Factors which impact the length of hospitalisation and death rate of COVID-19 patients based on initial triage using capillary blood gas tests: a single centre study.

An important element in the effective treatment of patients infected with the SARS-CoV-2 virus during the pandemic is an effective early triage to determine patient allocation and in-patient therapy. This paper assesses the prognostic value of capillary blood gas tests in predicting extended hospitalisation and death due to COVID-19. This retrospective statistical research is based on a group of 200 patients, hospitalised from 15 October 2020 to 08 March 2021. The study utilised the treatment documentation of these patients hospitalised due to COVID-19 at the Pulmonology and Thoracic Surgery Centre in Bystra (Southern Poland) during this period. The hospital has 50 beds with access to oxygen for COVID-19 patients and a five-bed intensive care unit. On the basis of the obtained results, conclusions were drawn that the need for early oxygen therapy with an oxygen mask and low pH values in capillary blood are significant risk factors for prolonging hospitalisation due to COVID-19. Age, the need for early oxygen mask therapy and low oxygen saturation are important risk factors for death from COVID-19. Capillary blood gas analysis is a simple and effective method of early in-patient segregation of COVID-19 patients.

Orbital reconstruction - applied materials, therapeutic agents and clinical problems of restoration of defects.

Reconstruction of large cavities in the skull and facial regions is important not only to restore health but also for the correction of facial distortions. Every visible deformity in the facial region of the patient affects their mental wellness and perception by society, entailing both, deterioration of health, but also a decrease in the performance in society, which translates into its productivity. With the progressive degradation of the natural environment, cancer, in the coming years, will be on the leading causes of morbidity and mortality. The review focuses on two main aspects: (i) the causes of injuries leading to the necessity of removal of orbital cavities occupied by the tumor and then their reconstruction, with the focus on the anatomical structure of the orbital cavity, (ii) the materials used to reconstruct the orbital cavities and analyze their advantages and disadvantages. The manuscript also underlines the not yet fully met challenges in the area of facial- and craniofacial reconstruction in people affected by cancer.

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.

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.

The Effect of Lauric Acid on Pathogens Colonizing the Burn Wound: A Pilot Study.

OBJECTIVE: Some of the most serious complications of burns include septic infections. Instead of fulfilling the function of a protective barrier, tissues damaged by high temperature create a niche that serves as an environment and source of nourishment for pathogens. An accepted practice is to use antibiotics to inhibit development of pathogens. Taking into consideration the characteristics of the burn wound and increasing antibiotic resistance, the search for new substances that have both antimicrobial and regenerative effects seems justified. The aim of the study was to determine the influence of lauric acid on bacteria-colonizing tissue samples taken during surgical treatment of burns. METHODS: Lauric acid was combined with 5 different ointment bases: Anhydrous Eucerin DAB, Anhydrous Eucerin II, Hydrophilic Vaseline, White Vaseline, and Lekobaza. The content of lauric acid in the ointment bases was 10% to 20% w/w. The preparations were applied onto samples of burnt skin collected during surgery. The samples were subsequently subjected to a microbiological test with the use of model strains of Pseudomonas aeruginosa, Staphylococcus aureus, and Escherichia coli. RESULTS: With one exception (White Vaseline), lauric acid showed a more pronounced effect on bacteria in 20% w/w concentration. In a 10% lauric acid concentration, no effect on bacteria was observed on the Hydrophilic Vaseline ointment base. Lauric acid had the strongest inhibiting effect on microbial growth of Gram-positive Staphylococcus aureus. Satisfactory zones of inhibition were also observed in the case of Escherichia coli. Growth inhibition of Pseudomonas aeruginosa was observed only when pure lauric acid was used. CONCLUSIONS: Due to its aseptic and regenerative effect on chemically damaged tissues, lauric acid can be a promising modifier of the burn healing process.

A Histologic Perspective on Electrical and Thermal Burn-Injured Human Skin.

OBJECTIVE: To analyze specific spectroscopic (FT-Raman) and thermal (limiting oxygen index) aspects of skin samples exposed to electrical injury compared with thermal injury. METHODS: An observational case-control study was conducted at the Dr Stanislaw Sakiel Center for Burns Treatment in Siemianowice, Silesia, Poland. A scanning electron microscope was used to diagnose and illustrate the topography of skin samples from electrical and thermal burns and the morphologic effects on damaged versus undamaged skin surfaces. In particular, researchers attempted to detect spectroscopic and thermal changes at the molecular level, namely, specific biomarkers of tissue degeneration and their regeneration under the influence of the applied modifiers (antioxidants and orthosilicic acid solutions). RESULTS: Modification with L-ascorbic acid and hydrogel of orthosilicic acid caused an increase in the intensity of the amide I Raman peaks, whereas modification with sodium ascorbate and orthosilicic acid resulted in the separation of the band protein side chains (1,440-1,448 cm), which is a part of tissue regeneration. The best result was obtained when the skin was treated with 7% orthosilicic acid (limiting oxygen index, 26%). CONCLUSIONS: Antioxidant treatment may be advantageous in minimizing injury in patients with thermal burns but not always in electrical burns.

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.

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.

Isolation, culturing and preparation for transplantation of amniotic mesenchymal stem cells Repetitive and reproducible laboratory, technical protocol.

To date, different methods of isolation of amniotic stem cells have been developed. Our previous studies have demonstrated that there are significant differences in viability and efficiency of the isolation and culture process depending on the enzyme and medium used. The aim of this study was to present efficient protocol, which can be used within good manufacturing practise conditions. Amniotic membranes were obtained from ten woman 31-39 years old who signed informed constent. GMP regulations are applicable. The described protocol aims to obtain a clinically significant cell yield (>1*108). The cells may be maintained in the growth phase even for 2 months. The mesenchymal cells constitute about 75-95% of the cells in primary culture. Supervisory authorities require repetitive and reproducible laboratory protocol for stem cells culture. Presented protocol allow achieving clinically significant cell yield (>1*108) in 4-5 weeks. Cells can be transplanted as suspension or cell sheet.

A New Triage Method for Burn Disasters: Fast Triage in Burns (FTB).

BACKGROUND There are few triage methods based on patient age and extent of burn. However, in case of mass casualties, age might be hard to define. Burn segregation in mass-casualty accidents requires an easy, fast, and effective method. Triage in burns should also segregate casualties requiring treatment in burn centers. The aim of this study was to create a proprietary segregation algorithm dedicated to mass-casualty incidents. MATERIAL AND METHODS A retrospective analysis of 939 burned patients admitted to the Clinical Department of Burns, Plastic and Reconstructive Surgery, Military Institute of Medicine (MIM) in Warsaw and to the Center for the Treatment of Burns (CTB) in Siemianowice Slaskie in 2012 and 2013 was performed. The aim was to reveal which early factors could be used during segregation of burn victims in mass-casualty incidents on the battlefield and in civilian circumstances. Only easy and quick-to-evaluate factors that can be examined without medical equipment and laboratory tests were used in creating the proprietary triage algorithm. RESULTS As a result of our study, we created an algorithm for fast triage in mass-casualty situations. The algorithm is based on parameters that can be easily evaluated without additional equipment. To create the algorithm, we used factors that had the strongest impact on mortality prediction in severely burned patients, in multifactor analysis: advanced age (p<0.001. OR=1.04), extent of the deep burn (p<0.001. OR=1.1), and low systolic arterial pressure (p<0.001. OR=0.96). CONCLUSIONS The FTB (Fast Triage in Burns) algorithm is a new triage method dedicated for massive burn events in civilian circumstances. The FTB algorithm is a simple, quick, and credible means of segregating burn victims. The algorithm is dedicated to use in pre-hospital care, during mass-casualty events both in civilian and battlefield circumstances. The aim is to be able to evaluate burn victims immediately, without access to medical equipment or additional tests and to evaluate indications for burn center care. It is a unique method designed to be used during segregation in isolated burn mass-casualty incidents.

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.

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.

Poly[tri(ethylene glycol) ethyl ether methacrylate]-coated surfaces for controlled fibroblasts culturing.

Well-defined thermosensitive poly[tri(ethylene glycol) monoethyl ether methacrylate] (P(TEGMA-EE)) brushes were synthesized on a solid substrate by the surface-initiated atom transfer radical polymerization of TEGMA-EE. The polymerization reaction was initiated by 2-bromo-2-methylpropionate groups immobilized on the surface of the wafers. The changes in the surface composition, morphology, philicity, and thickness that occurred at each step of wafer functionalization confirmed that all surface modification procedures were successful. Both the successful modification of the surface and bonding of the P(TEGMA-EE) layer were confirmed by X-ray photoelectron spectroscopy (XPS) measurements. The thickness of the obtained P(TEGMA-EE) layers increased with increasing polymerization time. The increase of environmental temperature above the cloud point temperature of P(TEGMA-EE) caused the changes of surface philicity. A simultaneous decrease in the polymer layer thickness confirmed the thermosensitive properties of these P(TEGMA-EE) layers. The thermosensitive polymer surfaces obtained were evaluated for the growth and harvesting of human fibroblasts (basic skin cells). At 37 °C, seeded cells adhered to and spread well onto the P(TEGMA-EE)-coated surfaces. A confluent cell sheet was formed within 24 h of cell culture. Lowering the temperature to an optimal value of 17.5 °C (below the cloud point temperature of the polymer, TCP, in cell culture medium) led to the separation of the fibroblast sheet from the polymer layer. These promising results indicate that the surfaces produced may successfully be used as substrate for engineering of skin tissue, especially for delivering cell sheets in the treatment of burns and slow-healing wounds.

In-vivo expression profiling of Pseudomonas aeruginosa infections reveals niche-specific and strain-independent transcriptional programs.

Pseudomonas aeruginosa is a threatening, opportunistic pathogen causing disease in immunocompromised individuals. The hallmark of P. aeruginosa virulence is its multi-factorial and combinatorial nature. It renders such bacteria infectious for many organisms and it is often resistant to antibiotics. To gain insights into the physiology of P. aeruginosa during infection, we assessed the transcriptional programs of three different P. aeruginosa strains directly after isolation from burn wounds of humans. We compared the programs to those of the same strains using two infection models: a plant model, which consisted of the infection of the midrib of lettuce leaves, and a murine tumor model, which was obtained by infection of mice with an induced tumor in the abdomen. All control conditions of P. aeruginosa cells growing in suspension and as a biofilm were added to the analysis. We found that these different P. aeruginosa strains express a pool of distinct genetic traits that are activated under particular infection conditions regardless of their genetic variability. The knowledge herein generated will advance our understanding of P. aeruginosa virulence and provide valuable cues for the definition of prospective targets to develop novel intervention strategies.

[Long-term results of airways' injury after a methane explosion]. / Odlegle skutki oparzenia dróg oddechowych w wyniku wybuchu metanu.

Injuries caused by thermal trauma more and more often affect people and they are an important problem of contemporary medicine. It is connected with the civilization development. Burn concerns not only the exterior integuments of the body but also airways, in those cases, the death rate among those who were injured with the thermal trauma increases. The treatment of both the burns and their complications is a long-lasting process, involving many specialists of various disciplines, and not always does it bring the wanted effects. The aim of the study was the assessment of the late morphological sequels inside the larynx and ventilation efficiency as a result of thermal inhalation trauma in the airways of the coal miners after the methane explosion. The methodology of examinations consisted of subjective evaluation of ventilation efficacy by MRC scale and ventilating rate measurement FEV1, FVC, FEV1%, PEF, MEF50, PIF, MIF 50, FIV. The morphological larynx evaluation was conducted based on videolaryngoscopy. 23 injured coal miners, who had been burnt at work in coal mines in 2003 in the methane explosion, were put under scrutiny. All the above mentioned examinations were done to the study group, but also to the specially selected control group, 23 coal miners. During the ventilating rates analysis in the study group, their lower values were noticed in comparison to the control group, however, the values were still within the clinical norms. Videolaryngoscopy showed hypertrophy of the laryngeal tissues.

Laser in the treatment of hypertrophic burn scars.

This prospective study looked at the outcome of laser (light amplification by stimulated emission of radiation) treatment for hypertrophic scarring. Dermatrade mark K laser (a set of combined lasers erbium:yttrium aluminium garnet/carbon dioxide, qualified as a class IV laser) was used. Between 21 June 2000 and 19 November 2002, at the Siemianowice Burn Center, Poland, 592 interventions, using laser, were performed on N= 327 patients (220 women and 107 men, aged between 3 and 80 years). The majority of cases [N= 223 (68.9%)] were patients with post-burn hypertrophic scars, and 104 cases (31.8%) had various types of hypertrophic scars. Evaluation took place using an adapted Vancouver Scar Scale and digital photographs as well as the patient's opinion. It was noted that after laser treatment, satisfactory results were achieved in 72% of cases. The scars had become less red (192/327 scored no redness at the end of the study versus 92/327 upon initial), less raised (272/327 scored a flat scar versus 72/327 upon initial) and demonstrated an improved viscoelasticity (192/327 scored a soft skin versus 62/327 upon initial). Laser treatment did not improve contractures in post-burn hypertrophic scars. Results were not confirmed using objective measurement tools, as these were not available to us.