Cirrhosis is a disease with poor prognosis that requires the development of a novel therapeutic approach alternative to liver transplantation. In this study, we focused on the placenta and aimed to clarify the effects of human placental extract (HPE) on cirrhosis.
Methods
A mouse model of carbon tetrachloride-induced cirrhosis was used to evaluate the effect of HPE administration subcutaneously and compared with the control group (n = 8 for each group). In vitro and in vivo, real time-PCR and immunostaining were performed for HPE mechanistic analysis. Spatial transcriptomics was also performed for detailed analysis of the effect of HPE on cirrhosis.
Results
HPE administration improved serum ALT levels compared to control mice. Furthermore, there was a decrease in the number of senescent cells in the liver and the mRNA levels of secrete senescence-associated secretory phenotype factors and Cdkn2a (p16). In vitro, HPE induced macrophage polarization to the anti-inflammatory M2 phenotype. Spatial transcriptomics was also performed to analyze the underlying anti-inflammatory mechanism. The results showed that HPE strongly polarized macrophages to the M2 phenotype, especially in macrophage-rich regions in the liver. Gene expression pathway analysis using spatial transcriptomics also revealed the possibility of improving senescent cell-derived inflammation via mitochondrial function.
Conclusions
HPE improves serum ALT levels via anti-inflammatory mechanisms in macrophages and senescent cells. HPE serves as a novel agent for cirrhosis treatment.
{"title":"Human placental extract improves liver cirrhosis in mice with regulation of macrophages and senescent cells","authors":"Natsuki Ishikawa , Yusuke Watanabe , Yuichirou Maeda , Tomoaki Yoshida , Naruhiro Kimura , Hiroyuki Abe , Akira Sakamaki , Hiroteru Kamimura , Takeshi Yokoo , Kenya Kamimura , Atsunori Tsuchiya , Shuji Terai","doi":"10.1016/j.reth.2025.01.017","DOIUrl":"10.1016/j.reth.2025.01.017","url":null,"abstract":"<div><h3>Introduction</h3><div>Cirrhosis is a disease with poor prognosis that requires the development of a novel therapeutic approach alternative to liver transplantation. In this study, we focused on the placenta and aimed to clarify the effects of human placental extract (HPE) on cirrhosis.</div></div><div><h3>Methods</h3><div>A mouse model of carbon tetrachloride-induced cirrhosis was used to evaluate the effect of HPE administration subcutaneously and compared with the control group (n = 8 for each group). In vitro and <em>in vivo</em>, real time-PCR and immunostaining were performed for HPE mechanistic analysis. Spatial transcriptomics was also performed for detailed analysis of the effect of HPE on cirrhosis.</div></div><div><h3>Results</h3><div>HPE administration improved serum ALT levels compared to control mice. Furthermore, there was a decrease in the number of senescent cells in the liver and the mRNA levels of secrete senescence-associated secretory phenotype factors <em>and Cdkn2a (p16)</em>. <em>In vitro,</em> HPE induced macrophage polarization to the anti-inflammatory M2 phenotype. Spatial transcriptomics was also performed to analyze the underlying anti-inflammatory mechanism. The results showed that HPE strongly polarized macrophages to the M2 phenotype, especially in macrophage-rich regions in the liver. Gene expression pathway analysis using spatial transcriptomics also revealed the possibility of improving senescent cell-derived inflammation via mitochondrial function.</div></div><div><h3>Conclusions</h3><div>HPE improves serum ALT levels via anti-inflammatory mechanisms in macrophages and senescent cells. HPE serves as a novel agent for cirrhosis treatment.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"28 ","pages":"Pages 509-516"},"PeriodicalIF":3.4,"publicationDate":"2025-02-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143092864","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Myocardial infarction (MI) is the most common cardiovascular disease (CVD) and the leading cause of mortality worldwide. Recent advancements have identified human endometrial mesenchymal stem cells (hEnMSCs) as a promising candidate for heart regeneration, however, challenges associated with cell-based therapies have shifted focus toward cell-free treatments (CFTs), such as exosome therapy, which show considerable promise for myocardial tissue regeneration. MI was induced in male Wistar rats by occluding the left anterior descending (LAD) coronary artery. The hEnMSCs-derived exosomes (hEnMSCs-EXOs) were encapsulated in injectable fibrin gel inside the cardiac tissue. The encapsulated hEnMSC-EXOs were administered, and their effects on myocardial regeneration, angiogenesis, and heart function were monitored for 30 days post-MI. The treatments were evaluated through histological analysis, echocardiographic parameters of left ventricular internal dimension at end-diastole (LVIDD) and end-systole (LVID), left ventricular end-diastole volume (LVEDV), left ventricular end-systole volume (LVESV), and left ventricular ejection fraction (LVEF) and molecular studies. Histological findings demonstrated significant fibrosis and left ventricular remodeling following MI. Treatment with fibrin gel-encapsulated hEnMSCs-EXOs substantially reduced fibrosis, enhanced angiogenesis, and prevented heart remodeling, leading to improved cardiac function. Notably, 30 days after encapsulated hEnMSCs-EXOs were delivered corresponded with a less inflammatory microenvironment, supporting cardiomyocyte retention in ischemic tissue. This study highlights the potential of encapsulated hEnMSCs-EXOs in fibrin gel as a novel therapeutic strategy for ischemic myocardium repair post-MI. The findings underscore the importance of biomaterials in advancing stem cell-based therapies and lay a foundation for clinical applications to mitigate heart injury following MI.
{"title":"Therapeutic potential of exosomes derived from human endometrial mesenchymal stem cells for heart tissue regeneration after myocardial infarction","authors":"Masoumeh Sepehri , Shahram Rabbani , Jafar Ai , Naghmeh Bahrami , Hossein Ghanbari , Mojdeh Salehi Namini , Majid Sharifi , Fatemeh Kouchakzadeh , Mohsen Abedini Esfahlani , Somayeh Ebrahimi-Barough","doi":"10.1016/j.reth.2025.01.007","DOIUrl":"10.1016/j.reth.2025.01.007","url":null,"abstract":"<div><div>Myocardial infarction (MI) is the most common cardiovascular disease (CVD) and the leading cause of mortality worldwide. Recent advancements have identified human endometrial mesenchymal stem cells (hEnMSCs) as a promising candidate for heart regeneration, however, challenges associated with cell-based therapies have shifted focus toward cell-free treatments (CFTs), such as exosome therapy, which show considerable promise for myocardial tissue regeneration. MI was induced in male Wistar rats by occluding the left anterior descending (LAD) coronary artery. The hEnMSCs-derived exosomes (hEnMSCs-EXOs) were encapsulated in injectable fibrin gel inside the cardiac tissue. The encapsulated hEnMSC-EXOs were administered, and their effects on myocardial regeneration, angiogenesis, and heart function were monitored for 30 days post-MI. The treatments were evaluated through histological analysis, echocardiographic parameters of left ventricular internal dimension at end-diastole (LVIDD) and end-systole (LVID), left ventricular end-diastole volume (LVEDV), left ventricular end-systole volume (LVESV), and left ventricular ejection fraction (LVEF) and molecular studies. Histological findings demonstrated significant fibrosis and left ventricular remodeling following MI. Treatment with fibrin gel-encapsulated hEnMSCs-EXOs substantially reduced fibrosis, enhanced angiogenesis, and prevented heart remodeling, leading to improved cardiac function. Notably, 30 days after encapsulated hEnMSCs-EXOs were delivered corresponded with a less inflammatory microenvironment, supporting cardiomyocyte retention in ischemic tissue. This study highlights the potential of encapsulated hEnMSCs-EXOs in fibrin gel as a novel therapeutic strategy for ischemic myocardium repair post-MI. The findings underscore the importance of biomaterials in advancing stem cell-based therapies and lay a foundation for clinical applications to mitigate heart injury following MI.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"28 ","pages":"Pages 451-461"},"PeriodicalIF":3.4,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143092851","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-31DOI: 10.1016/j.reth.2025.01.016
Enrico Ragni , Andrea Papait , Michela Maria Taiana , Paola De Luca , Giulio Grieco , Elsa Vertua , Pietro Romele , Cecilia Colombo , Antonietta Rosa Silini , Ornella Parolini , Laura de Girolamo
Introduction
Mesenchymal stromal cells (MSCs) gained attention for their anti-inflammatory and trophic properties, with musculoskeletal diseases and osteoarthritis (OA) being among the most studied conditions. Alongside cells, their released factors and extracellular vesicles (EVs), overall termed “secretome”, are actively sifted being envisioned as the main therapeutic actors. In addition to standard supplementation given by foetal bovine serum (FBS) or human platelet lysate (hPL), new good manufacturing practice (GMP)-compliant serum/xeno (S/X)-free media formulations have been proposed, although their influence on MSCs phenotype and potential is scarcely described. The aim of this study is therefore to evaluate, in the OA context, the differences in secretome composition and potential after adipose-MSCs (ASCs) cultivation in both standard (FBS and hPL) and two next generation (S/X) GMP-ready supplements.
Methods
Immunophenotype and secretory ability at soluble protein and EV-related levels, including embedded miRNAs, were analysed in the secretomes by means of flow cytometry, nanoparticle tracking analysis, high throughput ELISA and qRT-PCR arrays. Secretomes effect was tested in in vitro models of chondrocytes, lymphocytes and monocytes to mimic the OA microenvironment.
Results
Within a conserved molecular signature, a divergent fingerprint emerged for ASCs' secretomes collected after expansion in standard FBS/hPL or next-generation S/X formulations. Regarding soluble factors, a less protective feature for those in the secretome collected after ASCs were cultured in S/X media emerged. Moreover, the overall message for EV-miRNAs was characterized by a preponderance of protective signals in FBS and hPL conditions in a context of general safeguard given by ASCs released molecules. This dichotomy was reflected on secretomes’ potential in vitro, with expansion in hPL resulting in the most effective secretome for chondrocytes and in FBS for immune cells.
Conclusions
These data open the question about the implications from using new media for MSCs expansion for clinical application. Although the undeniable advantages for GMP compliant processes, this study results suggest that new media formulations would deserve a deep characterization to drive the choice of the most effective one tailored to each specific application.
{"title":"Cell culture expansion media choice affects secretory, protective and immuno-modulatory features of adipose mesenchymal stromal cell-derived secretomes for orthopaedic applications","authors":"Enrico Ragni , Andrea Papait , Michela Maria Taiana , Paola De Luca , Giulio Grieco , Elsa Vertua , Pietro Romele , Cecilia Colombo , Antonietta Rosa Silini , Ornella Parolini , Laura de Girolamo","doi":"10.1016/j.reth.2025.01.016","DOIUrl":"10.1016/j.reth.2025.01.016","url":null,"abstract":"<div><h3>Introduction</h3><div>Mesenchymal stromal cells (MSCs) gained attention for their anti-inflammatory and trophic properties, with musculoskeletal diseases and osteoarthritis (OA) being among the most studied conditions. Alongside cells, their released factors and extracellular vesicles (EVs), overall termed “secretome”, are actively sifted being envisioned as the main therapeutic actors. In addition to standard supplementation given by foetal bovine serum (FBS) or human platelet lysate (hPL), new good manufacturing practice (GMP)-compliant serum/xeno (S/X)-free media formulations have been proposed, although their influence on MSCs phenotype and potential is scarcely described. The aim of this study is therefore to evaluate, in the OA context, the differences in secretome composition and potential after adipose-MSCs (ASCs) cultivation in both standard (FBS and hPL) and two next generation (S/X) GMP-ready supplements.</div></div><div><h3>Methods</h3><div>Immunophenotype and secretory ability at soluble protein and EV-related levels, including embedded miRNAs, were analysed in the secretomes by means of flow cytometry, nanoparticle tracking analysis, high throughput ELISA and qRT-PCR arrays. Secretomes effect was tested in <em>in vitro</em> models of chondrocytes, lymphocytes and monocytes to mimic the OA microenvironment.</div></div><div><h3>Results</h3><div>Within a conserved molecular signature, a divergent fingerprint emerged for ASCs' secretomes collected after expansion in standard FBS/hPL or next-generation S/X formulations. Regarding soluble factors, a less protective feature for those in the secretome collected after ASCs were cultured in S/X media emerged. Moreover, the overall message for EV-miRNAs was characterized by a preponderance of protective signals in FBS and hPL conditions in a context of general safeguard given by ASCs released molecules. This dichotomy was reflected on secretomes’ potential <em>in vitro</em>, with expansion in hPL resulting in the most effective secretome for chondrocytes and in FBS for immune cells.</div></div><div><h3>Conclusions</h3><div>These data open the question about the implications from using new media for MSCs expansion for clinical application. Although the undeniable advantages for GMP compliant processes, this study results suggest that new media formulations would deserve a deep characterization to drive the choice of the most effective one tailored to each specific application.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"28 ","pages":"Pages 481-497"},"PeriodicalIF":3.4,"publicationDate":"2025-01-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143092863","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Intramuscular adipose tissue (IMAT) is frequently formed in certain pathological conditions, such as biological aging, and ectopic fat accumulation leads to muscle weakness and a subsequent decline in physical function. Although mesenchymal progenitors (MPs) are present in postnatal skeletal muscle and are the cells from which IMAT originates, the molecular mechanism by which MPs contribute to IMAT formation has not been completely elucidated. Recently, we found that PDZ domain-containing ring finger 3 (PDZRN3), an E3-ubiquitin ligase, was highly expressed in MPs. In this study, we aimed to clarify the functions of PDZRN3 in MPs and the roles of PDZRN3 in IMAT formation using in vitro and in vivo experiments.
Methods
Primary mouse MPs isolated from hindlimb muscles were applied to adipogenic differentiation conditions, and expression fluctuation of PDZRN3 was verified with adipogenic differentiation and Wnt signaling markers. The role of PDZRN3 on MP’s adipogenesis was evaluated in vitro by gene knock-down experiments. To evaluate the contribution of PDZRN3 to IMAT formation in vivo, tamoxifen-inducible MP-specific Pdzrn3 knockout (Pdzrn3MPcKO) mice were developed.
Results
PDZRN3 was more expressed in MPs than in muscle stem cells, and its expression profile of PDZRN3 fluctuated with the adipogenic differentiation of MPs. Our results revealed that PDZRN3 suppressed the adipogenesis of MPs in vitro through the activation of Wnt signaling and that a decrease in PDZRN3 accelerated adipogenesis. Indeed, IMAT significantly increased in the denervated muscles of Pdzrn3MPcKO mice.
Conclusions
Our findings suggest that PDZRN3 is a key molecule in regulating IMAT formation. Since ectopic fat accumulation is frequently found in the skeletal muscles of older adults and also muscular dystrophy patients, PDZRN3 and its related pathways may represent a novel therapeutic target for these muscle pathologies.
{"title":"PDZRN3 regulates adipogenesis of mesenchymal progenitors in muscle","authors":"Hiroki Iida , Minako Kawai-Takaishi , Yoshihiro Miyagawa , Yasuhiko Takegami , Akiyoshi Uezumi , Takeshi Honda , Shiro Imagama , Tohru Hosoyama","doi":"10.1016/j.reth.2025.01.018","DOIUrl":"10.1016/j.reth.2025.01.018","url":null,"abstract":"<div><h3>Introduction</h3><div>Intramuscular adipose tissue (IMAT) is frequently formed in certain pathological conditions, such as biological aging, and ectopic fat accumulation leads to muscle weakness and a subsequent decline in physical function. Although mesenchymal progenitors (MPs) are present in postnatal skeletal muscle and are the cells from which IMAT originates, the molecular mechanism by which MPs contribute to IMAT formation has not been completely elucidated. Recently, we found that PDZ domain-containing ring finger 3 (PDZRN3), an E3-ubiquitin ligase, was highly expressed in MPs. In this study, we aimed to clarify the functions of PDZRN3 in MPs and the roles of PDZRN3 in IMAT formation using <em>in vitro</em> and <em>in vivo</em> experiments.</div></div><div><h3>Methods</h3><div>Primary mouse MPs isolated from hindlimb muscles were applied to adipogenic differentiation conditions, and expression fluctuation of PDZRN3 was verified with adipogenic differentiation and Wnt signaling markers. The role of PDZRN3 on MP’s adipogenesis was evaluated <em>in vitro</em> by gene knock-down experiments. To evaluate the contribution of PDZRN3 to IMAT formation <em>in vivo</em>, tamoxifen-inducible MP-specific <em>Pdzrn3</em> knockout (<em>Pdzrn3</em><sup>MPcKO</sup>) mice were developed.</div></div><div><h3>Results</h3><div>PDZRN3 was more expressed in MPs than in muscle stem cells, and its expression profile of PDZRN3 fluctuated with the adipogenic differentiation of MPs. Our results revealed that PDZRN3 suppressed the adipogenesis of MPs <em>in vitro</em> through the activation of Wnt signaling and that a decrease in PDZRN3 accelerated adipogenesis. Indeed, IMAT significantly increased in the denervated muscles of <em>Pdzrn3</em><sup>MPcKO</sup> mice.</div></div><div><h3>Conclusions</h3><div>Our findings suggest that PDZRN3 is a key molecule in regulating IMAT formation. Since ectopic fat accumulation is frequently found in the skeletal muscles of older adults and also muscular dystrophy patients, PDZRN3 and its related pathways may represent a novel therapeutic target for these muscle pathologies.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"28 ","pages":"Pages 473-480"},"PeriodicalIF":3.4,"publicationDate":"2025-01-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143092852","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Cartilage tissue is important in the human body as a ubiquitous connective tissue. However, information on the limits of chondrocyte growth and the effects of basic FGF (bFGF) in vivo is scarce. This study aims to investigate the effects of bFGF on the growth rate, cytology, and morphology of implanted cultured epiphyseal cartilage-derived chondrocytes into NOD/Shi-scid IL-2Rγnull mice (NOG mice).
Methods
Chondrocytes were isolated from the epiphyseal cartilage derived from the digits of polydactyly patients and were cultivated in a medium supplemented with bFGF (bFGF-treated cells) and without bFGF (bFGF-untreated cells). These cultivated cells were subcutaneously implanted into the backs of NOG mice.
Results
bFGF-treated cells exhibited a higher growth rate than bFGF-untreated cells. Cartilage was formed after two weeks of implantation. The cartilages generated by bFGF-treated cells (denoted as “BT-cartilage” hereafter) were larger in size and heavier in weight than those cartilages generated by bFGF-untreated cells (denoted as “BUT-cartilage” hereafter). BT-cartilage grew exponentially after 10 weeks.
Conclusions
From these results, bFGF-treated cells exhibited increased ability for both proliferation and differentiation compared to bFGF-untreated cells. The results of this study may lead to the generation of new alternative therapies for bone and cartilage through the use of epiphyseal chondrocytes.
{"title":"Effects of basic fibroblast growth factor on cartilage to bone: Time-course histological analysis of in vivo cartilage formation from polydactyly-derived chondrocytes","authors":"Michiyo Nasu , Shinichiro Takayama , Rin Amagase , Akihiro Umezawa","doi":"10.1016/j.reth.2025.01.013","DOIUrl":"10.1016/j.reth.2025.01.013","url":null,"abstract":"<div><h3>Introduction</h3><div>Cartilage tissue is important in the human body as a ubiquitous connective tissue. However, information on the limits of chondrocyte growth and the effects of basic FGF (bFGF) <em>in vivo</em> is scarce. This study aims to investigate the effects of bFGF on the growth rate, cytology, and morphology of implanted cultured epiphyseal cartilage-derived chondrocytes into NOD/Shi-scid IL-2Rγ<sup>null</sup> mice (NOG mice).</div></div><div><h3>Methods</h3><div>Chondrocytes were isolated from the epiphyseal cartilage derived from the digits of polydactyly patients and were cultivated in a medium supplemented with bFGF (bFGF-treated cells) and without bFGF (bFGF-untreated cells). These cultivated cells were subcutaneously implanted into the backs of NOG mice.</div></div><div><h3>Results</h3><div>bFGF-treated cells exhibited a higher growth rate than bFGF-untreated cells. Cartilage was formed after two weeks of implantation. The cartilages generated by bFGF-treated cells (denoted as “BT-cartilage” hereafter) were larger in size and heavier in weight than those cartilages generated by bFGF-untreated cells (denoted as “BUT-cartilage” hereafter). BT-cartilage grew exponentially after 10 weeks.</div></div><div><h3>Conclusions</h3><div>From these results, bFGF-treated cells exhibited increased ability for both proliferation and differentiation compared to bFGF-untreated cells. The results of this study may lead to the generation of new alternative therapies for bone and cartilage through the use of epiphyseal chondrocytes.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"28 ","pages":"Pages 462-472"},"PeriodicalIF":3.4,"publicationDate":"2025-01-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143092850","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-28DOI: 10.1016/j.reth.2025.01.006
Shuwei Sun , Fengzheng Zhu , Qingling Xu , Xianglin Hou , Weihong Nie , Kaiyan Su , Li Wang , Zhuo Liu , Tao Shan , Chunying Shi
Myocardial infarction (MI) was a cardiovascular emergency that led to heart failure, arrhythmia, and sudden death. Basic fibroblast growth factor (bFGF) was revealed to promote angiogenesis and protect cardiomyocytes against ischemic injury. But conventional delivery of bFGF in an uncontrolled manner was inefficient and diffusive, limiting its application in MI therapy. Currently, stimuli-responsive drug delivery is emphasized in tissue regeneration. The present study constructed a CFBP-bFGF recombinant protein, which could specifically target upregulated connective tissue growth factor (CTGF) and release bFGF in ischemic myocardium. In a rat model with MI, intravenous administration of CFBP-bFGF significantly accumulated in ischemic myocardium by targeting with CTGF. The responsive release of CFBP-bFGF effectively enhanced blood vessel regeneration, decreased cardiomyocyte apoptosis, and improved cardiac function recovery. In addition, the molecular mechanism was further explored by RNA sequencing and transcriptome analysis. Besides activating the pathways and genes related to angiogenesis and cardiac protection, CFBP-bFGF also decreased the expression of fibrosis-related pathways and genes, such as TGF-β. These results demonstrated that the CTGF-responsive CFBP-bFGF was effective for targeting release that promoted the functional recovery of MI.
{"title":"Modified bFGF targeting connective tissue growth factor in the injured microenvironment improved cardiac repair after chronic myocardial ischemia","authors":"Shuwei Sun , Fengzheng Zhu , Qingling Xu , Xianglin Hou , Weihong Nie , Kaiyan Su , Li Wang , Zhuo Liu , Tao Shan , Chunying Shi","doi":"10.1016/j.reth.2025.01.006","DOIUrl":"10.1016/j.reth.2025.01.006","url":null,"abstract":"<div><div>Myocardial infarction (MI) was a cardiovascular emergency that led to heart failure, arrhythmia, and sudden death. Basic fibroblast growth factor (bFGF) was revealed to promote angiogenesis and protect cardiomyocytes against ischemic injury. But conventional delivery of bFGF in an uncontrolled manner was inefficient and diffusive, limiting its application in MI therapy. Currently, stimuli-responsive drug delivery is emphasized in tissue regeneration. The present study constructed a CFBP-bFGF recombinant protein, which could specifically target upregulated connective tissue growth factor (CTGF) and release bFGF in ischemic myocardium. In a rat model with MI, intravenous administration of CFBP-bFGF significantly accumulated in ischemic myocardium by targeting with CTGF. The responsive release of CFBP-bFGF effectively enhanced blood vessel regeneration, decreased cardiomyocyte apoptosis, and improved cardiac function recovery. In addition, the molecular mechanism was further explored by RNA sequencing and transcriptome analysis. Besides activating the pathways and genes related to angiogenesis and cardiac protection, CFBP-bFGF also decreased the expression of fibrosis-related pathways and genes, such as TGF-β. These results demonstrated that the CTGF-responsive CFBP-bFGF was effective for targeting release that promoted the functional recovery of MI.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"28 ","pages":"Pages 438-450"},"PeriodicalIF":3.4,"publicationDate":"2025-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143092849","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The histological grading of carcinoma has been one of the central applications of task-specific deep learning in pathology. The deep learning method has pushed away the regression approach, which has been exploited for two-class classification, to address multi-class classification. However, the applicability of the regression approach on multi-class carcinoma grading has not been extensively investigated. Here, we show that the regression approach is sufficiently compatible with classification regarding the four-class grading of clear cell renal cell carcinoma using 11,826 histological image patches from 16 whole slide images. Using convolutional neural network models (DenseNet-121 and Inception-v3), we found that regression models predict as accurately as classification models, achieving an accuracy of 0.990 at the highest, with fewer prediction errors by two or more grades. Furthermore, we found that the predictions by the regression models qualitatively capture intra-tumor heterogeneity of grades using the composite image patches. Our results demonstrate that the regression approach offers advantages in making a core of the multi-class grade prediction tools for practice.
{"title":"Applicability of the regression approach for histological multi-class grading in clear cell renal cell carcinoma","authors":"Mayu Shibata , Akihiro Umezawa , Saki Aoto , Kohji Okamura , Michiyo Nasu , Ryuichi Mizuno , Mototsugu Oya , Kei Yura , Shuji Mikami","doi":"10.1016/j.reth.2025.01.011","DOIUrl":"10.1016/j.reth.2025.01.011","url":null,"abstract":"<div><div>The histological grading of carcinoma has been one of the central applications of task-specific deep learning in pathology. The deep learning method has pushed away the regression approach, which has been exploited for two-class classification, to address multi-class classification. However, the applicability of the regression approach on multi-class carcinoma grading has not been extensively investigated. Here, we show that the regression approach is sufficiently compatible with classification regarding the four-class grading of clear cell renal cell carcinoma using 11,826 histological image patches from 16 whole slide images. Using convolutional neural network models (DenseNet-121 and Inception-v3), we found that regression models predict as accurately as classification models, achieving an accuracy of 0.990 at the highest, with fewer prediction errors by two or more grades. Furthermore, we found that the predictions by the regression models qualitatively capture intra-tumor heterogeneity of grades using the composite image patches. Our results demonstrate that the regression approach offers advantages in making a core of the multi-class grade prediction tools for practice.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"28 ","pages":"Pages 431-437"},"PeriodicalIF":3.4,"publicationDate":"2025-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143092847","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Elucidation of the paracrine interaction between chondrocytes and macrophages is useful for understanding the mechanisms of cartilage regeneration. Extracellular vesicles are granular substances with a diameter of approximately 150 nm, surrounded by a phospholipid bilayer membrane. In recent years, research has been conducted on clinical applications of extracellular vesicles. It has been shown that macrophages promote cartilage maturation, and macrophages acquire anti-inflammatory properties through cartilage, but the detailed mechanism of paracrine action involving extracellular vesicles remains unclear. Therefore, we focused on the effect of chondrocyte-derived extracellular vesicles on changes in macrophage characteristics.
Methods
Macrophages induced with granulocyte-macrophage colony stimulating factor (M1-like macrophages) and auricular chondrocytes were co-cultured using cell culture inserts and exosome inhibitors, and the expression of macrophage markers were analyzed. Next, extracellular vesicles separated from auricular chondrocytes were added to in vitro macrophage culture medium, and time-lapse observations of macrophage uptake of auricular chondrocyte-derived extracellular vesicles were performed. In addition, the effects of extracellular vesicles on the expression of macrophage markers were also analyzed.
Results
The expression of CD206, an M2 macrophage marker, was increased in macrophages due to the paracrine effect of chondrocytes, and CD206 expression was further increased by pharmacological inhibition of chondrocyte-derived exosomes. It was shown that chondrocyte-derived extracellular vesicles were taken up by macrophages and promoted the production of interleukin-10, an anti-inflammatory cytokine while reducing CD206 expression.
Conclusions
Auricular chondrocyte-derived extracellular vesicles promoted the production of interleukin-10 in bone marrow M1-like macrophages but reduced CD206 expression.
{"title":"Small extracellular vesicles derived from auricular chondrocytes promote secretion of interleukin 10 in bone marrow M1-like macrophages","authors":"Tetsuya Kobatake , Yoshiyuki Miyamoto , Yuko Fujihara , Hideto Saijo , Kazuto Hoshi , Atsuhiko Hikita","doi":"10.1016/j.reth.2025.01.009","DOIUrl":"10.1016/j.reth.2025.01.009","url":null,"abstract":"<div><h3>Introduction</h3><div>Elucidation of the paracrine interaction between chondrocytes and macrophages is useful for understanding the mechanisms of cartilage regeneration. Extracellular vesicles are granular substances with a diameter of approximately 150 nm, surrounded by a phospholipid bilayer membrane. In recent years, research has been conducted on clinical applications of extracellular vesicles. It has been shown that macrophages promote cartilage maturation, and macrophages acquire anti-inflammatory properties through cartilage, but the detailed mechanism of paracrine action involving extracellular vesicles remains unclear. Therefore, we focused on the effect of chondrocyte-derived extracellular vesicles on changes in macrophage characteristics.</div></div><div><h3>Methods</h3><div>Macrophages induced with granulocyte-macrophage colony stimulating factor (M1-like macrophages) and auricular chondrocytes were co-cultured using cell culture inserts and exosome inhibitors, and the expression of macrophage markers were analyzed. Next, extracellular vesicles separated from auricular chondrocytes were added to in vitro macrophage culture medium, and time-lapse observations of macrophage uptake of auricular chondrocyte-derived extracellular vesicles were performed. In addition, the effects of extracellular vesicles on the expression of macrophage markers were also analyzed.</div></div><div><h3>Results</h3><div>The expression of CD206, an M2 macrophage marker, was increased in macrophages due to the paracrine effect of chondrocytes, and CD206 expression was further increased by pharmacological inhibition of chondrocyte-derived exosomes. It was shown that chondrocyte-derived extracellular vesicles were taken up by macrophages and promoted the production of interleukin-10, an anti-inflammatory cytokine while reducing CD206 expression.</div></div><div><h3>Conclusions</h3><div>Auricular chondrocyte-derived extracellular vesicles promoted the production of interleukin-10 in bone marrow M1-like macrophages but reduced CD206 expression.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"28 ","pages":"Pages 421-430"},"PeriodicalIF":3.4,"publicationDate":"2025-01-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143092848","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-23DOI: 10.1016/j.reth.2025.01.010
Jiawei Wang , Chunzhong Qiao , Baoyang Luo , Lei Qin
Objective
Hepatocellular carcinoma (HCC) is a lethal malignancy in the world. LncRNA THUMPD3-AS1 is implicated in tumorigenesis and progression in various tumors. Therefore, this study was applied to investigate the action of THUMPD3-AS1 in HCC by regulating microRNA (miR)-4465 and KPNA2.
Methods
The clinical specimens of HCC were collected to determine THUMPD3-AS1, KPNA2, miR-4465, E-cadherin, Vimentin, N-cadherin, ZEB1 and SNAIL levels. HCC cells were screened and transfected with sh-THUMPD3-AS1 or miR-4465 mimic to explore their roles in HCC cell phenotype and epithelial-mesenchymal transition (EMT)-related factors. The involvement of miR-4465 in THUMPD3-AS1-mediated HCC was proved. The relationship of THUMPD3-AS1, KPNA2 and miR-4465 was verified.
Results
Overexpressed THUMPD3-AS1 and KPNA2 and reduced miR-4465 were present in HCC clinical tissues. THUMPD3-AS1 bound to miR-4465 to target KPNA2. Silencing of THUMPD3-AS1 or restoration of miR-4465 repressed HCC cell phenotypes and EMT in vitro. Inhibition of miR-4465 mitigated the role of silenced THUMPD3-AS1 in HCC.
Conclusion
This study stresses that THUMPD3-AS1 induces EMT in HCC cells and ultimately promotes HCC cell growth and migration by competitively inhibiting miR-4465 expression and thus upregulating KPNA2.
{"title":"LncRNA THUMPD3-AS1/microRNA-4465/KPNA2 axis impacts human hepatocellular carcinoma cell phenotypes","authors":"Jiawei Wang , Chunzhong Qiao , Baoyang Luo , Lei Qin","doi":"10.1016/j.reth.2025.01.010","DOIUrl":"10.1016/j.reth.2025.01.010","url":null,"abstract":"<div><h3>Objective</h3><div>Hepatocellular carcinoma (HCC) is a lethal malignancy in the world. LncRNA THUMPD3-AS1 is implicated in tumorigenesis and progression in various tumors. Therefore, this study was applied to investigate the action of THUMPD3-AS1 in HCC by regulating microRNA (miR)-4465 and KPNA2.</div></div><div><h3>Methods</h3><div>The clinical specimens of HCC were collected to determine THUMPD3-AS1, KPNA2, miR-4465, E-cadherin, Vimentin, N-cadherin, ZEB1 and SNAIL levels. HCC cells were screened and transfected with sh-THUMPD3-AS1 or miR-4465 mimic to explore their roles in HCC cell phenotype and epithelial-mesenchymal transition (EMT)-related factors. The involvement of miR-4465 in THUMPD3-AS1-mediated HCC was proved. The relationship of THUMPD3-AS1, KPNA2 and miR-4465 was verified.</div></div><div><h3>Results</h3><div>Overexpressed THUMPD3-AS1 and KPNA2 and reduced miR-4465 were present in HCC clinical tissues. THUMPD3-AS1 bound to miR-4465 to target KPNA2. Silencing of THUMPD3-AS1 or restoration of miR-4465 repressed HCC cell phenotypes and EMT <em>in vitro</em>. Inhibition of miR-4465 mitigated the role of silenced THUMPD3-AS1 in HCC.</div></div><div><h3>Conclusion</h3><div>This study stresses that THUMPD3-AS1 induces EMT in HCC cells and ultimately promotes HCC cell growth and migration by competitively inhibiting miR-4465 expression and thus upregulating KPNA2.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"28 ","pages":"Pages 413-420"},"PeriodicalIF":3.4,"publicationDate":"2025-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143092846","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-22DOI: 10.1016/j.reth.2025.01.003
Zhe Lu , Shifeng Ren , Bingjie Wang, Yajun Zhang, Xiaodong Mu, Zhihui Wang
Muse cell has become a promising source of cells for disease treatment due to its remarkable characteristics, including stress tolerance, low tumorigenicity, effective homing ability, and differentiation into histocompatibility cells after transplantation. However, there are some obvious obstacles that need to be overcome in the efficient expansion of Muse cells. We extracted mesenchymal stem cells (MSCs) from human umbilical cord and their MSCs phenotypes were verified by flow cytometry. Then, immune magnetic sorting was performed to obtain Muse cells, and the expression of pluripotency related factors and the ability to differentiate into three germ layers were verified with sorted Muse cells. We then tested a new 3D culture method with dynamic microsphere carrier to possibly expand Muse cells more efficiently. Finally, in vivo experiments were conducted to check the homing ability of Muse cells to muscle injury. Our results showed that, the cultivation and expansion of Muse cells can be more effectively achieved through dynamic microsphere carrier; compared to non-Muse cells, Muse cells have stronger pluripotency and differentiation ability, and their homing ability in the muscle injury mice model is superior to that of non-Muse cells. Therefore, with the method of immune magnetic sorting and dynamic microsphere carrier, highly regenerative Muse cells can be more effectively sorted and expanded from MSCs.
{"title":"3D dynamic culture of muse cells on a porous gelatin microsphere after magnetic sorting: Achieving high purity proliferation","authors":"Zhe Lu , Shifeng Ren , Bingjie Wang, Yajun Zhang, Xiaodong Mu, Zhihui Wang","doi":"10.1016/j.reth.2025.01.003","DOIUrl":"10.1016/j.reth.2025.01.003","url":null,"abstract":"<div><div>Muse cell has become a promising source of cells for disease treatment due to its remarkable characteristics, including stress tolerance, low tumorigenicity, effective homing ability, and differentiation into histocompatibility cells after transplantation. However, there are some obvious obstacles that need to be overcome in the efficient expansion of Muse cells. We extracted mesenchymal stem cells (MSCs) from human umbilical cord and their MSCs phenotypes were verified by flow cytometry. Then, immune magnetic sorting was performed to obtain Muse cells, and the expression of pluripotency related factors and the ability to differentiate into three germ layers were verified with sorted Muse cells. We then tested a new 3D culture method with dynamic microsphere carrier to possibly expand Muse cells more efficiently. Finally, in vivo experiments were conducted to check the homing ability of Muse cells to muscle injury. Our results showed that, the cultivation and expansion of Muse cells can be more effectively achieved through dynamic microsphere carrier; compared to non-Muse cells, Muse cells have stronger pluripotency and differentiation ability, and their homing ability in the muscle injury mice model is superior to that of non-Muse cells. Therefore, with the method of immune magnetic sorting and dynamic microsphere carrier, highly regenerative Muse cells can be more effectively sorted and expanded from MSCs.</div></div>","PeriodicalId":20895,"journal":{"name":"Regenerative Therapy","volume":"28 ","pages":"Pages 402-412"},"PeriodicalIF":3.4,"publicationDate":"2025-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143092845","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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