Adipose-derived Stem Cells Research Articles

Thermal-crosslinked acellular dermal matrix combined with adipose-derived stem cells to regenerate vascularized adipose tissue

The reconstruction of large-sized soft tissue defects remains a substantial clinical challenge, with adipose tissue engineering emerging as a promising solution. The acellular dermal matrix (ADM), known for its intricate spatial arrangement and active cytokine involvement, is widely employed as a scaffold in soft tissue engineering. Since ADM shares high similarity with decellularized adipose matrix, it holds potential as a substitute for adipose tissue. This study explores the adipogenic ability of a spongy material derived from ADM via vacuum-thermal crosslinking (T-ADM), characterized by high porosity, adjustable thickness, and suitable mechanical strength. Adipose-derived stem cells (ADSCs) are considered ideal seed cells in adipose tissue engineering. Nevertheless, whether pre-adipogenic induction is necessary before their incorporation remains debatable. In this context, ADSCs, both with and without pre-adipogenic induction, were seeded into T-ADM to regenerate vascularized adipose tissue. A comparative analysis of the two constructs was performed to evaluate angiogenesis and adipogenesisin vitro, and tissue regeneration efficacyin vivo. Additionally, RNA-seq analysis was utilized to investigate the potential mechanisms. The results showed that T-ADM exhibited good performance in terms of volume retention and maintenance of adipocyte phenotype, confirming its suitability as a scaffold for adipose tissue engineering.In-vitrooutcomes demonstrated that pre-adipogenic induction enhanced the adipogenic level of ADSCs, but reduced their ability to promote vascularization. Furthermore, constructs utilizing pre-induced ADSCs showed an insignificant superiority inin-vivofat formation, and neovascularization compared with those with non-induced ADSCs, which may be attributed to similar macrophage regulation, and balanced modulation of the proliferator-activated receptor-γand hypoxia-inducible factor 1αpathways. Consequently, the direct use of ADSCs is advocated to streamline the engineering process and reduce associated costs. The combined strategy of T-ADM with ADSCs proves to be feasible, convenient and effective, offering substantial potential for addressing large-sized tissue deficits and facilitating clinical applications.

Growth factors IGF-1 and KGF and adipose-derived stem cells promote migration and viability of primary human keratinocytes in an in vitro wound model

IntroductionIn the field of plastic surgery, epidermal transplantation is a potential treatment for chronic wounds that results in only minor donor site morbidity. Improving the regenerative capacities of epidermal grafts or single-cell suspensions and therefore accelerating healing processes would be of significant interest.MethodsIn the present study, we analyzed the effects of growth factors and adipose-derived stem cells (ADSCs) on keratinocyte properties. For optimum translation into the clinical setting, primary human keratinocytes and patient-matched ADSCs were isolated and used in an in vitro wound model.ResultsThe keratinocyte migration and viability increased after treatment with the growth factors insulin-like growth factor 1 (IGF-1) and keratinocyte growth factor (KGF). A similar effect was observed with the use of a concentrated ADSC-conditioned medium (ADSC-CM). It was further possible to isolate the keratinocytes in a xenogen-free medium, which is essential for clinical translation. Importantly, a patient-dependent influence on the effects of the growth factors and ADSC-CM was observed.DiscussionThis study provides potential for the improvement of epidermal transplantation in the treatment of chronic wounds using xenogen-free isolated and cultivated keratinocytes, growth factors, and ADSC. Translating these results into clinical application may help accelerate wound healing and shorten the time until patients can return to everyday life.

Early Autologous Fat Grafting for Facial Lipoatrophy Caused by Undifferentiated Connective Tissue Disease.

A variety of autoimmune disorders that do not fit the criteria for specific connective tissue diseases are included in the category of undifferentiated connective tissue disease (UCTD). Facial lipoatrophy is an underreported presenting characteristic, despite the prevalence of cutaneous symptoms. The aim of this article is to demistrate the potential of early autologous fat grafting (AFG) in treating facial depressions caused by UCTD. A 29-year-old woman with UCTD presented with progressive bilateral cheek depressions and was managed with tacrolimus lately. Sheet-like depressions can be found during physical examination, which are more pronounced on the left cheek. The patient underwent AFG surgery to the affected areas. During the surgery, a total of 30 mL of fat was harvested from the lower abdomen, and 5.0 mL of pure fat was eventually gained and grafted to the bilateral cheek depressions. Satisfying aesthetic outcomes were achieved with smooth facial contours. Through the use of adipose-derived stem cells, early AFG in the active stage of UCTD can enhance face appearance while also possibly having local anti-inflammatory and antifibrotic benefits. It also has important implications for improving the look and slowing the progression of other autoimmune diseases that affect the face.

The Potential Role of Adipose-Derived Stem Cells in Regeneration of Peripheral Nerves

The Potential Role of Adipose-Derived Stem Cells in Regeneration of Peripheral Nerves

Vitamin D Enhancement of Adipose Biology: Implications on Obesity-Associated Cardiometabolic Diseases

Vitamin D is activated into 1α,25(OH)2D through two hydroxylation steps that are primarily catalyzed by 25-hydroxylase in the liver and 1α-hydroxylase in the kidneys. The active form of vitamin D regulates myriads of cellular functions through its nuclear receptor, vitamin D receptor (VDR). Vitamin D metabolizing enzymes and VDR are expressed in adipose tissues and vitamin D regulates multiple aspects of adipose biology including the recruitment and differentiation of adipose stem cells into adipocytes and metabolic, endocrine, and immune properties. Obesity is associated with low vitamin D status, which is thought to be explained by its sequestration in large mass of adipose tissues as well as dysregulated vitamin D metabolism. Low vitamin D status in obesity may negatively impact adipose biology leading to adipose tissue dysfunctions, the major pathological factors for cardiometabolic diseases in obesity. In this review, the current understanding of vitamin D metabolism and its molecular mechanisms of actions, focusing on vitamin D–VDR regulation of adipose biology with their implications on obesity-associated diseases, is discussed. Whether improving vitamin D status leads to reductions in adiposity and risks for cardiometabolic diseases is also discussed.

Species-Specific Optimisation of Cryopreservation Media for Goat and Buffalo Adipose-Derived Mesenchymal Stem Cells.

Adipose-derived mesenchymal stem cells (ADSCs) are promising for clinical and veterinary applications due to their ease of isolation, high yield, and multilineage differentiation potential. Effective cryopreservation is vital to ensure their availability for large-scale applications. This study evaluated cryopreservation strategies for goat (gADSCs) and buffalo (bADSCs) ADSCs, using combinations of intracellular (dimethyl sulfoxide, DMSO) and exocellular cryoprotectants, including fetal bovine serum (FBS), polyethylene glycol (PEG), trehalose, bovine serum albumin (BSA), and dextran. Post-thaw parameters such as viability, recovery, metabolic activity, clonogenicity, oxidative stress, apoptosis, and senescence were assessed. Results revealed species-specific differences in cryopreservation requirements. gADSCs were optimally preserved in a medium with 5% DMSO, 3% FBS, 2% PEG, 3% trehalose, and 2% BSA, while bADSCs performed best in an FBS-free medium containing 5% DMSO, 2% PEG, 3% trehalose, and 2% BSA. DMSO-FBS formulations supported high recovery and metabolic activity but were associated with increased oxidative stress and apoptosis. Dextran-based cryomedia effectively preserved gADSCs but failed to maintain bADSC functionality. Biochemical composition analysis indicated significantly higher lipid content in bADSCs, likely influencing cryopreservation efficacy. These findings underscore the need for tailored cryopreservation strategies to address species-specific differences. Incorporating exocellular cryoprotectants reduced FBS dependency, minimised oxidative damage, and maintained functional attributes. This study highlights the potential for optimised, cost-effective biobanking solutions that accommodate species-specific requirements, advancing the use of ADSCs in veterinary and translational research.

Purified adipose tissue-derived extracellular vesicles facilitate adipose organoid vascularization through coordinating adipogenesis and angiogenesis.

One of the major challenges in the way of better fabricating vascularized adipose organoids is the destructive effect of adipogenic differentiation on preformed vasculature, which probably stems from the discrepancy between the in vivo physiological microenvironment and the in vitro culture conditions. As an intrinsic component of adipose tissue (AT), adipose tissue-derived extracellular vesicles (AT-EVs) have demonstrated both adipogenic and angiogenic ability in recent studies. However, whether AT-EVs could be employed to coordinate the angiogenesis and adipogenesis in the vascularization of adipose organoids remains largely unexplored. Herein, we present an efficient method for isolating higher-purity AT-EV preparations from lipoaspirates, and verify the superiority of AT-EV preparations' angiogenic and adipogenic capabilities over those from unpurified lipoaspirates. Next, in the spheroid culture model, it was discovered that the addition of AT-EVs could effectively improve the aggregation through enhancing intercellular adhesion of monoculture spheroids composed of human umbilical vascular endothelial cells (HUVECs), and helped produce vascularized adipose organoids with proper lipolysis and glucose uptake ability in the coculture spheroids comprised of adipose-derived stem cells (ADSCs) and HUVECs. Subsequently, it was observed that AT-EVs could exert a retaining effect on the vasculature of prevascularized coculture spheroids cultured in an adipogenic environment, compared to the reduced vascular networks where AT-EVs were absent. Altogether, these results indicate that AT-EVs, by means of releasing bioactive molecules that emulate the in vivo microenvironment, can modify non-replicative in vitro microenvironments, coordinate in vitro adipogenesis and angiogenesis, and facilitate the fabrication of vascularized adipose organoids.&#xD.

Recent advances in adipose-derived mesenchymal stem cell-derived exosomes for regulating macrophage polarization

Adipose-derived mesenchymal stem cells (ADSCs) exhibit superior immunomodulatory properties and have broad therapeutic applications. They induce macrophage M2 polarization for anti-inflammatory responses. Exosomes derived from ADSCs (ADSC-EXOs) exhibit biological functions similar to those of ADSCs but can circumvent the limitations associated with cellular injection therapies. Potent anti-inflammatory substances contained in exosomes include the glycoprotein MFGE8, the cytokines such as prostaglandin E2, IL-6, and IGF, as well as non-coding nucleotides (miR-451a, miR-23, miR-30d-5p, let-7, lncRNA DLEU2, circRps5, Circ-Ptpn4, and mmu_ circ_0001359). The anti-inflammatory and immunomodulatory properties of these exosomes provide new perspectives for therapeutic approaches for graft inflammation, bone healing, acute lung injury, kidney stones, myocardial infarction, and diabetes-related diseases. This review summarizes the contents and functions of ADSC-EXOs, outlines their properties and the characteristics of macrophage phenotypes, and emphasizes their impact on macrophage polarization and their contribution to immune-related diseases.

Development and Characterization of Magnetic Nanoemulsion-Based Senolytic Peptides for Osteoarthritis Treatment

The formulation and characterization of a novel nanoemulsion (NE) delivery system for senomodulator peptides aimed at enhancing the treatment of osteoarthritis (OA) are reported, in combination with magnetic nanoparticles (MNPs), for improving targeted delivery and traceability. Osteoarthritis, a prevalent degenerative joint disease associated with aging, is currently not effectively treated by disease-modifying therapies, posing a consistent health burden on individuals and healthcare systems worldwide. Existing treatments, such as nonsteroidal anti-inflammatory drugs and intra-articular injections, are limited by inadequate local drug concentrations and rapid clearance, often necessitating costly joint replacement. Lipid-based NE composed of biocompatible and biodegradable vitamin E and sphingomyelin, associated with the senolytic peptide NE:TUB1, is able to target senescent cells implicated in OA progression. Improved cellular retention and therapeutic effects of the associated TUB1 peptide, compared to its free form, have been demonstrated, suggesting a significant enhancement in therapeutic potential. The incorporation of MNPs to obtain NE:TUB1-MNP formulations offers the advantage of being traceable in vivo through clinically available imaging technologies, with the potential to enhance targeting capabilities through magnetic guidance. The characterization of NE:TUB1-MNPs involved the assessment of their physical and chemical properties, interaction with cells, cytotoxicity profile, and nanoparticle uptake in vitro using human primary adipose-derived stem cells. NE and NE:TUB1-MNP are shown to be stable, non-toxic, and capable of efficient intracellular uptake. The inclusion of MNPs not only supports cell viability and proliferation but also facilitates medium and long-term product traceability within joints, offering a promising approach for localized treatment. The enhanced anti-senescent role of NE:TUB1-MNP formulations are highlighted, suggesting their potential utility in mitigating OA progression and possibly other degenerative diseases. In conclusion, the study presents a novel therapeutic approach for OA, NE:TUB1-MNPs, leveraging the synergistic effects of peptide-functionalized nanoemulsions and magnetic nanoparticles to improve targeted delivery and therapeutic outcomes. This innovative formulation could pave the way for new treatments for OA and other joint-related conditions, offering significant advancements in regenerative medicine.

Fabrication and Characterization of Electrospun DegraPol® Tubes Releasing TIMP-1 Protein to Modulate Tendon Healing

Background: Tendon rupture repair can result from fibrotic scar formation through imbalanced ECM deposition during remodeling. The tissue inhibitors of matrix metalloprotease (TIMPs) not only decrease ECM degradation, regulated by matrix metalloproteases (MMPs), but also restrict TGF-β1 activation and thus diminish fibrosis. Methods: Rabbit tenocytes (rbTenocytes) and rabbit adipose-derived stem cells (rbASCs) were cultivated under different TIMP-1 concentrations. Proliferation and gene expression were assessed. TIMP-1 was incorporated into emulsion electrospun DegraPol® (DP) tubes that were characterized by SEM for fiber thickness, pore size, and wall thickness. Static and dynamic water contact angles, FTIR spectra, and TIMP-1 release kinetics were determined. Results: While the proliferation of rbTenocytes and rbACS was not affected by TIMP-1 supplementation in vitro, the gene expression of Col1A1 was increased in rbTenocytes, the gene expression of ki67 was increased in both cell types, the gene expression of tenomodulin was increased in both cell types at 100 ng/mL TIMP-1, and alkaline phosphatase expression ALP rose significantly in rbASCs. Electrospun TIMP-1/DP fibers had a ~5 μm diameter, a ~10 μm pore size, and a mesh thickness of ~200 μm. TIMP-1/DP meshes were more hydrophilic than pure DP meshes. TIMP-1 was released from the meshes with a sustained release of up to 7 days. Conclusions: TIMP-1/DP tubes may be used to modulate the fibrotic tissue reaction when applied around conventionally sutured tendon ruptures.

Enhancing fat graft survival: thymosin beta-4 facilitates mitochondrial transfer from ADSCs via tunneling nanotubes by upregulating the Rac/F-actin pathway.

Autologous fat grafting is a widely used technique in plastic and reconstructive surgery, but its efficacy is often limited by the poor survival rate of transplanted adipose tissue. This study aims to enhance the survival of fat grafts by investigating the role of thymosin beta-4 (Tβ4) in facilitating mitochondrial transfer from adipose-derived stem cells (ADSCs) to adipocytes and newly formed blood vessels within the grafts via tunneling nanotubes (TNTs). We demonstrate that Tβ4 upregulates the Rac/F-actin pathway, leading to an increased formation of TNTs and subsequent transfer of mitochondria from ADSCs. This process mitigates oxidative stress, reduces apoptosis, and promotes revascularization, thereby improving the quality and volume retention of fat grafts. Our findings provide a novel mechanistic insight into the enhancement of fat graft survival and suggest that mitochondrial transplantation and Tβ4 are potential therapeutic strategies to improve clinical outcomes in autologous fat transfer procedures.

PIEZO1-mediated mechanotransduction regulates collagen synthesis on nanostructured 2D and 3D models of fibrosis.

Progressive fibrosis can lead to tissue malfunction and organ failure due to the pathologic accumulation of a collagen-rich extracellular matrix. In vitro models provide useful tools for deconstructing the roles of specific biomechanical or biological mechanisms, such as substrate micro- and nanoscale architecture, in these processes for identifying potential therapeutic targets. Here, we investigated how the mechanosensitive ion channel PIEZO1 influences fibrotic gene and protein expression in adipose-derived stem cells (hASCs). Specifically, we examined the role of PIEZO1 and the mechanosensitive transcription factors YAP/TAZ in sensing aligned or non-aligned substrate architecture to regulate collagen formation. We utilized both 2D microphotopatterned substrates and 3D electrospun polycaprolactone (PCL) substrates to study the role of culture dimensionality. We found that PIEZO1 regulates collagen synthesis in hASCs in a manner that is sensitive to substrate architecture. Activation of PIEZO1 induced significant morphological changes in hASCs, particularly when cultured on aligned substrates, leading to a 30-40 % reduction in cell spreading area and increased cell elongation, in 3D-aligned cultures. Picrosirius Red staining and immunoblotting revealed that PIEZO1 activation reduced collagen accumulation in 3D culture. While YAP translocated to the cytoplasm following PIEZO1 activation, depleting YAP and TAZ did not change collagen expression significantly downstream of PIEZO1 activation, implying that YAP/TAZ translocation from the nucleus and decreased collagen synthesis may be independent consequences of PIEZO1 activation. Our studies demonstrate a role for PIEZO1 in cellular mechanosensing of substrate architecture and provide targetable pathways for treating fibrosis and for enhancing tissue-engineered and regenerative approaches for fibrous tissue repair. STATEMENT OF SIGNIFICANCE: This study examines how cells sense and respond to their physical environment via PIEZO1 mechanotransduction. We discovered that cells use PIEZO1 to detect the alignment of surrounding structures, influencing the production of collagen - a key component in fibrosis. Our study used both 2D and 3D models to mimic different tissue environments, providing new insights into how cellular responses change in more complex settings. Importantly, we found that activating PIEZO1 alters cell shape and collagen production, especially on aligned surfaces. Interestingly, while PIEZO1 activation caused YAP translocation to the cytoplasm, this translocation did not directly affect collagen production. This work advances our understanding of fibrosis development and identifies PIEZO1 as a potential target for new therapies.

Intra-Articular Stromal Vascular Fraction and Mesenchymal Stem Cell Injections Show Variable Efficacy and Higher Potential Complications Compared to Corticosteroid and Hyaluronic Acid in Treatment of Knee Osteoarthritis: A Meta-Analysis of Randomized Controlled Trials.

This study aimed to evaluate the clinical outcomes of intra-articular stromal vascular fraction (SVF) and adipose-derived mesenchymal stem cell (ASC) injections in patients with knee osteoarthritis (OA) through a meta-analysis of randomized clinical trials. PubMed, Embase, the Cochrane Library, and Google Scholar were systematically searched to identify level I studies that compared the clinical efficacy of SVF or ASC with that of other non-operative treatments. Clinical scores measured 3, 6 and 12 months post-injection were standardized to pain and functional scales for meta-analysis based on minimal clinically important differences. Follow-up MRI findings and safety-related data were also investigated. Nine studies involving 671 patients were included. SVF demonstrated superior pain and function score improvements compared to saline or hyaluronic acid (HA) at 3, 6, and 12 months post-injection. However, SVF was inferior to corticosteroid at 3 months, showed no difference at 6 months, and was comparable or slightly superior at 12 months. ASC consistently showed better pain and function score improvements compared to saline and conservative treatment at all time points, with no significant differences observed compared to HA. High heterogeneity was noted in SVF function score results, requiring cautious interpretation. No serious adverse events related to SVF or ASC were reported, although complications associated with liposuction were observed. SVF showed consistent superiority in pain and function scores compared to HA and saline but not corticosteroids, except for the 3-month function score. ASC improved pain and function scores compared to saline and conservative treatment, but was not superior to HA. While SVF and ASC significantly improved clinical scores, the potential complications from liposuction limit their ability to replace existing minimally invasive treatments for knee OA. I, meta-analyses of randomized controlled trials.

The Impact of Amotosalen Photochemical Pathogen Inactivation on Human Platelet Lysate.

Human Platelet Lysate (hPL) is a platelet-derived and growth factor-rich supplement for cell culture. It can be prepared from surplus platelet concentrates initially intended for transfusion. Amotosalen-based photochemical pathogen inactivation of platelet concentrates is used in a number of blood establishments worldwide to minimize the risk of pathogen transmission from donor to patient. This pathogen inactivation method has not been formally validated for direct use on hPL. Here, we have studied the impact of pathogen inactivation on hPL and compared it to untreated hPL prepared from pathogen-inactivated platelet concentrates or control hPL. We used mass spectrometry, ELISA, and in vitro mesenchymal stem cell culture for determining residual amotosalen, final growth factor content, and cell doubling, respectively. The data have shown amotosalen concentrations to be reduced a thousand-fold following pathogen inactivation, leaving trace quantities of photosensitizer molecules in the final hPL product. Some growth factors have been reported to be significantly more impacted in hPL that is directly pathogen-inactivated compared to both control conditions. This has no significant effect on the growth kinetics of adipose-derived mesenchymal stem cells. Direct amotosalen-based pathogen inactivation has a measurable impact on certain growth factors in hPL, but this does not outweigh the likely benefits of reducing the odds of donor-to-patient pathogen transmission.

High paracrine activity of hADSCs cartilage microtissues inhibits extracellular matrix degradation and promotes cartilage regeneration

Due to its unique structure, articular cartilage has limited self-repair capacity. Microtissues are tiny tissue clusters that can mimic the function of target organs or tissues. Using cells alone for microtissue construction often results in the formation of necrotic cores. However, the extracellular matrix (ECM) of native cartilage can provide structural support and is an ideal source of microcarriers. Autologous adipose-derived mesenchymal stem cells (ADSCs) and bone marrow mesenchymal stem cells (BMSCs) are widely used in cartilage tissue engineering. In this study, we fabricated microcarriers and compared the behavior of two homologous cell types in the microcarrier environment. The microcarrier environment highlighted the advantages of ADSCs and promoted the proliferation and migration of these cells. Then, ADSCs microtissues (ADSCs-MT) and BMSCs microtissues (BMSCs-MT) were fabricated using a three-dimensional dynamic culture system. In vitro and in vivo experiments verified that the cartilage regeneration ability of ADSCs-MT was significantly superior to that of BMSCs-MT. Transcriptomics revealed that ADSCs-MT showed significantly lower expression levels of ECM degradation, osteogenesis, and fibrocartilage markers. Finally, the protective effect of microtissues on inflammatory chondrocytes was validated. Overall, the ADSCs-MT constructed in this study achieved excellent cartilage regeneration and could be promising for the autologous application of cartilage microtissues.

Porcine pericardial decellularized matrix bilayer patch containing adipose stem cell-derived exosomes for the treatment of diabetic wounds.

Chronic hard-to-heal wounds pose a significant threat to patients' health and quality of life, and their clinical management remains a challenge. Adipose-derived stem cell exosomes (ADSC-exos) have shown promising results in promoting diabetic wound healing. However, effectively enhancing the retention of exosomes in wounds for treatment remains a key issue that needs to be addressed. There is a pressing need to develop new materials or methods to improve the bioavailability of exosomes. Porcine pericardium, an extracellular matrix-rich tissue, is easily obtainable and widely available. Decellularized porcine pericardium removes cellular components while retaining an extracellular matrix that supports cellular growth, making it an ideal raw material for preparing wound dressings. In this study, we developed porcine pericardial decellularized matrix bilayer patches loaded with ADSC-exos, which were transplanted into diabetic mouse skin wounds. Histological and immunohistochemical analyses revealed that these bilayer matrix patches accelerate wound healing by promoting granulation tissue formation, re-epithelialization, stimulating vascularization, and enhancing collagen production. In terms of the underlying biological mechanism, we found that decellularized extracellular matrix bilayer patches loaded with ADSC-exos enhanced the proliferation and migration of human dermal fibroblasts (HDFs) and HaCaT cells in vitro, and promoted tube formation in human umbilical vein endothelial cells (HUVECs). This research demonstrated that the porcine pericardial decellularized matrix is well-suited for exosome delivery and that these bilayer patches hold great potential in promoting diabetic wound healing, providing evidence to support the future clinical application of ADSC-exos.

Sprouting and network data assessed in the chicken aortic ring assay under stimulation with human adipose-derived stem cell secretomes harvested from single cells or spheroids of different sizes

Sprouting and network data assessed in the chicken aortic ring assay under stimulation with human adipose-derived stem cell secretomes harvested from single cells or spheroids of different sizes

Regulation of UCP1 expression by PPARα and pemafibrate in human beige adipocytes.

Thermogenic adipocytes are able to dissipate energy as heat from lipids and carbohydrates through enhanced uncoupled respiration, due to UCP1 activity. PPAR family of transcription factors plays an important role in adipocyte biology. The purpose of this work was to characterize the role of PPARα and pemafibrate in the control of thermogenic adipocyte formation and function. We used human multipotent adipose-derived stem cells and primary cultures of stroma-vascular fraction cells, transfected with siRNA against PPARα, differentiated into white or beige adipocytes, by the treatment of rosiglitazone or pemafibrate. The expression of key marker genes of adipogenesis and thermogenesis was determined using RT-qPCR and Western blotting. An RNAseq analysis was also performed. We show that inhibition of PPARα mRNA increases UCP1 mRNA and protein expression in beige adipocytes induced by rosiglitazone. Knock-down of PPARα also increases stimulated glycerol release. Pemafibrate, described as a selective PPARα modulator, induces adipogenesis and the expression of UCP1 in the absence of PPARα expression. These effects are inhibited by a specific PPARγ antagonist highly suggesting that the pemafibrate effects in adipogenesis and beiging were mediated by PPARγ. Conversion of white into thermogenic adipocytes is mainly due to the activation of PPARγ. Moreover, we show that PPARα seems to act as a hindrance for PPARγ-dependent beiging. Our data question the role of PPARα in human adipocyte browning and the specificity of pemafibrate in adipocytes.

Silanized acrylic graphene oxide nanocomposite reinforced mechanically tunable GelMA/HAMA printable bio-ink for adipose-derived stem cells differentiated mature smooth muscle cells

Silanized acrylic graphene oxide nanocomposite reinforced mechanically tunable GelMA/HAMA printable bio-ink for adipose-derived stem cells differentiated mature smooth muscle cells

PCL-fibrin-alginate hydrogel based cell co-culture system for improving angiogenesis and immune modulation in limb ischemia.

Stem cell therapy has demonstrated promise in regenerative medicine due to their ability to differentiate into various cell types and secrete growth factors. However, challenges such as poor survival rate of transplanted cells under ischemic and immune conditions limit its effectiveness. To address these issues, we developed a polycaprolactone (PCL)-fibrin-alginate matrix hydrogel, which combines adipose-derived stem cells and human umbilical vein endothelial cells with a PCL fiber, encapsulated within fibrin and alginate hydrogel to enhance cell survival, proliferation, and immune modulation. This structure offers protection to the encapsulated cells, supports angiogenesis, and modulates the immune response, significantly improving therapeutic outcomes in a mouse model of hindlimb ischemia. Our in vitro and in vivo results demonstrate the scaffold's ability to support cell viability, promote angiogenesis, and modulate inflammatory responses, indicating its potential as a promising platform for ischemic tissue repair and regenerative medicine. This innovative approach to cell-based therapy highlights the importance of scaffold design in enhancing the therapeutic efficacy of stem cell treatments for ischemic diseases.