concentrations were determined by macro‐broth dilution assay. The data were analysed using SPSS software (Duncan post hoc test). AgNPs@Falcaria revealed higher antibacterial and antifungal activities than many standard antibiotics (p ≤ 0.01). Also, AgNPs@Falcaria prevented the growth of all bacteria at 2–8 mg ml−1 concentrations and removed them at 4–16 mg ml−1 concentrations (p ≤ 0.01). AgNPs@Falcaria inhibited the growth of all fungi at 2–4 mg ml−1 concentrations and destroyed them at 4–8 mg ml−1 concentrations (p ≤ 0.01). For in vivo experiments, after creating a cutaneous wound, rats were randomly divided into six groups: untreated control, treatment with eucerin basal ointment, treatment with 3% tetracycline ointment, treatment with 0.2% AgNO3 ointment, treatment with 0.2% F. vulgaris ointment and treatment with 0.2% AgNPs@Falcaria ointment. Use of AgNPs@Falcaria ointment in the treatment groups substantially decreased (p ≤ 0.01) the wound area, total cells, neutrophils and lymphocytes and markedly raised (p ≤ 0.01) wound contracture, hydroxyproline, hexosamine, hexuronic acid, fibrocytes and fibrocyte/fibroblast ratio compared to the other groups. In summary, the synthesized AgNPs@Falcaria revealed non‐cytotoxicity and antioxidant, antibacterial, antifungal and cutaneous wound healing activities in a dose‐dependent manner.

Green synthesis of silver nanoparticles using aqueous extract of Stachys lavandulifolia flower, and their cytotoxicity, antioxidant, antibacterial and cutaneous wound‐healing properties

In a biological process where the herbal tea (Stachys lavandulifolia) aqueous extract was applied as a capping and reducing agent, nanoparticles (NPs) of silver (Ag) were synthesized. These AgNPs were characterized using Fourier transform‐infrared spectroscopy, field emission‐scanning electron microscopy, energy‐dispersive X‐ray spectroscopy, transmission electron microscopy and ultraviolet–visible spectroscopy. The synthesized AgNPs had great cell viability dose‐dependently [investigating the effect of the plant on human umbilical vein endothelial cell line] and indicated this method was non‐toxic. In this study, the 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) free radical scavenging test was carried out to examine antioxidant properties, which revealed similar antioxidant properties for AgNPs and butylated hydroxytoluene. Agar diffusion tests were applied to determine the antibacterial characteristics. The macro‐broth tube test was run to determine minimum inhibitory concentration. All data of antibacterial and cutaneous wound‐healing examinations were analyzed by SPSS 21 software (Duncan post hoc test). AgNPs showed higher antibacterial property than all standard antibiotics (p ≤ 0.01). Also, AgNPs prevented the growth of all bacteria at 2–8 mg/ml concentrations and destroyed them at 2–16 mg/ml concentrations (p ≤ 0.01). For the in vivo experiment, after creating the cutaneous wound, the rats were randomly divided into six groups: untreated control; treatment with Eucerin basal ointment; treatment with 3% tetracycline ointment; treatment with 0.2% AgNO3 ointment; treatment with 0.2% S. lavandulifolia ointment; and treatment with 0.2% AgNPs ointment. These groups were treated for 10 days. For histopathological and biochemical analysis of the healing trend, a 3 × 3‐cm section was prepared from all dermal thicknesses at day 10. Use of AgNPs ointment in the treatment groups substantially reduced (p ≤ 0.01) the wound area, total cells, neutrophil, macrophage and lymphocyte, and remarkably raised (p ≤ 0.01) the wound contracture, hydroxyl proline, hexosamine, hexuronic acid, fibrocyte and fibrocytes/fibroblast rate compared with other groups. Seemingly, AgNPs can be used as a medical supplement owing to their non‐cytotoxic, antioxidant, antibacterial and cutaneous wound‐healing properties.

Biochemical, histopathological, and pharmacological evaluations of cutaneous wound healing properties of Quercus brantii ethanolic extract ointment in male rats

The recent experiments have revealed the property of ethnomedicinal plants on the treatment of the cutaneous wound. The purpose of the experiment was to investigate the wound healing and antioxidant potentials of the internal layer of Quercus brantii ethanolic extract ointment. DPPH free radical scavenging test was carried out to examine the antioxidant effect of the internal layer of Q. brantii ethanolic extract, which indicated similar antioxidant activity with butylated hydroxy toluene (BHT) as a positive control. In vivo design, 120 Sprague Dawley male rats were used. After creating the cutaneous wound, the animals were randomly divided into four groups: untreated control, treatment with Eucerin ointment, treatment with 3% tetracycline ointment, and treatment with 3% Q. brantii ethanolic extract ointment (3 g of Q. brantii ethanolic extract + 97 g base ointment). At days 10, 20, and 30 after creating the wound, for histopathological and biochemical analysis of the cutaneous wound healing trend, a section was prepared from all dermal thicknesses. At days 10, 20, and 30, Q. brantii ethanolic extract ointment significantly (p > 0.05) enhanced wound contraction, fibrocyte count, hexuronic acid, and hydroxyproline levels and decreased the wound area, all cell, macrophage, lymphocyte, and neutrophil counts as compared with the basal ointment and control groups. In conclusion, the acquired findings showed the cutaneous wound healing potential of Q. brantii ethanolic extract ointment.

Antioxidant, cytotoxic, and wound healing effects of Allium saralicum R.M. Fritsch aqueous extract ointment in male rats: a gross, histological, and biochemical experiment

In recent years, cutaneous wound healing by medicinal plants has been on the raise. In recent research, we appraised antioxidant, cytotoxicity, and cutaneous wound healing activities of Allium saralicum R.M. Fritsch aqueous extract ointment in rats. In this study, 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay was carried out to test the antioxidant property, which revealed high antioxidant potential similar to butylated hydroxytoluene (BHT) as a positive control. Synthesizing the aqueous extract dose-dependently demonstrated great cell viability and that this method was non-toxic for synthesizing A. saralicum. In vivo experiment, after creating the cutaneous wound, the animals were randomly divided into four groups: untreated control, treatment with Eucerin basal ointment, treatment with 3% tetracycline ointment, treatment with 10% A. saralicum ointment. These groups were treated for 30 days. For biochemical and histopathological analysis of the healing trend, a 3 × 3-cm section was prepared from all dermal thicknesses on days 10, 20, and 30. The use of A. saralicum ointment in the treatment groups led to substantial diminish (p < 0.05) in the levels of wound area, total cells, neutrophil, macrophage, and lymphocyte, and remarkable raise (p < 0.05) in the levels of wound contracture, hydroxyl proline, hexosamine, hexuronic acid, fibrocyte, and the rate of fibrocyte to fibroblast as compared with the control and basal ointment groups. Adapting to the findings, A. saralicum ointment can cure the cutaneous wound without any cytotoxicity.

Pharmacological, biochemical, and histopathological evaluations of cutaneous wound healing property of Mentha piperita aqueous extract ointment

In our study, we evaluated cutaneous wound healing potential of Mentha piperita aqueous extract ointment in rats. In vivo design, after creating the cutaneous wound on the back of the rats, the animals were randomly divided into four groups: untreated control, treatment with Eucerin ointment, treatment with 3% tetracycline ointment, and treatment with 3% M. piperita aqueous extract ointment. The groups were treated for 30 days. For biochemical and histopathological analysis of the cutaneous wound healing trend, a 3 × 3 cm section was prepared from all dermal thicknesses on days 10, 20, and 30. The data were analyzed by SPSS-21 software. The use of M. piperita aqueous extract ointment in the treatment groups led to remarkable reduction (p < 0.05) in the levels of wound area, total cells, neutrophil, lymphocyte, and macrophage and remarkable increase (p < 0.05) in the levels of wound contracture, hydroxyl proline, hexosamine, hexuronic acid, fibrocyte, and the rate of fibrocyte to fibroblast compared with the control and basal ointment groups. According to the findings, M. piperita aqueous extract ointment can cure the cutaneous wound.

Experimental study on Wistar rats: the Allium eriophyllum Boiss aqueous extract ointment effectively treat induced cutaneous wound

Cutaneous wound healing by medicinal plants which have fewer side effects than chemical drugs have been on the increase. In our study, we evaluated cutaneous wound healing potential of Allium eriophyllum Boiss aqueous extract ointment in rats. In in vivo design, after creating the cutaneous wound on the back of the rats, the animals were randomly divided into four groups: untreated control, treatment with Eucerin ointment, treatment with 3% tetracycline ointment, treatment with 3% A. eriophyllum aqueous extract ointment. The groups were treated for 30 days. For biochemical and histopathological analysis of the cutaneous wound healing trend, a 3 × 3 cm section was prepared from all dermal thicknesses on days 10, 20, and 30. The data were analyzed by SPSS 21 software. The use of A. eriophyllum aqueous extract ointment in the treatment groups led to remarkable reduce (p < 0.05) in the levels of wound area, total cells, neutrophil, lymphocyte, and macrophage, and remarkable increase (p < 0.05) in the levels of wound contracture, hydroxyl proline, hexosamine, hexuronic acid, fibrocyte, and the rate of fibrocyte to fibroblast as compared to the control and basal ointment groups. According to the findings, A. eriophyllum aqueous extract ointment can cure the cutaneous wound.

Platelet glycoprotein VI and C-type lectin-like receptor 2 deficiency accelerates wound healing by impairing vascular integrity in mice.

Platelets promote wound healing by forming a vascular plug and by secreting growth factors and cytokines. Glycoprotein (GP)VI and C-type lectin-like receptor (CLEC)-2 signal through a (hem)-immunoreceptor tyrosine-based activation motif, which induces platelet activation. GPVI and CLEC-2 support vascular integrity during inflammation in the skin through regulation of leukocyte migration and function, and by sealing sites of vascular damage. In this study, we investigated the role of impaired vascular integrity due to GPVI and/or CLEC-2 deficiency in wound repair using a full-thickness excisional skin wound model in mice. Transgenic mice deficient in both GPVI and CLEC-2 exhibited accelerated skin wound healing, despite a marked impairment in vascular integrity. The local and temporal bleeding in the skin led to greater plasma protein entry, including fibrinogen and clotting factors, was associated with increased fibrin generation, reduction in wound neutrophils and M1 macrophages, decreased level of tumor necrosis factor (TNF)-α, and enhanced angiogenesis at day 3 after injury. Accelerated wound healing was not due to developmental defects in CLEC-2 and GPVI double-deficient mice as similar results were observed in GPVI-deficient mice treated with a podoplanin-blocking antibody. The rate of wound healing was not altered in mice deficient in either GPVI or CLEC-2. Our results show that, contrary to defects in coagulation, bleeding following a loss of vascular integrity caused by platelet CLEC-2 and GPVI deficiency facilitates wound repair by increasing fibrin(ogen) deposition, reducing inflammation, and promoting angiogenesis.

Assessment of antioxidant and cutaneous wound healing effects of Falcaria vulgaris aqueous extract in Wistar male rats

Treatment of wounds by ethnomedicinal plants which have fewer side effects than chemical drugs has been on the rise. In this experiment, we evaluated cutaneous wound healing potential of aqueous extract of Falcaria vulgaris in Wistar male rats. DPPH free radical scavenging test was used to examine the antioxidant effect of F. vulgaris aqueous extract, which indicated high antioxidant activity compared to butylated hydroxy toluene (BHT) as the positive control. In our study, after creating the cutaneous wound on the back of the rats, the animals were randomly divided into four groups; untreated control, treatment with Eucerin ointment, treatment with 3% tetracycline ointment, treatment with 3% F. vulgaris aqueous extract ointment. The groups were treated for 30 days. For histopathological and biochemical analysis of the cutaneous wound healing trend, a 3 × 3 cm section was prepared from all dermal thicknesses at days 10, 20, and 30. The use of F. vulgaris aqueous extract ointment in the treatment groups led to significant decrease (p < 0.05) in the levels of wound area, total cells, lymphocyte, neutrophil, macrophage, and significant enhance (p < 0.05) in the levels of wound contracture, hydroxyproline, hexosamine, fibrocyte, fibroblast, and the rate of fibrocyte to fibroblast as compared to the control and basal ointment groups. According to the results, F. vulgaris aqueous extract ointment can treat the cutaneous wound.

Live imaging of wound angiogenesis reveals macrophage orchestrated vessel sprouting and regression

Wound angiogenesis is an integral part of tissue repair and is impaired in many pathologies of healing. Here, we investigate the cellular interactions between innate immune cells and endothelial cells at wounds that drive neoangiogenic sprouting in real time and in vivo. Our studies in mouse and zebrafish wounds indicate that macrophages are drawn to wound blood vessels soon after injury and are intimately associated throughout the repair process and that macrophage ablation results in impaired neoangiogenesis. Macrophages also positively influence wound angiogenesis by driving resolution of anti‐angiogenic wound neutrophils. Experimental manipulation of the wound environment to specifically alter macrophage activation state dramatically influences subsequent blood vessel sprouting, with premature dampening of tumour necrosis factor‐α expression leading to impaired neoangiogenesis. Complementary human tissue culture studies indicate that inflammatory macrophages associate with endothelial cells and are sufficient to drive vessel sprouting via vascular endothelial growth factor signalling. Subsequently, macrophages also play a role in blood vessel regression during the resolution phase of wound repair, and their absence, or shifted activation state, impairs appropriate vessel clearance.

Expression of pro-inflammatory genes in lesions, spleens and blood neutrophils after burn injuries in mice treated with silver sulfodiazine.

Objective(s):It is now supposed that cytokines released during the burn injuries have a great impact on the immunological and pathological responses after the burn. The main objective of this study was to investigate the expression of some pro-inflammatory genes in the wound, spleen and blood neutrophils during the healing process of burn wounds in a murine model.Materials and Methods:The expression of ten pro-inflammatory genes were examined in wounds, spleens and blood neutrophils of mice with burn injuries treated with either silver sulfodiazine or phosphate-buffered saline (PBS) using RT-PCR at the end of the first and second weeks after injuries.Results:None of the pro-inflammatory genes were expressed in the skin, spleen and blood neutrophils of healthy mice. In the group control, IL-12P35, IL-12P40, CCR5, IL-1β and IFN-γ were expressed in the spleen and blood neutrophils in the first week. Instead, CCL5, CCR5, IL-1β and IFN-γ were expressed in the wound, but in the second week, the expression of the genes became similar. In the test group, in the first week, TNF-α, IL-12P35, IL-12P40 and IL-1β were expressed in the lesions, CCL4, IL-1α, IL-12P35, IL-12P40, CCR5 and IFN-γ were expressed in the spleen and no pro-inflammatory gene expression was detected in blood neutrophils.Conclusion:IL-1β and IFN-γ are expressed in wound, spleen and neutrophils of untreated mice, but not in silver sulfodiazine treated mice. Hence, treatment with silver sulfodiazine suppressed the expression of pro-inflammatory genes in some stages of healing.

Metabolic profiles of exudates from chronic leg ulcerations

Chronic leg ulceration is a disease usually associated with other comorbidities, and significantly reduces patient quality of life. Infected leg ulcers can lead to limb-threatening sequelae or mortality. Leg ulcerations are colonized by a number of microbes that are able to cause life-threating infections in susceptible patients. Wound exudate is a body fluid that collects metabolites from patient eukaryotic cells and from prokaryotic bacterial communities inhabiting the wound. This study aimed at identification of metabolites in exudates collected from chronic leg ulcers, and correlation of this metabolome with patient comorbidities and microbiological status of the wound. By means of NMR spectroscopy we detected 42 metabolites of microbial or patient origin. The metabolites that were in abundance in exudates analyzed were lactate, lysine, and leucine. Metabolites were associated with the presence of neutrophils in wounds and destruction of high quantities of microbes, but also with hypoxia typical for venous insufficiency. The combination of nuclear magnetic resonance spectroscopy technique and partial least squares discriminant analysis allowed us to further discriminate groups of metabolites with regards to potential clinical meaning. For example, to discriminate between S.aureus versus all other isolated microbial species, or between patients suffering from type I or II diabetes versus patients without diabetes. Therefore, wound exudate seems to be highly applicable material for discriminant analysis performed with the use of NMR technique to provide for rapid metabolomics of chronic wound status.