Novel lactoferrin-conjugated gallium complex to treat Pseudomonas aeruginosa wound infection.

Pseudomonas aeruginosa is one of the leading causes of opportunistic infections such as chronic wound infection that could lead to multiple organ failure and death. Gallium (Ga3+) ions are known to inhibit P. aeruginosa growth and biofilm formation but require carrier for localized controlled delivery. Lactoferrin (LTf), a two-lobed protein, can deliver Ga3+ at sites of infection. This study aimed to develop a Ga-LTf complex for the treatment of wound infection. The characterisation of the Ga-LTf complex was conducted using differential scanning calorimetry (DSC), Infra-Red (FTIR) and Inductive Coupled Plasma Optical Emission Spectrometry (ICP-OES). The antibacterial activity was assessed by agar disc diffusion, liquid broth and biofilm inhibition assays using the colony forming units (CFUs). The healing capacity and biocompatibility were evaluated using a P.aeruginosa infected wound in a rat model. DSC analyses showed thermal transition consistent with apo-lactoferrin; FTIR confirmed the complexation of gallium to lactoferrin. ICP-OES confirmed the controlled local delivery of Ga3+. Ga-LTf showed a 0.57 log10 CFUs reduction at 24 h compared with untreated control in planktonic liquid broth assay. Ga-LTf showed the highest antibiofilm activity with a 2.24 log10 CFUs reduction at 24 h. Furthermore, Ga-LTf complex is biocompatible without any adverse effect on brain, kidney, liver and spleen of rats tested in this study. Ga-LTf can be potentially promising novel therapeutic agent to treat pathogenic bacterial infections.

In vitro-in vivo wound healing efficacy of Tridax Procumbens extract loaded Carboxymethylcellulose film.

In this work, Tridax Procumbens Extracts (TPE) were blended with Carboxymethylcellulose (CMC), and film was developed through the casting method. The phytochemical screening of the TPE/CMC film was carried out and found the presence of carbohydrates, tannins, saponins, and cardiac glycosides. The presence of elements such as C, O, Na, P, Cl, K, Ca, Mn, and Nb in TP/CMC film was confirmed through EDX. The melting point of the film was found around 291 ± 0.5 °C which was determined through the DSC curve. The maximum tensile strength of the TPE/CMC was found as 14 ± 0.5 MPa. The film showed antibacterial activity against Escherichia coli, Streptococcus pyogenes, Staphylococcus aureus, and Vibrio cholera compared to the control. Cell viability study exhibited 95 % and 98 % cell proliferation for the test film after interacting with the L929 cell line for 18 h and 24 h. The optical contact angle of the TPE/CMC film was also determined. The in-vivo, wound healing studies on adult mice showed healing within 10 days only and the histopathological results revealed the maximum number of fibroblasts with a high density of collagen fibers in the test group indicating that the prepared film can be an effective wound dressing material.

κ-Carrageenan-essential oil loaded composite biomaterial film facilitates mechanosensing and tissue regenerative wound healing.

Polysaccharides κ-carrageenan (κ-Car) have become a predominant source in developing bioactive materials. We aimed to develop biopolymer composite materials of κ-Car with coriander essential oil (CEO) (κ-Car-CEO) films for fibroblast-associated wound healing. Initially, we loaded the CEO in to κ-Car and CEO through homogenization and ultrasonication to fabricate composite film bioactive materials. After performing morphological and chemical characterizations, we validated the developed material functionalities in both in vitro and in vivo models. The chemical and morphological analysis with physical structure, swelling ratio, encapsulation efficiency, CEO release, and water barrier properties of films examined and showed the structural interaction of κ-Car and CEO-loaded into the polymer network. Furthermore, the bioactive applications of CEO release showed initial burst release followed by controlled release from the κ-Car composite film with fibroblast (L929) cell adhesive capabilities and mechanosensing. Our results proved that the CEO-loaded into the κ-Car film impacts cell adhesion, F-actin organization, and collagen synthesis, followed by in vitro mechanosensing activation, further promoting wound healing in vivo. Our innovative perspectives of active polysaccharide (κ-Car)-based CEO functional film materials could potentially accomplish regenerative medicine.

LC-MS/MS metabolomics profiling of Glechoma hederacea L. methanolic extract; in vitro antimicrobial and in vivo with in silico wound healing studies on Staphylococcus aureus infected rat skin wound.

LC-MS/MS analysis of Glechoma hederacea L. methanolic extract (GHME), revealed the identification of 25 metabolites. Ursolic acid (1), 2α-hydroxyursolic acid or corosolic acid (2), 2ß-hydroxyursolic acid or epi-corosolic (3), luteolin 7-O-ß-D-glucopyranoside (4) and rosmarinic acid (5) were isolated and identified using spectroscopy. Antibacterial activity of GHME against multi drug resistance Staphylococcus aureus clinical isolates was measured. Minimum inhibitory concentrations (MICs) were ranged from 62.5 to 500 µg/ml. In vivo wound healing potential of 2%, and 5% GHME prepared hydrogels were criticized on Staphylococcus aureus infected wound rat model. 5% GHME prepared hydrogel treated group showed significant (p < 0.05) shrinkage of their colony forming unit/ml (CFU/ml) values in comparison with standard Fucidin. Meanwhile, wound closure associated with full re-epithelization and hair follicles proliferation was noticed after ten days of treatment. Finally, among the GHME isolated compounds, luteolin 7-O-ß-D-glucopyranoside (4) exhibited the highest molecular docking score (-9.6 kcal/mol) against matrix metalloproteinase-8 target (MMP-8).

Polysucrose hydrogel and nanofiber scaffolds for skin tissue regeneration: Architecture and cell response.

Scaffolds capable of mediating overlapping multi-cellular activities to support the different phases of wound healing while preventing scarring are essential for tissue regeneration. The potential of polysucrose as hydrogels and electrospun mats for wound healing was evaluated in vitro by seeding fibroblasts, endothelial cells and macrophages either singly or in combination. It was found that the scaffold architecture impacted cell behaviour. Electrospun mats promoted fibroblasts flattened morphology while polysucrose methacrylate (PSucMA) hydrogels promoted fibroblast spheroids formation, accentuated in the presence of endothelial cells. Hydrogels exhibited lower inflammatory response than mats and curcumin loaded scaffolds reduced TNF-α production. In vivo biocompatibility of the hydrogels tested on Wistar rats was superior to electrospun mats. In vivo wound healing studies indicated that PSucMA hydrogels integrated the surrounding tissue with better cellular infiltration and proliferation throughout the entire wound region. PSucMA hydrogels led to scarless wound closure comparable with commercially available gels.

Multifunctional hydrogels based on oxidized pectin and gelatin for wound healing improvement.

Gelatin (G) cross-linked with oxidized pectin (OP) was studied as a potential scaffold material for tissue engineering. The effect of oxidation on the chemical properties of pectin was investigated by determining the carbonyl and carboxyl amounts. The OP treatment led to a significant decrease of all values (Mn, Mw, [η] and Rh) determined by size exclusion chromatography (SEC) coupled on line with multiangle light scattering and viscometer detectors. Cross-linking parameters were elucidated by FTIR and TNBS assay. In general, the degree of crosslinking increased with the oxidation of pectin. It was found that the presence of the crosslinking agents caused a reduction in swelling and in the gelatin release which was determined by the BCA kit assay. From the hemolysis test, the membrane of red blood cells was not disrupted by the contact of films and the rate of release of hemoglobin was lower than 5%. The coagulation properties were evaluated by the dynamic blood clotting test. The G/OP hydrogels manifested a good activity of wound healing in the albino rats' model. Moreover, the films did not produce any unwilling symptoms. So, it was concluded that studied films have the potentiality to be used as wound healing biomaterials.

Interpenetrating Polymer Network Hydrogels Formed Using Antibiotics as a Dynamic Crosslinker for Treatment of Infected Wounds.

Antibacterial hydrogels, particularly antibiotic-loaded hydrogels, are promising wound dressing materials for treatment of bacteria-infected wound. However, it is challenging to achieve sustained release of antibiotics from hydrogels through physical encapsulation of the antibiotics. Herein, an interpenetrating polymer network P(AA-co-HEMA)Gen hydrogel is reported with double crosslinking formed by free radical polymerization of 2-hydroxyethyl methacrylate (HEMA) and acrylic acid (AA), while using the antibiotic gentamicin (Gen) as the dynamic physical crosslinker. Gentamicin is incorporated into the hydrogel networks via electrostatic interaction between the carboxyl groups of poly(acrylic acid) and the amino groups of gentamicin, which leads to pH-responsive drug release and a significant increase in mechanical strength (i.e., elastic modulus, viscous modulus, and compressive modulus). More importantly, the hydrogels with optimal compositions demonstrate long-lasting antibacterial activity against both Gram-positive bacteria (Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli) over 28 d. The in vivo studies that are conducted in an S. aureus-infected full-thickness skin wound model demonstrate that the double crosslinking hydrogels loaded with gentamicin eliminate bacteria in the wounds more effectively and significantly accelerate wound healing as compared to 3M dressing and the control without any treatment. Taken together, this antibiotic-loaded interpenetrating polymer network hydrogel is potentially a promising wound dressing material for the treatment of bacteria-infected wound.

In vivo wound healing effect of Italian and Algerian Pistacia vera L. resins.

Pistacia vera oleoresin is one of the natural products used traditionally for the management of wounds. However, there were no scientific reports documented so far on the wound healing activities to substantiate the claim. This study assesses the potential of the oleoresin of P. vera collected in Italy and Algeria for wound healing efficacy via in vivo circular wound excision model. Italian and Algerian oleoresins were subjected to purification and successive fractionation to obtain three matrices. The fractions have been characterized using GC-FID and GC-MS analyses. Oleoresins mixed with vaseline (5% w/w) were topically applied on wound excision induced on the dorsum of rabbits. Wound healing effects were evaluated by percent of wound contraction. Biopsies performed after healing were histologically assessed. Phytochemical results showed a high content of terpenoids components inducing an efficient wound healing effect determined by an in vivo study. Italian and Algerian oleoresins ointments showed significant wound contraction from day 8 to day 16 as compared to the negative control. The two ointments have not showed statistically difference as compared to Cicatryl, reference drug. These results have also been confirmed by the histological evaluation of the tissues involved. The absence of signs of toxicity on the skin of rabbits indicated the safety of the ointments. The study showed that both oleoresins have a very high effectiveness as wound healing agents and appear to justify their traditional use in wound healing in several countries and offer a scientific support to the treatment of traditional healers.

In Vitro and In Vivo Characterization Methods for Evaluation of Modern Wound Dressings.

Chronic wound management represents a major challenge in the healthcare sector owing to its delayed wound-healing process progression and huge financial burden. In this regard, wound dressings provide an appropriate platform for facilitating wound healing for several decades. However, adherent traditional wound dressings do not provide effective wound healing for highly exudating chronic wounds and need the development of newer and innovative wound dressings to facilitate accelerated wound healing. In addition, these dressings need frequent changing, resulting in more pain and discomfort. In order to overcome these issues, a wide range of affordable and innovative modern wound dressings have been developed and explored recently to accelerate and improve the wound healing process. However, a comprehensive understanding of various in vitro and in vivo characterization methods being utilized for the evaluation of different modern wound dressings is lacking. In this context, an overview of modern dressings and their complete in vitro and in vivo characterization methods for wound healing assessment is provided in this review. Herein, various emerging modern wound dressings with advantages and challenges have also been reviewed. Furthermore, different in vitro wound healing assays and in vivo wound models being utilized for the evaluation of wound healing progression and wound healing rate using wound dressings are discussed in detail. Finally, a summary of modern wound dressings with challenges and the future outlook is highlighted.

Enhanced antimicrobial and full-thickness wound healing efficiency of hydrogels loaded with heparinized ZnO nanoparticles: In vitro and in vivo evaluation.

Nanotechnology-based fabricated wound dressings are known as appropriate substrates to enhance healing in both acute and chronic wounds. These types of materials have the ability to deliver therapeutic agents. In this study, a wound dressing including heparinized zinc oxide nanoparticles in combination with chitosan and poly(vinyl alcohol) was developed to investigate its antibacterial and regenerative properties in a rat model of full thickness skin wounds. By adding nanoparticles, the mechanical strength increased up to twice as compared to the sample without nanoparticles. In addition, heparin release profile follows the Hixson-Crowell release kinetic. Protein adsorption enhanced by adding nanoparticles in hydrogels and the prepared wound dressings were completely biocompatible. In terms of antibacterial activity, the minimum inhibitory concentration decreased by conjugation of heparin on the surface of zinc oxide nanoparticles compared to the non-functionalized nanoparticles, and, this shows the increased antibacterial synergistic effect by adding heparin to nanoparticles. Furthermore, it was found that the heparinized zinc oxide nanoparticles effectively accelerate wound closure, re-epithelialization and decrease collagen deposition compared to other groups after implantation. Hence, the prepared wound dressings have the capacity to significantly enhance healing of acute wounds.

Formulation of Chitosan Stabilized Silver Nanoparticle-Containing Wound Healing Film: In Vitro and In Vivo Characterization.

A wound indicates a discontinuity in the epithelial integrity of the skin along with structural and functional disruption of the underlying normal tissue. The study focused on chitosan stabilized silver nanoparticles (CH-AgNP) further incorporated in a chitosan-based (CH-AgNP-CHF) film for wound healing. Dual advantages of chitosan as a wound-healing agent in addition to the antimicrobial property of CH-AgNP nanoparticles was explored. Based on preliminary trials, 1-2% w/v chitosan as film former, 15-25% w/v glycerin as plasticizer and Teflon as casting surface was selected. The optimized CH-AgNP-CHF had tensile strength 1.39 ± 0.009 N/mm2, % Elongation 33.33 ± 1.634, 76.66 ± 0.584% degree of swelling, WVTR of 2024.43 ± 32.78 gm.m-2 day-1 and 1144.57 ± 13.45 gm.m-2 day-1 after 24 h and after 21 days respectively. The CH-AgNP-CHF reported highest % inhibition of 62.22 ± 0.91 against Escherichia coli as compared to chitosan solution, chitosan film and CH-AgNP solution. Based on in vivo animal study, CH-AgNP-CHF showed highest wound closure rate at 3rd, 5th, 7th, 14th and 21st day to indicate better and faster wound healing compared to marketed SilverKind® Nanofine gel, blank chitosan film and sterile gauze treated group (Control). Thus, CH-AgNP-CHF is more effective alternative for wound healing.

Formulation and comparative characterization of nanoparticles of curcumin using natural, synthetic and semi-synthetic polymers for wound healing.

AIM: The present research work aimed to prepare and characterize nanoparticles of curcumin using polymers from different sources like natural, synthetic and semi-synthetic, and compare their activity for wound healing. Curcumin, BCS class II drug, is a polyphenol with proven wound healing activity. MATERIAL AND METHODS: The curcumin-loaded chitosan and carboxymethylcellulose (CMC) nanoparticles were prepared by ionotropic gelation method. In contrast, poly-lactic co-glycolic acid (PLGA) nanoparticles were prepared by a double emulsion solvent evapouration method. The different sources of polymers include natural (chitosan), synthetic (PLGA) and semi-synthetic CMC were used for the preparation of nanoparticles. KEY FINDINGS: The percentage entrapment efficiency of curcumin-loaded nanoparticles was found to be in the order of polymers PLGA>chitosan>CMC. The in-vitro release study of carboxymethyl cellulose -curcumin nanoparticles was found to be 74.96% for 24 h. The presence of a specific peak of curcumin in all the polymeric nanoparticles specifies drug incorporation in the polymeric matrix. The in-vivo study revealed that curcumin-loaded chitosan nanoparticles fasten the healing process of the wound due to the synergistic effect produced by using a combination of curcumin and chitosan. SIGNIFICANCE: Curcumin-loaded nanoparticles showed significant enhancement in wound healing action by lowering the dose of curcumin and effecting synergistically due to the use of chitosan.

In vivo efficacy of biocompatible silver nanoparticles cream for empirical wound healing.

Wound healing is a complicated process that begins at the onset of injury and a continued process till complete healing. The emergence of nanotechnology has provided a new therapeutic modality to silver nanoparticles in treatment of wounds. However, the safety of these silver nanoparticles in the process of wound healing is yet to be elucidated; nevertheless, biocompatibility is the primary concern. Biosynthesis of silver nanoparticles was synthesized using aqueous clove extract and silver nitrate solution under microwave and the obtained particles size were 30-60 nm and roughly spherical in shape. The present study focused on the efficacy of biocompatible silver nanoparticles in vivo wound healing process. Consequently, this study supported the incorporation of biosynthesized silver nanoparticles in wound dressings as a cream formulation for improved healthcare.

Antioxidant, antibacterial and in vivo wound healing properties of laminaran purified from Cystoseira barbata seaweed.

Laminaran, a polysaccharide extracted from marine algae, exhibits attractive properties being non-toxic, hydrophilic and biodegradable. The aim of this study was to investigate the effectiveness of a cream based on the brown seaweed Cystoseira barbata laminaran (CBL) for healing full thickness wounds induced on rats. The antibacterial activity of CBL was evaluated against Gram positive and Gram negative bacteria and the antioxidant properties were assessed using five different assays. To highlight the healing effectiveness of CBL-based cream, the response to treatment was assessed by macroscopic, histologic and biochemical parameters and was compared to controls. CBL, recovered (7.27%) by ultrafiltration (1-10 kDa) and predominantly consisted of (1 → 3)-linked ß-d-glucopyranose residues with a small level of (1 → 6)-glycosidic bonds, showed noticeable antioxidant and antibacterial properties. The cream containing CBL as an active ingredient exerted a promoting healing effect. The wound contraction reached 98.57 ±â€¯1.31% after thirteen days of treatment. The derma in CBL treated group is properly arranged and revealed an improved collagen deposition and an increased fibroblast and vascular densities compared with the control groups. Overall, these results established, for the first time, a scientific in vivo evidence of the efficiency of CBL as a wound healing agent of interest in modern medicine.

Coumestrol/hydroxypropyl-ß-cyclodextrin association incorporated in hydroxypropyl methylcellulose hydrogel exhibits wound healing effect: in vitro and in vivo study.

Several beneficial effects on the skin have been reported for coumestrol (COU), such as protection against photoaging and improvement of skin elasticity and thickness in postmenopausal women. However no reports on the effect of COU on wound healing were found. Nevertheless, COU has low aqueous solubility, which is a crucial limitation for biological tests. The present study was designed as a two-step experiment to evaluate the wound healing effect of COU. First, we used fibroblasts and the experimental in vitro artificial wound model, scratch assay, to compare the effects of COU free, dissolved in dimethyl sulfoxide (DMSO) or Dulbecco's modified Eagle's medium (DMEM), or associated with hydroxypropyl-ß-cyclodextrin (HPßCD). The 50 µM (66.1%) and 10 µM (56.3%) COU/HPßCD association induced cell proliferation and migration in inflicted wounds. Subsequently, the in vivo wound healing experimental model (Wistar rats) revealed that COU/HPßCD incorporated into hypromellose (HPMC) hydrogel had similar efficacy in wound healing in comparison to the positive control (Dersani®), with the advantage that 50% wound healing was achieved within a shorter period. In summary, the results successfully demonstrated, for the first time, the wound healing effect of COU/HPßCD incorporated into HPMC hydrogel and describe the feasibility of the biological tests with the use of HPßCD instead DMSO.

Novel asymmetric chitosan/PVP/nanocellulose wound dressing: In vitro and in vivo evaluation.

The present study was to develop a novel chitosan based symmetric and asymmetric bionanocomposite for potential wound dressing application. Chitosan (C)/Poly (vinyl pyrrolidone) (P)/nanocellulose (NC) membrane were fabricated by salt leaching method with the addition of 3% and 5% wt of nanocellulose. To obtain asymmetric material one side of the membrane was coated by stearic acid (S) which could form hydrophobic surface and another side acts as a hydrophilic surface. Nanocellulose of size 2-10nm was synthesized and characterized by TEM analysis. SEM showed the hydrophilic surface of asymmetric bionanocomposite consists of porous structure and hydrophobic surface is smooth and homogeneous. The results revealed that the Chitosan/PVP/Nanocellulose 3%-Stearic acid (CPNC3%-S) had a moderate swelling ratio, porosity, barrier and mechanical properties. Incorporation of nanocellulose into chitosan/PVP matrix could enhance the antibacterial activity. The hydrophobic surface of the CPNC3%-S bionanocomposite shows water repellent and antiadhesion properties towards E. coli bacteria and also the hydrophilic surface exhibit excellent antibacterial property and cytotoxicity towards bacterial pathogens. In vivo wound healing test shows better re-epithelialization and wound contraction compared with control and Chitosan/PVP-stearic acid (CP-S) bionanocomposite. Asymmetric bionanocomposite Chitosan/PVP/Nanocellulose coated with 3%-Stearic acid (CPNC3%-S) exhibited very good invitro cytocompatibility and enabled a faster wound healing than symmetric dressing, hence showing great potential to be applied as wound dressings.

Chitosan-hyaluronic acid composite sponge scaffold enriched with Andrographolide-loaded lipid nanoparticles for enhanced wound healing.

In this work chitosan-hyaluronic acid composite sponge scaffold enriched with Andrographolide (AND) lipid nanocarriers was developed. Nanocarriers were prepared using solvent diffusion method by applying 23 factorial design. NLC4 had the highest desirability value (0.882) and therefore, it was chosen as an optimal nanocarrier. Itexhibited spherical shape with 253nm, 83.04% entrapment efficiency and a prolonged release of AND up to 48-h. Consecutively, NLC4 was incorporated in a chitosan-hyaluronic acid gel and lyophilized for 24h to obtain chitosan-hyaluronic acid/NLC4 nanocomposite sponge to enhance AND delivery to wound sites. The morphology of sponge was characterized by scanning electron microscopy. Nanocomposites showed a porosity of 56.22% with enhanced swelling. The in vivo evaluation in rats reveals that the chitosan-hyaluronic acid/NLC4 sponge enhances the wound healing with no scar and improved tissue quality. These results strongly support the possibility of using this novel chitosan-hyaluronic acid/NLC4 sponge for wound care application.

In situ functionalized nanobiocomposites dressings of bamboo cellulose nanocrystals and silver nanoparticles for accelerated wound healing.

An innovative approach was adopted where in situ synthesized silver nanoparticles (AgNPs) from leaf extract mediated reduction of AgNO3 were simultaneously impregnated into the matrix of cellulose nanocrystals (CNCs) isolated from Dendrocalamus hamiltonii and Bambusa bambos leaves, for formation of nanobiocomposites (NCs) in film and ointment forms. Here, use of plant CNCs was chosen as an alternate to bacterial cellulose for wound dressings. NCs possessing water absorption capacity and strong antibacterial activity showed synergistic effect on in vivo skin wound healing and documented faster and significant wound closure in treated mice. NCs exhibited lesser inflammation and early vasculogenesis at day 3 coupled with increased fibroblasts and collagen content at day 8 leading to faster neo-epithelization by day 14. Highly effective, biocompatible, and easy to apply NCs wound dressings (ointment and films) containing low amounts of Ag (0.05±0.01wt%) are potential candidates for effective skin repair.

Evaluation of in situ injectable hydrogels as controlled release device for ANXA1 derived peptide in wound healing.

In this paper, for the first time, hydrogels containing Annexin A1 N-terminal derived peptide, Ac2-26, as a novel dressing were successfully developed for dermal wound repair application. High mannuronic (M) content alginate and low molecular weight chitosan have been used as hydrogel carrier. Peptide recovery analyses, FTIR studies and molecular modelling highlighted chemical interactions between peptide and hydrogel polymers. Ac2-26 resulted entrapped into chitosan hydrogel matrix that prevented its release, whereas such interaction in alginate hydrogel slowed down peptide diffusion enabling its sustained release till 72 h. In vivo wound healing studies conducted on mice dorsal wounds indicate that after the 9th day of post wounding Ac2-26/alginate hydrogels could significantly accelerate wound healing, with complete closure of the wound on day 14th. Therefore, these results suggest that the developed of Ac2-26 high M content alginate hydrogel could be a promising wound dressing with potential application in dermal wound healing.