Chitosan based ethanolicAllium Sativumextract hydrogel film: a novel skin tissue regeneration platform for 2nd degree burn wound healing.

This study investigated the potential of ethanolic garlic extract-loaded chitosan hydrogel film for burn wound healing in an animal model. The ethanolic garlic extract was prepared by macerating fresh ground garlic cloves in ethanol for 24 h, followed by filtration and concentration using a rotary evaporator. Hydrogels were then prepared by casting a chitosan solution with garlic extract added at varying concentrations for optimization and, following drying, subjected to various characterization tests, including moisture adsorption (MA), water vapor transmission rate (WVTR), and water vapor permeability rate (WVPR), erosion, swelling, tensile strength, vibrational, and thermal analysis, and surface morphology. The optimized hydrogel (G2) was then analyzedin vivofor its potential for healing 2nd degree burn wounds in rats, and histological examination of skin samples on day 14 of the healing period. Results showed optimized hydrogel (G2; chitosan: 2 g, garlic extract: 1 g) had MA of 56.8% ± 2.7%, WVTR and WVPR of 0.00074 ± 0.0002, and 0.000 498 946 ± 0.0001, eroded up to 11.3% ± 0.05%, 80.7% ± 0.04% of swelling index, and tensile strength of 16.6 ± 0.9 MPa, which could be attributed to the formation of additional linkages between formulation ingredients and garlic extract constituents at OH/NH and C=O, translating into an increase in transition melting temperature and enthalpy (ΔT= 238.83 °C ± 1.2 °C, ΔH= 4.95 ± 0.8 J g-1) of the chitosan moieties compared with blank. Animal testing revealed G2 formulation significantly reduced the wound size within 14 d of the experiment (37.3 ± 6.8-187.5 ± 21.5 mm2) and had significantly higher reepithelization (86.3 ± 6.8-26.8 ± 21.5 and 38.2% ± 15.3%) compared to untreated and blank groups by hastening uniform and compact deposition of collagen fibers at the wound site, cementing developed formulation a promising platform for skin regeneration.

Collaterally Sensitive ß-Lactam Drugs as an Effective Therapy against the Pre-Existing Methicillin Resistant Staphylococcus aureus (MRSA) Biofilms.

Methicillin-resistant Staphylococcus aureus (MRSA) is among the leading causes of nosocomial infections and forms biofilms, which are difficult to eradicate because of their increasing resistance to antimicrobial agents. This is especially true for pre-existing biofilms. The current study focused on evaluating the efficacy of three ß-lactam drugs, meropenem, piperacillin, and tazobactam, alone and in combination against the MRSA biofilms. When used individually, none of the drugs exhibited significant antibacterial activity against MRSA in a planktonic state. At the same time, the combination of meropenem, piperacillin, and tazobactam showed a 41.7 and 41.3% reduction in planktonic bacterial cell growth, respectively. These drugs were further assessed for biofilm inhibition and removal. The combination of meropenem, piperacillin, and tazobactam caused 44.3% biofilm inhibition, while the rest of the combinations did not show any significant effects. Results also revealed that piperacillin and tazobactam exhibited the best synergy against the pre-formed biofilm of MRSA, with 46% removal. However, adding meropenem to the piperacillin and tazobactam combination showed a slightly reduced activity towards the pre-formed biofilm of MRSA and removed 38.7% of it. Although the mechanism of synergism is not fully understood, our findings suggest that these three ß-lactam drugs can be used in combination as very effective therapeutic agents for the treatment of pre-existing MRSA biofilms. The in vivo experiments on the antibiofilm activity of these drugs will pave the way for applying such synergistic combinations to clinics.

Microwave-Treated Physically Cross-Linked Sodium Alginate and Sodium Carboxymethyl Cellulose Blend Polymer Film for Open Incision Wound Healing in Diabetic Animals-A Novel Perspective for Skin Tissue Regeneration Application.

This study aimed at developing the microwave-treated, physically cross-linked polymer blend film, optimizing the microwave treatment time, and testing for physicochemical attributes and wound healing potential in diabetic animals. Microwave-treated and untreated films were prepared by the solution casting method and characterized for various attributes required by a wound healing platform. The optimized formulation was tested for skin regeneration potential in the diabetes-induced open-incision animal model. The results indicated that the optimized polymer film formulation (MB-3) has significantly enhanced physicochemical properties such as high moisture adsorption (154.6 ± 4.23%), decreased the water vapor transmission rate (WVTR) value of (53.0 ± 2.8 g/m2/h) and water vapor permeability (WVP) value (1.74 ± 0.08 g mm/h/m2), delayed erosion (18.69 ± 4.74%), high water uptake, smooth and homogenous surface morphology, higher tensile strength (56.84 ± 1.19 MPa), and increased glass transition temperature and enthalpy (through polymer hydrophilic functional groups depicting efficient cross-linking). The in vivo data on day 16 of post-wounding indicated that the wound healing occurred faster with significantly increased percent re-epithelialization and enhanced collagen deposition with optimized MB-3 film application compared with the untreated group. The study concluded that the microwave-treated polymer blend films have sufficiently enhanced physical properties, making them an effective candidate for ameliorating the diabetic wound healing process and hastening skin tissue regeneration.

Novel Curcumin-Encapsulated α-Tocopherol Nanoemulsion System and Its Potential Application for Wound Healing in Diabetic Animals.

Objective: This project was aimed at formulating a novel nanoemulsion system and evaluating it for open incision wound healing in diabetic animals. Methods: The nanoemulsions were characterized for droplet size and surface charge, drug content, antioxidant and antimicrobial profiling, and wound healing potential in diabetic animals. The skin samples excised were also analyzed for histology, mechanical strength, and vibrational and thermal analysis. Results: The optimized nanoemulsion (CR-NE-II) exhibited droplet size of26.76 ± 0.9 nm with negative surface charge (-10.86 ± 1.06 mV), was homogenously dispersed with drug content of68.05 ± 1.2%, released almost82.95 ± 2.2%of the drug within first 2 h of experiment with synergistic antioxidant (95 ± 2.1%) and synergistic antimicrobial activity against selected bacterial strains in comparison to blank nanoemulsion, and promoted significantly fast percent reepithelization (96.47%). The histological, vibrational, thermal, and strength analysis of selected skin samples depicted a uniform and even distribution of collagen fibers which translated into significant increase in strength of skin samples in comparison to the control group. Conclusions: The optimized nanoemulsion system significantly downregulated the oxidative stress, enhanced collagen deposition, and precluded bacterial contamination of wound, thus accelerating the skin tissue regeneration process.

Physicochemical Characterization of Chitosan-Decorated Finasteride Solid Lipid Nanoparticles for Skin Drug Delivery.

Finasteride is considered the drug of choice for androgenic alopecia and benign prostate hyperplasia. The aim of the study was to formulate nanodrug carriers of finasteride with enhanced retentive properties in the skin. The finasteride was formulated as solid lipid nanoparticles that were decorated with different concentrations of chitosan for improved retentive properties. Solid lipid nanoparticles (SLNs) were synthesized by "high-speed homogenization technique" using stearic acid as a solid lipid while PEG-6000 and Tween-80 were used as surfactants. The SLNs were evaluated for particle size, polydispersity index (PDI), zeta potential, drug entrapment efficiency, and drug release behavior. The mean particle size of SLNs was in the range of 10.10 nm to 144.2 nm. The PDI ranged from 0.244 to 0.412 while zeta potential was in the range of 8.9 mV to 62.6 mV. The drug entrapment efficiency in chitosan undecorated formulations was 48.3% while an increase in drug entrapment was observed in chitosan-decorated formulations (51.1% to 62%). The in vitro drug release studies of SLNs showed an extended drug release for 24 hours after 4 hours of initial burst release. The extended drug release was observed in chitosan-coated SLNs in comparison with uncoated nanoparticles. The permeation and retention study revealed higher retention of drug in the skin and low permeation with chitosan-decorated SLNs that ranged from 39.4 µg/cm2 to 13.2 µg/cm2. TEM images depicted spherical shape of SLNs. The stability study confirmed stable formulations in temperature range of 5°C and 40°C for three months. It is concluded from this study that the SLNs of finasteride were successfully formulated and chitosan decoration enhanced the drug retention in the skin layers. Therefore, these formulations could be used in androgenic alopecia and benign prostate hyperplasia to avoid the side effects, drug degradation, and prolonged use of drug with conventional oral therapy.

An Investigation for Skin Tissue Regeneration Enhancement/Augmentation by Curcumin-Loaded Self-Emulsifying Drug Delivery System (SEDDS).

Diabetes, one of the global metabolic disorders, is often associated with delayed wound healing due to the elevated level of free radicals at the wound site, which hampers skin regeneration. This study aimed at developing a curcumin-loaded self-emulsifying drug delivery system (SEDDS) for diabetic wound healing and skin tissue regeneration. For this purpose, various curcumin-loaded SEDDS formulations were prepared and optimized. Then, the SEDDS formulations were characterized by the emulsion droplet size, surface charge, drug content/entrapment efficiency, drug release, and stability. In vitro, the formulations were assessed for the cellular uptake, cytotoxicity, cell migration, and inhibition of the intracellular ROS production in the NIH3T3 fibroblasts. In vivo, the formulations' wound healing and skin regeneration potential were evaluated on the induced diabetic rats. The results indicated that, after being dispersed in the aqueous medium, the optimized SEDDS formulation was readily emulsified and formed a homogenous dispersion with a droplet size of 37.29 ± 3.47 nm, surface charge of -20.75 ± 0.07 mV, and PDI value of less than 0.3. The drug content in the optimized formulation was found to be 70.51% ± 2.31%, with an encapsulation efficiency of 87.36% ± 0.61%. The SEDDS showed a delayed drug release pattern compared to the pure drug solution, and the drug release rate followed the Fickian diffusion kinetically. In the cell culture, the formulations showed lower cytotoxicity, higher cellular uptake, and increased ROS production inhibition, and promoted the cell migration in the scratch assay compared to the pure drug. The in vivo data indicated that the curcumin-loaded SEDDS-treated diabetic rats had significantly faster-wound healing and re-epithelialization compared with the untreated and pure drug-treated groups. Our findings in this work suggest that the curcumin-loaded SEDDS might have great potential in facilitating diabetic wound healing and skin tissue regeneration.

Microwave-Assisted Physically Cross-Linked Chitosan-Sodium Alginate Hydrogel Membrane Doped with Curcumin as a Novel Wound Healing Platform.

This project purposes to develop chitosan and sodium alginate-based hydrogel membranes loaded with curcumin through microwave-based physical cross-linking technique and its evaluation for wound healing potential. For the purpose, curcumin-loaded chitosan and sodium alginate membranes were developed using microwave at fixed frequency of 2450 MHz, power 350 W for 60 s, and tested for their physicochemical attributes like swelling, erosion, surface morphology, drug content, and in vitro drug release. The membranes were also subjected to tensile strength and vibrational and thermal analysis followed by testing in vivo on animals. The results indicated that microwave treatment significantly enhanced the swelling ability, reduced the erosion, and ensured smooth surface texture with optimal drug content. The drug was released in a slow fashion releasing total of 41 ± 4.2% within 24-h period with a higher tensile strength of 16.4 ± 5.3 Mpa. The vibrational analysis results revealed significant fluidization of hydrophilic domains and defluidization of hydrophobic domains which translated into a significant rise in the melting temperature and corresponding enthalpy which were found to be 285.2 ± 3.2 °C and 4.89 ± 1.4 J/g. The in vivo testing revealed higher percent re-epithelialization (75 ± 2.3%) within 14 days of the treatment application in comparison to only gauze and other treatments applied, with higher extent of collagen deposition having well-defined epidermis and stratum corneum formation. The microwave-treated chitosan-sodium alginate hydrogel membranes loaded with curcumin may prove to be another alternative to treat skin injuries. Graphical Abstract.

Microwave Enabled Physically Cross Linked Sodium Alginate and Pectin Film and Their Application in Combination with Modified Chitosan-Curcumin Nanoparticles. A Novel Strategy for 2nd Degree Burns Wound Healing in Animals.

This study reports microwave assisted physically cross-linked sodium alginate and pectin film and their testing in combination with modified chitosan-curcumin nanoparticles for skin tissue regeneration following 2nd degree burn wound. Film was formulated by solution casting method and physically cross-linked using microwave irradiation at frequency of 2450 MHz, power 750 Watt for different time intervals for optimization. The optimized formulation was analyzed for various physiochemical attributes. Afterwards, the optimized film and optimized modified chitosan-curcumin nanoparticles were tested in combination for skin regeneration potential following burn wound in vivo and skin samples extracted and tested for different attributes. The results indicated that the optimized film formulation (5 min microwave treatment) physicochemical attributes significantly enhanced addressing the properties required of a wound healing platform. The vibrational analysis indicated that the optimized film experienced significant rigidification of hydrophilic domains while the hydrophobic domains underwent significant fluidization which also resulted in significant increase in the transition temperatures and system enthalpies of both polymer moieties with microwave treatment. The combined film and nanoparticles application significantly increased protein content in the wounds which were evident from higher absorbance ratios of amide-I and amide-II (2.15 ± 0.001), significantly higher melting transition temperature and enthalpy (∆T = 167.2 ± 15.4 °C, ∆H = 510.7 ± 20.1 J/g) and higher tensile strength (14.65 ± 0.8 MPa) with significantly enhanced percent re-epithelization (99.9934 ± 2.56) in comparison to other treatments. The combined application of film and nanoparticles may prove to be a new novel treatment strategy for 2nd degree burn wound healing.

Physicochemical Characterization of Finasteride Nanosystem for Enhanced Topical Delivery.

INTRODUCTION: The current work aimed to formulate a novel chitosan-based finasteride nanosystem (FNS-NS) for skin delivery to optimize the drug availability in skin for a longer time and enhance ex vivo performance of finasteride against androgenic alopecia. METHODS: Both undecorated and chitosan decorated FNS-NSs were synthesized by a high energy emulsification technique. All the prepared nanosystems were further subjected to physicochemical characterizations like pH, viscosity, encapsulation efficiency, surface morphology and in vitro drug release behavior. The influence of the nanosystem on the drug permeation and retention in rat skin was examined using Franz diffusion cell apparatus. RESULTS: The droplet size of developed nanosystems ranged from 41 to 864 nm with a low polydispersity index. The zeta potential of the nanosystems was between -10 mV and +56 mV. This chitosan decorated nanosystem exhibited controlled drug release, ie about 78-97% in 24 h. Among all the nanosystems, our chitosan decorated formulation (F5) had low drug permeation (16.35 µg/cm2) and higher drug retention (10.81 µg/cm2). CONCLUSION: The abovementioned results demonstrate satisfactory in vitro drug release, skin retention profiles and ex vivo performance with chitosan decorated FNS-NS (F5). This optimized formulation could increase drug availability in skin and could become a promising carrier for topical delivery to treat androgenic alopecia.

Formulation and Evaluation of Microwave-Modified Chitosan-Curcumin Nanoparticles-A Promising Nanomaterials Platform for Skin Tissue Regeneration Applications Following Burn Wounds.

Improved physicochemical properties of chitosan-curcumin nanoparticulate carriers using microwave technology for skin burn wound application are reported. The microwave modified low molecular weight chitosan variant was used for nanoparticle formulation by ionic gelation method nanoparticles analyzed for their physicochemical properties. The antimicrobial activity against Staphylococcus aureus and Pseudomonas aeruginosa cultures, cytotoxicity and cell migration using human dermal fibroblasts-an adult cell line-were studied. The microwave modified chitosan variant had significantly reduced molecular weight, increased degree of deacetylation and decreased specific viscosity. The nanoparticles were nano-sized with high positive charge and good dispersibility with entrapment efficiency and drug content in between 99% and 100%, demonstrating almost no drug loss. Drug release was found to be sustained following Fickian the diffusion mechanism for drug release with higher cumulative drug release observed for formulation (F)2. The microwave treatment does not render a destructive effect on the chitosan molecule with the drug embedded in the core of nanoparticles. The optimized formulation precluded selected bacterial strain colonization, exerted no cytotoxic effect, and promoted cell migration within 24 h post application in comparison to blank and/or control application. Microwave modified low molecular weight chitosan-curcumin nanoparticles hold potential in delivery of curcumin into the skin to effectively treat skin manifestations.

Smart nanocrystal of indomethacin: Nanonization and characterization through top down method of media milling.

Indomethacin is potent and effective drug belongs to NSAID group having low bioavailability. To address this issue the novel method is Nanosuspensions which can be achieved through bottom up and top down methods. The drug concentration, batch size and crystallinity retention are the problems associated with bottom up method consequently top down method was applied. In current project batch size of 350 ml was prepared by mixing 3.5% of Indomethacin with polymer solution. Then it was introduced into Dena⌖ having 0.2µm yttrium reinforced zirconium beads. The effect of milling time was observed for sixty minutes. Stable nanocrystals with particle size of 161nm ±1.90 with PDI of 0.229 ±0.06 were produced. The DSC and PXRD confirmed the crystallinity of created nanocrystals. The pattern of particle size reduction was initially abrupt and then gradual. The two months Stability studies at 4°C and at 25°C revealed that polymers combination (PVP-K30, HPMC-6cps, SDS) were effective in marinating the stability. The SEM and TEM studies unfastened that nanocrystals were homogenously distributed with discrete crystalline morphology. The fabricated nanocrystals demonstrated marked dissolution rate compared to the raw and marketed formulations. It is demonstrated that it is useful for industry due to high drug concentration, large batch size and retention of distinct characteristics.

Microwave assisted chitosan-polyethylene glycol hydrogel membrane synthesis of curcumin for open incision wound healing.

Chitosan and polyethylene glycol hydrogel membranes containing curcumin were synthesized using microwave technology at fixed frequency, power and time of 2450 MHz, 500 Watt and 120 s. Polymers were solubilized separately, combined with drug and mixed in two different ratios i.e. F1=80:20 and F2=85:15. The untreated and microwave treated hydrogel membranes were analyzed for degree of swelling, degree of degradation, tensile strength, surface morphology, vibrational and thermal analysis and in vitro drug release. Results indicated that F2(micro) showed a significantly high degree of swelling (96.49±1.21 %), low degradation (9.88±1.68 %), sustained drug release through slow erosion (55.1±3.11 %) via non-Fickian diffusion. The vibrational and thermal analysis revealed rigidification of hydrophilic domains of the polymers by formation of hydrogen bonds between chitosan and PEG moieties (OH/NH) and elasticity of hydrophobic domains (asymmetric and symmetric CH moieties and/or C=O moieties) which not only significantly increased the transition temperature and enthalpy (297.2±3.2 °C and 4.24±1.4 J/g) of the chitosan moiety but also resulted in enhanced tensile strength (18.2±1.3 Mpa). In vivo wound healing study revealed significantly faster wound healing in the F2(micro) treated animal group in comparison to a control animal group where at day 14, a significant re-epithelization (87.26 %) with smaller wound size was observed. Hence microwave assisted chitosan-PEG hydrogel membrane of curcumin is advocated to be a suitable plate form for wound healing applications.

Binary inclusion complexes of diflunisal with ß-cyclodextrin and hydroxypropyl-ß-cyclodextrin: Preparation and characterization.

Low aqueous solubility and bioavailability is the limiting factor to achieve desired therapeutic efficacy for variety of new and existing drug moieties. The goal of the present study was to explore the effects of ß-cyclodextrin (ßCD) and hydroxypropyl-ß-cyclodextrin (HPßCD) on the solubility and dissolution profile of diflunisal (DIF) prepared by using two different methods (physical mixing and solvent evaporation) at DIF-cyclodextrins weight ratios of 1:1, 1:2 and 1:4. The phase solubility studies demonstrated that DIF solubility increased proportionally with an increase in ßCD and HPßCD concentration. The inclusion complexes were subjected to characterization of scanning electron microscopy (SEM), fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC) and X-ray diffractometry (XRD). Solvent evaporation yielded higher DIF solubility than physical mixing method. HPßCD-DIF inclusion complexes yielded higher dissolution profile than ßCD complexes when prepared under same experimental design. FTIR, DSC and XRD confirmed the successful inclusion of DIF into cyclodextrin (ßCD/HPßCD) by both preparation methods with enhanced water solubility and drug release in comparison with pure drug.

Simulation Studies and Dynamic Interaction of Venom Peptides with Ion Channels.

BACKGROUND: Arthropods such as scorpion, snake, insects, and spider as well as the marine animals like sea anemone and cone snails are venomous animals producing venoms with a complex mixture of peptide, poly peptides and small proteins. The disulfide rich peptides isolated from these animals are potent substances which specifically and selectively modulate different ion channels. The significant characteristics of these distinctive pharmacologically potent compounds highlights the molecular details of their peptide-ion channels interactions as well as provides the opportunities for the development of novel and natural therapeutic agents to treat various diseases including neurological disorders also. A good deal is going into the understanding of their therapeutic applications by unrevealing their mode of action. CONCLUSION: In this review, an attempt is made to summarizes the molecular behavior of these venom peptides, their pattern of interactions that how molecular simulation studies are used to investigate the dynamic interaction between these peptides and ion channels, structural prediction of peptide channel complex and calculation of binding free energy.

5-Fluorouracil ethosomes - skin deposition and melanoma permeation synergism with microwave.

This study focuses on the use of ethosome and microwave technologies to facilitate skin penetration and/or deposition of 5-fluorouracil in vitro and in vivo. Low ethanol ethosomes were designed and processed by mechanical dispersion technique and had their size, zeta potential, morphology, drug content and encapsulation efficiency characterized. The skin was pre-treated with microwave at 2450 MHz for 2.5 min with ethosomes applied topically and subjected to in vitro and in vivo skin drug permeation as well as retention evaluation. The drug and/or ethosomes cytotoxicity, uptake and intracellular trafficking by SKMEL-28 melanoma cell culture were evaluated. Pre-treatment of skin by microwave promoted significant drug deposition in skin from ethosomes in vitro while keeping the level of drug permeation unaffected. Similar observations were obtained in vivo with reduced drug permeation into blood. Combination ethosome and microwave technologies enhanced intracellular localization of ethosomes through fluidization of cell membrane lipidic components as well as facilitating endocytosis by means of clathrin, macropinocytosis and in particularly lipid rafts pathways. The synergistic use of microwave and ethosomes opens a new horizon for skin malignant melanoma treatment.

Microwave-aided skin drug penetration and retention of 5-fluorouracil-loaded ethosomes.

OBJECTIVES: Skin drug retention is required in local treatment of skin cancer. This study investigated the interplay effects of ethosomes and microwave in transdermal drug delivery. Skin pre-treatment by microwave and applied with liquified medicine is deemed to 'cement' the skin thereby raising skin drug deposition. METHODS: 5-fluorouracil-loaded ethosomes were prepared and subjected to size, zeta potential, morphology, drug content, drug release and skin permeation tests. The molecular characteristics of untreated, microwave and/or ethosome-treated skins were examined by Fourier transform infrared and raman spectroscopy, thermal and electron microscopy techniques. RESULTS: The skin drug retention was promoted using larger ethosomes with negative zeta potentials that repelled anionic lipids of skin and hindered vesicle permeation into deep layers. These ethosomes had low ethanol content. They were less able to fluidize the lipid and defluidize the protein domains at epidermis to enlarge aqueous pores for drug permeation. Pre-treatment of skin by 2450 MHz microwave for 2.5 min further increased skin drug penetration and retention of low ethanol ethosomes and provided lower drug permeation than cases treated for 1.15 min and 5 min. A 2.5 min treatment might be accompanied by specific dermal protein fluidization via C=O moiety which translated to macromolecular swelling, narrowing of intercellular spaces at lower skin layers, increased drug retention and reduced drug permeation. CONCLUSION: Ethosomes and microwave synergized to promote skin drug retention.

Formulation and in vitro evaluation of clotrimazole gel containing almond oil and tween 80 as penetration enhancer for topical application.

Achieving a desirable percutaneous absorption of drug molecule is a major concern in formulating dermal and transdermal products. The use of penetration enhancers could provide a successful mean for this purpose. The aim of this study was to develop Clotrimazole gel and to evaluate the effect of almond oil and tween 80 (in different concentrations), on the permeation of drug through rabbit skin in vitro. In order to investigate the effect of penetration enhancers used in this study on the permeation of Clotrimazole through sections of excised rabbit skin, Franz diffusion cell was employed. Sample solution was withdrawn at specific time interval up to 24 h. Significant difference in permeation among the eight formulations was seen in the study. The permeation profile of various formulations also showed that the added enhancers in individual batches affected the permeation of the drug. Drug permeation increased with increased concentration of Tween 80 and decreased concentration of almond oil. Furthermore, almond oil combined with tween 80 showed synergistic effect. The clotrimazole gels were successfully formulated and could be beneficial for topical use.