Ascorbate formulation improves healing efficacy in excisional wound mice model through interplay between pro and anti-inflammatory cytokines and angiogenic markers.

BACKGROUND AND OBJECTIVE: Biomedical research in regenerative medicine prompts researchers to formulate cost-effective therapeutics for wound healing. The present study was conducted to characterize the ascorbate based formulation vis-a-vis investigating the molecular dynamics of the formulation. MATERIALS AND METHODS: To characterize the formulation, particle size, zeta potential, thermal stability, compatibility, anti-oxidant, and permeation prospective were measured using standard protocols. The in-vitro healing potential and safety formulae were evaluated using the L929 cell line. For molecular unravelling of the pharmacodynamics of formulation, an excision wound model was used, and 54 mice were randomly and equally divided into three groups, i.e., untreated, betadine-treated, and formulation-treated, to ascertain the interplay between cytokines and chemokines and their culminative role in the release of growth factors. RESULTS: The ascorbate formulae were found to be amorphous, biocompatible, safe, and long-lasting, with particle sizes and zeta potentials of 389.7 ± 0.69 nm and -38.1 ± 0.65 mV, respectively, and anti-oxidative potential. An in-vitro study revealed that the formulation has a significant (p<0.05) migration potential and is non-toxic. Expression profiling of TGF-ß, FGF-2, VEGF, and collagen III & I showed significant (p<0.05) up-regulation, whereas significant (p<0.05) down-regulation of pro-inflammatory genes like IL-1α, IL-1ß, TNF-α, IL-6, and temporal change in CCR-5 was observed in formulae-treated animals as compared to other groups. CONCLUSION: By up-regulating angiogenic and collagen-promoting growth factor gene expression while down-regulating pro-inflammatory gene expression, ascorbate formulation promotes wound healing via extracellular matrix and granulation tissue deposition with significant improvement in tensile strength.

Early cutaneous inflammatory response at different degree of burn and its significance for clinical diagnosis and management.

A complete characterization of the burn wound based on cutaneous architectural changes and inflammatory response is extremely important to provide evidence for progressive changes in the burn wound. Burn wounds are highly susceptible to conversion into deeper wounds, which need special care and attention; thereby, the complete characterization of burn wound type and their subsequent inflammatory status in the cutaneous system at the earliest is of paramount importance. Inflammatory markers at different degrees will help clinicians devise better and more specific treatment strategies for each burn type. The present study is carried out to profile pro-inflammatory gene expression along with immune cell quantification, vascular perfusion, and histopathological assessment in the cutaneous system of murine models. The study revealed that burn injury caused an immediate increase in vascular perfusion in superficial and partial-thickness burns, whereas there was a decrease in vascular perfusion in full-thickness burns. An influx of lymphocytes at the edges of burn wounds in each type of burn injury was well-orchestrated with the event of vascular perfusion. Further, pro-inflammatory gene expression profiling revealed significant upregulation vis-à-vis upregulation of TNF-α and MCP-1 genes, with an increase in the number of neutrophils following 72 h of injury that evidently cemented the conversion of superficial burn into partial-thickness burn. The molecular findings were profoundly supported by the histopathological changes. Thus, our foundational studies show distinct characteristic cutaneous changes correlated with the expression of key pro-inflammatory genes in three different types of burn injuries. Characterization of these cutaneous inflammatory responses provides a promising future for medical interventions involved with different degrees of burn injury, and it will also help in the pre-clinical testing of therapies for burn injury.

Combined radiation burn injuries: A note.

Combined radiation injury occurs when radiation is accompanied by any other form of trauma. The past experiences of Hiroshima, Nagasaki, and Chernobyl have revealed that a large number of victims of such nuclear accidents or attacks suffer from combined radiation injuries. The possibility of a nuclear attack seems very far-fetched, but the destruction that would occur in such an event would be massive, with a huge lossof lives. Therefore, preparedness for the same should be done beforehand. The severity of combined radiation depends upon various factors, such as radiation dose, type, tissues affected, and traumas. The article focuses on combined radiation burn injury (CRBI) which may arise due to the combination of ionising radiation with thermal burns. CRBI can have varied effects on different organs like the hematopoietic, digestive, lymphatic, cardiovascular, and respiratory systems. Some of the most profound lethal effects are hematopoietic dysfunction, gastrointestinal leakage, bacterial translocation to other organ sites, pulmonary fibrosis, and pneumonitis. In this article, we have attempted to accumulate the knowledge of ongoing research on the functioning of different organ systems, which are affected due to CRBI and possible countermeasures to minimize the effects, thus improving survival.

Vascular perfusion: A predictive tool for thermal burn injury.

Damage to the blood vascular system and their altered permeability is a prominent pathological event in case of dermal injury, particularly in burn trauma situations. Prediction of vascular perfusion at the site of damage could be an attractive tool in the estimation of thermal burn injury. A very few reports are available with reference to this tool in defining the thermal injury status. We have used the vascular perfusion estimation method as a tool in assessing the severity of thermal damage to the animal skin. To validate this tool, the mice were subjected to the thermal burn at 90 °C for 10, 20 & 30 s and excised burned skin samples were analyzed for vascular perfusion 24hr post-burn treatment. The vascular perfusion was significantly altered in a time-dependent fashion. This method also provided information regarding blood vessel damage at varied time points. The results of this study clearly indicate the severity of skin damage by the thermal burn. The finding of the present study could have greater implications in predicting the degree of burn. This method is very simple and cost-effective compared to other available modalities used for the estimation of thermal burn injury. The method certainly has the benefit of the estimation of burn injury in the animal models.

Alleviation of radiation combined skin injury in rat model by topical application of ascorbate formulation.

PURPOSE: This research endeavor was undertaken to elucidate the impact of an innovative ascorbate formulation on the regeneration process of full-thickness excision wounds in a rat model exposed to whole-body gamma irradiation, replicating conditions akin to combat or radiation emergency scenarios. MATERIALS AND METHODS: We established a comprehensive rat model by optimizing whole body γ-radiation doses (5-9 Gy) and full-thickness excision wound sizes (1-3 cm2) to mimic radiation combined injury (RCI). The developed RCI model was used to explore the healing potential of ascorbate formulation. The study includes various treatment groups (i.e., sham control, radiation alone, wound alone, radiation + wound, and radiation + wound + formulation). The ascorbate formulation was applied twice daily, with a 12-hour gap between each application, starting 1 hour after the initiation of the wound. The healing potential of the formulation in the RCI context was evaluated over 14 days through hematological, molecular, and histological parameters. RESULTS: The combination of a 5 Gy radiation dose and a 1 cm2 wound was identified as the optimal setting to develop the RCI model for subsequent studies. The formulation was used topically immediately following RCI, and then twice daily until complete healing. Treatment with the ascorbate formulation yielded noteworthy outcomes and led to a substantial reduction (p < .05) in the wound area, accelerated epithelialization periods, and an increased wound contraction rate. The formulation's localized healing response improved organ weights, normalized blood parameters, and enhanced hematopoietic and immune systems. A gene expression study revealed the treatment up-regulated TGF-ß and FGF, and down-regulated PDGF-α, TNF-α, IL-1ß, IL-6, MIP-1α, and MCP-1 (p < .05). Histopathological assessments supported the formulation's effectiveness in restoring cellular architecture and promoting tissue regeneration. CONCLUSION: Topical application of the ascorbate formulation in RCI resulted in a significant improvement in delayed wound healing, leading to accelerated wound closure by mitigating the expression of inflammatory responses.

Evaluation of Decontamination Efficacy of Electrolytically Generated Hypochlorous Acid for the Vesicating Agent: A Multimodel Study.

BACKGROUND: Sulfur Mustard is a strong vesicant and chemical warfare agent that imposes toxicity to the lungs, eyes, and skin after accidental or intended exposure. OBJECTIVES: The current study was intended to explore in vitro and in vivo decontamination properties of electrolytically generated HOCl (hypochlorous acid) against CEES (2-chloroethyle ethyle sulphide), a known sulfur mustard simulant & vesicating agent. METHODS: in vitro studies were carried out using UV spectroscopy and GC-MS methods. In vivo studies were performed in Strain A and immune-compromised mice by subcutaneous as well as prophylactic topical administration of HOCl pretreated CEES. The blister formation and mortality were considered as end-point. Histopathological study was conducted on skin samples by H & E method. DNA damage studies measuring γ-H2AX and ATM have been carried out in human blood using flow cytometry. Anti-bacterial action was tested by employing broth micro dilution methods. A comparative study was also carried out with known oxidizing agents. RESULTS: The topical application of pre-treated CEES at 5, 30 min and 1 h time points showed significant (p<0.001) inhibition of blister formation. DNA damage study showed reduced mean fluorescence intensity of DSBs nearly 17-20 times, suggesting that HOCl plays a protective role against DNA damage. Histopathology showed no sign of necrosis in the epidermis upto 5 min although moderate changes were observed at 30 min. Pretreated samples were analyzed for detection of reaction products with m/z value of 75.04, 69.08, 83.93, 85.95, 123.99, 126.00, and 108.97. HOCl showed a strong bactericidal effect at 40 ppm. The absorbance spectra of HOCl treated CEES showed lowered peaks in comparison to CEES alone and other oxidizing agents. CONCLUSION: In a nutshell, our results signify the decontamination role of HOCl for biological surface application.

Challenges with Wound Infection Models in Drug Development.

Wound research is an evolving science trying to unfold the complex untold mechanisms behind the wound healing cascade. In particular, interest is growing regarding the role of microorganisms in both acute and chronic wound healing. Microbial burden plays an important role in the persistence of chronic wounds, ultimately resulting in delayed wound healing. It is therefore important for clinicians to understand the evolution of infection science and its various etiologies. Therefore, to understand the role of bacterial biofilm in chronic wound pathogenesis, various in vitro and in vivo models are required to investigate biofilms in wound-like settings. Infection models should be refined comprising an important signet of biofilms. These models are eminent for translational research to obtain data for designing an improved wound care formulation. However, all the existing models possess limitations and do not fit properly in the model frame for developing wound care agents. Among various impediments, one of the major drawbacks of such models is that the wound they possess does not mimic the wound a human develops. Therefore, a novel wound infection model is required which can imitate the human wounds. This review article mainly discusses various in vitro and in vivo models showing microbial colonization, their advantages and challenges. Apart from these models, there are also present ex vivo wound infection models, but this review mainly focused on various in vitro and in vivo models available for studying wound infection in controlled conditions. This information might be useful in designing an ideal wound infection model for developing an effective wound healing formulation.

Can miRNAs Serve as Potential Markers in Thermal Burn Injury: An In Silico Approach.

Burn injury has been a major cause of morbidity at global levels. They can occur by multiple agents, such as thermal radiation and chemicals. Among all, thermal burn is predominant and may require specialized treatment in some patients. Although various biomarkers are reportedly used in thermal burn for understanding the pathophysiology of the injury, their limitations prompt for the search of suitable markers that can address the depth and severity of the burn. MicroRNAs (miRNAs) are conserved noncoding molecules that seem to be the promising marker due to their role in multiple pathways and participation in different physiological processes of the body. The present review highlights the role of miRNAs in the repair of the wound and their interaction with specific genes in response to burn stress. Key miR candidates include miR-21, miR-29a, miR-378a-5p, miR-100, miR-27b, miR-200c, miR-150, miR-499-5p, miR-92a, miR-194, and miR-146b, which are identified for their respective targets involved in wound repair. Furthermore, bioinformatics and computational tools were used to confirm the miRNAs and their specific targets. Gene and miRNA expression data sets were downloaded from Research Collaboratory for Structural Bioinformatics Protein Data Bank Database and RNAComposer, respectively, and docked by PatchDock. The possible implications of the identified miRNAs could be in understanding the mechanism of burn injury. These can also be studied with the available drugs being used for burn injury. Apart from that, new intended molecules may also be tested for their effect on these miRNAs.

Creation of rapid and reproducible burn in animal model with a newly developed burn device.

BACKGROUND: The currently available practices for creation of burns in the animals are mostly manual which may lead to lack of uniform wound. There is a need to develop a suitable device that could reproduce and uniformly create burn wound in animal models without the procedural variations and human variability. Present study deals with development of a burn device which has been designed for creation of animal models for burn injury. METHODS: The designed burn device comprises of two main components a heating metal stylus and the thermal sensor. Metal stylus consists of two parts with top part acts as handle and bottom part contains the aluminum probe which quickly heats and cool. The temperature monitoring sensor is attached adjacent to the tip of the probe. The temperature and timer are digitally displayed and can be adjusted as per the requirement. This device is tested for creation of uniform burn in the mice model. Animals were divided into different groups and thermal burn was generated for 60 °C, 80 °C & 100 °C respectively. Burn wound was generated dorsally on shaved skin with contact time of 40 s. Skin biopsies of burn wound were collected after 24 h for histopathology analysis to determine the burn characteristics. Blood vessels damage in the skin was detected by transillumination and digital segmentation using VesSeg tool. RESULTS: The device is able to deliver the different temperature to the animal skin. After reaching the 60 °C, 80 °C & 100 °C for 40 s respectively electronic relays shut down the device. The different groups of the animals showed significant difference in burn morphology in temperature dependent manner. Non significant variation in the burn area of different experimental groups animals was observed. All three zones vis-a-vis coagulation, stasis and hyperaemia were observed at 100 °C whereas found indistinct in 80 °C and 60 °C treated groups. Histopathological studies clearly demonstrated the differences in damage induction in stratum corneum, epidermis, dermis, collagen and hair follicle at selected thermal points. Severe blood vessels damage was observed only at 100 °C. The vascular perfusion was recorded 14% and 57% higher in 60 °C and 80 °C treated animals respectively when compared to control animals. However, at 100 °C due to highest vessel damage the perfusion was reduced to 53% compared to control. CONCLUSION: Present study demonstrates that the device is able to generate precise and uniform burn wound in mice model. The device may be useful for burn related studies and validation of burn wound care products.