Adipose Tissue-derived Cells Research Articles

Current status of nanofat in the management of knee osteoarthritis: A systematic review

BACKGROUND Osteoarthritis (OA) is a prevalent joint disorder requiring innovative treatment approaches. AIM To evaluate the use of nanofat, a specialized form of adipose tissue-derived cells, in the treatment of OA, by examining its efficacy, safety profile, mechanisms of action, comparative effectiveness, and long-term outcomes. METHODS A comprehensive review of preclinical studies, clinical trials, and in vitro investigations was conducted. The included studies provided insights into the potential role of nanofat in OA treatment, addressing its efficacy, safety profile, mechanisms of action, comparative effectiveness, and long-term outcomes. RESULTS Clinical studies consistently reported the efficacy of nanofat in providing pain relief and functional improvement in patients with OA. Local adverse events were limited to the injection site, such as localized pain and inflammation, and resolved within a few days to weeks. Systemic adverse events were rare, and no significant long-term complications were observed. Mechanistically, nanofat was found to enhance chondrocyte proliferation, reduce inflammation, and promote angiogenesis, thereby contributing to its therapeutic effects. CONCLUSION Nanofat therapy holds promise as a therapeutic option for managing OA, providing pain relief, functional improvement, and potential tissue regeneration. The safety profile of nanofat treatment appears favorable, but long-term data are still limited. Standardized protocols, larger randomized controlled trials, longer follow-up periods, and cost-effectiveness evaluations are warranted to establish optimal protocols, comparative effectiveness, and long-term outcomes. Despite current limitations, nanofat therapy demonstrates translational potential and should be considered in clinical practice for OA treatment, with careful patient selection and monitoring.

Development of a yeast-based sensor platform for evaluation of ligands recognized by the human Free Fatty Acid 2 Receptor.

Yeast-based sensors have shown great applicability for deorphanization of G protein-coupled receptors (GPCRs) and screening of ligands targeting these. A GPCR of great interest is free fatty acid 2 receptor (FFA2R), for which short-chain fatty acids such as propionate and acetate are agonists. FFA2R regulates a wide array of downstream receptor signaling pathways in both adipose tissue and immune cells and has been recognized as a promising therapeutic target, having been implicated in several metabolic and inflammatory diseases. While research aiming to identify ligands recognized by FFA2R for translational applications is ongoing, screening is complicated by the complex regulatory and cell-specific responses mediated by the receptor. To simplify screening towards identification of novel ligands, heterologous platforms are valuable tools that offer efficient identification of ligand activity in the absence of regulatory mechanisms. Here, we present a yeast-based sensor designed to evaluate G protein α i1 mediated FFA2R signaling, with an assay time of 3 h. We verify this platform towards the natural agonists, propionate and acetate, and show applicability towards evaluation of synthetic agonists, antagonists, and allosteric agonists. As such, we believe that the developed yeast strain constitutes a promising screening platform for effective evaluation of ligands acting on FFA2R.

Lipocalin 2 in Obesity and Diabetes: Insights into Its Role in Energy Metabolism

Background: Lipocalin 2 (LCN2), also known as neutrophil gelatinase-associated lipocalin, is a 25 kDa protein involved in immune defense, inflammation, and metabolism. Results: LCN2 is widely expressed across various tissues, including immune cells, bone, adipose tissue, liver, kidneys, lung, spleen, and epithelial cells, and exhibits sex- and fat depot-specific expression patterns. Structurally, LCN2 contains a hydrophobic lipid-binding pocket and glycosylation sites, enabling it to interact with diverse ligands and form dimers. In innate immunity, LCN2 plays a critical role by sequestering iron-laden siderophores, thereby restricting bacterial growth. Beyond its role in infection control, LCN2 is implicated in metabolic inflammation and diseases such as obesity and diabetes. Recent research has highlighted a pivotal role for LCN2 in mitochondrial phospholipid metabolism and mitochondrial function. In metabolic diseases and mitochondrial metabolism, LCN2 appears to display paradoxical effects. While some studies link it to improved insulin sensitivity, glucose regulation, and mitochondrial function, others associate it with insulin resistance, obesity, and mitochondrial dysfunction. These inconsistencies may arise from differences in experimental conditions and study populations. Conclusions: This review provides an up-to-date summary of LCN2’s multifaceted roles in obesity, diabetes, energy balance, and mitochondrial function, emphasizing its context-dependent effects. LCN2 appears to have dual roles, exerting both protective and detrimental outcomes depending on the physiological or pathological context, sex, cell types, and experimental conditions. Further research is necessary to unravel its complex functions and resolve conflicting findings, particularly in metabolic disorders.

Obesity reshapes regulatory T cells in the visceral adipose tissue by disrupting cellular cholesterol homeostasis.

Regulatory T cells (Tregs) accumulate in the visceral adipose tissue (VAT) to maintain systemic metabolic homeostasis but decline during obesity. Here, we explored the metabolic pathways controlling the homeostasis, composition, and function of VAT Tregs under normal and high-fat diet feeding conditions. We found that cholesterol metabolism was specifically up-regulated in ST2hi VAT Treg subsets. Treg-specific deletion of Srebf2, the master regulator of cholesterol homeostasis, selectively reduced ST2hi VAT Tregs, increasing VAT inflammation and insulin resistance. Single-cell RNA/T cell receptor (TCR) sequencing revealed a specific loss and reduced clonal expansion of ST2hi VAT Treg subsets after Srebf2 deletion. Srebf2-mediated cholesterol homeostasis potentiated strong TCR signaling, which preferentially promoted ST2hi VAT Treg accumulation. However, long-term high-fat diet feeding disrupted VAT Treg cholesterol homeostasis and impaired clonal expansion of the ST2hi subset. Restoring Treg cholesterol homeostasis rescued VAT Treg accumulation in obese mice, suggesting that modulation of cholesterol homeostasis could be a promising strategy for Treg-targeted therapies in obesity-associated metabolic diseases.

The culture of equine adipose tissue-derived mesenchymal cells in serum-free media

Mesenchymal stem/stromal cells (MSCs) isolated from equine adipose tissue (AT) represent a promising material for the creation of bioveterinary products for the prevention and treatment of many diseases. The production of these cells for clinical use requires improved serum-free culture conditions. The microenvironment can influence the properties of MSCs. It is believed that the requirements for culture conditions without animal blood serum are species specific. The purpose of this study was to evaluate the commercially available serum-free media (SFM) MesenCult (STEMCELL Technologies, USA), created for human MSCs, for the cultivation of equine MSC(AT). One part of the cells was propagated for 10 passages in the standard DMEM medium with a low glucose content (1 g/l) and 10% fetal bovine serum (FBS), and the second in SFM. The results show that the propagation of equine MSCs in MesenCult serum free, intended for the cultivation of human MSCs, is possible, since the cells adapt well to it and retain properties characteristic of cells that are cultured in DMEM with FBS: morphology, growth rate, doubling time and mitotic index, clone-forming abilities, diploid set of chromosomes, a large number of cells with the CD90 phenotype (90.8%) and low with the CD31 (0.8%), CD34 (0.9%) phenotype, as well as the potency for induction of differentiation into adipo-, osteo- and chondrogenic directions. Equine MSC(AT) showed stable characteristics after being cultured for 10 passages in SFM, providing a promising basis for their further use. Our results demonstrate that MesenCult media may be an alternative for serum-free culture of equine MSC(AT) for expansion in preclinical studies.

Ischemia-induced expression status of cofilin 1, CRSP2, HSP90, HSP27, and IL8 in epicardial adipose tissue and single cell transcriptomic profiling of stromal cells.

Epicardial adipose tissue (EAT) is a rich source of EAT-derived stromal cells (EATDS), which possess regenerative potential. CRSP2, HSP27, IL8, HSP90, and Cofilin 1 were detected in the secretome of left ventricular stromal cells under ischemia challenge. However, the association of these genes in the EAT and EATDS remain understudied. We aim to assess the status of cofilin 1, CRSP2, HSP27, IL8, and HSP90 in the EAT of myocardial infarction (MI) and coronary artery bypass graft (CABG) swine models and in vitro stimulated ischemic EATDS. Expression status of these proteins in EAT were assessed by immunostaining, and in EATDS using qRT-PCR, immunostaining, and Western blot. EATDS phenotyping was performed using sc-RNAseq analysis. Cofilin 1 was increased while the other four genes were decreased in the CABG. IL8 and HSP90 were increased, while CRSP2, HSP27, and cofilin 1 were decreased in the MI group. Similar trend was displayed in the expression of these genes in EATDS. Additionally, EATDS displayed versatile phenotypes at single cell resolution based on the differential expression of various gene signatures. The findings revealed novel insights into EAT/EATDS biology and further understanding regarding the EATDS sub-phenotypes would open novel avenues in translational cardiology.

Диагностическая эффективность и прогностическая значимость антропометрических параметров как маркеров гиперлептинемии у подростков

Leptin is an energy homeostasis marker synthesized by cells of adipose tissue (adipocytes) and gastric mucosa. The search for clinical and anthropometric parameters associated with energy homeostasis disorders in excess body weight (BW) and obesity is necessary for stratification of patients at risk for early implementation of energy homeostasis disorders. The purpose of this re-search was to evaluate the relationship between serum leptin concentration and anthropometric parameters reflecting the topography of subcutaneous fat in adolescents of different ethnicities with excess BW and obesity in order to identify the most informative anthropometric parameters of energy homeosta-sis disorders. Materials and methods used: a single-center cross-sectional study of 362 adolescents aged 11 to 17 y/o residing in the Republic of Buryatia and the Irkutsk Oblast of Russia was conducted in 2015-2016 that included the participants as follows: 186 Caucasians (46 boys/140 girls) and 176 Mongoloids (96 boys/80 girls); 136 adolescents in the control group (38%) and 226 adolescents with excess BW and obesity (62%). All par-ticipants had their linear height, BW, waist (WC) and hip (HC) circumference, skinfold thickness measured as well as the body mass index (BMI, in kg per m2) calculated. The concentrations of glucose, total cholesterol, high-density, low-density and very low-density lipoprotein cholesterol, triglycerides and in-sulin were determined. The HOMA index was calculated. Serum leptin was de-termined by enzyme immunoassay using DBC Leptin ELISA kit (Diagnostics Biochem Canada Inc., Canada) with detection using BioTek Elx808 Microplate Spectrophotometer (Agilent Technologies, Inc., USA). Results: a strong posi-tive correlation was found between serum leptin levels and SDS BMI and trip-onderal BMI in adolescents of both ethnic groups: in Caucasian boys r=0.747 (p<0.001), r=0.767 (p<0.001), in Caucasian girls r=0.687 (p<0.001) and r=0.695 (p<0.001); in Mongoloid boys r=0.765 (p<0.001), r=0.852 (p<0.001), in Mongoloid girls r=0.612 (p<0.001), r=0.667 (p<0.001), respectively. Ac-cording to the results of ROC analysis, AUC WC/height showed good diagnos-tic significance for detecting hyperleptinemia (AUC=0.8-0.9) in adolescents from all the studied groups. Conclusion: the triponderal index and the WC/height ratio are informative anthropometric indicators for energy metabo-lism disorders and hyperleptinemia in Caucasian and Mongoloid adolescents of both genders with excess BW and obesity.

Obesity as the main factor of metabolic syndrome influencing on respiratory function

In the review, a generalized analysis of current scientific data explaining the physiological mechanisms of the influence of obesity on respiratory system is carried out. The multifactorial nature of the respiratory effect of obesity, including mechanical and inflammatory effects, is emphasized. The consequences of restrictive and obstructive changes in the biomechanics of respiration, changes in the topographic distribution of lung ventilation, mismatch of ventilation and perfusion, and a decrease in the efficiency of the respiratory muscles are considered. Elucidation of the central mechanisms of the respiratory action of proinflammatory mediators expressed by adipose tissue cells is recognized as a promising area of research. Special attention is paid to the action of leptin, which is the main regulator of metabolism and respiratory control in obesity. Its ability to modulate the central respiratory chemosensitive is discussed. It is assumed that an increase in pulmonary ventilation due to an increase in leptin production in obesity has a compensatory character and allows obese patients to maintain normocapnia despite an increase in mechanical load on respiration system. Whereas leptin resistance and suppressed hypercapnic ventilation response play a key role in the development of obesity–hypoventilation syndrome. It is concluded that it is necessary to further study the physiological mechanisms of the influence of obesity on the respiratory function in order to find new effective therapeutic methods for the treatment of diseases associated with obesity, which is the main factor in the development of metabolic syndrome.

Disulfiram-Loaded Nanoparticles Inhibit Long-Term Proliferation on Preadipocytes.

Disulfiram (DSF) reduces insulin resistance and weight gain in obese mice. However, the effect on adipose tissue is unexplored due to their high instability under physiological conditions, limiting clinical applications. Thus, it is meaningful to develop a DSF carrier for sustained release to adipose tissue. We optimized the synthesis of poly-ε-caprolactone (PCL) nanoparticles (NPs) loaded with DSF and analyzed their effect on adipose tissue cells in vitro. The NPs were synthesized by nanoprecipitation method, varying its solvent, either acetone or acetone/dichloromethane (60:40) (v/v), and ratio PCL:DSF (w/w) 1:2, 1:1, 2:1 and, 1:0; finding the best condition was obtained with acetone/dichloromethane solvent mixture and 2:1 PCL:DSF. Then, NPs toxicity was analyzed on adipose cells (preadipocytes, white-like adipocytes, and macrophages) assessing association and internalization, cell viability, and cell death mechanism. NPs were spherical with a particle size distribution of 203.2 ± 29.33 nm, a ζ-potential of -20.7 ± 4.58 mV, a PDI of 0.296 ± 0.084, and a physical drug loading of 18.6 ± 5.80%. Sustained release was observed from 0.5 h (10.94 ± 2.38%) up to 96h (91.20 ± 6.03%) under physiological conditions. NPs internalize into macrophages, white-like adipocytes and preadipocytes without modifying cell viability on white-like adipocytes and macrophages. Preadipocytes reduce cell viability, inducing mitochondrial damage, increased mitochondrial reactive oxygen species production and loss of mitochondrial membrane potential, leading to effector caspases 3/7 cleaved, resulting in apoptosis. Finally, long-term proliferation inhibition was observed, highlighting the bioequivalent effect of PCL-DSF NPs compared to free DSF. Our data demonstrated the biological interaction of PCL NPs with adipose cells in vitro. The selective cytotoxicity of DSF towards preadipocytes resulted in milder effects when it was delivered nanoencapsulated compared to the free drug. These results suggest promising pharmacological alternatives for DSF long-term delivery on adipose tissue.

Essential role of Card11 in airway hyperresponsiveness in high-fat diet-induced obese mice

A high-fat diet (HFD) can induce airway hyperresponsiveness (AHR) in obese mice, independent of allergic sensitization. This study aimed to identify the key molecules related to AHR in HFD-induced obese mice. In a cluster analysis of time series gene expression in the adipose and lung tissues of HFD-induced obese mice, we identified the Caspase Recruitment Domain Family Member 11 (Card11) gene as an essential molecule. We measured CARD11 expression in peripheral blood mononuclear cells (PBMCs) from obese individuals with asthma and performed Card11 signal inhibition in HFD-induced obese mice via Card11 siRNA. Card11 expression was significantly increased in M1 macrophages (IL-1β+CD11c+CD206- in CD11b+) in adipose tissue and in ILC3s (RORγt+ in IL7R+ of Lin-) in lung tissue from HFD-induced obese mice. In addition, CARD11+ populations among ILC3s and LPS-stimulated IL-1β+CD16+ monocytes from the PBMCs of obese individuals with asthma were significantly greater than those from obese controls or nonobese individuals with asthma. AHR in HFD-induced obese mice disappeared when we inhibited the Card11 signaling pathway by administering Card11 siRNA during the first or last seven weeks of the 13-week HFD feeding. Finally, we confirmed that Card11 siRNA decreased the number of M1 macrophages in adipose tissue and the number of ILC3s in lung tissue in vitro. Card11 significantly contributes to the development of AHR in HFD-induced obese mice by affecting immune cells in both adipose and lung tissues. The middle stage of HFD feeding seemed to be critical for these processes.

A Lipoma of Large Proportion in the Oral Cavity: A Case Report

Background Lipoma is a benign neoplasm that can affect the whole body and rarely occurs in the oral cavity. It is slow-growing and asymptomatic, and conservative surgical treatment with total removal of the lesion is the treatment of choice. It originates from epithelial cells of mature adipose tissue. Case Presentation The aim of this study was to report a clinical case of a large lipoma in a 21-year-old male patient with melanoderma who presented a nodule in the region of the buccal vestibule extending from the region of the second molar to the ipsilateral canine, with an approximate size of 40 mm in its largest diameter in the anteroposterior direction and approximately 20 mm in diameter in the buccolingual direction. The initial diagnostic hypothesis was based on an ultrasound of the face, which detected a solid, circumscribed, echogenic nodule compatible with a lipoma of the face. Surgical treatment with excisional biopsy was chosen. The histopathological examination revealed fragments with the presence of discrete acanthosis and clear hyperkeratosis in the epithelium, along with the proliferation of mature adipocytes constituting a neoplastic parenchyma surrounded by a stroma. Conclusion The findings have confirmed the diagnosis of oral lipoma. The patient has evolved with a good prognosis and remained under follow-up without recurrence or sequelae. The present report has corroborated what has been observed in similar cases described in the literature regarding recurrence and excellent prognosis.

CLINICAL EFFECTIVENESS AND FEATURES OF THE POSTOPERATIVE PERIOD IN PATIENTS AFTER OSTEOSYNTHESIS OF THE MANDIBLE USING CELL TECHNOLOGIES

Due to a signifi cant increase in the frequency and complexity of maxillofacial injuries, one of the priorities in the development of modern surgical dentistry is the search for new eff ective methods of treating facial bone fractures. In recent decades, the attention of clinicians has been focused on biological treatment methods aimed at restoring bone defects using cell technologies and biological transplants.The aim of the study was to determine the eff ect of using osteoplastic materials based on multipotent adipose- derived mesenchymal stromal cells in dental patients in the surgical treatment of mandibular fractures.Materials and methods. The clinical material included 56 patients, 13 of whom underwent mandibular osteosynthesis with hydroxyapatite bone graft substitute (subgroup 1I), 25 of whom underwent surgery with a combination of multipotent adipose tissue mesenchymal stromal cells, hydroxyapatite bone graft substitute and platelet-rich plasma (subgroup 1B), and 18 of whom underwent spontaneous healing of the fracture line (subgroup 1C). Patients were clinically evaluated on a daily basis. Clinical parameters were assessed on days 1, 3, 5, 7, 10, 14 and 28 after surgery. The postoperative pain syndrome was assessed using the Numerical Rating Scale (NRS), considering the subjective signs of pain in the operated patients.The statistical processing of the study results was performed using the statistical analysis software for biomedical research ‘Microsoft Excel 21’ and ‘Statistica’ (licence number STA862D175437Q). In the statistical processing of the results, we used the variational analysis of series – calculation of the arithmetic mean and its standard error (Mim) – and assessed the signifi cance of diff erences in the results obtained in the comparison groups using the Student’s t-test method. p<0.05 was considered a signifi cant diff erence. [17]. The study was conducted in accordance with the basic provisions of the «Rules of Ethical Principles for Scientifi c Medical Research Involving Human Subjects» approved by the Declaration of Helsinki (1964-2013), ICH GCP (1996), EEC Directive 609 (24.11.1986), Orders of the Ministry of Health of Ukraine ¹ 690 of 23.09.2009, ¹ 944 of 14.12.2009, ¹ 616 of 03.08.2012. The Commission on Biomedical Ethics of the Bukovinian State Medical University found no violations of ethical and moral standards in the conduct of the research (ProtocolNo. 6, 16.03.2023). Patients signed informed consent to participate in this study and all measures were taken to ensure their anonymity.The results. The comparative analysis of the clinical eff ectiveness of the use of various osteoplastic materials in the performance of osteosynthesis surgery of the lower law confi rmed the advantages of the use of bone tissue substitute based on hydroxyapatite and its combination with multipotent adipose tissue mesenchymal stromal cells before spontaneous augmentation. It is confi rmed by the data of subjective and objective symptoms of patients during the postoperative period.Conclusions. Our data show that osteoplastic material based on multipotent mesenchymal stromal cells of adipose tissue, bone substitute based on hydroxyapatite and platelet-rich blood plasma is a promising biological material. Its application improves the regenerative and reparative properties of bone tissue, facilitates the postoperative period and shortens the duration of hospitalization, promotes the development of innovative ways of reconstructive osteogenesis in modern maxillofacial surgery.

Methylation of the ESR1 promoters in visceral adipose tissue and its relationship with obesity.

Obesity is associated with decreased ESR1 expression level in visceral adipose tissue. However, it is unclear exactly what mechanisms are responsible for this decline. The aim of this study was to investigate the impact of aberrant methylation of the ESR1 alternative promoters on decreased ESR1 expression and its connection to obesity. Visceral adipose tissues and peripheral blood cells were obtained from 21 patients (non-obese and obese) undergoing inguinal hernia or gallbladder removal. Alternative promoter regions, C, E2 and F of the ESR1 gene, were analyzed by Methylation-Specific PCR (MSP) and mRNA levels were measured by quantitative real-time PCR (qPCR) in both visceral adipose tissue and peripheral blood cells. All statistical analyses were performed by SPSS (23.0). The methylation percentage in the three promoter regions of ESR1 was not different in obese individuals compared to non-obese individuals. We observed that promoter C had the highest methylation frequency in obese patients, although it was not statistically significant. Additionally, we observed that the hypermethylation of ESR1's promoter C was significantly associated with lower mRNA expression level in obesity (p = 0.020). This study suggests that methylation of ESR1 promoter C may be a factor in the development of obesity or a consequence of obesity. Further studies with advanced methods and larger study groups are needed to clarify this issue.

Advanced Non-Operative Interventions for Anterior Knee Pain.

This review presents evidence for advanced non-operative interventions, including extracorporeal shockwave therapy (ESWT), prolotherapy, platelet-rich plasma (PRP), adipose tissue-derived cells, bone marrow aspirate concentrate, various additional non-corticosteroid injectates, and needle-based interventions for common causes of anterior knee pain in the adult population. These etiologies include osteoarthritis of the knee, patellofemoral pain syndrome, chondromalacia patella, Hoffa fat pad impingement syndrome, patellar/quadriceps tendinopathy, and prepatellar bursitis. This review discusses patient care options using a case-based understanding of interventions by condition while recognizing strength of evidence. ESWT and PRP are the most robustly studied and have greatest evidence for treating tibiofemoral osteoarthritis and for long-term benefit in treating patellar tendinopathy. PRP may have evidence for treatment of chondromalacia and prolotherapy for management of tibiofemoral arthritis; both have limited evidence. Botulinum neurotoxin type A has strong evidence to support use in treating patellofemoral pain syndrome. There is limited evidence to support the use of viscosupplementation, percutaneous needle tenotomy, and medicinal signaling cell-based therapies beyond platelet-rich plasma for anterior knee pain. There is limited research on the management of quadriceps tendinopathy, prepatellar bursitis, patellofemoral osteoarthritis, and Hoffa's fat pad impingement syndrome. Further research and standardization of protocols are necessary to fully assess these treatments' efficacy. ESWT, cell-based, and needle-based interventions, may serve as effective treatment options for patients with anterior knee pain. Selection of each intervention requires understanding the evidence, level of risk, and appropriate application based on a patient's level of activity to enable clinicians to enhance patient outcomes and quality of life.

Adipocyte FGF21 Signaling Defect Aggravated Adipose Tissue Inflammation in Gestational Diabetes Mellitus.

Gestational diabetes mellitus (GDM) is associated with increased inflammation in adipose tissues. Fibroblast growth factor 21 (FGF21) is an endocrine hormone which signals to multiple tissues to regulate metabolism. However, its role in GDM remains largely unknown. In this study, we found that impaired FGF21 signaling in GDM correlates with worsened inflammation and insulin resistance in white adipose tissues in mice. Mechanistically, the pregnancy-related upregulation of FGF21 signaling in adipocytes promotes the differentiation of regulatory T cells (Tregs), which are critical for reducing pregnancy-induced adipose tissue inflammation. The anti-inflammatory effects of FGF21 may involve linolenic acid-mediated PGE2 synthesis in adipocytes. These findings underscore FGF21's role in mediating crosstalk between mature adipocytes and immune cells in white adipose tissue and suggest that targeting FGF21 signaling and its downstream metabolites could offer a potential therapeutic approach for GDM in humans.

A Single Center Experience of Special Cases: Isolated Adrenal Myelolipoma and Adrenocortical Adenoma with Myelolipomatous Component.

Isolated adrenal myelolipoma (IAM) is an uncommon, nonfunctioning tumour of the adrenal gland, primarily composed of adipose tissue and hematopoietic trilinear cells. The etiopathogenesis and clinical relevance of this neoplasm remain poorly understood. However, similar myelolipomatous alterations can also occur within adrenocortical adenomas. This report presents our findings on IAM and adrenocortical adenomas with a myelolipomatous component (AMC), focusing on comparing these cases' clinical, demographic, and pathological characteristics. Data from twenty patients were retrospectively analyzed, including twelve diagnosed with IAM and eight with AMC, all of whom underwent adrenalectomy. In the IAM cohort, surgical intervention was primarily indicated based on tumour size and/or related symptoms, while in AMC cases, the indication was often driven by hormonal activity. Patients with IAM had an average age of 52.5 years, with a predominance of female patients (75%). A significant proportion of this group was obese, with concomitant hypertension (HT) and/or type 2 diabetes mellitus (DM). The tumours were predominantly right-sided, with a median size of 69.0 ± 40.0 mm. Notably, 58% of patients with IAM presented with flank or abdominal pain attributed to the mass effect. None of the IAM cases exhibited hormonal activity. Conversely, the AMC group had a younger average age of 46 years, with a high prevalence also in female patients (63%). The tumours were generally smaller, with a median size of 40.0 ± 16.0 mm, and were mostly left-sided. All patients in this group had a history of HT and/or DM, with six exhibiting hormonally active tumours, which manifested as various clinical syndromes, including Cushing syndrome, pheochromocytoma, and Conn syndrome. While IAM and AMC share several common features, they also demonstrate distinct differences. The presence of endocrinological syndromes was more frequent in AMC cases, whereas IAM cases rarely showed hormonal activity. The most pronounced difference between the two groups was the tumour size at diagnosis, which contributed to the varying clinical presentations upon hospital admission. Furthermore, the high prevalence of obesity, HT, and DM in both groups suggests that these comorbidities may play a role in the development of myelolipomatous patterns observed in these tumours.

Overexpression of Chromatin Remodeling Factor SRG3 Down-Regulates IL1β-Expressing M1 Macrophages and IL17-Producing T Cells in Adipose Tissues.

The SWItch3-related gene (SRG3) is a core component of ATP-dependent SWI/SNF complexes, which are crucial for regulating immune cell development and function (e.g., macrophages and CD4+ T cells), embryonic development, and non-immune cell differentiation. Notably, SRG3 overexpression has been shown to polarize macrophages in the central nervous system toward an anti-inflammatory M2 phenotype, thereby protecting against the development of experimental autoimmune encephalomyelitis in mice. However, the effect of SRG3 on immune responses in adipose tissues remains unclear. To address this issue, we examined the cellularity and inflammatory status of adipose tissue in B10.PL mice overexpressing the SRG3 gene under the ubiquitous β-actin promoter (SRG3β-actin). Interestingly, SRG3 overexpression significantly reduced adipocyte size in both white and brown adipose tissues, without affecting the overall adipose tissue weight. Such phenotypic effects might be associated with the improved glucose tolerance observed in SRG3β-actin B10.PL mice. Moreover, we found that SRG3 overexpression down-regulates IL1β-expressing M1 macrophages, leading to a significant decrease in the M1/M2 macrophage ratio. Additionally, SRG3β-actin B10.PL mice showed a dramatic reduction in neutrophils as well as IL1β- and IL17-producing T cells in adipose tissues. Taken together, our results indicate that SRG3 plays a vital role in maintaining immune homeostasis within adipose tissues.

Origin of dedifferentiated adipocyte-derived cells (DFAT) during ceiling culture in an Adiponectin Cre-Recombinase mouse model.

Dedifferentiated adipose tissue-derived (DFAT) cells represent an attractive source of stem cells for tissue engineering and the potential treatment of several clinical conditions. Our objective was to determine whether DFAT cells originate from mature adipocytes and address whether contamination from the stromal vascular fraction (SVF) could be as a source for these cells. A murine adiponectin-creERT; mT/mG model was used with the excision of the cassette induced by tamoxifen injection for the cells expressing adiponectin (adipoq). This model allows distinguishing mature adipocytes (green fluorescence) from other SVF cell types (red fluorescence) based on the fluorescent protein expressed. Mature adipocytes and SVF cells were isolated from adipose tissues by collagenase digestion. Ceiling cultures were imaged by time-lapse microscopy. Confocal microscopy was used to follow cells over 21 days. Time-lapse microscopy experiments showed liposecretion occurring in mature adipocytes displaying green fluorescence. Confocal imaging allowed the identification of a heterogeneous cell population expressing green but also red fluorescence after 21 days of culture. Asymmetrical division of mature adipocytes was not observed. In conclusion, liposecretion of mature adipocytes is a phenomenon that can be observed in vitro and DFAT cells do originate from mature adipocytes. However, the population of DFAT cells is heterogenous.

Genetically Engineered Clostridium Prevents Perivascular Adipose Tissue and Endothelial Senescence and Dysfunction in Aging

Abstract Introduction (Peri)vascular senescence is a bona fide risk for several aging-associated diseases that threatens healthspan. Gut microbiome may be one of triggers that promotes vascular diseases as host ages. However, the interplay between vascular senescence and gut microbiome is largely unknown. Our recent studies revealed an increase in gut-derived phenylacetic acid (PAA) in the aging population (TwinsUK Aging Cohort, n=7,303) and aged mice. Our preliminary shotgun metagenomics also showed that ppfor-harboring Clostridium sp. ASF356 age-dependently generates PAA, which induces perivascular adipose tissue (PVAT) and endothelial cell (EC) senescence and dysfunction in old mice. Yet, it remains unclear whether and how genetic engineering of Clostridium sp. ASF356 (by depleting ppfor gene) can prevent aortic PVAT-EC senescence and restore vascular function in aging. Methods For bacterial genetic engineering, we depleted ppfor gene in Clostridium ASF356 by ClosTron technique and further mono-colonized aged mice (24-months old) and germ-free mice, as control, to reduce plasma PAA and thereby prevent aortic PVAT-EC senescence and improve endothelial dysfunction. Results As a proof of the concept, our studies exhibited that mono-colonization of germ-free mice with wild-type Clostridium sp. ASF356 markedly elevates plasma PAA levels. It was accompanied by hallmarks of aortic PVAT-EC senescence, including increased DNA damage (↑ γ-H2A.X phosphorylation), heightened SASP (↑ IL-6 and VCAM1), and proliferative arrest (↑ p16INK4a and p21WAF1/Cip1). Additionally, our findings revealed upreglation of Notch1 in PVAT, contributing to reduced energy supply (↓ UCP1) and augmented senescence-messaging secretome (↑ IL-6) towards ECs. Our further studies revealed that mono-colonization of aged mice (pre-treated with antibiotics) with Δppfor Clostridium ASF356 mutants intriguingly resulted in decreased plasma PAA levels. This intervention led to the rescues of cellular senescence in both aortic PVAT and ECs, consequently improving vascular function. Specifically, mice subjected to this treatment exhibited a significant reduction in SA-β-galactosidase+ cells, as well as decreased levels of γ-H2A.X phosphorylation, p16INK4a, p21WAF1/Cip1, and IL-6 in their aortic PVAT and ECs compared to wild-type aged mice. Conclusions The current study suggests that genetically engineered Δppfor Clostridium ASF356 mutants reverse cellular senescence and restore function in aortic PVAT and endothelial cells, highlighting the potential for targeted gut microbiome manipulation as an innovative senotherapy to combat vascular aging.

MicroRNA Expression in Chronic Venous Leg Ulcers and Implications for Wound Healing: A Scoping Review.

Purpose: Chronic venous leg ulcers (CVLUs) comprise the majority of lower-extremity wounds, yet their pathophysiology is not fully understood. While research has shown that microRNAs are an important component of wound inflammation, few have explored the role of microRNAs (miRNAs) in the healing of CVLUs. This scoping review examines miRNAs in CVLUs and the association with wound healing. Methods: In December 2023, we searched MEDLINE/PubMed, Embase, Scopus, and CINAHL for studies published in 2013-2023 examining miRNAs in CVLU healing. Results: Six studies met inclusion criteria. MicroRNAs were extracted from various specimens including serum, skin biopsy samples, and adipose tissue-derived mesenchymal cells from individuals with CVLUs. Overexpression of miR-221, miR-222, miR-92a, and miR-301a-3p hindered angiogenesis, while overexpression of miR-296, miR-126, miR-378, and miR-210 facilitated angiogenesis. Overexpression of miR-34a/c, miR-301a-3p, miR-450-5p, miR-424-5p, miR-516-5p, and miR-7704 increased local inflammatory responses and inhibited keratinocytes proliferation, impairing healing, while overexpression of miR-19a/b and miR-20 downregulated keratinocytes' inflammatory response, promoting healing. Downregulation of miR-205, miR-96-5p, and miR-218-5p enhanced cellular proliferation and promoted wound healing. Downregulation of miR-17-92 was linked with impaired healing. Discussion: MicroRNAs play a role in regulating angiogenesis, inflammatory responses, and cell migration in chronic-wound healing. However, studies of miRNAs in CVLUs are limited and lack a standardized approach to measurement and quantification. Further research is warranted to elucidate the mechanisms underlying microRNA involvement in CVLU healing to better understand the pathophysiology and for the future development of targeted therapies.