Experimental evidence for Parthanatos-like mode of cell death of heat-damaged human skin fibroblasts in a cell culture-based in vitro burn model.

The cellular mechanisms of burn conversion of heat damaged tissue are center of many studies. Even if the molecular mechanisms of heat-induced cell death are controversially discussed in the current literature, it is widely accepted that caspase-mediated apoptosis plays a central role. In the current study we wanted to develop further information on the nature of the mechanism of heat-induced cell death of fibroblasts in vitro. We found that heating of human fibroblast cultures (a 10 s rise from 37 °C to 67 °C followed by a 13 s cool down to 37 °C) resulted in the death of about 50% of the cells. However, the increase in cell death started with a delay, about one hour after exposure to heat, and reached the maximum after about five hours. The lack of clear evidence for an active involvement of effector caspase in the observed cell death mechanism and the lack of observation of the occurrence of hypodiploid nuclei contradict heat-induced cell death by caspase-mediated apoptosis. Moreover, a dominant heat-induced increase in PARP1 protein expression, which correlated with a time-delayed ATP synthesis inhibition, appearance of double-strand breaks and secondary necrosis, indicate a different type of cell death than apoptosis. Indeed, increased translocation of Apoptosis Inducing Factor (AIF) and Macrophage Migration Inhibitory Factor (MIF) into cell nuclei, which correlates with the mentioned enhanced PARP1 protein expression, indicate PARP1-induced, AIF-mediated and MIF-activated cell death. With regard to the molecular actors involved, the cellular processes and temporal sequences, the mode of cell death observed in our model is very similar to the cell death mechanism via Parthanatos described in the literature.

How did the COVID-19 pandemic affect burn centres in German-speaking countries?

The exponential growth of COVID-19 cases in early 2020 presented a massive challenge for healthcare systems and called for the adaptation of emergency care routines and intensive care capacities. We, therefore, analyzed a possible impact of the COVID-19 pandemic on the general structure and emergency preparedness of burn centers in German-speaking countries through a cross-sectional descriptive survey questionnaire. The survey was conducted for the first time in January 2019 by Al-Shamsi et al. before the beginning of the COVID-19 pandemic. It was performed for a second time in November 2020 during the second wave of COVID-19 infections in German-speaking countries. We noticed a pronounced increase in the preparation for a great number of patients in need of intensive care including the enlargement of overall capacity when necessary. We also showed a notable decrease in the specific preparation for burn disasters and also reduced communication with first responders and other burn centers. To what extent these alterations were caused by the impact the pandemic had on healthcare systems could not be determined in this study and should be the subject of future research.

Which Moisturizer to Use in Scar Therapy after Burn Injuries? A Subjective and Objective Skin and Scar Evaluation after Topical Treatment with Dexpanthenol, Aloe Vera, and Plant Oil.

Background and Objectives: Good scar management in burn care is essential. Nevertheless, there are no consistent recommendations regarding moisturizers for scar management. Our aim was to investigate and compare the effects of commonly used products on normal skin and burn scars. Materials and Methods: A total of 30 skin-healthy (control group) and 12 patients with burn scars were included in this study. For an intraindividual comparison, each participant received creams consisting of dexpanthenol (P), aloe vera (A), and a natural plant oil (O) with instructions to apply them daily to a previously defined area for at least 28 days. Objective scar evaluation was performed with Visioscan®; Tewameter®; Cutometer®, and the Oxygen To See® device. Subjective evaluation was performed with an "application" questionnaire, the Patient and Observer Scar Assessment Scale (POSAS), and with the "best of three" questionnaire. Results: After (A) a high trend of amelioration of +30%, TEWL was detected on the scar area. Blood flow increased slightly on healthy skin areas after (A) application to +104%. The application of (A) on healthy skin demonstrated a positive effect on the parameters of scaliness (+22%, p < 0.001), softness (+14%, p = 0.046), roughness R1 (+16%, p < 0.001) and R2 (+17%, p = 0.000), volume (+22%, p < 0.001), and surface area (+7%, p < 0.001) within the control group. After (P), a significant improvement of the baseline firmness parameter of +14.7% was detected (p = 0.007). (P) also showed a beneficial effect on the parameters of R1 (+7%, p = 0.003), R2 (+6%, p = 0.001), and volume (+17%, p = 0.001). (O) lead to a statistically significant improvement of volume (+15%, p = 0.009). Overall, most study participants stated (A) to be the "best of three". Conclusions: (A) performed statistically best, and is a well-tolerated moisturizing product. However, further quantitative studies are needed to provide statistically significant clarification for uniform recommendations for scar therapy.

Comparative Clinical Study of Suprathel® and Jelonet® Wound Dressings in Burn Wound Healing after Enzymatic Debridement.

Following the enzymatic debridement of deep dermal burns, the choice of wound dressing is crucial for providing an adequate environment and suitable conditions for rapid wound healing. As Suprathel® and fatty gauze (Jelonet®) are the most commonly used dressings in burn centers, the aim of this study is to compare Suprathel® and Jelonet® in the treatment of deep dermal burns after enzymatic debridement with respect to wound healing, patient comfort, and pain. A total of 23 patients with deep dermal burns of the hand or foot (mean total body surface area of 4.31%) were included in this prospective, unicentric, open, comparative, and intra-individual clinical study. After enzymatic debridement, wounds were divided into two areas: one was treated with Suprathel® and the other with Jelonet®. Suprathel® remained on the wounds without dressing changes while Jelonet® was regularly changed. Wound healing, infection, bleeding, exudation, time for dressing changes, and pain were documented (from days 2 to 48) after injury. Satisfactory results were obtained in 22 cases; only one patient had to undergo a second debridement followed by skin grafting. No significant difference in time to final wound healing could be observed (18-19 d). Patients reported significantly less pain during the dressing changes for Suprathel® compared to Jelonet®. Furthermore, the wound areas treated with Suprathel® showed significantly less exudation and bleeding. Wound infections rarely occurred in both groups. In conclusion, the authors found that both wound dressings could be used to achieve safe and rapid wound healing after the enzymatic debridement of deep dermal burns of the hands and feet. However, compared to Jelonet®, Suprathel® showed superior results in terms of patient comfort and pain reduction.

The performance of clinical risk scores in the diagnosis of necrotising fasciitis.

OBJECTIVE: Necrotising fasciitis (NF) is a quickly progressing and potentially life-threatening infection, involving the fascia and subcutaneous tissues. The diagnosis of this disease is challenging, especially due to a lack of specific clinical signs. In order to ensure a better and quicker identification of NF patients, a laboratory risk indicator score has been developed for NF (LRINEC). A variant has widened this score by adding clinical parameters (modified LRINEC). This study shows current outcomes of NF and compares the two scoring systems. METHODS: This study was conducted between 2011 and 2018, and included patient demographics, clinical presentations, sites of infection, comorbidities, microbiological and laboratory findings, antibiotic therapies and LRINEC as well as modified LRINEC scores. The primary outcome was in-hospital mortality. RESULTS: A cohort of 36 patients, diagnosed with NF, were included in this study. The mean hospital stay was 56 days (±38.2 days). The mortality rate in the cohort was 25%. The sensitivity of the LRINEC score was 86%. Calculation of the modified LRINEC score showed an improvement of the sensitivity to 97%. The average LRINEC score and modified LRINEC score for patients who died and who survived were equal (7.4 versus 7.9 and 10.4 versus 10.0, respectively). CONCLUSION: The mortality rate of NF remains high. The modified LRINEC score increased the sensitivity in our cohort to 97%, and this scoring system could be supportive in the diagnosis of NF for early surgical debridement.

Nexobrid Treatment for Burn Injuries in Patients With Chronic Obstructive Pulmonary Disease and Home Oxygen Therapy.

There is an increased risk for burn injuries associated with home oxygen therapy of patients with chronic obstructive pulmonary disease (COPD) since 10% to 50% of these patients continue to smoke. Enzymatic eschar removal of facial burns is gaining popularity but intubation of this specific patient group often leads to prolonged weaning and can require tracheostomy. This study dealt with the question if enzymatic debridement in these patients can also be performed in analgosedation. A selective review of the literature regarding burn trauma associated with home oxygen use in patients with COPD was performed, as well as a retrospective analysis of all patients with burn injuries associated with home oxygen use and COPD that were admitted to the study clinic. In the literature, 1746 patients with burns associated with home oxygen use are described, but none of them received enzymatic debridement. In this study, 17 patients were included. All three patients in this study with facial full-thickness burn injuries received enzymatic debridement. The mortality rate in this cohort was 17.6% (3/17). Up to date, there is limited experience performing regional anesthesia debridement in patients with COPD. This is the first manuscript describing the use of enzymatic debridement in patients with COPD and home oxygen therapy. We could confirm other studies that intubation of these patients leads to prolonged ventilation hours and increases the probability for poor prognosis. Therefore, we described the treatment of enzymatic debridement in analgosedation without intubation.

Lactate-Based Scoring System in the Diagnosis of Necrotizing Fasciitis.

Necrotizing fasciitis (NF) is a rare and quickly progressing infection and leads to 100% mortality if untreated. Quick diagnosis and an early and radical surgical treatment are essential for stopping bacterial progression. Unfortunately, the absence of clear clinical signs makes the diagnosis often challenging. Therefore, we searched for easy determinable predictive laboratory markers for NF. This is the first study which includes lactate values in a new score. A retrospective analysis of patients with NF (n = 44) and patients with erysipelas (n = 150) was performed. Lactate values, patients' demographics, clinical presentations, site of infection, comorbidities, microbiological and laboratory findings, antibiotic therapies, and LRINEC and modified LRINEC scores were analyzed. Logistic regression analysis was used to derive adjusted weights, and final simple point score was assessed with a ROC curve analysis. Patients with NF had a mean age of 57 years and patients with erysipelas 65 years. The median hospital length of stay was 8 and 49 days in patients with erysipelas and NF, respectively. Although only one patient (0.7%) in the group of erysipelas died, the mortality rate of patients with NF was 9/44 (20.5%). The lactate values were statistically significant higher in the NF group, 4.1 vs 2.0 mmol/l (P < .001). The new created CologNe-FaDe-score shows the highest AUC-value with 0.907. With the help of lactate values, the CologNe-FaDe-score consists of easily practicable and highly available parameters, which could sensitize diagnosis.

Is the Parkland formula still the best method for determining the fluid resuscitation volume in adults for the first 24 hours after injury? - A retrospective analysis of burn patients in Germany.

BACKGROUND: R Rapid fluid resuscitation is a crucial therapy during the treatment of patients with extensive burns. In 1968, the Parkland Formula was introduced for the calculation of the estimated volume of the resuscitation fluid. Since then, different methods for the calculation of fluid resuscitation volume have been developed. We aimed to evaluate if the Parkland formula is still the most effective method for fluid resuscitation volume calculation in burn patients. METHODS: In the period between January 2015 and January 2019, data from 569 patients over 16 years old with burns of more than 20% total body surface area (TBSA) and at least 15% TBSA full thickness burns were entered in the German burn registry. The patients were divided into 5 groups (0, +1, -1, +2, -2) according to the volume of the resuscitation fluid they received. Group 0 patients received the amount of fluid calculated according to the Parkland formula (n = 83). The 4 other groups received reduced (-1, -2) or increased (+1, +2) fluid volumes in comparison to the value obtained by the Parkland formula. RESULTS: Patients in Group 0 presented a significantly lower mortality in the first week (4.5%) compared to groups -2 (16.7%) and group +2 (19.5%) (p = 0.021). Furthermore, the mean number of operations in group +2 (5.81) was higher than in group -2 (3.81). Surviving patients from group +2 presented a longer hospital stay (68.1 days) compared to the other groups. Additionally, the logistic regression analysis showed a higher survival of patients in groups -2 and -1 (regression coefficients -0.11 and -0.086; Odds Ratio 0.896 and 0.918; 95% Confidence Interval (CI) 0,411-1.951 and 0.42-2.004). CONCLUSION: In this retrospective study, register based analysis a restrictive fluid regime was associated with a higher survival compared to the liberal Parkland guided fluid regime.

[Where there´s smoke - there´s no fire? - Burns from E-Cigarette explosions]. / Viel Rauch und sonst nichts? ­ Verbrennungsfolgen durch E-Zigaretten-Gebrauch.

INTRODUCTION: The popularity of E-Cigarettes is increasing. Besides addiction and pulmonary health damage, reports of burn injuries from e-cigarette explosions are also increasing. Mostly, explosions of e-cigarettes are attributed to its lithium-ion battery. Due to increasing cases and missing guidelines we want to present three cases of our hospital and publish recommendations for the management of burn injuries caused by e-cigarette explosions. PATIENTS/MATERIAL AND METHODS: Three cases of e-cigarette explosions which occurred between 2016 and 2019, are presented. RESULTS: All three e-cigarette explosions occurred in the trouser pockets. Two patients were male one patient was female. The age ranged from 24 to 64 years, the burned total body surface area (TBSA) from 3 % to 12.5 %. All three patients required skin grafting and the length of stay in hospital ranged from five to eleven days. CONCLUSION: In the synopsis of recent literature, we recommend the following management of burns due to e-cigarette explosions. The guidelines of the Advanced Trauma Life Support should be followed, signs of an inhalation trauma should be checked and litmus test should be performed prior to irrigation with aqueous solutions to prevent exothermic reactions with remaining metals. If litmus test shows alkali pH wounds should be irrigated by mineral oil.

Are burn centers in German-speaking countries prepared to respond to a burn disaster? Survey-based study.

Burn disasters present a challenge not only to burn centers but the entire healthcare system. Most burn centers worldwide are unprepared to deal with a burn disaster as it is an uncommon event. We investigated the status of burn center preparedness in German-speaking countries to respond to a burn disaster. Self-administered survey questionnaires were sent to the directors of burn centers; the questions of survey used before in a similar way in Belgium were translated into German language. Of the 46 questioned burn centers, 32 (78%) responded, including all of the German adult burn centers. A clear difference in the preparation status of the burn centers in the three countries was observed due to geopolitical factors such as decentralized healthcare systems. However, the healthcare system is generally well-prepared concerning command, transfer, and capacity to provide sustained supplies to handle a massive influx of patients. Nevertheless, there are some gaps in the areas of planning and preparation, funding for disaster activities, and regular training of staff for burn disasters. We call for a unified burn disaster plan and increased cooperation between burn centers and civil defense regarding communication and training. We strongly recommend the implementation of a special disaster fund and telemedicine in disaster management to circumvent shortages in burn staff.

Vitamin D deficiency may stimulate fibroblasts in Dupuytren's disease via mitochondrial increased reactive oxygen species through upregulating transforming growth factor-ß1.

Dupuytren's disease, a benign fibroproliferative disorder of the palmar fascia, represents an ideal model to study tissue fibrosis. Transforming growth factor-ß1 (TGF-ß1) and its downstream Smad signaling system is well established as a keyplayer during fibrogenesis. Vitamin D has been extensively studied as an anti-fibrotic agent in malignant chronic diseases. A number of studies have shown that myofibroblasts are main target cells of 1,25(OH)2D3 inhibitory action. The myofibroblast in the palmar aponeurosis of patients in different stages of Dupuytren's disease was found by electron microscopy to contain a large number of mitochondria. Mitochondria play a critical role in cell metabolism being the major source of reactive oxygen species (ROS) in cells. TGF-ß1 has been shown to increase mitochondrial ROS production in different cell types, which mediate fibrosis related gene expression and myofibroblast differentiation. TGF-ß1 increases mitochondrial ROS production in patients with Dupuytren's contracture potentially in consequence of Vitamin D deficiency, leading to myofibroblast differentiation. Thus, targeting this basic pathomechanism seems suitable to establish new treatment strategies.

Smooth and textured silicone surfaces of modified gel mammary prostheses cause a different impact on fibroproliferative properties of dermal fibroblasts.

Our study addressed the question of whether surface properties of modern standard gel prostheses may have a measurable impact on the fibrogenic properties of cultured human dermal fibroblasts. Fibroblasts were isolated from breast implants by using the explant culture technique and incubated either on smooth or on textured silicone elastomere surfaces. Fibroblast growth was observed 4 weeks following incubation. Expression of transforming growth factor (TGF)-ß1 was measured after cell culture. Incubated fibroblasts on textured surfaces showed a fivefold lower growth rate during all experiments. TGF-ß1 expression was lowered in smooth surface fibroblasts compared with textured surface cultures. Our results show that smooth and textured silicone surfaces of modified gel breast implants have a different impact on the fibroproliferative properties of dermal fibroblasts. These preliminary results seem promising and we aim to further perform qualitative and quantitative analyses of the inflammatory processes in the environment of the implant and their link to the TGF-ß pathway.

Increased metabolic activity of fibroblasts derived from cords compared with nodule fibroblasts sampling from patients with Dupuytren's contracture.

BACKGROUND: The pathogenesis of Dupuytren's contracture is characterized by progress in several stages. The disease process includes two structurally distinct fibrotic elements, the so-called nodule and the cord. Several hypotheses have been proposed regarding the cause. Until now, there has been ongoing discussion regarding whether cords or nodules are the primary source of the disease. METHODS: Tissues from 24 patients with Dupuytren's disease were obtained during hand surgery, providing samples from nodules and cords for cell culture. Tissue from the flexor retinaculum excised from healthy patients during surgical procedures on the hand served as control. By using the explant culture technique, fibroblasts were isolated and grown in standard medium. Besides morphological analysis, XTT proliferation tests were performed at various time periods to measure the metabolic activity of the cultivated fibroblasts. RESULTS: Statistical analysis revealed significant differences of nodule- and cord-derived fibroblasts after 1 hour (p = 0.0150), 8 hours (p < 0.0001), and 16 hours (p < 0.0001). CONCLUSIONS: The authors suggest that distinct proliferative properties of Dupuytren-derived fibroblasts are dependent on the cells' origin. Conclusions about the cause or pathogenesis cannot yet be drawn. Further investigation concerning their apparently different fibroproliferative properties is necessary.

N-acetyl-L-cysteine abrogates fibrogenic properties of fibroblasts isolated from Dupuytren's disease by blunting TGF-beta signalling.

Dupuytren's disease, a benign fibroproliferative disorder of the palmar fascia, represents an ideal model to study tissue fibrosis. Transforming growth factor-beta1 (TGF-beta1) and its downstream Smad signalling system is well established as a key player during fibrogenesis. Thus, targeting this basic pathomechanism seems suitable to establish new treatment strategies. One such promising treatment involves the substance N-acetyl-L-cysteine (NAC), shown to have antifibrotic properties in hepatic stellate cells and rat fibroblasts. In order to investigate antifibrotic effects of N-acetyl-L-cysteine (NAC), fibroblasts were isolated from surgically resected fibrotic palmar tissues (Dupuytren fibroblasts, DF) and exposed to different concentrations of NAC and recombinant TGF-beta1. Fibroblasts isolated from tendon pulleys served as controls (control fibroblasts, CF). Smad signalling was investigated by a Smad binding element driven reporter gene analysis. Both cell types express TGF-beta1, indicating autocrine signalling in DF and CF. This was confirmed by comparing reporter gene activity from LacZ and Smad7 adenovirus infected cells. NAC treatment resulted in abrogation of Smad mediated signalling comparable to ectopically overexpressed Smad7, even when the cells were stimulated with recombinant TGF-beta1 or ectopically expressed a constitutively active TGF-beta receptor type I. Additionally, NAC dose-dependently decreased expression of three major indicators of impaired fibrotic matrix turnover, namely alpha-smooth muscle actin (alpha-SMA), alpha 1 type I procollagen (Col1A1), and plasminogen activator inhibitor-type I (PAI-1). Our results suggest that TGF-beta signalling and subsequent expression of fibrogenesis related proteins in Dupuytren's disease is abrogated by NAC thus providing a basis for a therapeutic strategy in Dupuytren's disease and other fibroproliferative disorders.

Accelerated wound healing by in vivo application of keratinocytes overexpressing KGF.

Epidermal regeneration is a complex process, strongly influenced by growth factors, including keratinocyte growth factor (KGF). The objective of this study was to establish immortalized HaCaT keratinocytes and KMST-6-fibroblasts stably expressing KGF. Transfection efficiency, genomic integration, and functionality of the transgene were determined by ELISA and PCR, and KGF-expressing clones were selected using an air-liquid interface test system. HaCaT cells displayed stronger transgene expression compared to transfected fibroblasts, and the most effective HaCaT clone was incubated on a membrane carrier to form a "membrane cell graft." Twenty-one superficial second-degree burn wounds were created in each of three pigs, and wound healing capacity of the generated "polypeptide cell delivery system" after grafting was examined. Untransfected HaCaT keratinocytes and membrane-covered and untreated burn wounds served as controls. Histological and macroscopical follow-up revealed that grafting of transfected HaCaT cells resulted in complete reepithelialization within 5 days, while wounds covered with untransfected cells needed 2 days longer. At untreated sites, a thin epithelium was detectable after 10 days. The results indicate that wound healing processes can be stimulated distinctly by growth factors secreted from HaCaT cells, with a prominent role for transgenic KGF.