A 3D In Vitro Model for Burn Wounds: Monitoring of Regeneration on the Epidermal Level

Burns affect millions every year and a model to mimic the pathophysiology of such injuries in detail is required to better understand regeneration. The current gold standard for studying burn wounds are animal models, which are under criticism due to ethical considerations and a limited predictiveness. Here, we present a three-dimensional burn model, based on an open-source model, to monitor wound healing on the epidermal level. Skin equivalents were burned, using a preheated metal cylinder. The healing process was monitored regarding histomorphology, metabolic changes, inflammatory response and reepithelialization for 14 days. During this time, the wound size decreased from 25% to 5% of the model area and the inflammatory response (IL-1β, IL-6 and IL-8) showed a comparable course to wounding and healing in vivo. Additionally, the topical application of 5% dexpanthenol enhanced tissue morphology and the number of proliferative keratinocytes in the newly formed epidermis, but did not influence the overall reepithelialization rate. In summary, the model showed a comparable healing process to in vivo, and thus, offers the opportunity to better understand the physiology of thermal burn wound healing on the keratinocyte level.

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119 A Nitric Oxide Releasing Bio-active Wound Dressing for Burn Wound Infection Treatment

Journal of Burn Care & Research ◽

10.1093/jbcr/iraa024.122 ◽

2020 ◽

Vol 41 (Supplement_1) ◽

pp. S79-S80

Author(s):

Jahnabi Roy ◽

Lori Estes ◽

Robert J Christy ◽

Hitesh Handa ◽

Shanmugasundaram Natesan

Keyword(s):

Wound Healing ◽

Wound Infection ◽

Wound Dressing ◽

Healing Process ◽

Fibroblast Proliferation ◽

Burn Wound ◽

Burn Wounds ◽

Log Reduction

Abstract Introduction The future of multi-domain battlefield operation requires wound dressings to prevent infection at the point of injury. Majority of antimicrobial agents only target wound infection while other healing events are left to their natural fates. Nitric oxide (NO) acts against both gram-positive and -negative bacteria and has the potential to positively affect wound healing. In this study we have developed a novel wound dressing integrated with a NO donor - S-nitroso- glutathione (GSNO) in a hybrid formulation of alginate and poly(vinyl alcohol) (PVA) to prevent/treat burn wound infection. Methods The NO releasing wound dressing was fabricated using PVA, alginate, and glycerol, crosslinked with CaCl2 incorporating GSNO. Thereafter, release kinetics were measured up to 4 days. The antibacterial efficacy was determined against both P. aeruginosa and S. aureus. Then the biocompatibility of the NO wound dressing was assessed using in vitro fibroblast proliferation and wound healing assay. Finally, the efficacy of the wound dressing was assessed in vivo using a 3-cm diameter porcine burn wound infection model. Results The Alginate-PVA-GSNO dressing showed a desired physiological level NO flux of 4.42 × 10-10 mol cm-2 min-1 for 72 hours. Alginate−PVA−GSNO dressings showed ~3 log reduction in S. aureus and ~2 log reduction in P. aeruginosa CFU/mg when compared to control. The NO-releasing dressing improved fibroblast proliferation and migration resulting in complete closure of the wound within 48 h in vitro. The safety and efficacy of NO-releasing dressing were successfully established in the both P. aeruginosa and S. aureus infected porcine burn wounds. Histological assessments are carried out to determine the effect of NO-releasing dressing on overall healing process. Conclusions This study shows Alginate−PVA-GSNO wound dressing provides antimicrobial and wound healing properties in vitro. Preliminary in vivo wound healing studies established the safety and efficacy profile of NO-releasing dressing to treat burn wounds. Applicability of Research to Practice An easy to apply, field deployable and effective antimicrobial wound dressing is still a major requisite for combat burn wounds. NO delivering alginate-PVA based wound dressing may be an ideal candidate to inhibit infection as well as promote the wound healing process.

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The Efficacy of Silver-Based Electrospun Antimicrobial Dressing in Accelerating the Regeneration of Partial Thickness Burn Wounds Using a Porcine Model

Polymers ◽

10.3390/polym13183116 ◽

2021 ◽

Vol 13 (18) ◽

pp. 3116

Author(s):

Thien Do ◽

Tien Nguyen ◽

Minh Ho ◽

Nghi Nguyen ◽

Thai Do ◽

...

Keyword(s):

Wound Healing ◽

Burn Injury ◽

Healing Process ◽

Bactericidal Effect ◽

Wound Healing Process ◽

Burn Wounds ◽

Partial Thickness Burn ◽

Tissue Damages

(1) Background: Wounds with damages to the subcutaneous are difficult to regenerate because of the tissue damages and complications such as bacterial infection. (2) Methods: In this study, we created burn wounds on pigs and investigated the efficacy of three biomaterials: polycaprolactone-gelatin-silver membrane (PCLGelAg) and two commercial burn dressings, Aquacel® Ag and UrgoTulTM silver sulfadiazine. In vitro long-term antibacterial property and in vivo wound healing performance were investigated. Agar diffusion assays were employed to evaluate bacterial inhibition at different time intervals. Minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and time-kill assays were used to compare antibacterial strength among samples. Second-degree burn wounds in the pig model were designed to evaluate the efficiency of all dressings in supporting the wound healing process. (3) Results: The results showed that PCLGelAg membrane was the most effective in killing both Gram-positive and Gram-negative bacteria bacteria with the lowest MBC value. All three dressings (PCLGelAg, Aquacel, and UrgoTul) exhibited bactericidal effect during the first 24 h, supported wound healing as well as prevented infection and inflammation. (4) Conclusions: The results suggest that the PCLGelAg membrane is a practical solution for the treatment of severe burn injury and other infection-related skin complications.

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Injectable Nanocurcumin-Formulated Chitosan-g-Pluronic Hydrogel Exhibiting a Great Potential for Burn Treatment

Journal of Healthcare Engineering ◽

10.1155/2018/5754890 ◽

2018 ◽

Vol 2018 ◽

pp. 1-14 ◽

Cited By ~ 18

Author(s):

Le Hang Dang ◽

Thi Hiep Nguyen ◽

Ha Le Bao Tran ◽

Vu Nguyen Doan ◽

Ngoc Quyen Tran

Keyword(s):

Wound Healing ◽

Wound Repair ◽

Healing Process ◽

Nanocomposite Hydrogel ◽

Burn Wound ◽

Burn Treatment ◽

Nanocomposite Hydrogels ◽

Burn Wound Healing

Burn wound healing is a complex multifactorial process that relies on coordinated signaling molecules to succeed. Curcumin is believed to be a potent antioxidant and anti-inflammatory agent; therefore, it can prevent the prolonged presence of oxygen free radicals which is a significant factor causing inhabitation of optimum healing process. This study describes an extension of study about the biofunctional nanocomposite hydrogel platform that was prepared by using curcumin and an amphiphilic chitosan-g-pluronic copolymer specialized in burn wound healing application. This formular (nCur-CP, nanocomposite hydrogel) was a free-flowing sol at ambient temperature and instantly converted into a nonflowing gel at body temperature. In addition, the storage study determined the great stability level of nCur-CP in long time using UV-Vis and DLS. Morphology and distribution of nCur in its nanocomposite hydrogels were observed by SEM and TEM, respectively. In vitro studies suggested that nCur-CP exhibited well fibroblast proliferation and ability in antimicrobacteria. Furthermore, second- and third-degree burn wound models were employed to evaluate the in vivo wound healing activity of the nCur-CP. In the second-degree wound model, the nanocomposite hydrogel group showed a higher regenerated collagen density and thicker epidermis layer formation. In third degree, the nCur-CP group also exhibited enhancement of wound closure. Besides, in both models, the nanocomposite material-treated groups showed higher collagen content, better granulation, and higher wound maturity. Histopathologic examination also implied that the nanocomposite hydrogel based on nanocurcumin and chitosan could enhance burn wound repair. In conclusion, the biocompatible and injectable nanocomposite scaffold might have great potential to apply for wound healing.

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Biodegradable Electrospun Nonwovens Releasing Propolis as a Promising Dressing Material for Burn Wound Treatment

Pharmaceutics ◽

10.3390/pharmaceutics12090883 ◽

2020 ◽

Vol 12 (9) ◽

pp. 883

Author(s):

Mateusz Stojko ◽

Jakub Włodarczyk ◽

Michał Sobota ◽

Paulina Karpeta-Jarząbek ◽

Małgorzata Pastusiak ◽

...

Keyword(s):

Active Compound ◽

Healing Process ◽

Mechanical Damage ◽

Degradation Process ◽

Molar Ratio ◽

Wound Healing Process ◽

Burn Wound ◽

Burn Wounds

The selection of dressing is crucial for the wound healing process. Traditional dressings protect against contamination and mechanical damage of an injured tissue. Alternatives for standard dressings are regenerating systems containing a polymer with an incorporated active compound. The aim of this research was to obtain a biodegradable wound dressing releasing propolis in a controlled manner throughout the healing process. Dressings were obtained by electrospinning a poly(lactide-co-glycolide) copolymer (PLGA) and propolis solution. The experiment consisted of in vitro drug release studies and in vivo macroscopic treatment evaluation. In in vitro studies released active compounds, the morphology of nonwovens, chemical composition changes of polymeric material during degradation process, weight loss and water absorption were determined. For in vivo research, four domestic pigs, were used. The 21-day experiment consisted of observation of healing third-degree burn wounds supplied with PLGA 85/15 nonwovens without active compound, with 5 wt % and 10 wt % of propolis, and wounds rinsed with NaCl. The in vitro experiment showed that controlling the molar ratio of lactidyl to glycolidyl units in the PLGA copolymer gives the opportunity to change the release profile of propolis from the nonwoven. The in vivo research showed that PLGA nonwovens with propolis may be a promising dressing material in the treatment of severe burn wounds.

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Mechanism of Wound-Healing Activity ofHippophae rhamnoidesL. Leaf Extract in Experimental Burns

Evidence-based Complementary and Alternative Medicine ◽

10.1093/ecam/nep189 ◽

2011 ◽

Vol 2011 ◽

pp. 1-9 ◽

Cited By ~ 51

Author(s):

Nitin K. Upadhyay ◽

Ratan Kumar ◽

M. S. Siddiqui ◽

Asheesh Gupta

Keyword(s):

Wound Healing ◽

Wound Repair ◽

Leaf Extract ◽

Lipid Peroxide ◽

Enzymatic Antioxidants ◽

Burn Wound ◽

Burn Wounds ◽

Aqueous Leaf Extract

The present investigation was undertaken to evaluate the healing efficacy of lyophilized aqueous leaf extract of Sea buckthorn (Hippophae rhamnoidesL., family Elaeagnaceae) (SBT) and to explore its possible mechanism of action on experimental burn wounds in rats. The SBT extract, at various concentrations, was applied topically, twice daily for 7 days. Treatment with silver sulfadiazine (SSD) ointment was used as reference control. The most effective concentration of the extract was found to be 5.0% (w/w) for burn wound healing and this was further used for detailed study. The SBT-treated group showed faster reduction in wound area in comparison with control and SSD-treated groups. The topical application of SBT increased collagen synthesis and stabilization at the wound site, as evidenced by increase in hydroxyproline, hexosamine levels and up-regulated expression of collagen type-III. The histological examinations and matrix metalloproteinases (MMP-2 and -9) expression also confirmed the healing efficacy of SBT leaf extract. Furthermore, there was significant increase in levels of endogenous enzymatic and non-enzymatic antioxidants and decrease in lipid peroxide levels in SBT-treated burn wound granulation tissue. The SBT also promoted angiogenesis as evidenced by anin vitrochick chorioallantoic membrane model andin vivoup-regulated vascular endothelial growth factor (VEGF) expression. The SBT leaf extract had no cytotoxic effect on BHK-21 cell line. In conclusion, SBT aqueous leaf extract possesses significant healing potential in burn wounds and has a positive influence on the different phases of wound repair.

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RELEVANCE AND PERSPECTIVES OF EXPERIMENTAL WOUND MODELS IN WOUND HEALING RESEARCH

Asian Journal of Pharmaceutical and Clinical Research ◽

10.22159/ajpcr.2017.v10i7.18276 ◽

2017 ◽

Vol 10 (7) ◽

pp. 57 ◽

Cited By ~ 1

Author(s):

Santram Lodhi ◽

Gautam P Vadnere

Keyword(s):

Wound Healing ◽

Search Engines ◽

Wound Repair ◽

Healing Process ◽

Chorioallantoic Membrane ◽

Tube Formation ◽

Wound Healing Process ◽

Burn Wound

The wound healing process consists of four highly integrated and overlapping phases: Hemostasis, inflammation, proliferation, and tissue remodeling. These phases and their biophysiological functions must occur in the proper sequence, at a specific time and continue for a specific duration at an optimal intensity. There are many factors that can affect wound healing which interferes with one or more phases in this process, thus causing improper or impaired tissue repair. This review was aimed to collect data and made a critical analysis. This will provide concise information regarding different models and parameters used for wound healing study. The data related to different wound models are collected using popular search engines as well as relevant science search engines and database including Google Scholar, Science Direct, and PubMed. A new drug substance can be evaluated for wound healing activity using different in vitro models such as cell culture, chick chorioallantoic membrane model, tube formation on metrigel and capillary growth model. The in vivo wound models such as incision, excision, dead space, burn wound, ischemic wound, and diabetic wound models are frequently used. Each model has specific importance. The limitations and advantages of each are described in this review. Although animal wound repair is an imperfect reflection of human wound healing and its clinical challenges, these models can be fundamental tools for the development of new approaches to rational wound therapy.

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Fabrication and In-Vivo Study of Micro-Colloidal Zanthoxylum acanthopodium-Loaded Bacterial Cellulose as a Burn Wound Dressing

Polymers ◽

10.3390/polym12071436 ◽

2020 ◽

Vol 12 (7) ◽

pp. 1436 ◽

Cited By ~ 1

Author(s):

Khatarina Meldawati Pasaribu ◽

Saharman Gea ◽

Syafruddin Ilyas ◽

Tamrin Tamrin ◽

Appealwan Altruistis Sarumaha ◽

...

Keyword(s):

Bacterial Cellulose ◽

Wound Dressing ◽

Healing Process ◽

Three Dimensional ◽

Antimicrobial Activities ◽

Sem Analysis ◽

Wound Healing Process ◽

Burn Wound

Bacterial cellulose (BC) is a biopolymer commonly used for wound dressing due to its high biocompatible properties either in-vitro or in-vivo. The three-dimensional fiber structure of BC becomes an advantage because it provides a template for the impregnation of materials in order to improve BC’s properties as a wound dressing, since BC has not displayed any bioactivity properties. In this study, micro-colloidal Zanthoxylum acanthopodium (MZA) fruit was loaded into BC fibers via an in-situ method. Z. acanthopodium is known to have anti-inflammatory, antioxidant and antimicrobial activities that can support BC to accelerate the wound healing process. The FTIR, XRD and SEM analysis results showed that the loading process of MZA and the composite fabrication were successfully carried out. The TGA test also showed that the presence of MZA in BC fibers decreased Tmax composite from BC, from 357.8 to 334.5 °C for BC-MZA3. Other aspects, i.e., water content, porosity, hemocompatibility and histology studies, also showed that the composite could potentially be used as a wound dressing.

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Development of a Topical Insulin Polymeric Nanoformulation for Skin Burn Regeneration: An Experimental Approach

International Journal of Molecular Sciences ◽

10.3390/ijms22084087 ◽

2021 ◽

Vol 22 (8) ◽

pp. 4087

Author(s):

Maria Quitério ◽

Sandra Simões ◽

Andreia Ascenso ◽

Manuela Carvalheiro ◽

Ana Paula Leandro ◽

...

Keyword(s):

Wound Healing ◽

Healing Process ◽

In Vitro Release ◽

Peptide Hormone ◽

Plga Nanoparticles ◽

Wound Healing Process ◽

Target Area ◽

Encapsulation Process

Insulin is a peptide hormone with many physiological functions, besides its use in diabetes treatment. An important role of insulin is related to the wound healing process—however, insulin itself is too sensitive to the external environment requiring the protective of a nanocarrier. Polymer-based nanoparticles can protect, deliver, and retain the protein in the target area. This study aims to produce and characterize a topical treatment for wound healing consisting of insulin-loaded poly-DL-lactide/glycolide (PLGA) nanoparticles. Insulin-loaded nanoparticles present a mean size of approximately 500 nm and neutral surface charge. Spherical shaped nanoparticles are observed by scanning electron microscopy and confirmed by atomic force microscopy. SDS-PAGE and circular dichroism analysis demonstrated that insulin preserved its integrity and secondary structure after the encapsulation process. In vitro release studies suggested a controlled release profile. Safety of the formulation was confirmed using cell lines, and cell viability was concentration and time-dependent. Preliminary safety in vivo assays also revealed promising results.

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In Vivo Testing of Sericin-Polyurethane Nanofiber Mats for Wound Healing in Rats

Key Engineering Materials ◽

10.4028/www.scientific.net/kem.751.581 ◽

2017 ◽

Vol 751 ◽

pp. 581-585 ◽

Cited By ~ 1

Author(s):

Piyaporn Kampeerapappun ◽

Pornpen Siridamrong

Keyword(s):

Wound Healing ◽

Healing Process ◽

Active Agent ◽

L929 Cell ◽

Wound Healing Process ◽

Original Size ◽

Nanofiber Mats ◽

Dressing Materials

The objective of this study was to investigate sericin-polyurethane nanofiber cover (SUC) for wound dressing materials in a rat skin. Sericin-polyurethane blended nanofibers were fabricated by using electrospinning. The composition of 3%w/v polyurethane in ethanol and 19% w/v sericin were blended and electrospun at 15 kV, 20 cm from tip to collector with a feed rate of 6.2 ml/hr. The mats, approximately 1.5 mm thick, were sterile by gamma irradiation with a radiation dose of 15 kGy. The samples of in vitro and in vivo testing were separated into three groups; gauze, polyurethane nanofiber cover (UC), and SUC. In vitro cultured L929 cell lines were investigated with inverted microscope. It was found that cells migrated to SCU. For in vivo tests, the remaining wound in rats was measured on day 2-14 after excision. Compared to original size of wound samples, the size of the wound remained 24% for SUC, 33% for gauze, and 34% for UC at day 8. The sericin, an active agent, contained in SUC mats was about 5 µl at 1.5 ×1.5 cm. It can be concluded that sericin is non-toxic to cells and can promote wound healing process in rats.

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670 Delayed Enzymatic Debridement of Burn Wounds using the Proteolytic Gel SN514

Journal of Burn Care & Research ◽

10.1093/jbcr/irab032.316 ◽

2021 ◽

Vol 42 (Supplement_1) ◽

pp. S191-S192

Author(s):

Angela R Jockheck-Clark ◽

Randolph Stone ◽

Michelle Holik ◽

Lucy Schaffer ◽

Shanmugasundaram Natesan ◽

...

Keyword(s):

Burn Injury ◽

Vehicle Control ◽

Surrounding Tissue ◽

Burn Wound ◽

Burn Wounds ◽

Delayed Treatment ◽

Medical Evacuation ◽

Contamination Levels

Abstract Introduction Thermal burns account for 5–10% of casualties sustained in present-day conflicts and are expected to be one of the most common wounds to occur in future conflicts. In prolonged field care (PFC) situations, medical evacuation could be delayed for days. During this time, burn wounds can become infected, detrimentally impact neighboring tissue, and cause systemic immune responses. Therefore, it is essential to test and evaluate non-surgical debridement agents that could be implemented prior to reaching a Role 3 military treatment facility. This work details how the proprietary proteolytic gel SN514 impacts burn debridement when applied within a PFC-like timeline. SN514 contains an enzyme formulation that is thermostable, easy to apply, and selectively degrades non-viable tissue in vitro and in vivo. Methods Deep-partial thickness contact burns were created using an established porcine model and covered with gauze or an antimicrobial incise drape. Four days later, the burns were treated with one of five treatments: 0.2% SN514, 0.8% SN514, a vehicle control, gauze, or an antimicrobial silver dressing. Treatments were re-applied every 24 hours for 72 to 96 hours. The effects of the treatment regiments were compared histologically. Biopsies were also taken to monitor bacterial contamination levels. Results Burns treated with SN514 were partially debrided and visually distinct from those treated with gauze, the silver dressing, or the vehicle control. Preliminary analyses suggest that SN514-treated burns that had been covered with “dry” gauze had a much lower debridement efficiency than those treated with the incise drape. This suggests that SN514 debridement efficiency may depend on the presence of a moist eschar. Preliminary analyses also suggest that there was little difference in burn wound bacterial counts among the five treatment groups. Conclusions SN514 is able to debride burns that experienced delayed treatment, without any evidence of harm to the surrounding tissue or evidence of exacerbating the original burn injury. SN514-treated wounds displayed little to no blood loss and did not increase burn wound infection levels compared to wounds treated with gauze or an antimicrobial silver dressing.

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