Larval Wound Therapy: Possibilities and Potential Limitations-A Literature Review.

Patient-centered care (PCC) is recognized as a standard in healthcare for determining high quality. Honoring patients' values, experiences, needs, and preferences in devising, coordinating, and delivering care underscores the enhancement of the therapeutic rapport between patients and healthcare providers. Wound treatment involves a multi-stage process encompassing diagnostics and local wound dressing, which reduces the risk of infection through a coordinated interdisciplinary team. Within this team, nurses undertake specific professional functions and roles. The implementation of local therapy using innovative and scientifically substantiated methods may be hindered by a deficit of knowledge or inappropriate knowledge among staff and patients themselves. This study presents the challenges concerning the care of patients with chronic wounds treated using Lucilia sericata larvae, based on a review of the current scientific literature. A critical analysis of the literature spanning from 2002 to 2022 was conducted using the Medline, PubMed, Cochrane, and Termedia databases, employing keywords such as "maggot debridement therapy" in relation to acceptance and perception. As a result of the preliminary selection, 472 papers were identified, of which 12 publications were included in the development of this concept. The acquired data were organized and presented in the concluding section in the form of tables, accompanied by descriptions and references to individual studies. Negative psychological and somatic sensations were among the most prominent challenges among patients treated with Maggot Debridement Therapy (MDT). Pain related to peripheral ischemia or infection in this group of patients requires pain prophylaxis, including hyperalgesia and allodynia, in order to improve method tolerance. On the other hand, augmenting patients' understanding of MDT diminishes negative emotions, reinforces positive behaviors, and mitigates anxiety levels. MDT constitutes an effective and safe method. Its widespread use for chronic wounds requires substantial knowledge among healthcare professionals and patient education, along with that of their caregivers, to develop a positive attitude.

Immunohistochemical Expression of Upregulated Gene 4 Protein Expression (URG4/URGCP) and Its Association with 5-Year Survival in Patients with Colon Adenocarcinoma.

(1) Background: Colorectal cancer (CRC) is the third most common cancer in terms of incidence and mortality. Approximately 90% of all colorectal cancer cases are adenocarcinomas, originating from epithelial cells of the colorectal mucosa. Upregulated gene 4 (URG4) is an oncogene involved in cancer development. The aim of the study was to assess the immunohistochemical expression of URG4 protein expression in Polish patients with colon adenocarcinoma who were not treated with any therapy before radical surgery. (2) Methods: The study used colon tissue samples taken from people with a confirmed diagnosis of colorectal adenocarcinoma after a thorough histopathological examination. The associations between the immunohistochemical expression of URG4 and clinical parameters were analyzed by the Chi2 test or Chi2Yatesa test. The study conducted an analysis of the correlation between the expression of URG4 and the five-year survival rate of patients through the application of the Kaplan-Meier analysis and the log-rank statistical test. The intracellular localization of URG4 was identified through the utilization of transmission electron microscopy (TEM) methodology. (3) Results: In univariate Cox regression analyses, immuno-histochemical expression of URG4, grade of histological differentiation, depth of invasion, angioinvasion, PCNA expression, stage of disease and lymph node involvement were found to be significant prognostic factors. Within our patient cohort, it was observed that the degree of tumour differentiation and URG4 expression were found to be distinct prognostic factors in regard to the 5-year survival rates of those with colon adenocarcinoma. (4) Conclusions: High immunohistochemical expression of URG4 correlates with poor prognosis in patients with colon adenocarcinoma.

The Clinical Application of Immunohistochemical Expression of Notch4 Protein in Patients with Colon Adenocarcinoma.

The Notch signalling pathway is one of the most conserved and well-characterised pathways involved in cell fate decisions and the development of many diseases, including cancer. Among them, it is worth noting the Notch4 receptor and its clinical application, which may have prognostic value in patients with colon adenocarcinoma. The study was performed on 129 colon adenocarcinomas. Immunohistochemical and fluorescence expression of Notch4 was performed using the Notch4 antibody. The associations between the IHC expression of Notch4 and clinical parameters were analysed using the Chi2 test or Chi2Yatesa test. The Kaplan-Meier analysis and the log-rank test were used to verify the relationship between the intensity of Notch4 expression and the 5-year survival rate of patients. Intracellular localisation of Notch4 was detected by the use of the immunogold labelling method and TEM. 101 (78.29%) samples had strong Notch4 protein expression, and 28 (21.71%) samples were characterised by low expression. The high expression of Notch4 was clearly correlated with the histological grade of the tumour (p < 0.001), PCNA immunohistochemical expression (p < 0.001), depth of invasion (p < 0.001) and angioinvasion (p < 0.001). We can conclude that high expression of Notch4 is correlated with poor prognosis of colon adenocarcinoma patients (log-rank, p < 0.001).

The Prognostic Significance of Apoptotic Protease Activating Factor (Apaf-1) Protein Expression in Colon Adenocarcinoma Tissue-Preliminary Report.

BACKGROUND: The Apoptotic protease activating factor 1 (Apaf-1) protein, as one of the factors involved in the activation of the mitochondrial apoptotic pathway, plays an important role in cancer biology. Apaf-1 expression in tumour cells has been shown to be downregulated, with significant implications for tumour progression. Hence, we investigated the expression of Apaf-1 protein in the Polish population of patients with colon adenocarcinoma without any therapy prior to radical surgery. Moreover, we assessed the relation between Apaf-1 protein expression and the clinicopathological factors. The prognostic activity of this protein was analyzed in relation to 5-year survival of patients. In order to show the localization of Apaf-1 protein at the cellular level, the immunogold labelling method was used. METHODS: The study was conducted using the colon tissue material from patients with histopathologically confirmed colon adenocarcinoma. Immunohistochemical expression of Apaf-1 protein was performed using Apaf-1 antibody at dilution 1:600. The associations between the immunohistochemistry (IHC) expression of Apaf-1 and clinical parameters were analyzed using the Chi2 test and Chi2Yatesa test. Kaplan-Meier analysis and the log-rank test were used to verify the relationship between the intensity of Apaf-1 expression and 5-year survival rate of patients. The results were considered statistically significant when p < 0.05. RESULTS: Apaf-1 expression was evaluated by immunohistochemical staining in whole tissue sections. Thirty-nine (33.23%) samples had strong Apaf-1 protein expression and 82 (67.77%) samples were characterized by low expression. The high expression of Apaf-1 was clearly correlated with the histological grade of the tumour (p = 0.001), proliferating cell nuclear antigen (PCNA) immunohistochemical expression (p = 0.005), age (p = 0.015), depth of invasion (p < 0.001) and angioinvasion (p < 0.001). The 5-year survival rate was significantly higher in the group of patients with high expression of this protein (log-rank, p < 0.001). CONCLUSIONS: We can conclude that Apaf-1 expression is positively correlated with reduced survival of colon adenocarcinoma patients.

The prognostic activity of acylglycerol kinase immunohistochemical expression in colon adenocarcinoma patients.

Introduction: Adenocarcinoma of the colon and rectum (COAD) is one of the most commonly diagnosed cancers of the gastrointestinal system. Acylglycerol kinase (AGK) is a known lipid kinase producing lysophosphatidic acid (LPA) from monoacylglycerol. It is widely expressed in the heart, brain, kidney, and muscle. Moreover, AGK is a significant cancer-related gene and is upregulated in many human malignancies, e.g. prostate cancer, breast cancer, oral squamous cell carcinoma, hepatocellular carcinoma, and renal carcinoma. However, the expression pattern and clinical significance of AGK in colon adenocarcinoma patients, especially in individuals living in Europe, remain unclear. Aim: The current study investigated the expression of AGK protein in colon adenocarcinoma samples to assess its prognostic significance by correlating its immunohistochemical expression with the clinicopathological variables and survival of individuals living in Poland. Material and methods: Tissue specimens were received from 110 colon adenocarcinoma patients who underwent surgical resection at the Municipal Hospital in Jaworzno in 2013-2015. The paraffin-embedded specimens were cut into 4-µm-thick sections and incubated with rabbit polyclonal antibody to AGK (final dilution 1 : 500) (Invitrogen; cat. number PA5-28566). Results and conclusions: AGK was strongly expressed in colon adenocarcinoma tissues in comparison to non-pathological colon specimens. The high level of AGK immunoexpression was demonstrated to be clearly correlated with the malignancy-related clinicopathological factors and 5-year overall survival rate of patients.

Analysis of ghrelin, leptin, and interleukin-6 salivary concentration among children aged 7-10 years and its relationship with nutritional status and some anthropometric data.

INTRODUCTION: Obesity is a complex condition with multifactorial aetiopathogenesis. Adipose tissue is reservoir of many adipokines which play a great role in proinflammatory response in obesity. Aim of the study: Comparative assessment of ghrelin, leptin, and interleukin-6 (IL-6) salivary concentration among children having proper and excess of body mass. Analysis of the interrelationship between the obtained concentrations of substances and selected anthropometric parameters and blood pressure values in the studied children. MATERIAL AND METHODS: The study group comprised 102 children aged 7-10 years. The nutritional status of children was assessed by the use of the BMI index. The control group (n  = 74) comprised children with proper body mass, and the study group (n = 28) contained children having overweight/obesity. Saliva samples were taken from all children at school. Subsequently, some anthropometric parameters and blood pressure values of the children were measured. The laboratory assessment of substances was made by ELISA method. Next, statistical analysis of all obtained results was performed using professional software. RESULTS: Salivary ghrelin, leptin, and IL-6 concentrations were statistically significantly higher in the study group than in the control group (p = 0.001). The study revealed a positive correlation between salivary ghrelin concentration and BMI in the whole study population (p = 0.001), and between ghrelin concentration and body weight, waist circumference, hip circumference, and waist-to-hip ratio in all subjects. In the study group, the BMI value was positively correlated only with IL-6 saliva concentration (p = 0.005). CONCLUSIONS: The study revealed significant differences between saliva ghrelin, leptin, and IL-6 concentration between the control group and the study group. The above findings can be a good predictor with which to detect co-existing metabolic alternations in obese patients.

The Analysis of the Nutritional Status and Dietary Habits among Children Aged 6-10 Years Old Attending Primary Schools in Poland.

A high prevalence of obesity among children is influenced by serious implications. Obesity mainly results from behavioral factors, such as improper dietary habits. This study aims to evaluate the nutritional status and dietary habits of children aged 6-10 (n = 908) attending primary schools in Poland, Europe. The research tool was a questionnaire that was completed by one of the children's parents. A statistical analysis was made using statistical software. The value of p = 0.05 was considered to be statistically significant. A total of 74.7% of children surveyed have a normal body mass. As many as 91.7% and 76.6% of children, respectively, eat a first and second breakfast daily. Nearly half of parents (48.9%) state that their child consumes milk or other dairy products daily. A total of 74.3% of children drink water daily. A total of 27.6% eats fish less frequently than once a week. A total of 7.6% of children eat fish several times a week. As many as 20.6% of the respondents state that their child eats brown bread several times a week, whereas 19.9% state that their child never eats brown bread. A total of 55.1% of children eat fruits and/or vegetables daily. A total of 14.1% of children surveyed consume sweets daily. The study revealed a positive correlation between BMI and the frequency of mineral water consumption (p = 0.013) in 9 y.o. girls. It was also revealed that the number of consumed fruit/vegetables increases with the BMI value among 10 y.o. boys (p = 0.044). Conclusions: The dietary habits of the investigated children are still improper. There is a great need for education on this issue, but family involvement is also required.

A Questionnaire-Based Study to Obtain a Consensus from 5 Polish Burns Centers on Eschar Removal by Bromelain-Based Enzymatic Debridement (Nexobrid®) in Burns Following the 2020 Updated European Consensus Guidelines.

BACKGROUND The supplementary treatment of burns with enzymatic debridement with Nexobrid® was approved in Europe in 2013. The 2017 European consensus guidelines on the removal of eschar in burns by bromelain-based enzymatic debridement were updated in 2020. This questionnaire-based study aimed to obtain a consensus from 5 Polish burns centers on eschar removal by Nexobrid® in burns following the 2020 updated European consensus guidelines. MATERIAL AND METHODS A panel of 5 experts representing the leading burn treatment centers in Poland (Cracow, Gryfice, Siemanowice Slaskie, Poznan, and Leczna) was convened. A modified Delphi process was implemented with panel member selection, literature review, 2 rounds of voting in which panelists were asked to evaluate the European consensus and Polish consensus building by data analysis, statements preparation, final voting, and manuscript drafting. RESULTS The knowledge and experience of experts from Poland's leading burn centers resulted in the development of guidelines, formulated as 24 statements representing the following areas: indications and usage, pain management, application principles, post-enzymatic debridement wound dressing, and early and long-term outcomes. An analysis of the 7-point Likert scale polls revealed that 23 of the 24 statements achieved 100% consensus. CONCLUSIONS The findings from this survey from 5 major centers in Poland supported the main recommendations from the 2020 updated European consensus guidelines on the removal of eschar in burns by Nexobrid® and may serve as a practical guide for surgeons who care for patients with burns in this country.

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.

Preliminary Research on the Effect of Hyperbaric Oxygen Therapy in Patients with Post-COVID-19 Syndrome.

Negative consequences and medical complications of COVID-19 can persist for up to several months after initial recovery. These consequences can include stroke, diabetes, decreased lung diffusing capacity, sleep apnea, pulmonary fibrosis, arrhythmia, myocarditis, fatigue, headaches, muscle aches, heart rate fluctuations, sleep problems, memory problems, nervousness, anxiety, and other neurological disorders. Thirty-one patients who reported symptoms related to previous COVID-19 disease of both sexes were enrolled in the initial program. The patients underwent compression sessions in a multiplace hyperbaric chamber. Each patient underwent a cycle of 15 compressions. Before the first session, each participant completed a venous blood gas test, a Fullerton test, and two spirometry tests (one before the Fullerton test and one after the test). Patients completed psychotechnical tests, a questionnaire on quality of life (Polish version of EQ-5D-5L), and a questionnaire on specific symptoms accompanying the disease and post-infection symptoms. The results showed significant improvements in areas such as quality of life, endurance and strength, some spirometric parameters, the anion gap and lactate levels, working memory, and attention in the group of treated patients. In contrast, there were no changes in pH, pO2, pCO2, glucose, and excess alkaline values. A follow-up interview confirmed that the beneficial effects were maintained over time. Considering the results obtained, including the apparent improvement in the patient's clinical condition, it can be concluded that the use of 15 compression sessions was temporarily associated with a noticeable improvement in health and performance parameters as well as improvement in certain blood gas parameters.

Using Maggot Debridement Therapy in Treatment of Necrosis in the Forearm Caused By Docetaxel Extravasation: A Case Report.

Extravasation of doxorubicin, vincristine or vinblastine leads to necrosis, damage of the muscles and nerves, deep ulceration, as well as limb dysfunction. Necrosis and deep ulcers develop within 7 to 28 days. Like necrotomy, Lucilia sericata maggot therapy is recognised as a method enabling effective, safe and quick removal of necrotic tissue. The purpose of the study was to present local treatment of hypodermic necrosis caused by docetaxel extravasation in course of systemic cancer therapy. A woman, 59 years of age, in course of systemic therapy due to advanced cancer of the left breast (T2N1M1 CS IV) with confirmed metastases within the body of the fourth lumbar vertebra and in the liver, receiving a combination treatment with pertuzumab, trastuzumab, and docetaxel. During the therapy, a conservative treatment was applied due to extravasation for over three months. Effects in the right forearm included swelling, redness, signs of 4x10cm inflammatory infiltrate, with 1x4cm necrotic crust visible in the central region. Hypodermic necrosis was debrided using L. sericata maggots, and subsequently specialist dressings were applied to promote granulation and healing. In the case discussed here, effectiveness of MDT was rather poor, however the treatment minimised the risk of infection associated with evacuation of necrosis. Attempts to use MDT should be continued to enable more comprehensive understanding of problems related to management of necrosis in wounds developing during cancer therapy.

The use of biostatic human amnion and platelet-rich plasma in topical treatment of toxic epidermal necrolysis-A case report.

BACKGROUND: Toxic epidermal necrolysis (TEN) (also known as Lyell syndrome) and Stevens-Johnson syndrome (SJS) are life-threatening mucocutaneous blistering diseases. They are characterized by generalized blisters and epidermal inflammation, most likely resulting from the administration or interaction of medicines. AIMS: To report potential new method in the treatment of TEN. PATIENTS/METHODS: This article presents a case report of a 35-year-old man suffering from TEN covering about 95% of his body surface. Lesions occurred in the patient during antiepileptic therapy, after taking simultaneously amoxicillin (with clavulanic acid) and naproxen followed by lamotrigine treatment. Standard general treatment was performed. Intravenous feeding was necessary. Due to acute respiratory failure, the patient required mechanical ventilation. Two methods were combined in topical treatment: application of platelet-rich plasma (PRP) and a simultaneous biostatic human amnion transplant. RESULTS: In the presented case, the combination of both methods contributed to a significant acceleration of wound healing. After the application of PRP and biostatic amnion transplantation, the healing of wounds on the back and posterior surfaces of the legs was completed after six days. The surgical treatment most probably contributed to a significant acceleration of wound healing. CONCLUSION: The case report shows that topical TEN/SJS treatment with biostatic human amnion and PRP has a positive clinical effect and may be a new method of treatment of TEN.

Historical and modern research on propolis and its application in wound healing and other fields of medicine and contributions by Polish studies.

ETHNOPHARMACOLOGICAL RELEVANCE: The history of medical application of propolis (also known as bee glue) dates back to the times of ancient Greeks, Romans, Persians and Egyptians. Honey and other bee products, including propolis, occupy an important place in Polish folk medicine. Scientific research on propolis in Poland began in the early 1960s in Zabrze and continues until now. AIM OF THE REVIEW: The aim of this review is to provide an overview of information on Polish research on propolis and its medical application with particular emphasis on studies concerning wound healing. Consequently, our goal is also to shed a new light on therapeutic potential of Polish propolis in order to support future research in the field. MATERIALS AND METHODS: A systematic review of scientific literature on propolis and its medical application was performed by using the literature databases (PubMed, Web of Science, Google Scholar). We paid special attention to papers describing the effect of propolis on skin wound healing as well as to Polish contribution to research on propolis. RESULTS: Professor Stan Scheller was the first Polish scientist dealing with propolis and its medical potential. His legacy was continued by several research teams that studied the topic in various aspects. They analyzed propolis composition, its antioxidant, anti-inflammatory, antimicrobial, antiapoptotic and anticancer properties as well as its application in dentistry and wound treatment. Burn wound healing physiology after propolis administration was thoroughly studied on pig model, whereas research on patients proved the efficacy of propolis in chronic venous leg ulcer treatment. CONCLUSION: Polish scientists have made a significant contribution to the research on propolis, its biological properties and influence on wound healing. Propolis ointments can effectively accelerate the healing process and improve healing physiology, so they can be recommended as a promising topical medication for wound treatment in the future clinical and preclinical trials.

Negative Pressure Wound Therapy in Pyoderma Gangrenosum Treatment.

BACKGROUND Pyoderma gangrenosum (PG) is a rare, non-infectious, fulminant dermatosis of the skin. The lack of objective diagnostic criteria requires differential diagnosis and exclusion of extensive ulcerative skin diseases. Currently, treatment includes a combination of systemic steroids, immunosuppressants, and topical agents, but after decades of research, no clear scientific evidence exists for a criterion standard treatment. The use of NPWT (negative pressure wound therapy) effectively reduces the wound area, eliminates exudate, and reduces bacterial titer, thereby stimulating neoangiogenesis. CASE REPORT We present the case of an 83-year-old man with confirmed pyoderma gangrenosum. In the examination, a pink-red wound was observed, measuring 5×15 cm, II/III° according to National Pressure Ulcer Advisory Panel (NPUAP), covered with a non-physiological, crusty (scar-like) epidermis, from under which a foul purulent discharge emerged when pressed. NPWT therapy was started in August. During this period, wound healing was observed, without signs of undermining. After 52 days of treatment, NPWT was completely discontinued. Further local actions were then carried out using specialized dressings and periodic tacrolimus. CONCLUSIONS We observed the positive effect of NPWT on the wound, which, together with doxycycline treatment, led to the elimination of purulent exudate from the wound and inhibition of wound enlargement. The use of NPWT as a supportive method in the treatment of PG wounds is safe and effective, and it can be successfully used in nursing.

Preparation of placental tissue transplants and their application in skin wound healing and chosen skin bullous diseases - Stevens-Johnson syndrome and toxic epidermal necrolysis treatment.

Unique properties of amniotic membrane make it a promising source for tissue engineering and a clinically useful alternative for patients suffering from chronic wounds including, for example, ulcers, burns, ocular surface damages and wounds occurring in the course of bullous diseases like stevens-johnson syndrome and toxic epidermal necrolysis. Its use has many advantages over standard wound care, as it contains pluripotent cells, nutrients, anti-fibrotic and anti-inflammatory cytokines, growth factors and extracellular matrix (ECM) proteins. Placental tissues can be prepared as a medical component, an advanced therapy medicinal product or a tissue graft. In addition to basic preparation procedures such as washing, rinsing, cutting, drying and sterilisation, there are many optional steps such as perforation, crosslinking and decellularisation. Finally, transplants should be properly stored-in cryopreserved or dehydrated form. In recent years, many studies including basic science and clinical trials have proven the potential to expand the use of amniotic membrane and amnion-derived cells to the fields of orthopaedics, dentistry, surgery, urology, vascular tissue engineering and even oncology. In this review, we discuss the role of placental tissues in skin wound healing and in the treatment of various diseases, with particular emphasis on bullous diseases. We also describe some patented procedures for placental tissue grafts preparation.

The clinical significance of Notch1 immunoexpression in Caucasian patients with colorectal adenocarcinoma.

INTRODUCTION: Colorectal cancer (CRC) is traditionally regarded as the most commonly diagnosed gastrointestinal malignant disease. Nevertheless, despite advances in diagnosis and novel therapeutic options, the clinical outcomes of patients are still not satisfactory. AIM: To investigate the clinicopathological and prognostic roles of Notch1 expression, the immunohistochemical investigation was performed in samples of CRC tumour tissues, adjacent non-pathological mucosa, and metastatic foci in regional lymph nodes in Caucasian patients. MATERIAL AND METHODS: Paraffin-embedded adenocarcinoma samples were assessed immunohistochemically for Notch1 protein and scored according to the percentage of cells with a positive reaction combined with staining intensity. Connections between Notch1 immunoexpression and clinicopathological factors including the 5-year overall survival (OS) were evaluated. RESULTS: The level of the Notch1 immunohistochemical reactivity was correlated with the grade of the histological differentiation, size of the primary tumour, regional lymph node involvement, and perineural invasion (all p < 0.001). Kaplan-Meier survival analysis showed that the survival time for patients with a low expression of Notch1 was significantly longer than that for patients with moderate or strong level of Notch1 immunoreactivity (p < 0.001). CONCLUSIONS: The enhanced level of Notch1 immunoexpression was significantly associated with malignancy-related clinicopathological factors and reduced the 5-year overall survival in CRC patients.

Catalase immunoexpression in colorectal lesions.

INTRODUCTION: It is generally accepted that the gastrointestinal tract, and especially the colon, is constantly exposed to reactive oxygen species (ROS) that may be responsible for the appearance of genetic mutations. To keep a steady-state control over ROS production-detoxification, organisms have evolved a defensive system. Nevertheless, many reports have described decreased level of antioxidant enzymes, especially catalase (CAT), in cancer tissues. AIM: In this work we try to assess the immunohistochemical expression of CAT protein in colorectal adenoma and adenocarcinoma samples. MATERIAL AND METHODS: This study was performed on resected specimens obtained from 122 patients who had undergone surgical resection for colorectal cancer, and from 120 patients who had undergone colonoscopy. Paraffin- embedded, 4 µm-thick tissue sections were stained for rabbit polyclonal anti CAT antibody obtained from GeneTex (cat. no. GTX110704). RESULTS: In adenoma strong immunoexpression was detected mainly in infiltrating mononuclear cells within lamina propria. High expression of CAT was significantly associated with grade of dysplasia (high grade vs. low grade, p = 0.037). In adenocarcinoma samples, the high level of CAT immunoexpression was significantly correlated with histological grade of tumour (G1 vs. G2 vs. G3, p = 0.001) and depth of invasion (T1 vs. T2 vs. T3 vs. T4, p = 0.003). CONCLUSIONS: Development of colorectal cancer is associated with increased expression of CAT in the stage of adenoma and decreased expression in the stage of adenocarcinoma.

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.

Heterogeneous Mixture of Amniotic Cells is Likely a Better Source of Stem Cells than Adipose Tissue.

Stem cells are increasingly being used in the course of burn treatment. As several different types of stem cells are available for the purposes, it is important to chose the most efficient and the most practicable stem cell type. The aim of this study was to compare the potential of heterogeneous amnion cell mixture with the presently used standard therapy, the adipose tissue-derived stem cells. The placenta was collected during a Cesarean section procedure. Adipose tissue tissue-derived cells were isolated using the Cytori's Celution® System. Cells were tested for fulfillment of the minimum criteria for stem cells. The efficiency of cell cultures was tested by an analysis of population doubling, cell proliferation, cell cycle and cell migration. Amniotic cells presented a higher ability for differentiation to chondrocytes and osteocytes than adipose-derived regenerative cells but a lower ability for differentiation toward adipocytes. Additionally, in vitro experiments have demonstrated a higher applicability of amniotic cells than adipose tissue-derived stem cells. Amniotic cells show several advantages: easy access to placenta, low costs and a lack of ethical dilemmas related to stem cell harvesting. The main disadvantage is, however, their availability, as isogenic treatment would only be possible for women around children-bearing age, unless personalized banks for amniotic cells would be established.

Amniotic cells share clusters of differentiation of fibroblasts and keratinocytes, influencing their ability to proliferate and aid in wound healing while impairing their angiogenesis capability.

An alternative to cultured skin cell grafts usage in burn treatment is the graft of allogenic stem cells. We verified whether amniotic stem cells are better than the present therapeutic standard: grafts of autologous keratinocytes and fibroblasts along with autologous adipose-derived stem cells, and whether amniotic stem cells can support the growth of autologous keratinocytes and fibroblasts in the culture. The study was performed on the material from 18 amnia. Skin cells were obtained from 3 patients. In order to assess the influence of stem cells on keratinocytes and fibroblasts, the following experiments were performed: impact on viability and cell cycle, wound healing capability, angiogenesis capability, influence on the proliferation speed and capability to differentiate into skin cells. We demonstrated that human amniotic membrane-derived mesenchymal stem cells (hAMMSCs) share amniotic proteins with skin cells. Amniotic stem cells may replace skin fibroblasts in grafts due to the close similarity in their surface antigens, with significantly larger proliferative potential and ability to stimulate wound healing. It was shown that adding amniotic cells to both keratinocytes and fibroblast cultures accelerates directional migration by ≥ 40%. We confirmed in this study the influence of amniotic cells on the proliferation and cell cycle of fibroblasts and keratinocytes. Amniotic stem cells can be successfully used not only as a first choice graft but also to replace 3T3 line cells, supporting the proliferation of the cells during the culturing, as well as a supplementary graft supporting an autologous graft of keratinocytes and fibroblasts.