American journal of physiology. Cell physiology最新文献

{"title":"PLGA nanoparticles restore acidic pH and degradative function to compromised lysosomes with Cy3-labeling providing enhanced tracking to lysosomes.","authors":"Jiaqi Li, Tianchen Wang, Wennan Lu, Davit Jishkariani, Andrew Tsourkas, Simon Kaja, Kyle H Vining, Jedtanut Thussananutiyakul, Ashley Spence, Rohini M Nair, Joshua L Dunaief, Claire H Mitchell","doi":"10.1152/ajpcell.00494.2025","DOIUrl":"10.1152/ajpcell.00494.2025","url":null,"abstract":"<p><p>Lysosomal dysfunction and elevated lysosomal pH are hallmark features of age-related neurodegenerative diseases including age-related macular degeneration (AMD), Alzheimer's disease (AD), and Parkinson's disease (PD). Restoring lysosomal acidity is important for maintaining enzymatic degradation, preventing protein aggregation, and reducing cellular waste accumulation in degenerating tissues. Acidic nanoparticles represent a promising therapeutic strategy to normalize lysosomal pH; however, accurate monitoring of their delivery, retention, and dosage is critical for rigorous evaluation. To address this, we developed fluorescently labeled poly(d,l-lactide-co-glycolide) (PLGA) nanoparticles conjugated with Cyanine3 amine (Cy3). Nanoparticle uptake was systematically optimized, achieving over 90% delivery to lysosomes of induced pluripotent stem cell-derived retinal pigment epithelial (iPS-RPE) cells, although uptake rates varied among adjacent cells. Once internalized, nanoparticles demonstrated remarkable stability, with no detectable change in concentration, distribution, or size for at least 28 days. iPS-RPE cells exhibited higher nanoparticle internalization compared with the ARPE-19 cell line and optic nerve head astrocytes. The capacity of the nanoparticles to restore function to stressed lysosomes was confirmed by their ability to reacidify lysosomes, restore cathepsin B activity, and increase the levels of active cathepsin D. The nanoparticles also reduced the levels of LC3II in astrocytes treated with chloroquine, indicating that they can also restore autophagy rates. In summary, this study demonstrates the value of Cy3 labeling for enhanced nanoparticle tracking to lysosomes. The findings also identify PLGA nanoparticles as powerful tools for restoring degradative lysosomal function and autophagy in cells undergoing lysosomal stress.<b>NEW & NOTEWORTHY</b> Tools that restore acidic pH in compromised lysosomes can enhance autophagy and waste clearance in degenerative disorders characterized by excessive accumulation. Here, we describe the synthesis of lysosome-targeted nanoparticles composed of poly(d,l-lactide-co-glycolide) (PLGA) polymers covalently bound to the fluorescent dye Cyanine3 amine (Cy3). These Cy3-PLGA nanoparticles enable precise tracking of lysosomal delivery and demonstrate sustained long-term retention within lysosomes, supporting their potential for future applications aimed at restoring lysosomal pH in aging and degenerating diseases.</p>","PeriodicalId":7585,"journal":{"name":"American journal of physiology. Cell physiology","volume":" ","pages":"C509-C523"},"PeriodicalIF":4.7,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145964933","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Retinoic acid promotes expression of inflammatory factors in proliferative adult human heart cells.","authors":"Ian M Gans, Amanda J Lessard, Sergey V Ryzhov, Calvin P Vary, Douglas B Sawyer","doi":"10.1152/ajpcell.00696.2025","DOIUrl":"10.1152/ajpcell.00696.2025","url":null,"abstract":"<p><p>Retinoid signaling is increased in the hearts of patients with coronary artery disease and during acute myocardial infarction (MI). The effects of retinoids on cardiac repair after injury remain incompletely understood. Our laboratory has derived proliferative cardiac cell clones from adult human left ventricle biopsies and is investigating how these cells might participate in cardiac repair in heart failure. We treated clones isolated from unique individuals with retinoic acid (RA) and performed unbiased proteomics, bioinformatic analyses, and targeted follow-up experiments to identify and confirm RA-regulated factors and processes. RA increased the expression of well-known proinflammatory proteins including interleukin-1 (IL1A and B) and inducible cyclooxygenase 2 (COX2), while decreasing the expression of extracellular matrix (ECM) factors such as thrombospondin 1 and collagens. Additionally, we found that basal expression of retinoid metabolizing enzymes (e.g., ALDH1A3) significantly correlated with expression of cytokines and inflammatory mediators including IL1A/B and COX2 across clones from different donors. Secretion of IL1B by clones was found to respond to physiological and pharmacological doses of RA, and monocyte migration in vitro responded to secretions from RA-treated clones. Our findings suggest a mechanism by which retinoids promote inflammation and contribute to adverse cardiac remodeling in the injured heart, providing a potential avenue to regulate myocardial inflammation and remodeling processes.<b>NEW & NOTEWORTHY</b> Within the injured heart, cells are exposed to elevated retinoic acid signaling resulting from mobilized stores of its precursor, vitamin A, and increased cardiac expression of synthesizing enzymes. This study investigated the effects of retinoic acid, a potent regulator of cell fate and function, on human proliferative cardiac cell clones derived from left ventricle biopsies. The results show an increase in inflammatory factor secretion, immune cell activation, and decreased extracellular matrix expression.</p>","PeriodicalId":7585,"journal":{"name":"American journal of physiology. Cell physiology","volume":" ","pages":"C367-C378"},"PeriodicalIF":4.7,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12990825/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145780010","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The roles of Tenascin C in kidney diseases.","authors":"Tim Caspers, Peter Boor, Barbara Mara Klinkhammer","doi":"10.1152/ajpcell.00514.2025","DOIUrl":"10.1152/ajpcell.00514.2025","url":null,"abstract":"<p><p>Tenascin C (TNC), an extracellular matrix glycoprotein, is crucial for embryonic development and tissue repair, inflammation, extracellular matrix remodeling, and fibrosis, particularly in kidney diseases. Although its expression is typically low in healthy adult kidneys, TNC is upregulated in various kidney disease conditions, including acute kidney injury (AKI) and chronic kidney disease (CKD). TNC influences fibroblast activation, and elevated TNC levels correlate with CKD severity, highlighting its potential as a biomarker for diagnosis and monitoring of fibrogenesis. TNC's multifaceted role offers opportunities for therapeutic interventions. Here, we provide an overview of TNC's structural and functional attributes, its regulatory mechanisms, and its multifactorial role in kidney disease development and progression. We also discuss recent approaches aiming to use TNC as a target for diagnostic and therapeutic purposes.</p>","PeriodicalId":7585,"journal":{"name":"American journal of physiology. Cell physiology","volume":" ","pages":"C552-C569"},"PeriodicalIF":4.7,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145793028","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Plectin associates with focal adhesions and contributes to cytoskeletal organization and mechanical properties of astrocytes.","authors":"Borut Furlani, Maja Potokar, Victorio Martin Pozo Devoto, Dolores Pérez-Sala, Gerhard Wiche, Robert Zorec, Jernej Jorgačevski","doi":"10.1152/ajpcell.00425.2025","DOIUrl":"10.1152/ajpcell.00425.2025","url":null,"abstract":"<p><p>Reactive astrogliosis, a hallmark of central nervous system pathologies, involves a spectrum of astrocyte responses, including morphological remodeling and the upregulation of intermediate filaments such as vimentin and glial fibrillary acidic protein (GFAP). Changes in astrocyte shape are driven by cytoskeletal dynamics and are important for interactions with the surrounding microenvironment. Focal adhesions (FAs), which serve as physical and signaling links between the cytoskeleton and the extracellular matrix, play a central role in these structural adaptations. Here, we identify plectin, a versatile cytoskeletal linker, as an important modulator of FA-associated processes in cultured mouse astrocytes. We demonstrate that plectin localizes to FAs in astrocytes, and its deficiency is associated with changes in their number, maturation, and turnover. Plectin also displays polarization within FAs, depending on their maturation state, and it contributes to the recruitment of key cytoskeletal elements, particularly vimentin, to FAs. In plectin-deficient astrocytes, the vimentin and GFAP network exhibits impaired connectivity, accompanied by altered viscoelastic properties of the cells. Compared with astrocytes maintained in serum-free neurobasal medium, astrocytes cultured in serum-containing medium, which resemble reactive astrocytes, exhibit elevated plectin levels along with an increased number and size of FAs, supporting the involvement of plectin in pathological conditions.<b>NEW & NOTEWORTHY</b> Plectin contributes to FA dynamics in astrocytes and exhibits spatial polarization within individual FAs as revealed by superresolution microscopy (SIM and STED). Atomic force microscopy demonstrated that plectin deficiency alters cell viscoelasticity, unveiling the role of plectin in the mechanical properties of astrocytes. Plectin expression, along with the FA protein vinculin, is upregulated in astrocytes cultured under serum-containing conditions-an in vitro model of reactive astrocytes-compared with serum-free, native-like conditions.</p>","PeriodicalId":7585,"journal":{"name":"American journal of physiology. Cell physiology","volume":" ","pages":"C327-C344"},"PeriodicalIF":4.7,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145739980","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Orai channels in proliferation, invasion, and chemoresistance of tumor cells.","authors":"Isaac Jardín, Alvaro Macias-Díaz, Vanesa Jimenez-Velarde, Tarik Smani, Juan A Rosado","doi":"10.1152/ajpcell.00907.2025","DOIUrl":"10.1152/ajpcell.00907.2025","url":null,"abstract":"<p><p>Calcium signaling via store-operated calcium entry (SOCE) is critical for cellular functions implicated in cancer progression. Alterations in Orai channel isoforms, particularly Orai1 and Orai3, modulate SOCE and influence tumor cell proliferation, invasion, and survival. Here, we review and synthesize current evidence showing how Orai1 and Orai3 isoforms modulate oncogenic calcium signals through pathways such as phosphatidylinositol 3-kinase (PI3K)/Akt, ERK1/2, and NF-κB, contributing to tumor progression and chemoresistance by regulating apoptosis, autophagy, and oxidative stress responses. This isoform-specific remodeling enables tumor cells to adapt to therapeutic challenges and oxidative environments. Emerging data suggest that modulating Orai channel function and isoform composition may sensitize some cancer cells to apoptosis and attenuate invasive behavior, at least in specific experimental models. Taken together, available studies support a role for Orai channels as important regulators of tumor-associated Ca²⁺ signaling and highlight their potential as context-dependent targets to modulate survival and invasive behavior in cancer models.</p>","PeriodicalId":7585,"journal":{"name":"American journal of physiology. Cell physiology","volume":" ","pages":"C496-C508"},"PeriodicalIF":4.7,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145987560","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cystamine is a redox-dependent inverse agonist of the histamine H₄ receptor.","authors":"Takeshi Honda, Tatsuya Sakaguchi, Atsuo Kuramasu","doi":"10.1152/ajpcell.00891.2025","DOIUrl":"10.1152/ajpcell.00891.2025","url":null,"abstract":"<p><p>Cystamine, the oxidized dimer of cysteamine, has been reported to exert anti-inflammatory actions, but the underlying molecular mechanism remains unclear. Because the histamine H₄ receptor (H4R) is a key regulator of mast cell chemotaxis and inflammatory signaling, we examined whether cystamine directly modulates H4R activity. Cystamine potently inhibited histamine-induced mast cell migration (IC₅₀ = 440 nM) without affecting stem cell factor-induced migration, indicating pathway specificity. Although cystamine suppressed transglutaminase activity, this required millimolar concentrations and did not account for its effect on migration. At low micromolar concentrations, cystamine attenuated histamine-dependent activation of Rac1 and Rac2 GTPases and extracellular signal-regulated kinase (ERK). In H4R-expressing HEK293A cells, cystamine reduced basal and agonist-induced cyclic AMP response element reporter activity, demonstrating competitive antagonism and inverse agonism. Molecular docking supported direct binding of cystamine-but not its reduced monomer, cysteamine-to H4R. These findings identify cystamine as a redox-dependent inverse agonist of the H4R and provide a mechanistic explanation for its reported anti-inflammatory properties. Notably, cystamine generated endogenously or reformed locally from cysteamine under oxidative conditions may act on H4R in a redox-dependent and microenvironment-specific manner. Together, these insights suggest the potential for developing redox-dependent, tissue-selective H4R modulators for inflammatory diseases.<b>NEW & NOTEWORTHY</b> This study identifies cystamine as a previously unrecognized modulator of H4R. Cystamine inhibits mast cell migration and suppresses H4R-mediated Rac-ERK signaling at concentrations far below those required to inhibit transglutaminase, and it exhibits inverse agonist activity. Docking simulations show that cystamine, but not reduced cysteamine, engages the receptor's orthosteric site, revealing a redox-dependent difference in receptor interaction.</p>","PeriodicalId":7585,"journal":{"name":"American journal of physiology. Cell physiology","volume":" ","pages":"C390-C395"},"PeriodicalIF":4.7,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145888476","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Endotrophin as a biomarker and mediator in cardiovascular-kidney-metabolic syndrome: current insights and remaining questions.","authors":"Federica Genovese, Morten Karsdal, Hanne Devos, Cecilie Bager, Julio Nuñez, Antoni Bayes-Genis","doi":"10.1152/ajpcell.00769.2025","DOIUrl":"10.1152/ajpcell.00769.2025","url":null,"abstract":"<p><p>Endotrophin, a biologically active fragment derived from the α3 chain of collagen type VI, has emerged as both a risk biomarker and a potential pathogenic factor in cardiovascular-kidney-metabolic (CKM) syndrome. Over the past decade, research has shed light on its role in various noncommunicable diseases, emphasizing its signaling properties and diagnostic potential. Despite these advances, significant gaps remain in our understanding of how endotrophin contributes to CKM pathophysiology and whether targeting it therapeutically could modify disease progression. This narrative review synthesizes current evidence on endotrophin biological functions and clinical associations, drawing from both experimental and clinical studies. In addition, it identifies critical areas where further investigation is required, including the molecular mechanisms linking endotrophin to CKM-related tissue dysfunction and its causal role in disease development. By mapping current knowledge and highlighting research priorities, this review aims to advance the field toward a more complete understanding of endotrophin as a potential therapeutic target.</p>","PeriodicalId":7585,"journal":{"name":"American journal of physiology. Cell physiology","volume":" ","pages":"C409-C420"},"PeriodicalIF":4.7,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145713012","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"PIEZO1 channel mechanosensing in hepatobiliary physiology and disease.","authors":"Charalampos Konstantinou, Shaili K Patel, Patricia Lalor, J Bernadette Moore, Lee D Roberts, Naima Endesh, Tze M Wah, Adel Samson, Abdul R Hakeem, Kondragunta R Prasad, David J Beech, Laeticia Lichtenstein","doi":"10.1152/ajpcell.00782.2025","DOIUrl":"10.1152/ajpcell.00782.2025","url":null,"abstract":"<p><p>The hepatobiliary system is constantly exposed to dynamic mechanical forces, including fluid shear stress, bile canaliculi pressure, and extracellular matrix stiffness. Although traditionally studied for its metabolic and detoxifying functions, it is now increasingly recognized as a mechanosensitive organ. This review focuses on PIEZO1 mechanically gated ion channels that transduce physical cues into calcium-dependent signaling events. PIEZO2, the only other PIEZO isoform, is not known to be relevant in the hepatobiliary system. We examine the current knowledge on PIEZO1 in liver physiology, highlighting its roles in liver sinusoidal endothelial cells, hepatocytes, and macrophages. In health, PIEZO1 regulates key processes such as bile acid synthesis (through nitric oxide-mediated suppression of <i>CYP7A1</i>), bile flow, antioxidant defense, and iron homeostasis. In disease, PIEZO1 activity is linked to pathological processes such as inflammation, fibrosis, and angiogenesis in the context of cirrhosis and hepatocellular carcinoma. We discuss the idea that the liver alternates between two functional states depending on portal vein flow: a high-flow state favoring detoxification and metabolism, and a low-flow state that prioritizes bile acid production. Understanding how PIEZO1 contributes to these transitions offers new insights into liver's ability to adapt its function and metabolism. Further research on hepatobiliary PIEZO1 will advance the understanding of how physical exercise promotes health and opens new opportunities for enhancing liver regeneration after surgical resection and liver function in chronic diseases such as fibrosis and cirrhosis.</p>","PeriodicalId":7585,"journal":{"name":"American journal of physiology. Cell physiology","volume":" ","pages":"C421-C433"},"PeriodicalIF":4.7,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145853147","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Selective modulation of murine intestinal M₁ and M₃ muscarinic receptor expression has divergent effects on specialized epithelial cells and body weight.","authors":"Natalia Sampaio Moura, Alyssa Schledwitz, Kunrong Cheng, Yang Song, Min Seong Kwon, Cassandra A Cairns, Lea-Pearl Njei, Benjamin Raufman, Cinthia B Drachenberg, Jian-Ying Wang, Bing Ma, Jean-Pierre Raufman","doi":"10.1152/ajpcell.00739.2025","DOIUrl":"10.1152/ajpcell.00739.2025","url":null,"abstract":"<p><p>M₁ and M₃ muscarinic receptors encoded by <i>CHRM1</i> and <i>CHRM3</i> mediate neuronal and non-neuronal cholinergic signaling. Mice with global M₃R deficiency reportedly weigh less than controls, but the cell type(s) involved are unknown. As the intestinal epithelium modulates nutrient absorption, we asked whether deleting M₁R and M₃R only from intestinal epithelial cells would alter the distribution of specialized small intestinal epithelial cells or body weight. We reviewed reports of global M₁R and M₃R deficiency and body weight, used single-cell RNA sequencing (scRNA-Seq) to assess <i>Chrm1</i> and <i>Chrm3</i> expression by small intestinal epithelial cells, created mice with conditional intestinal epithelial cell M₁R and M₃R deletion (CKO mice), and compared the distribution of specialized intestinal epithelial cells and body weights of CKO and control mice, and the development of enteroids. Prior weight comparisons commonly used only male mice, frequently without comparison with littermate controls. scRNA-Seq analysis of tissues from M₁R and M₃R floxed mice revealed robust <i>Chrm1</i> and <i>Chrm3</i> expression by enteric goblet cells. CKO mice with selective mucosal depletion of <i>Chrm1</i> and <i>Chrm3</i> RNA were viable, fertile, and had fewer small intestinal goblet cells than controls. M₃R CKO mice had more tuft cells than controls. Although female mice weighed ∼20% less than males, we detected no weight differences between M₁R and M₃R CKO and control mice; enteroids derived from these mice developed at the same pace. Intestinal epithelial cell M₁R and M₃R deficiency impacts the distribution of specialized intestinal epithelial cells but not murine body weight.<b>NEW & NOTEWORTHY</b> Small intestinal goblet cells robustly express both <i>Chrm1</i> and <i>Chrm3</i>; mucosal immune cells express primarily <i>Chrm3</i>. Mice with intestinal epithelial cell (IEC) <i>Chrm3</i>/M₃R deletion have fewer small intestinal goblet cells but more tuft cells. The increase in tuft cells, which produce acetylcholine, may represent a feedback mechanism to compensate for reduced muscarinic receptor (MR) expression. IEC-selective deletion of MRs does not impact murine weight or enteroid development, suggesting that MRs beyond the intestinal epithelium regulate weight.</p>","PeriodicalId":7585,"journal":{"name":"American journal of physiology. Cell physiology","volume":" ","pages":"C525-C539"},"PeriodicalIF":4.7,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12840908/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145832896","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Lyophilized mesenchymal stem cell-derived extracellular vesicles improve outcomes in acute liver failure models.","authors":"Fumiya Suzuki, Hiroaki Haga, Tatsutoshi Inuzuka, Kyoko Hoshikawa, Tomohiro Katsumi, Keita Maki, Fumi Uchiyama, Yoshiyuki Ueno","doi":"10.1152/ajpcell.00361.2025","DOIUrl":"10.1152/ajpcell.00361.2025","url":null,"abstract":"<p><p>Mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) are promising for the treatment of liver diseases, including acute liver failure (ALF). However, efficient preservation methods suitable for clinical use remain under investigation. In this study, we evaluated the preservation efficacy and therapeutic effects of lyophilized MSC-EVs in a mouse model of ALF. EVs were isolated from bone marrow-derived MSCs and allocated to four preservation conditions: nonlyophilized (fresh), phosphate-buffered saline (PBS), 1% sucrose, and 5% sucrose. EVs were characterized by nanoparticle tracking analysis, Bioanalyzer, absorbance measurements, RNA sequencing, and transmission electron microscopy (TEM). ALF was induced by d-galactosamine and TNF-α, and mice were treated with PBS, empty EVs (m-Encapsome), nonlyophilized EVs, or lyophilized EVs (5% sucrose). Among the preservation conditions, the 5% sucrose group retained the highest EV yield, exhibited a unimodal particle size distribution, and preserved EV morphology, whereas the PBS and 1% sucrose groups showed structural damage and multimodal particle size distributions. Total RNA and protein levels were comparable among groups; however, miRNA sequencing demonstrated a strong correlation between nonlyophilized EVs and 5% sucrose-lyophilized EVs. In the ALF model, 5% sucrose-lyophilized EVs significantly reduced serum alanine transaminase (ALT) levels, inflammatory cytokines, hepatocyte necrosis, TUNEL-positive cells, PCNA-positive cells, and Ki-67-positive cells, with effects comparable to those of nonlyophilized EVs. These findings demonstrate that lyophilization with 5% sucrose effectively preserves MSC-EV integrity and therapeutic potential, supporting future clinical applications of MSC-EVs for liver disease treatment.<b>NEW & NOTEWORTHY</b> This study establishes lyophilization with 5% sucrose as an effective preservation method for mesenchymal stem cell-derived extracellular vesicles (MSC-EVs). Preserved EVs maintained structural integrity and miRNA profiles comparable to fresh EVs. In a mouse model of acute liver failure, they demonstrated equivalent therapeutic efficacy. These findings overcome storage limitations, facilitating the clinical translation of MSC-EVs as a stable treatment for liver diseases.</p>","PeriodicalId":7585,"journal":{"name":"American journal of physiology. Cell physiology","volume":" ","pages":"C540-C551"},"PeriodicalIF":4.7,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145987555","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}