Abdominoplasty Skin-Based Dressing for Deep Wound Treatment-Evaluation of Different Methods of Preparation on Therapeutic Potential.

The management of hard-to-heal wounds is a significant clinical challenge. Acellular dermal matrices (ADMs) have been successfully introduced to enhance the healing process. Here, we aimed to develop protocol for the preparation of novel ADMs from abdominoplasty skin. We used three different decellularization protocols for skin processing, namely, 1M NaCl and sodium dodecyl sulfate (SDS, in ADM1); 2M NaCl and sodium dodecyl sulfate (SDS, in ADM1); and a combination of recombinant trypsin and Triton X-100 (in hADM 3). We assessed the effectiveness of decellularization and ADM's structure by using histochemical and immunochemical staining. In addition, we evaluated the therapeutic potential of novel ADMs in a murine model of wound healing. Furthermore, targeted transcriptomic profiling of genes associated with wound healing was performed. First, we found that all three proposed methods of decellularization effectively removed cellular components from abdominoplasty skin. We showed, however, significant differences in the presence of class I human leukocyte antigen (HLA class I ABC), Talin 1/2, and chondroitin sulfate proteoglycan (NG2). In addition, we found that protocols, when utilized differentially, influenced the preservation of types I, III, IV, and VII collagens. Finally, we showed that abdominoplasty skin-derived ADMs might serve as an effective and safe option for deep wound treatment. More importantly, our novel dressing (ADM1) improves the kinetics of wound closure and scar maturation in the proliferative and remodeling phases of wound healing. In conclusion, we developed a protocol for abdominoplasty skin decellularization suitable for the preparation of biological dressings. We showed that different decellularization methods affect the purity, structure, and therapeutic properties of ADMs.

Skin Substitute Preparation Method Induces Immunomodulatory Changes in Co-Incubated Cells through Collagen Modification.

Chronic ulcerative and hard-healing wounds are a growing global concern. Skin substitutes, including acellular dermal matrices (ADMs), have shown beneficial effects in healing processes. Presently, the vast majority of currently available ADMs are processed from xenobiotic or cadaveric skin. Here we propose a novel strategy for ADM preparation from human abdominoplasty-derived skin. Skin was processed using three different methods of decellularization involving the use of ionic detergent (sodium dodecyl sulfate; SDS, in hADM 1), non-ionic detergent (Triton X-100 in hADM 2), and a combination of recombinant trypsin and Triton X-100 (in hADM 3). We next evaluated the immunogenicity and immunomodulatory properties of this novel hADM by using an in vitro model of peripheral blood mononuclear cell culture, flow cytometry, and cytokine assays. We found that similarly sourced but differentially processed hADMs possess distinct immunogenicity. hADM 1 showed no immunogenic effects as evidenced by low T cell proliferation and no significant change in cytokine profile. In contrast, hADMs 2 and 3 showed relatively higher immunogenicity. Moreover, our novel hADMs exerted no effect on T cell composition after three-day of coincubation. However, we observed significant changes in the composition of monocytes, indicating their maturation toward a phenotype possessing anti-inflammatory and pro-angiogenic properties. Taken together, we showed here that abdominoplasty skin is suitable for hADM manufacturing. More importantly, the use of SDS-based protocols for the purposes of dermal matrix decellularization allows for the preparation of non-immunogenic scaffolds with high therapeutic potential. Despite these encouraging results, further studies are needed to evaluate the beneficial effects of our hADM 1 on deep and hard-healing wounds.

Differential Response of MDA-MB-231 and MCF-7 Breast Cancer Cells to In Vitro Inhibition with CTLA-4 and PD-1 through Cancer-Immune Cells Modified Interactions.

Drugs targeting immune checkpoint molecules have been found effective in melanoma, lung cancer, and other malignancies treatment. Recent studies on breast cancer demonstrated the significance of inhibitory anti-CTLA-4 and anti-PD-1 in the regulation of disease progression. However, seemingly the same types of breast cancer do not always respond unambiguously to immunotherapy. Thus, here we set out to analyze the in vitro effects of inhibiting CTLA-4 and PD-1 on interactions between co-cultured lymphocytes and two selected breast adenocarcinoma cell lines. Breast cancer cells were co-cultured with lymphocytes to evaluate the effects of CTLA-4 and PD-1 inhibition. Proliferation, cell cycle, and viability assessment were measured in cancer cells. IFN-gamma, IL-10, perforin, granzyme B production, and CTLA-4 and PD-1 expression were analyzed in lymphocytes. We found that administration of anti-CTLA-4 improved the anti-cancer activity of T cells with reduced proliferation and viability of MDA-MB-231. Lack of response was observed in the context of MCF-7. In addition, differential expression of checkpoint proteins was found between studied cancer cells lines. Inhibition of molecules was followed by IL-10 and IFN-gamma decrease in lymphocytes co-cultured with MDA-MB-231, not demonstrated in reference to MCF-7. Furthermore, CTLA-4 blockage was associated with reduction of CTLA-4+ and PD-1+ lymphocytes in MDA-MB-231, with a significant increase in MCF-7, reduced by anti-PD-1. Altogether, our study revealed that anti-CTLA-4 and anti-PD-1 treatment can improve lymphocytes effects on breast cancer cells. Favorable effects seemed to be related to breast cancer cells features as differential responses were reported. Novel blocking antibodies strategies should be tested for more effective cancer inhibition.

The influence of patient's age on metabolic and bariatric results of laparoscopic sleeve gastrectomy in 2-year observation.

BACKGROUND: The incidence of obesity has been constantly growing and bariatric procedures are considered to be the most effective treatment solution for morbidly obese patients. The results of laparoscopic sleeve gastrectomy (LSG) may differ depending on patient's age, gender, preoperative body mass index (BMI) and physical activity. METHODS: The aim of this study was to evaluate age-related differences in the outcome of LSG in terms of weight loss parameters, lipid and carbohydrate profile. The retrospective analysis of 555 patients who had undergone LSG was performed to compare the metabolic outcomes of surgery in individuals < 45 and ≥ 45 years old. Evaluation of weight loss parameters along with selected laboratory data was performed to demonstrate the results of LSG in 2 years follow-up. RESULTS: Overall, 238 males and 317 females (43%/57%) with median age of 43 years and median preoperative BMI of 46.41 (42.06-51.02) kg/m2 were analyzed. Patients in both groups presented significant weight loss at 24 months after the surgery with comparable percentage of total weight loss (40.95% in < 45 years old group and 40.44% in ≥ 45 years old group). The percentage of excess weight loss (78.52% vs. 74.53%) and percentage of excess BMI loss (91.95% vs. 88.01%) were higher in patients < 45 years old. However, the differences were not statistically significant (p = 0.662, p = 0.788 respectively). Patients under 45 years old experienced faster decrease in fasting glucose level that was observed after only 3 months (109 mg/dl to 95 mg/dl in < 45 years old group vs. 103.5 mg/dl to 99.5 mg/dl in ≥ 45 years old group, p < 0.001). Both groups presented improvement of lipid parameters during the observation. However, patients < 45 years old achieved lower values of LDL at 3 and 12 months follow-up (115 mg/dl vs. 126 mg/dl, p = 0.010; 114.8 mg/dl vs. 122 mg/dl, p = 0.002). Younger group of patients also showed superior improvement of triglycerides level. CONCLUSIONS: LSG results in significant weight loss in all patients regardless age. In turn, superior and faster improvement in lipid and carbohydrate profile is achieved in patients under 45 years old.

Gender-related metabolic outcomes of laparoscopic sleeve gastrectomy in 6-month follow-up.

INTRODUCTION: Laparoscopic sleeve gastrectomy (LSG) is recently a leading method in surgical treatment of morbid obesity. The metabolic outcome of intervention may be a result of many factors such as age, gender, preoperative weight loss and dietary restrictions. AIM: To evaluate gender-related differences in the results of LSG in 6-month follow-up. MATERIAL AND METHODS: The study included 101 patients who underwent LSG at the University Clinical Hospital of Bialystok. Patients were divided and analyzed in 2 groups: males (n = 48) and females (n = 53). The primary analysis included the influence of gender on postoperative weight loss calculated using the percentage of excess weight loss (%EWL) and excess BMI loss (%EBMIL). For secondary outcomes the levels of glucose, insulin, glycated hemoglobin, aspartate transaminase, alanine transaminase, total cholesterol, high-density lipoprotein (HDL), low-density lipoprotein (LDL), triglycerides and C-reactive protein were analyzed. RESULTS: A significant influence of patients' gender was proved for both %EWL (p = 0.026) and %EBMIL (p = 0.001). Females had significantly higher %EWL in 6-month follow-up than males (p = 0.0034). The analysis also showed significantly higher %EBMIL for women at 3 and 6 months observation (p = 0.022 and p < 0.001 respectively). CONCLUSIONS: Laparoscopic sleeve gastrectomy is an effective method of obesity treatment especially in terms of postoperative weight loss. Females seem to benefit more from the procedure when analyzing the parameters of body mass reduction. However, further research is needed to provide strong evidence of an association between gender and the results of LSG.

Different surgical approaches in laparoscopic sleeve gastrectomy and their influence on metabolic syndrome: A retrospective study.

Obesity is a growing health, social, and economic issue and became an epidemic, according to recent report of World Health Organization.The only method with scientifically proved efficiency of body mass loss is a surgical treatment. Laparoscopic sleeve gastrectomy (LSG) is recently a leading method in metabolic surgery. There are no standards of operative technique for LSG so far. The influence of technique modification on metabolic effect has not been described clearly.The aim of this study was to evaluate metabolic effects in patients with morbid obesity who underwent various surgical approaches of LSG.The study included 120 patients who were randomly divided into 3 groups: Group I, where bougie size was 32 French (Fr), Group II-36 Fr and Group III-40 Fr. Each group was divided into 2 subgroups, based on the distance of resection beginning from the pylorus-2 or 6 cm. Statistical analysis of: body mass index (BMI), the Percentage of Excess Weight Loss (%EWL), the Percentage of Excess BMI Loss (%EBMIL), levels of glucose and insulin on an empty stomach, glycated hemoglobin (HbA1c), insulin resistance (Homeostatic Model Assessment of Insulin Resistance Index-HOMA-IR), aspartate transaminase (AST), alanine transaminase (ALT), total cholesterol, high-density lipoprotein (HDL), low-density lipoprotein (LDL), triglycerides (TG), and C-reactive protein (CRP) were under investigation.Statistically significant decrease in body mass, BMI, %EWL, %EBL, glucose, and insulin concentrations has been observed in all studied groups. It was the highest when the smallest calibration tube has been used (32 Fr). Similar results were observed in HOMA-IR and HbA1c levels. Statistically significant decrease of total cholesterol, LDL, and TG concentrations have been observed. Significant increase of HDL in every group has been also noted. Postoperative CRP values were the lowest when the smallest bougie was used.LSG is effective method of obesity treatment. Metabolic effects of LSG are the most noticeable when a small bougie size is used.

Peroneal artery-vein index as a potential factor of thrombosis occurrence in free osteocutaneous fibula flap.

BACKGROUND: Despite high popularity and great success rates of free osteocutaneous fibula flaps, the flap failure caused by vascular thrombosis is still a challenging problem. The authors present their evaluation of a potential thrombosis risk factor - a peroneal artery-vein index. METHODS: The authors evaluated the diameters of peroneal vessels and peroneal artery-vein indexes based on the computed tomography angiographies in 10 patients who underwent a mandible reconstruction with a free fibula flap and compared the results with clinical outcome. RESULTS: In one case the flap was lost, because of thrombosis in the donor vein. This patient presented superficial varicose veins of both lower extremities. Peroneal vein diameters in this patient ranged from 5,05 mm to 6,68 mm and were higher than in patients without complications. The peroneal artery-vein index in the patient with thrombosis ranged from 0,37 to 0,50 with median 0,40 and was lower than in patients without complications. CONCLUSIONS: High disproportion between peroneal artery and concomitant veins might be a potential risk factor of the occurrence of venal thrombosis. Detailed perioperative examination of peroneal veins in patients with varicosities should be considered.