A 2000 patient retrospective assessment of a new strategy for burn wound management in view of infection prevention and treatment.

Infections in burn patients are still the principal cause of complications in burn injuries. The aim of this study is to assess a new strategy for burn wound management in view of infection prevention and treatment in the experience of the Burn Treatment Center in Siemianowice Slaskie. The applied methodology involved the analysis of patient records describing the hospital's epidemiological situation between 2014 and 2016. The analysis also included the use and cost of antibiotics, silver-containing dressings, and other antiseptics relative to the number of sepsis cases, including those caused by Pseudomonas aeruginosa, as well as the mortality ratio. The total costs of prevention and treatment of infections were reduced, while the use of silver-containing dressings and antiseptics increased. The number of patients with sepsis decreased, including cases caused by P. aeruginosa, and the mortality ratio was reduced. Introducing a strategy for burn wound-oriented infection prevention and treatment in burn patients provides a number of benefits. It is also cost-effective. Using locally applied active dressings and antiseptics can be a welcome choice for often-unnecessary antibiotic therapy of a suspected or existing burn wound infection.

A new option for definitive burn wound closure - pair matching type of retrospective case-control study of hand burns in the hospitalised patients group in the Dr Stanislaw Sakiel Centre for Burn Treatment between 2009 and 2015.

Nearly 80% of all burns include the hands of affected individuals. Skin grafting is the gold standard in burns treatment, but in the case of the burn wound bed, it may require the necessity of utilising skin substitutes to facilitate closure. The aim of this study is to assess the impact of a porcine-derived wound dressing (Oasis™) for application to hand burns compared to a synthetic dressing (Suprathel™). Comparative assessments were made, including the time to heal, quality of healing and pain intensity. A retrospective, unblinded, matching pair case-control of hand burns was performed. A control group of 24 patients was treated with Suprathel dressing, and a study group of six patients underwent application of the Oasis dressing. The wound healing process was evaluated by taking histopathological specimens and also utilising the Bates-Jensen Wound Assessment Tool. A 10-cm Visual Analogue Scale (VAS) was used for pain assessment. Other parameters measured included dressing loss because of infection and the need of rehabilitation. The progress of wound healing on the fourth day in the study group was 30%. A decrease in the level of pain was recorded on the fourth day after surgery. There was a decrease of 5% in the risk of rehabilitation in the treatment group.

Transfer of fibroblast sheets cultured on thermoresponsive dishes with membranes.

In cell or tissue engineering, it is essential to develop a support for cell-to-cell adhesion, which leads to the generation of cell sheets connected by extracellular matrix. Such supports must be hydrophobic and should result in a detachable cell sheet. A thermoresponsive support that enables the cultured cell sheet to detach using only a change in temperature could be an interesting alternative in regenerative medicine. The aim of this study was to evaluate plates covered with thermoresponsive polymers as supports for the formation of fibroblast sheets and to develop a damage-free procedure for cell sheet transfer with the use of membranes as transfer tools. Human skin fibroblasts were seeded on supports coated with a thermoresponsive polymer: commercial UpCell™ dishes (NUNC™) coated with thermoresponsive poly(N-isopropylacrylamide) (PNIPAM) and dishes coated with thermoresponsive poly(tri(ethylene glycol) monoethyl ether methacrylate) (P(TEGMA-EE)). Confluent fibroblast sheets were effectively cultured and harvested from both commercial PNIPAM-coated dishes and laboratory P(TEGMA-EE)-coated dishes. To transfer a detached cell sheet, two membranes, Immobilon-P(®) and SUPRATHEL(®), were examined. The use of SUPRATHEL for relocating the cell sheets opens a new possibility for the clinical treatment of wounds. This study established the background for implementing thermoresponsive supports for transplanting in vitro cultured fibroblasts.

The Impact of Telemedicine on Patients with Hidradenitis Suppurativa in the COVID-19 Era.

(1) Background: Hidradenitis suppurativa is a chronic, inflammatory skin disease. It is characterized by the transformation of normal skin into skin with abscesses, nodules, tunnels, and scars. The most commonly affected areas are the armpits, groins, buttocks, and subscapular area. Patients with HS require constant care under the supervision of the outpatient clinic. Due to the COVID-19 pandemic, consultations have been introduced in the form of telemedicine. The aim of this study was to evaluate the availability of HS treatment during the COVID-19 pandemic and to assess patient satisfaction, problems with access to medical care, and the impact of the pandemic on the course of the disease. (2) Methods: An internet survey with an anonymous questionnaire was used to assess the effectiveness of telemedicine consultations. The survey consisted of 25 closed questions, and responses were kept fully anonymous. (3) Results: Most respondents reported minor problems with accessing specialized HS medical care during the COVID-19 pandemic (n = 25, 35.71%). However, 35.71% (n = 25) of them reported major problems with appointments for specialized ambulatory treatment during the last few months of the pandemic, mainly due to delayed appointments. Almost half of the respondents had been diagnosed with COVID-19 (n = 34, 48.57%), and 58.57% (n = 41) of respondents did not see a correlation between COVID-19 infection and HS progression. (4) Conclusions: Our study showed that the pandemic significantly limited access to medical advice, and patients with hidradenitis suppurativa prefer standard consultations.

A Survey of 1000 Respondents on the Polish Population's Knowledge and Attitudes about Tissue/Organ Donation and Transplantation in Times of Allogeneic Tissue Shortage.

Tissue-engineered human allogeneic skin grafts retrieved from a deceased donor play an important role in the therapy of extensive and deeply burned patients. However, there is a vital deficit of allogeneic skin donors, and the reserves of human allogeneic skin grafts are not sufficient. The goal of this work was to analyze the level of knowledge and attitudes of Polish society in the field of transplantation, with particular emphasis on allogeneic skin transplantation. The study used a self-made questionnaire comprised of 23 questions. 1000 respondents took part in this research. The respondents were a diverse group in terms of age, sex, education, and place of residence. The obtained results show a general positive attitude of the respondents towards the idea of transplantology. However, people with lower education presented a more negative attitude towards the donation of tissues and organs. Additionally younger people were not able to clearly declare readiness for organ procurement. What is more data analysis revealed certain gaps in more detailed knowledge and surprising attitudes. In that respect, the lack of awareness about the criteria for determining brain death could be mentioned. There was also a lack of acceptance for skin procurement in specific population groups. It can therefore be concluded that a key role in the success of the idea of transplantation in Poland is the broad and systematic education of the society.

Influence of electron beam irradiation on extracellular matrix of the human allogeneic skin grafts.

The nonviable allogeneic human skin grafts might be considered as the most suitable skin substitutes in the treatment of extensive and deep burns. However, in accordance to biological security such grafts require the final sterilization prior to clinical application. The aim of the study was to verify the influence of electron beam irradiation of three selected doses: 18, 25, and 35 kGy on the extracellular matrix of human skin. Prior to sterilization, the microbiological tests were conducted and revealed contamination in all examined cases. Individual groups were subjected to single electron beam radiation sterilization at proposed doses and then subjected to microbiological tests again. The results of microbiological testing performed for all irradiation doses used were negative. Only in the control group was a growth of microorganisms observed. The FTIR spectrometry tests were conducted followed by the histological evaluation and mechanical tests. In addition, cost analysis of radiation sterilization of individual doses was performed. The results of spectroscopic analysis, mechanical tests, and histological staining showed no significant changes in composition and characteristics of tested tissues after their irradiation, in comparison to control samples. The cost analysis has shown that irradiation with 18 kGy is the most cost-effective and 35 kGy is the least favorable. However, according to biological risk reduction, the recommended sterilization dose is 35 kGy, despite the higher price compared to the other doses tested.

The Causes for the Loss of Cellular Material at the Qualification Stage and the Influence of the Patient's Flora on Culture Infection.

There are several causes leading to the loss of cellular material earmarked for transplantation. This paper aims to evaluate the number of lost donors and lost cells in the culture by means of verifying both the results of the qualification tests and the presence of microorganisms in the cellular material. The analysis involved 86 donors hospitalized for thermal burns, from whom cells were harvested for keratinocyte and fibroblast cultures in the years 2011 to 2015. All potential donors underwent qualification tests: Anti-HIV-1,2; HBsAg; Anti-HCV-Ab; HBc, and a specific test for syphilis. In the case of skin fragments collected for culturing, the microbiological tests included the carrying fluid, the medium after 1 change, and the medium during culturing and before transplantation. Skin donors for cell cultures were assigned to the groups based on if the skin was collected up to 7 days following the burn or later. On average, 12% of the disqualifications were reported among donors for cell culturing. The most frequent cause of donor disqualification (54% of all disqualifications) was a positive HBc(+). The occurrence of fungal infections detected in the cellular material was over 30%. Establishing the culture after day 7 following the injury immediately increases the risk of infection by 25% in comparison to those cultures established before or on day 7 following the injury. Proper disinfection of donor place is crucial, but sometimes insufficient for maintenance sterility in cell culture. The risk of infection increases 25% after 7 following the injury.

Amniotic and Umbilical Cord of Transgenic Pigs as an Alternative Source of Stem Cells.

BACKGROUND: The placenta is worthy of consideration as a source of tissues for transplantation. Porcine material preparation allows significantly more grafts to be obtained than human material. Amniotic cells are a very practicable stem cell type and easy to obtain with great potential, compared with, for instance, adipose-derived stem cells. The aim of this paper was to verify if porcine transgenic amnion and umbilical cord could be as effective for xenotransplantation of stem cells as human material. METHODS: Oxytocin was administered to the sows on the day the material was obtained. The obtained material was divided by weight into 4 relatively equal parts, which were transferred into 1 of 4 containers. The containers had different transport media and a concentration of antibiotics. After cell isolation by homogenization, the number of live, dead, and apoptotic cells was assessed. RESULTS: Maintaining the sterility when obtaining material from breeding pigs was the biggest problem. Transport medium, despite the addition of antibiotics, was in most cases infected. Tests revealed that porcine cells have a tendency to leave tissue material and adhere to plastic as human cells do. Porcine cells are also fibroblast like, which can suggest that these cells might be tested for the presence of mesenchymal stem cells. However, some differences are visible in all parameters, which can result from contamination of material or improper transport medium. There were significant differences between viability and apoptotic cell number in human and transgenic pig cells isolated from both sources---amniotic membrane and umbilical cord.

Porcine Transgenic, Acellular Material as an Alternative for Human Skin.

Although new therapeutic approaches for burn treatment have made progress, there is still need for efficient coverage of donor fields. Promising dressing for skin graft donor site should be biocompatible, attach easily to the wound bed, remain in place until donor site has renewed, and decrease morbidity at the site. Porcine skin may be applied as a dressing for severe burns. Therefore pig skin xenografts can be used also as donor field coverage. In the Burn Treatment Centre, we used gauze soaked in Vaseline to secure donor fields. The aim of the study was to check if transgenic porcine skin is better than standard in donor site coverage used in our center. We showed that dressing reduces pain experienced by patients. The dressing leads to a reduction of hospitalization time by an average of 8 days. The dressing is as safe as the gold standard. Securing the donor field reduces the risk of colonization of the wound in the second smear after application by 60%. The disadvantage of the dressing is the inability to absorb blood; the use of hemostatic ointments in combination with the skin of transgenic pigs should be considered in the future.

A new approach to the production of a biovital skin graft based on human acellular dermal matrix produced in-house, in vitro revitalized internally by human fibroblasts and keratinocytes on the surface.

Patients with extensive and deep burns who do not have enough donor sites for autologous skin grafts require alternative treatment methods. Tissue engineering is a useful tool to solve this problem. The aim of this study was to find the optimal method for the production of a biovital skin substitute based on acellular dermal matrix (ADM) and in vitro cultured fibroblasts and keratinocytes. In this work, nine methods of ADM production were assessed. The proposed methods are based on the use of the following enzymes: Dispase II, collagenase I/ethylenediaminetetraacetic acid (EDTA), collagenase II/EDTA, and mechanical perforation using DermaRoller and mesh dermatome. The obtained ADMs were examined (both on the side of the basement membrane and on the "cut-off" side) by means of scanning electron microscopy, immunohistochemistry tests and strength tests. ADM was revitalized with human fibroblasts and keratinocytes. The ability of in-depth revitalization of cultured fibroblasts and their ability to secrete collagen IV was examined. The obtained results indicate that the optimal method of production of live skin substitutes is the colonization of autologous fibroblasts and keratinocytes on the scaffold obtained using two-step incubation method: Trypsin/EDTA and dispase II.

Amniotic Stem Cells Cultured on Thermoresponsive Polymers Allow Obtaining a Full Cell Sheet.

Amniotic stem cells promote adhesion and migration of epithelial cells. Obtaining a full sheet containing amniotic stem cells seems to be the best solution for the treatment of burn wounds. The main advantage of this method is obtaining a full sheet of cells by lowering the temperature below the transition temperature, which does not affect extracellular matrix. The purpose of this work was to produce a skin substitute-a full sheet consisting of amniotic mesenchymal stem cells-and compare with well-known fibroblast sheet. Amniotic membrane cells revealed better tendency to full sheet detachment than fibroblasts. Confluence after 24 hours was always higher on polymer-coated dishes than on normal polypropylene dishes. Also viability was higher than on the control culture dish, while the number of apoptotic cells was always highest on polypropylene (control). Ile-Lys-Val-ala-Val (IKVAV) 0.28 addition to poly (poly [ethylene glycol] ethyl methacrylate) (PTEGMA) caused best cell confluence and highest percentage of cells in mitosis phase of cell cycle, but also worst cell detachment was observed in both cell types on PTEGMA IKVAV 0.28. Viability of cells transferred in cell sheet form onto a new culture dish was higher than when detached as suspension enzymatically. Additionally, percentage of apoptotic cells transferred in cell sheet form onto a new culture dish was always lower than when detached as suspension enzymatically. Culturing of PTEGMA, PTEGMA IKVAV 0.28 and PTEGMA IKVAV 0.14 have a stimulating effect on number of cells in mitosis in amniotic cell culture even after cell sheet transfer onto a new dish, whereas such effect with fibroblast was not observed.

Thermal imaging and planimetry evaluation of the results of chronic wounds treatment with hyperbaric oxygen therapy.

BACKGROUND: One of the methods to treat chronic wounds is the use of hyperbaric oxygen (HBO). Objective measurement of the wound surface is an important element in the process of monitoring and predicting the progress of treatment. OBJECTIVES: The aim of the study was to evaluate the effect of hyperbaric oxygen therapy (HBOT) on ulcer wound healing in patients with chronic venous insufficiency ulcers and diabetic foot syndrome using thermal imaging and computerized planimetry. MATERIAL AND METHODS: During a 3-year period, 284 digital computer planimetry measurements were gathered from 142 patients treated for leg ulcers caused by chronic venous insufficiency and ulcers from diabetic foot syndrome at HBOT Unit of the Dr Stanislaw Sakiel Centre for Burns Treatment in Siemianowice Slaskie (Poland). Each patient took 30 HBOT sessions using a Haux multiplace HBO chamber at a pressure of 2.5 atmospheres absolute (ATA). The results of the treatment were monitored by thermovision and computerassisted planimetry measurements performed before and after HBOT. RESULTS: Both groups of patients exhibited a reduction in the surface and perimeter of the wound after HBOT. The treatment effects were also confirmed by thermal imaging. The areas calculated from thermal imaging and planimetry are different but correlated. CONCLUSIONS: It seems that a combination of thermal imaging and planimetry may enhance the diagnosis as well as provide the physician with more information about therapy effects.

Does the transplantation of keratinocytes really reduce the risk of death? Survival analysis of patients hospitalized at the Dr Stanislaw Sakiel Centre for Burns Treatment in 2008-2015.

BACKGROUND: Keratinocyte transplantation is an adjuvant procedure in the extensive burn therapy method. However, it must be taken into consideration that clinical results of keratinocyte transplantation are ambiguous and progress achieved in this method is still being verified, especially due to the high cost of cultured epithelial autograft (CEA) transplants. OBJECTIVES: The aim of this study was to verify the impact of cultured keratinocyte application on patients' survival. This study included a group of patients with the highest chance for a successful outcome of the therapy and excluded patients with no compelling reason to apply for such an expensive therapy. MATERIAL AND METHODS: This study included all the patients with burns diagnosed between January 1, 2008 and January 1, 2016, who were treated with cultured skin cells. Patients' age and gender, percentage of total body surface area (TBSA) affected, percentage of burn depth of the 3rd/4th degree, number of days between admission and surgery, and need for rehabilitation were analyzed. RESULTS: The cultured cell application did not significantly affect the risk of death (p > 0.05). CONCLUSIONS: Keratinocytes should be applied as an adjunctive method for the treatment of burns with at least 40% TBSA affected, but with a maximal burn depth of the 2nd degree. In the group of patients below 50 years of age, a higher number of transplants with a cell population above 20 million/mL and a significantly lower mortality rate were observed, which means that in the mentioned age group, this graft was more effective. It has been suggested that patients older than 50 years of age with burns deeper than of the 2nd degree should be treated with more advanced methods like, e.g., the application of stem cells.

Fibroblast and keratinocyte crosstalk: the effect of a poly(tri[ethylene glycol] ethyl ether methacrylate) thermoresponsive surface on short-term co-culture.

The treatment of difficult-to-treat wounds can be challenging. Although a number of approaches have been investigated, the healing process may be slow and unsatisfactory. An alternative approach is the use of a continuous sheet of skin cells applied over a wound which may improve cell implantation and patient recovery. To analyse the gene expression profile of fibroblast/keratinocyte co-culture on poly(tri[ethylene glycol] ethyl ether methacrylate) (P[TEGMA-EE]), a thermoresponsive biocompatible surface. Cultures were grown for 72 hours as a continuous layer on P(TEGMA-EE). Assays for genotoxicity, cell morphology, and fluorescence-assisted flow cytometry were performed to exclude adverse effects. A gene expression profile related to the extracellular matrix was investigated by microarray analysis. For fibroblast monocultures and fibroblast/keratinocyte co-cultures maintained for 72 hours on P(TEGMA-EE), no change in morphology or specific surface markers, or DNA damage (comet assay) was observed, relative to control surface. Moreover, no detrimental impact was ascertained based on microarray analysis. In response to lowered temperature, the detachment of a continuous cell layer sheet from the thermoresponsive surface was observed. When gene expression was compared between fibroblasts cultured alone and co-cultured with keratinocytes on P(TEGMA-EE), 10 genes were shown to be differentially expressed. Of these genes, six were significantly differentially expressed between cultures grown on P(TEGMA-EE) and human skin samples. Our results indicate that P(TEGMA-EE) is fully biocompatible and is therefore a suitable surface for successful preparation and recovery of two-layered fibroblast/keratinocyte co-culture as a continuous sheet of cells.

Towards understanding Pseudomonas aeruginosa burn wound infections by profiling gene expression.

Pseudomonas aeruginosa is a key opportunistic pathogen causing severe acute and chronic nosocomial infections in immunocompromised or catheterized patients. It is prevalent in burn wound infections and it is generally multi-drug resistant. Understanding the genetic programs underlying infection is essential to develop highly needed new strategies for prevention and therapy. This work reviews expression profiling efforts conducted worldwide towards gaining insights into pathogenesis by P. aeruginosa, in particular in burn wounds. Work on various infection models, including the burned mouse model, has identified several direct virulence factors and elucidated their mode of action. In vivo gene expression experiments using In vivo Expression Technology (IVET) ascertained distinct regulatory circuits and traits that have helped explain P. aeruginosa s success as a general pathogen. The sequencing of the whole genome from a number of P. aeruginosa strains and the construction of genome-wide microarrays have paved the road to the several insightful studies on the (interacting) traits underlying infection. A series of in vitro and initial in vivo gene expression studies revealed specific traits pivotal for infection, such as quorum sensing systems, iron acquisition and oxidative stress responses, and toxin production among others. The data sets obtained from global transcriptional profiling provide insights that will be essential for the development of new targets and options for prevention and intervention.

Computerized planimetry evaluation of hyperbaric oxygen therapy in the treatment of diabetic foot.

BACKGROUND: Diabetic foot ulcer is one of the major complications of diabetes mellitus in adults. OBJECTIVES: The aim of the study was to conduct a planimetry evaluation of the effectiveness of hyperbaric oxygen therapy (HBOT) in the treatment of patients with vascular disorders caused by diabetic foot. MATERIAL AND METHODS: The study included 94 patients, 30 females (32%) and 64 males (68%), aged 33-76 years, with diabetes lasting 1.5-32 years, who underwent HBOT due to diabetic foot. All patients from that group underwent vascular procedures prior to HBOT. In qualifying patients for hyperbaric oxygen therapy, transcutaneous oximetry method was applied (30-60 exposures in hyperbaric oxygen at pressure of 2.5 ATA). Progress in wound healing was evaluated by computerized planimetry system IRIS 4. RESULTS: In 26 patients the wounds were completely closed and in 37 patients the topical state was significantly improved - the wound surface decreased by 34% in average. During the treatment, in 11 patients amputation of fingers and metatarsal necrotic bones was performed, while in 9 patients amputation was prevented. CONCLUSIONS: A planimetry evaluation showed that the application of HBOT in the treatment of diabetic foot enhances foot ulcer healing, reduces tissue damage, contributes to the reduction of complications related to soft tissue and bone infections.

Atomic force microscopy in the production of a biovital skin graft based on human acellular dermal matrix produced in-house and in vitro cultured human fibroblasts.

The most efficient method in III° burn treatment is the use of the autologous split thickness skin grafts that were donated from undamaged body area. The main limitation of this method is lack of suitable donor sites. Tissue engineering is a useful tool to solve this problem. The goal of this study was to find the most efficient way of producing biovital skin substitute based on in house produced acellular dermal matrix ADM and in vitro cultured fibroblasts. Sixty samples of sterilized human allogeneic skin (that came from 10 different donors) were used to examine the influence of decellularizing substances on extracellular matrix and clinical usefulness of the test samples of allogeneic human dermis. Six groups of acellular dermal matrix were studied: ADM-1 control group, ADM-2 research group (24 h incubation in 0.05% trypsin/EDTA solution), ADM-3 research group (24 h incubation in 0.025% trypsin/EDTA solution), ADM-4 research group (24 h incubation in 0.05% trypsin/EDTA solution and 4 h incubation in 0,1% SDS), ADM-5 research group (24 h incubation in 0.025% trypsin/EDTA solution and 4 h incubation in 0,1% SDS), and ADM-6 research group (24 h incubation in 0,1% SDS). Obtained ADMs were examined histochemically and by atomic force microscopy (AFM). ADMs were settled by human fibroblasts. The number of cultured cells and their vitality were measured. The obtained results indicated that the optimal method for production of living skin substitutes is colonization of autologous fibroblasts on the scaffold prepared by the incubation of human allogeneic dermis in 0.05% trypsin/EDTA. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 726-733, 2018.

A review of decellurization methods caused by an urgent need for quality control of cell-free extracellular matrix' scaffolds and their role in regenerative medicine.

The natural extracellular matrix (ECM),thanks to its specific properties (e.g., collagenous lattice, a reservoir of growth factors, ECM-cell anchoring areas, an optimal pH and CO2 ),ensures an optimal microenvironment for homeostatic and regenerative cell development. In the context of regenerative medicine, ECM is a lair for residual and infiltrative cells. The aim of the clinical usage of cell-free ECM scaffolds is the enhancement of tissue regeneration with possible minimization of an adverse host reaction on allogeneic or xenogeneic biomaterial. Thus, the objective of decellularization is to obtain acellular grafts characterized by optimal biological properties, such as a lack of remaining cellular elements (e.g., cell membrane phospholipids and proteins, nucleic acids, mitochondria), lack of immunogenicity, lack of calcification promotion and lack of cytotoxicity (e.g., in unrinsed detergents). Furthermore, cell-free ECM scaffolds should present the optimal mechanical and structural properties that may ensure the biocompatibility of the graft. The maintenance of the ultrastructure composition of the ECM is one of the most important goals of decellularization. All physical, chemical, and biological methods proposed (used separately or in combination to extract cells from tissues/organs) are not 100% effective in cell removal and always cause a disruption of the ECM texture, as well as a probable loss of important structure components. Although cell-free ECM scaffolds are generally classified as medical devices, there are no widely accepted or legally defined criteria for quality control/evaluation methods of obtained matrices. Such criteria must be provided. Some of them have been proposed in this manuscript. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 909-923, 2018.

The role of dimethyl sulfoxide (DMSO) in ex-vivo examination of human skin burn injury treatment.

Dimethyl sulfoxide (DMSO) is one of the most versatile solvents in biological science, therefore it is frequently used as a solvent in biological studies and as a vehicle for drug therapy. DMSO readily penetrates, diffuses through biological membranes and ipso facto increases fluidity of liposomal membranes modelling stratum corneum. Thermal injury is associated with the appearance of lipid peroxidation products in the burned skin. The influence of DMSO on protein structure and stability is concentration and temperature dependant. The aim of this study was to assess the impact of DMSO on human burn wounds and examine the interactions between DMSO and skin surface. The real problem in burn treatment is hypoalbuminemia. At the level of the laboratory studies there was an attempt at answering the question of whether the DMSO will modify the standard serum solution. In the case of the incubation of skin fragments in 1%-100% DMSO, the following findings were reported: modification of the serum, appearance of low molecular weight oligomer bands, disappearance of albumin bands or reconstruction of native serum bands during incubation in antioxidant solutions. The result of the modification is also the exposure of FTIR 1603 and 1046cm-1 bands observed in frozen serum solutions. In the case of modification of the burned skin by DMSO solutions or antioxidants - frequency shifts, an increase in the intensity of amide I band as well as the appearance of the 1601cm-1 band can be specific biomarkers of the tissue regeneration process. In this study the areas 1780-1580cm-1 and 1418-1250cm-1 on the Raman spectra are particularly rich in spectral information.

The Isolation and Production of the Ready-to-Use Product (the Amniotic Stem Cell Culture) in Accordance with Good Manufacturing Practice Regulations.

According to the Committee for Advanced Therapies, amniotic stem cells were classified as an advanced therapy medicinal product. This work aims to standardize the isolation of amniotic stem cells and the selection of the optimal time of transplantation and cell application methods in burn patients according to the guidelines of the Good Manufacturing Practice. The placenta used in the study was sourced during a Cesarean section. The remnants of the amnion preparation were placed in a sterile container and transferred to a class B environment, where the primary cultures began. The highest average number of cells was obtained by tissue homogenization and culture growth on the AmnioGrow medium. The isolation of the pure monoculture should be performed using the antibodies against CD105. On the basis of an analysis of population doubling, the aging of a population, the cells' viability, and the severity of injury, the cells should be used between passages 3 and 6. Significant differences were found in the number and viability of cells that were transferred as a full sheet, depending on the transfer method. To sum up, amniotic cells are a promising source in the treatment of burns and can be used as a hospital exemption.