Continuous protein-density gradients: A new approach to correlate physical cues with cell response

To assess cellular behavior within heterogeneous tissues, such as bone, skin, and nerves, scaffolds with biophysical gradients are required to adequately replicate the in vivo interaction between cells and their native microenvironment. In this study, we introduce a strategy for depositing ultrathin films comprised of laminin-111 with precisely controlled biophysical gradients onto planar substrates using the Langmuir–Blodgett (LB) technique. The gradient is created by controlled desynchronization of the barrier compression and substrate withdrawal speed during the LB deposition process. Characterization of the films was performed using techniques such as atomic force microscopy and confocal fluorescence microscopy, enabling the comprehensive analysis of biophysical parameters along the gradient direction. Furthermore, human adipose-derived stem cells were seeded onto the gradient films to investigate the influence of protein density on cell attachment, showing that the distribution of the cells can be modulated by the arrangement of the laminin at the air–water interface. The presented approach not only allowed us to gain insights into the intricate interplay between biophysical cues and cell behavior within complex tissue environments, but it is also suited as a screening approach to determine optimal protein concentrations to achieve a target cellular output

Cell-Enriched Lipotransfer (CELT) Improves Tissue Regeneration and Rejuvenation without Substantial Manipulation of the Adipose Tissue Graft

The good availability and the large content of adult stem cells in adipose tissue has made it one of the most interesting tissues in regenerative medicine. Although lipofilling is one of the most frequent procedures in plastic surgery, the method still struggles with high absorption rates and volume losses of up to 70%. Therefore, many efforts have been made to optimize liposuction and to process the harvested tissue in order to increase fat graft retention. Because of their immunomodulatory properties, their cytokine secretory activity, and their differentiation potential, enrichment with adipose tissue-derived stem cells was identified as a promising tool to promote transplant survival. Here, we review the important parameters for lipofilling optimization. Finally, we present a new method for the enrichment of lipoaspirate with adipose tissue-derived stem cells and discuss the parameters that contribute to fat graft survival

Adipose Tissue-Derived Stem Cell Yield Depends on Isolation Protocol and Cell Counting Method

In plastic surgery, lipofilling is a frequent procedure. Unsatisfactory vascularization and impaired cell vitality can lead to unpredictable take rates in the fat graft. The proliferation and neovascularization inducing properties of adipose tissue-derived stem cells may contribute to solve this problem. Therefore, the enrichment of fat grafts with stem cells is studied intensively. However, it is difficult to compare these studies because many factors—often not precisely described—are influencing the results. Our study summarizes some factors which influence the cell yield like harvesting, isolation procedure and quantification. Stem cells were isolated after liposuction. Quantification was done using a cell chamber, colony counting, or flow cytometry with changes to one parameter, only, for each comparison. Quantification of cells isolated after liposuction at the same harvesting site from the same patient can vary greatly depending on the details of the isolation protocol and the method of quantification. Cell yield can be influenced strongly by many factors. Therefore, a comparison of different studies should be handled with care

Peripheral Nerve Regeneration–Adipose-Tissue-Derived Stem Cells Differentiated by a Three-Step Protocol Promote Neurite Elongation via NGF Secretion

The lack of supportive Schwann cells in segmental nerve lesions seems to be one cornerstone for the problem of insufficient nerve regeneration. Lately, adipose-tissue-derived stem cells (ASCs) differentiated towards SC (Schwann cell)-like cells seem to fulfill some of the needs for ameliorated nerve recovery. In this study, three differentiation protocols were investigated for their ability to differentiate ASCs from rats into specialized SC phenotypes. The differentiated ASCs (dASCs) were compared for their expressions of neurotrophins (NGF, GDNF, BDNF), myelin markers (MBP, P0), as well as glial-marker proteins (S100, GFAP) by RT-PCR, ELISA, and Western blot. Additionally, the influence of the medium conditioned by dASCs on a neuron-like cell line was evaluated. The dASCs were highly diverse in their expression profiles. One protocol yielded relatively high expression rates of neurotrophins, whereas another protocol induced myelin-marker expression. These results were reproducible when the ASCs were differentiated on surfaces potentially used for nerve guidance conduits. The NGF secretion affected the neurite outgrowth significantly. It remains uncertain what features of these SC-like cells contribute the most to adequate functional recovery during the different phases of nerve recovery. Nevertheless, therapeutic applications should consider these diverse phenotypes as a potential approach for stem-cell-based nerve-injury treatment

Serum Metabolomic Profiling of Patients with Lipedema

Lipedema is a chronic condition characterized by disproportionate and symmetrical enlargement of adipose tissue, predominantly affecting the lower limbs of women. This study investigated the use of metabolomics in lipedema research, with the objective of identifying complex metabolic disturbances and potential biomarkers for early detection, prognosis, and treatment strategies. The study group (n = 25) comprised women diagnosed with lipedema. The controls were 25 lean women and 25 obese females, both matched for age. In the patients with lipedema, there were notable changes in the metabolite parameters. Specifically, lower levels of histidine and phenylalanine were observed, whereas pyruvic acid was elevated compared with the weight controls. The receiver operating characteristic (ROC) curves for the diagnostic accuracy of histidine, phenylalanine, and pyruvic acid concentrations in distinguishing between patients with lipedema and those with obesity but without lipedema revealed good diagnostic ability for all parameters, with pyruvic acid being the most promising (area under the curve (AUC): 0.9992). Subgroup analysis within matched body mass index (BMI) ranges (30.0 to 39.9 kg/m2) further revealed that differences in pyruvic acid, phenylalanine, and histidine levels are likely linked to lipedema pathology rather than BMI variations. Changes in low-density lipoprotein (LDL)-6 TG levels and significant reductions in various LDL-2-carried lipids of patients with lipedema, compared with the lean controls, were observed. However, these lipids were similar between the lipedema patients and the obese controls, suggesting that these alterations are related to adiposity. Metabolomics is a valuable tool for investigating lipedema, offering a comprehensive view of metabolic changes and insights into lipedema’s underlying mechanisms

Первичные результаты применения стромально-васкулярной фракции аутологичной жировой ткани при стрессовом недержании мочи у мужчин

Introduction. Male stress urinary incontinence (SUI), which affects 10 % of the population, is most commonly caused by the effects of operations on the prostate gland. Despite the presence of a large number of surgical and conservative methods to treat this disease, the problem of choosing treatment tactics is still relevant. There are limited literature data on the effectiveness of using the stromal-vascular fraction (SVF) of autologous fat in the treatment of stress urinary incontinence. The aim of this study is to analyse the usage of SVF in the treatment of SUI patients.Materials and methods. A randomised study was carried out on a group of 8 patients with stress urinary incontinence of mild and moderate severity more than one year after radical prostatectomy or transurethral resection of the prostate. The patients received stromal-vascular fractions of autologous fat into the bladder sphincter area during the period November 2017 — June 2018.Results. After 4 weeks, the Pad Test and ICIQ-SF QoL assessments showed the first positive clinical results and improving quality of life. Checkpoints of 3, 6 and 12 months were selected for measuring results. Cytological analysis of the stromalvascular fraction of autologous fat revealed the expression of mesenchymal stem cell markers CD44, CD73, CD90, which may explain the method’s high efficiency.Conclusion. This work supports the hypothesis that transplantation of SVF from autologous adipose tissue is an effective and safe method for treating patients with incontinence.Введение. Стрессовое недержание мочи (СНМ) встречается у 10 % мужчин. Наиболее частой причиной возникновения СНМ у мужчин являются последствия операций на предстательной железе. Несмотря на наличие большого количества хирургических и консервативных методов лечения данного заболевания, проблема выбора тактики лечения до сих пор остается актуальной. Имеются ограниченные данные литературы об эффективности использования стромально-васкулярной фракции (СВФ) аутологичного жира в лечении стрессового недержания мочи. Целью данного исследования является анализ эффективности применения СВФ в лечении пациентов СНМ.Материалы и методы. Проводится открытое проспективное рандомизированное исследование. Исследуемая группа включала 8 пациентов со стрессовым недержанием мочи легкой и средней степени тяжести более одного года после радикальной простатэктомии или трансуретральной резекции простаты. Данным пациентам в период с ноября 2017 по июнь 2018 г. было выполнено введение стромально-васкулярной фракции аутологичного жира в зону сфинктера мочевого пузыря.Результаты. На основании Pad test и опросников ICIQ-SF, QoL через 4 недели получены первые положительные клинические результаты и улучшение качества жизни. Контрольными точками для оценки результатов выбраны 3, 6 и 12 месяцев. Цитологический анализ стромально-васкулярной фракции аутологичного жира выявил экспрессию маркеров мезенхимальных стволовых клеток CD44, CD73, CD90, что, вероятнее всего, и обуславливает высокую эффективность данного метода.Заключение. В данной работе отмечено, что трансплантация СВФ из аутологичной жировой ткани представляется эффективным и безопасным методом лечения пациентов с недержанием мочи.Введение. Стрессовое недержание мочи (СНМ) встречается у 10 % мужчин. Наиболее частой причиной возникновения СНМ у мужчин являются последствия операций на предстательной железе. Несмотря на наличие большого количества хирургических и консервативных методов лечения данного заболевания, проблема выбора тактики лечения до сих пор остается актуальной. Имеются ограниченные данные литературы об эффективности использования стромально-васкулярной фракции (СВФ) аутологичного жира в лечении стрессового недержания мочи. Целью данного исследования является анализ эффективности применения СВФ в лечении пациентов СНМ.Материалы и методы. Проводится открытое проспективное рандомизированное исследование. Исследуемая группа включала 8 пациентов со стрессовым недержанием мочи легкой и средней степени тяжести более одного года после радикальной простатэктомии или трансуретральной резекции простаты. Данным пациентам в период с ноября 2017 по июнь 2018 г. было выполнено введение стромально-васкулярной фракции аутологичного жира в зону сфинктера мочевого пузыря.Результаты. На основании Pad test и опросников ICIQ-SF, QoL через 4 недели получены первые положительные клинические результаты и улучшение качества жизни. Контрольными точками для оценки результатов выбраны 3, 6 и 12 месяцев. Цитологический анализ стромально-васкулярной фракции аутологичного жира выявил экспрессию маркеров мезенхимальных стволовых клеток CD44, CD73, CD90, что, вероятнее всего, и обуславливает высокую эффективность данного метода.Заключение. В данной работе отмечено, что трансплантация СВФ из аутологичной жировой ткани представляется эффективным и безопасным методом лечения пациентов с недержанием мочи.

Use of stromal-vascular fraction from autological fat tissue under stressed urinary disease in men

The aim of the work is to develop a minimally invasive method for restoring the function of the sphincter of the urinary bladder using autologous fat in the stromal-vascular fraction (SVF). Materials and methods. Patients with a diagnosis of stress urinary incontinence of moderate severity were given a transurethral injection of an autologous SVF into the zone of the external sphincter of the bladder. Isolation of SVF was performed by a nonenzymatic method using tumescent lipoaspiration. The resulting material was sent to the laboratory for the study of cellular composition. Results. Based on the Pad test and ICIQ-SF questionnaires, Qol received the first positive clinical results and improved quality of life after 4 weeks. Conclusions. In this work, it is noted that transplantation of SVF from autologous adipose tissue appears to be an effective and safe method of treatment of patients with incontinence.Цель работы - разработка малоинвазивного метода восстановления функции сфинктера мочевого пузыря с применением стромально-васкулярной фракции(СВФ) аутологичного жира. Материалы и методы. Пациентам с диагнозом стрессовое недержание мочи легкой и средней степеней тяжести выполнено трансуретральное введение аутологичной СВФ в зону наружного сфинктера мочевого пузыря. Выделение СВФ производилось неферментным методом при помощи тумесцентной липоаспирации. Полученный материал был направлен в лабораторию для исследования клеточного состава. результаты. На основании Pad test и опросников ICIQ-SF, Qol через 4 недели получены первые положительные клинические результаты и улучшение качества жизни. Выводы. В данной работе отмечено, что трансплантация СВФ из аутологичной жировой ткани представляется эффективным и безопасным методом лечения пациентов с недержанием мочи

Cattle genome-wide analysis reveals genetic signatures in trypanotolerant N'Dama

Abstract Background Indigenous cattle in Africa have adapted to various local environments to acquire superior phenotypes that enhance their survival under harsh conditions. While many studies investigated the adaptation of overall African cattle, genetic characteristics of each breed have been poorly studied. Results We performed the comparative genome-wide analysis to assess evidence for subspeciation within species at the genetic level in trypanotolerant N’Dama cattle. We analysed genetic variation patterns in N’Dama from the genomes of 101 cattle breeds including 48 samples of five indigenous African cattle breeds and 53 samples of various commercial breeds. Analysis of SNP variances between cattle breeds using wMI, XP-CLR, and XP-EHH detected genes containing N’Dama-specific genetic variants and their potential associations. Functional annotation analysis revealed that these genes are associated with ossification, neurological and immune system. Particularly, the genes involved in bone formation indicate that local adaptation of N’Dama may engage in skeletal growth as well as immune systems. Conclusions Our results imply that N’Dama might have acquired distinct genotypes associated with growth and regulation of regional diseases including trypanosomiasis. Moreover, this study offers significant insights into identifying genetic signatures for natural and artificial selection of diverse African cattle breeds