In Vitro Cellular & Developmental Biology. Animal最新文献_第2页

LncRNA TP53TG1 promotes the growth and osteo/dentinogenic differentiation of dental pulp stem cells by activating the Smad3 and JNK1/2 pathway. LncRNA TP53TG1通过激活Smad3和JNK1/2通路促进牙髓干细胞的生长和成骨/牙本质分化。

IF 1.7 4区生物学

In Vitro Cellular & Developmental Biology. Animal Pub Date : 2025-09-08 DOI: 10.1007/s11626-025-01086-3

Tingyue Li, Zihan Dai, Zhihua Wang, Minghao Wang, Chengxiong Cai, Xiaoru Zhu, Yang Zhao, Paul Roy Cooper, Shengchao Wang, Wenxi He

{"title":"LncRNA TP53TG1 promotes the growth and osteo/dentinogenic differentiation of dental pulp stem cells by activating the Smad3 and JNK1/2 pathway.","authors":"Tingyue Li, Zihan Dai, Zhihua Wang, Minghao Wang, Chengxiong Cai, Xiaoru Zhu, Yang Zhao, Paul Roy Cooper, Shengchao Wang, Wenxi He","doi":"10.1007/s11626-025-01086-3","DOIUrl":"https://doi.org/10.1007/s11626-025-01086-3","url":null,"abstract":"TP53TG1 is a long non-coding RNA related to the TP53 gene, which plays an important role in various biological processes such as tumorigenesis, cell cycle regulation, and DNA damage repair. In recent years, researchers have begun to explore the role of TP53TG1 in dental pulp biology, especially its potential impact on pulpitis and other pulp-related diseases. However, the role of TP53TG1 in human dental pulp stem cells (hDPSCs) remains unclear. In this study, we obtained TP53TG1 knockdown dental pulp stem cells by plasmid transfection to determine the biological role of TP53TG1 in DPSCs. We found that the expression of TP53TG1 increased significantly during odontogenic differentiation of DPSCs. SiRNA knockdown of TP53TG1 expression resulted in inhibition of proliferation of hDPSCs. During odontogenic differentiation, downregulation of TP53TG inhibited the expression of multiple differentiation-related indices, and alkaline phosphatase activity and the formation of mineralized nodules were also inhibited. In addition, Western blot found that knockdown of TP53TG1 also weakened SMAD3 and JNK1/2 signaling in DPSCs. In conclusion, our study revealed the differentiation-inducing role of TP53TG1 in DPSCs, which plays an important role in dental pulp repair and regeneration and provides new insights and approaches for the prevention and treatment of dental pulp diseases.","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145023182","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Inhibition of cGAS-STING signaling pathway alleviates high glucose-induced mesothelial-mesenchymal transition in human peritoneal mesothelial cell line HMrSV5. 抑制cGAS-STING信号通路可缓解高糖诱导的人腹膜间皮细胞系HMrSV5间质间质转化。

IF 1.7 4区生物学

In Vitro Cellular & Developmental Biology. Animal Pub Date : 2025-08-28 DOI: 10.1007/s11626-025-01107-1

Fuxing Dong, Luli Zheng, Fuyuan Hong

{"title":"Inhibition of cGAS-STING signaling pathway alleviates high glucose-induced mesothelial-mesenchymal transition in human peritoneal mesothelial cell line HMrSV5.","authors":"Fuxing Dong, Luli Zheng, Fuyuan Hong","doi":"10.1007/s11626-025-01107-1","DOIUrl":"https://doi.org/10.1007/s11626-025-01107-1","url":null,"abstract":"The mesothelial-mesenchymal transition (MMT) of peritoneal mesothelial cells is a critical factor contributing to the progression of peritoneal fibrosis. This study aimed to explore the effect of cGAS-STING signaling pathway on the MMT process in peritoneal mesothelial cells. The expressions of cGAS, STING, α-SMA, and Vimentin in HMrSV5 cells treated with high glucose were analyzed using WB. Subsequently, si-cGAS and the cGAS inhibitor RU.521 were employed to intervene in HMrSV5 cells. qPCR was utilized to evaluate the expression levels of genes involved in the cGAS-STING signaling pathway (cGAS, STING, IRF3, TBK1) and MMT-related genes (E-cadherin, Vimentin, α-SMA, TGF-β1). The protein expressions of the cGAS-STING signaling pathway and MMT-related proteins were detected by WB. The invasive capacity of cells in each cell was assessed using a Transwell assay, and the levels of pro-inflammatory cytokines (IL-6, TNF-α) in the supernatants of each cell were measured by ELISA. In the present study, we found that the expressions of cGAS, p-STING/STING, p-IRF3/IRF3, and p-TBK1/TBK1 proteins were significantly upregulated in HG-treated HMrSV5 cells. Furthermore, the activation of the cGAS-STING signaling pathway could be effectively suppressed in HMrSV5 cells transfected with si-cGAS or treated with RU.521. Additionally, treatment with si-cGAS or RU.521 not only attenuated the invasive capacity of HMrSV5 cells but also decreased the levels of pro-inflammatory cytokines and inhibited the expression of MMT-related markers. Suppression of the cGAS-STING signaling pathway mitigates HG-induced MMT in the human peritoneal mesothelial cell line HMrSV5.","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144952324","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Fetal bovine serum in cryomedia protects sheep spermatogonial stem cells and preserves stemness characteristics during cryopreservation. 低温培养基中的胎牛血清对绵羊精原干细胞具有保护作用，并在低温保存过程中保留了干细胞的特性。

IF 1.7 4区生物学

In Vitro Cellular & Developmental Biology. Animal Pub Date : 2025-08-21 DOI: 10.1007/s11626-025-01100-8

Tomy A Tomcy, Balakrishnan Binsila, Muhammed Sadikh, Balaganur Krishnappa, Natesan Ramachandran, Arunachalam Arangasamy, Veeramani Aranganathan, Sellappan Selvaraju

引用次数: 0

Therapeutic targeting of Nrf2/HO-1/NF-κB signaling axis with casticin mitigates intervertebral disc degeneration: in vitro and in vivo investigations. 用蓖麻素治疗靶向Nrf2/HO-1/NF-κB信号轴减轻椎间盘退变：体外和体内研究

IF 1.7 4区生物学

In Vitro Cellular & Developmental Biology. Animal Pub Date : 2025-08-19 DOI: 10.1007/s11626-025-01108-0

Long Wu, Zhanghong Wang, Zhipeng Wu, Yifan Wu

{"title":"Therapeutic targeting of Nrf2/HO-1/NF-κB signaling axis with casticin mitigates intervertebral disc degeneration: in vitro and in vivo investigations.","authors":"Long Wu, Zhanghong Wang, Zhipeng Wu, Yifan Wu","doi":"10.1007/s11626-025-01108-0","DOIUrl":"10.1007/s11626-025-01108-0","url":null,"abstract":"As a persistent osteoarticular degenerative condition, intervertebral disc deterioration (IDD) has been established as a principal causative element in lumbar spine discomfort development. The present investigation seeks to assess the protective effects of casticin against IDD progression and elucidate associated molecular pathways. The CCK8 kit was used to assess the cytotoxicity of casticin on rat nucleus pulposus cells (NPCs). Western blot assay, qRT-PCR, enzyme-linked immunosorbent assay, reactive oxygen species assay, and immunofluorescence were used to detect the expression levels of inflammatory mediators and ROS production between different groups. The nuclear translocation of NF-κB p65 and expression of Nrf2/HO-1 signal pathway in lipopolysaccharide (LPS)-induced NPCs were detected by confocal microscopy. Moreover, histological analysis was used to evaluate the degree of disc degeneration in rats. Casticin treatment inhibited the production of oxygen free radicals and inflammatory mediators induced by LPS, such as ROS, TNF-α, IL-1β, and PGE2. Not only that, we also found that casticin retained the content of type II collagen and aggrecan in NPCs and inhibited the expression of MMP-13 and ADAMTS-5. Moreover, casticin treatment activated the Nrf2/HO-1 signal axis and inhibited nuclear translocation of NF-κB p65 in LPS-exposed NPCs. Histological analysis found that the treatment of casticin in rat IDD models prevented the loss of notochordal cells and the disordered arrangement of fiber loops. Casticin inhibits LPS-stimulated oxidative stress, inflammatory response, and ECM degradation by activating the Nrf2/HO-1 signaling axis and indirectly blocking the NF-κB pathway.","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-08-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144872963","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

ATF4 regulates PI3K/AKT signaling axis to promote angiogenesis after myocardial infarction. ATF4调节PI3K/AKT信号轴促进心肌梗死后血管生成。

IF 1.7 4区生物学

In Vitro Cellular & Developmental Biology. Animal Pub Date : 2025-08-14 DOI: 10.1007/s11626-025-01085-4

Pingping He, Weirong Zeng, Jiao Li, Yu Zhang, Ranzun Zhao, Weiwei Liu, Yongchao Zhao, Zhijiang Liu, Changyin Shen, Wei Chen, Yan Wang, Bei Shi

{"title":"ATF4 regulates PI3K/AKT signaling axis to promote angiogenesis after myocardial infarction.","authors":"Pingping He, Weirong Zeng, Jiao Li, Yu Zhang, Ranzun Zhao, Weiwei Liu, Yongchao Zhao, Zhijiang Liu, Changyin Shen, Wei Chen, Yan Wang, Bei Shi","doi":"10.1007/s11626-025-01085-4","DOIUrl":"https://doi.org/10.1007/s11626-025-01085-4","url":null,"abstract":"Effective neovascularization is critical for tissue repair and the enhancement of cardiac function following myocardial infarction (MI). However, the hypoxic microenvironment post-MI significantly impedes neovascular formation. Although ATF4 has been implicated in heart failure and myocardial cell regeneration and repair, its role in angiogenesis remains unclear. This study utilized both in vitro and in vivo models to investigate the role of ATF4 in neovascularization after MI. In hypoxia-cultured murine endothelial cells (ECs), hypoxia was observed to inhibit EC proliferation, migration, and tube formation. In contrast, overexpression of ATF4 ameliorated these hypoxia-induced impairments. Conversely, inhibition of ATF4 further exacerbated the reduction in EC proliferation, migration, and tube formation induced by hypoxia. Notably, the beneficial effects of ATF4 were reversed by the PI3K/AKT inhibitor LY294002. Under hypoxic conditions, ATF4 overexpression significantly upregulated phosphorylated (p)-PI3K, p-AKT (T308), and p-AKT (S473) in ECs. LY294002, however, markedly reduced the expression of p-PI3K, p-AKT (T308), and p-AKT (S473) in hypoxic ECs overexpressing ATF4. In a murine MI model, ATF4 overexpression partially mitigated cardiac dysfunction and promoted neovascularization, effects that were significantly attenuated by LY294002. These findings suggest that ATF4 plays a crucial role in endothelial cell-mediated neovascularization under post-MI hypoxia by modulating the PI3K/AKT signaling pathway.","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144855125","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

ALDH4A1 knockdown inhibits in vitro atherosclerosis model by modulating Trim28-mediated P53 ubiquitination to suppress ferroptosis of vascular endothelial cells. ALDH4A1敲低通过调节trim28介导的P53泛素化抑制血管内皮细胞铁凋亡，抑制体外动脉粥样硬化模型。

IF 1.7 4区生物学

In Vitro Cellular & Developmental Biology. Animal Pub Date : 2025-08-13 DOI: 10.1007/s11626-025-01102-6

Xiaoyong Xu, Xiaorong Xu, Wangzhuo Zhou, Wenwen Wang, Bin Lin, Xumei Huang, Shan Chen

{"title":"ALDH4A1 knockdown inhibits in vitro atherosclerosis model by modulating Trim28-mediated P53 ubiquitination to suppress ferroptosis of vascular endothelial cells.","authors":"Xiaoyong Xu, Xiaorong Xu, Wangzhuo Zhou, Wenwen Wang, Bin Lin, Xumei Huang, Shan Chen","doi":"10.1007/s11626-025-01102-6","DOIUrl":"https://doi.org/10.1007/s11626-025-01102-6","url":null,"abstract":"Atherosclerosis (AS) is a primary contributor to cardiovascular disease (CVD), resulting in high mortality. Ferroptosis, triggered by lipid peroxidation, contribute to AS development. This study aimed to explore the regulatory relationships of Trim28, ALDH4A1, P53, and ferroptosis in the pathogenesis of AS. The AS cell model was constructed by treating HUVECs with oxidized low-density lipoprotein (ox-LDL). The roles of Trim28 overexpression in regulating AS development, P53 ubiquitination, and ferroptosis of vascular endothelial cells were investigated. Moreover, the interaction between Trim28 and ALDH4A1 was explored, followed by analyzing the effect of ALDH4A1 knockdown on P53 ubiquitination. Additionally, the impact of ALDH4A1 knockdown and P53 overexpression on AS development and ferroptosis of vascular endothelial cells was explored. Reduced Trim28 expression and increased ALDH4A1 and P53 expression were observed in HUVECs after treatment with ox-LDL. Overexpression of Trim28 mitigated AS development, promoted P53 ubiquitination, and suppressed ferroptosis of vascular endothelial cells. Additionally, ALDH4A1 could interact with Trim28, and ALDH4A1 knockdown enhanced P53 ubiquitination. Moreover, P53 overexpression reversed the inhibitory effects of ALDH4A1 knockdown on AS development and ferroptosis of vascular endothelial cells. Our findings indicate that Trim28, ALDH4A1, and P53 may be key regulators in AS development. Silencing of ALDH4A1 may alleviate AS development through regulating Trim28-mediated P53 ubiquitination to inhibit ferroptosis of vascular endothelial cells. These molecules may by promising therapeutic targets for AS and related CVD.","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144834927","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

KHDRBS3-mediated upregulation of circ_0024107 in gastric cancer cells and GC-MSCs synergistically drives gastric cancer cell migration and invasion. khdrbs3介导的胃癌细胞和GC-MSCs中circ_0024107的上调协同驱动胃癌细胞的迁移和侵袭。

IF 1.7 4区生物学

In Vitro Cellular & Developmental Biology. Animal Pub Date : 2025-08-04 DOI: 10.1007/s11626-025-01104-4

Feng Huang, Lin Wang, Xiang Wang, Jing Wen, Mei Wang

{"title":"KHDRBS3-mediated upregulation of circ_0024107 in gastric cancer cells and GC-MSCs synergistically drives gastric cancer cell migration and invasion.","authors":"Feng Huang, Lin Wang, Xiang Wang, Jing Wen, Mei Wang","doi":"10.1007/s11626-025-01104-4","DOIUrl":"https://doi.org/10.1007/s11626-025-01104-4","url":null,"abstract":"Gastric cancer is a significant global health concern due to its high morbidity and mortality. Gastric cancer-associated mesenchymal stem cells (GC-MSCs) significantly contribute to its progression, with circ_0024107 being notably elevated in these cells and essential for their tumor-promoting activities. However, the expression and function of this circRNA in gastric cancer cells as well as its upstream regulators remain unclear. qPCR was used to assess circ_0024107 expression levels. Gain- and loss-of-function experiments evaluated its roles. Transwell assays measured cell migration and invasion. KHDRBS3 was predicted and validated through database analysis and qPCR, and its effects on circ_0024107 were analyzed using qPCR and transwell assays. The expression and clinical implications of KHDRBS3 in gastric cancer were evaluated using the TCGA-STAD database. circ_0024107 expression was elevated in gastric cancer cells, where it promotes migration and invasion. GC-MSCs further enhanced these capabilities by upregulating circ_0024107. KHDRBS3 was identified and validated as a regulator of circ_0024107 expression in both gastric cancer cells and GC-MSCs. Knocking down KHDRBS3 significantly reduced circ_0024107 levels, hindering gastric cancer cell migration and invasion, and weakening the influence of GC-MSCs on tumor cells. KHDRBS3 was abnormally elevated in gastric cancer tissues and correlated with patients' poor prognosis. KHDRBS3-mediated upregulation of circ_0024107 in gastric cancer cells and GC-MSCs synergistically enhances gastric cancer progression. This elucidates novel molecular interactions between GC-MSCs and gastric cancer cells, thereby presenting a promising therapeutic target for effectively mitigating gastric cancer metastasis.","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-08-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144784233","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Optimization of a protocol for the high-yield isolation of active muscle stem cells from bovine skeletal muscle tissue. 从牛骨骼肌组织中高效分离活性肌肉干细胞的方案优化。

IF 1.7 4区生物学

In Vitro Cellular & Developmental Biology. Animal Pub Date : 2025-08-01 DOI: 10.1007/s11626-025-01072-9

Jeong Min Lee, Hyun Lee, Ye Rin Jeon, Keun Cheon Kim, Young Jae Lee, Ha Rin Namkung, So Yeon Nam, Min Seong Kim, Hee Ho Park, Seung Tae Lee

{"title":"Optimization of a protocol for the high-yield isolation of active muscle stem cells from bovine skeletal muscle tissue.","authors":"Jeong Min Lee, Hyun Lee, Ye Rin Jeon, Keun Cheon Kim, Young Jae Lee, Ha Rin Namkung, So Yeon Nam, Min Seong Kim, Hee Ho Park, Seung Tae Lee","doi":"10.1007/s11626-025-01072-9","DOIUrl":"https://doi.org/10.1007/s11626-025-01072-9","url":null,"abstract":"Beef is primarily made up of skeletal muscle tissue. Therefore, the cultivation of bovine muscle stem cells (MSCs) to provide a consistent supply of muscle cells would enhance the sustainability of the cultured beef industry. Here, we report a high-yield, simple, economic, and convenient protocol for the isolation of active MSCs from bovine skeletal muscle tissue. We optimized the enzymatic tissue dissociation protocol and the composition of the medium used for differential plating (DP) to enhance the purity of active MSCs isolated from primary cells derived from the tissue. In addition, the optimal source of bovine muscle tissue for the isolation of active MSCs was determined. The yield of active MSCs was maximized by incubating round area-derived skeletal muscle tissue for 30 min in 0.2% (w/v) collagenase type II in high-glucose DMEM (HG-DMEM), followed by 1% (w/v) pronase in HG-DMEM for 5 min, and conducting DP of the enzymatically dissociated skeletal muscle tissue-derived primary cells in HG-DMEM supplemented with 10% (v/v) FBS and 5 ng/mL bFGF. In conclusion, we established a simple, convenient, and inexpensive protocol for the high-yield isolation of active MSCs from bovine skeletal muscle tissue. This protocol could overcome the technical challenges that hamper the large-scale production of bovine muscle cells, thereby enabling the commercialization of cultured beef.","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144764882","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Non-animal testing of Iranian enterotoxemia vaccine: cell culture assay for Clostridium perfringens epsilon toxin. 伊朗肠毒血症疫苗的非动物试验：产气荚膜梭菌毒素的细胞培养试验。

IF 1.7 4区生物学

In Vitro Cellular & Developmental Biology. Animal Pub Date : 2025-08-01 Epub Date: 2025-07-02 DOI: 10.1007/s11626-025-01069-4

Anahita Emadi, Lida Abdolmohammadi Khiav, Sina Soleimani, Mohsen Lotfi, Faranak Abnaroodheleh, Maryam Dadar

{"title":"Non-animal testing of Iranian enterotoxemia vaccine: cell culture assay for Clostridium perfringens epsilon toxin.","authors":"Anahita Emadi, Lida Abdolmohammadi Khiav, Sina Soleimani, Mohsen Lotfi, Faranak Abnaroodheleh, Maryam Dadar","doi":"10.1007/s11626-025-01069-4","DOIUrl":"10.1007/s11626-025-01069-4","url":null,"abstract":"Epsilon toxin produced by Clostridium perfringens type D is the third most potent clostridial toxin. It causes enterotoxemia in sheep and lambs. The clostridial vaccine has been used against clostridial disease, and its efficacy is evaluated using the serum neutralization (SN) assay as a gold standard. Researchers are concerned about replacing in vivo tests with in vitro tests. Our study aimed to evaluate the cell culture assay to measure the neutralizing antibodies against Clostridium perfringens epsilon toxin as an alternative SN assay. Madin-Darby canine kidney (MDCK) and African green monkey kidney (Vero) cell lines were used to monitor the cell line response after treatment with purified epsilon toxin through microscopic examination and 3-[4,5-imethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) staining. Antibodies were calculated in cell culture assays, and SN results were analyzed using Pearson's correlation. Based on our results, only MDCK was sensitive to the epsilon toxin. The cytopathic effect in this cell culture was rounded. The relationship between toxin concentration and cell viability showed that increasing toxin concentrations significantly decreased cell viability. Good correlation coefficients were obtained between SN and the in vitro assay (r = 0.987) (p < 0.01). The antibody titers obtained by SN were within the range of the cytotoxicity assay and had high reproducibility. Therefore, cell culture may be a suitable alternative for SN assays. Cell culture is one of the tools used in toxicity testing, resulting in consistent and reproducible results.","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"862-870"},"PeriodicalIF":1.7,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144553400","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Therapeutic potential of miR-10a overexpressing mesenchymal stem cell-derived extracellular vesicles in modulating inflammation in collagen-induced arthritis. 过表达miR-10a的间充质干细胞来源的细胞外囊泡在调节胶原诱导关节炎炎症中的治疗潜力。

IF 1.7 4区生物学

In Vitro Cellular & Developmental Biology. Animal Pub Date : 2025-08-01 DOI: 10.1007/s11626-025-01098-z

Yaohui Bai, Jian Zhao, Mohammad Abtahi, Xiaohui Liu

{"title":"Therapeutic potential of miR-10a overexpressing mesenchymal stem cell-derived extracellular vesicles in modulating inflammation in collagen-induced arthritis.","authors":"Yaohui Bai, Jian Zhao, Mohammad Abtahi, Xiaohui Liu","doi":"10.1007/s11626-025-01098-z","DOIUrl":"https://doi.org/10.1007/s11626-025-01098-z","url":null,"abstract":"Rheumatoid arthritis (RA) is a chronic autoimmune condition that leads to joint damage. Mesenchymal stem cells (MSCs) are being recognized as a promising treatment option because of their capacity to modulate immune responses. Their therapeutic effects are mediated by released extracellular vesicles (EVs) which contain microRNAs known to influence inflammatory processes. This research focused on the impact of bone marrow MSC (BM-MSC)-derived EVs overexpressing miR-10a on cytokine production in a mouse model of collagen-induced arthritis (CIA). miR-10a was overexpressed in MSCs derived from bone marrow using Transfectamin. EVs were then isolated from the culture media of both miR-control and miR-10a-modified MSCs. Immunizing mice established the CIA model with type II collagen, after which they received either miR-control or miR-10a-enriched MSC-EVs. The severity of arthritis was evaluated through joint swelling measurements, and the concentrations of pro-inflammatory cytokines (such as interleukin (IL)-17a, interferon (IFN)-γ, and tumor necrosis factor (TNF)-α) alongside anti-inflammatory cytokines (including transforming growth factor (TGF)-β, IL-10, and IL-4) in the joints and serum were assessed using real-time PCR and enzyme-linked immunosorbent assay (ELISA), respectively. Our results indicated that treatment with miR-10a MSC-EVs led to a notable decrease in arthritis severity and joint damage in CIA mice. Furthermore, these EVs were found to lower levels of pro-inflammatory cytokines while enhancing anti-inflammatory cytokines compared to those treated with miR-control MSC-EVs. This study highlights how enhancing miR-10a expression can improve the therapeutic efficacy of MSC-EVs by altering the cytokine environment in CIA models.","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144764883","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0