In Vitro Cellular & Developmental Biology. Animal最新文献

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 1.5 4区生物学

In Vitro Cellular & Developmental Biology. Animal Pub 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":"https://doi.org/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":""},"PeriodicalIF":1.5,"publicationDate":"2025-07-02","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

Overexpression of MEOX2 inhibits breast cancer cell metastasis by targeting oxidative stress-induced RGS5. MEOX2过表达通过靶向氧化应激诱导的RGS5抑制乳腺癌细胞转移。

IF 1.5 4区生物学

In Vitro Cellular & Developmental Biology. Animal Pub Date : 2025-07-02 DOI: 10.1007/s11626-025-01066-7

Yujun Tang, Jie Luo, Bin Jiang, Jian Deng, Jiehua Li, Liuqing Qin

{"title":"Overexpression of MEOX2 inhibits breast cancer cell metastasis by targeting oxidative stress-induced RGS5.","authors":"Yujun Tang, Jie Luo, Bin Jiang, Jian Deng, Jiehua Li, Liuqing Qin","doi":"10.1007/s11626-025-01066-7","DOIUrl":"https://doi.org/10.1007/s11626-025-01066-7","url":null,"abstract":"This study aimed to investigate the role of mesenchymal homeobox 2 (MEOX2) on breast cancer cell metastasis and its underlying mechanism. Overexpression of MEOX2 in human lymphatic endothelial cell (HLEC) lines was established to assess the adhesion and transendothelial migration of MCF7 and MDA-MB-231 cells to the HLEC cells. After being treated with the oxidative stress inducer H2O2 and the antioxidant N-acetylcysteine (NAC), cell viability, reactive oxygen species (ROS) levels, adhesion, and transendothelial migration of MCF7 and MDA-MB-231 cells to HLEC cells were detected. Tumor volume changes were observed in the xenograft model. The expression of C-X-C chemokine receptor type 4 (CXCR4), C-C chemokine receptor type 7 (CCR7), MEOX2, and G protein signal transduction regulator 5 (RGS5) in tumor tissues and ROS levels were detected. MEOX2 was lowly expressed in breast cancer tissues. Upregulated MEOX2 inhibited the proliferation of lymphatic endothelial cells and the adhesion and transendothelial migration of MCF7 and MDA-MB-231 cells to HLEC cells. After MCF7 and MDA-MB-231 cells were treated with oxidative stress inducer H2O2, ROS levels increased, and cell viability and MEOX2 expression decreased. After NAC or overexpressed MEOX2 treatment, MEOX2 expression increased, ROS and RGS5 levels, adhesion, and transendothelial migration ability decreased in HLEC cells. Overexpression of MEOX2 resulted in smaller tumor volume, lower ROS levels, and lower CXCR4 and CCR7 expression levels. MEOX2 and RGS5 are pivotal in regulating breast cancer metastasis, offering valuable insights into potential therapeutic strategies for breast cancer metastasis.","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144553401","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

Ginsenoside Rb1 inhibits M1 macrophages-induced IGFBP2-mediated endothelial-mesenchymal transition to alleviate myocardial fibrosis in mice with chronic heart failure. 人参皂苷Rb1抑制M1巨噬细胞诱导的igfbp2介导的内皮-间质转化，减轻慢性心力衰竭小鼠心肌纤维化。

IF 1.5 4区生物学

In Vitro Cellular & Developmental Biology. Animal Pub Date : 2025-07-02 DOI: 10.1007/s11626-025-01060-z

Yang Jiang, Qi Zhao, Ting Zhang, Songbo Lan, Xu Yan, Qi Chen

{"title":"Ginsenoside Rb1 inhibits M1 macrophages-induced IGFBP2-mediated endothelial-mesenchymal transition to alleviate myocardial fibrosis in mice with chronic heart failure.","authors":"Yang Jiang, Qi Zhao, Ting Zhang, Songbo Lan, Xu Yan, Qi Chen","doi":"10.1007/s11626-025-01060-z","DOIUrl":"https://doi.org/10.1007/s11626-025-01060-z","url":null,"abstract":"Ginsenoside Rb1 ameliorates renal fibrosis, yet its effects on myocardial fibrosis (MF) remain unclear. In this study, we aimed to explore the role of ginsenoside Rb1 in chronic heart failure (CHF) and MF. To explore the correlation between endothelial-mesenchymal transition (EndMT) in endothelial cells and IGFBP2 expression in M1 macrophages, M1 macrophages were polarized and co-cultured with myocardial microvascular endothelial cells (MMVECs). IGFBP2 levels in the macrophages and levels of endothelial-specific markers and EndMT-related indexes in MMVECs were measured. Additionally, we treated the macrophages with ginsenoside Rb1. The CHF mice model was established using transverse aortic constriction (TAC) and then treated with ginsenoside Rb1. The effects of Rb1 on cardiac function, MF, and cardiomyocyte hypertrophy in CHF mice were assessed. We observed the successful differentiation of M1 macrophages using in vitro experiments. M1 macrophages co-cultured with MMVECs demonstrated the ability to enhance the EndMT effect in MMVECs, as evidenced by elevated levels of IGFBP2 in the macrophages and a reduction in the viability of MMVECs. This decrease in cell viability was mitigated following the knockdown of IGFBP2. Rb1 treatment significantly suppressed the expression of IGFBP2 and inhibited the occurrence of the EndMT in MMVECs. The in vivo experiment findings showed that ginsenoside Rb1 notably enhanced cardiac function, attenuated cardiomyocyte hypertrophy, and alleviated MF in CHF mice. Furthermore, ginsenoside Rb1 inhibited M1 macrophage polarization, reduced IGFBP2 expression in the myocardium, and suppressed the EndMT effect of MMVECs in mice. Ginsenoside Rb1 alleviated MF in mice with CHF by inhibiting M1 macrophage IGFBP2-mediated EndMT.","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144553399","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

LncRNA TDRG1 facilitates high glucose-induced retinal microvascular endothelial cell injury via miR-7-5p/G3BP2. LncRNA TDRG1通过miR-7-5p/G3BP2促进高糖诱导的视网膜微血管内皮细胞损伤。

IF 1.5 4区生物学

In Vitro Cellular & Developmental Biology. Animal Pub Date : 2025-07-01 DOI: 10.1007/s11626-025-01056-9

Shuying Xie, Zehong Liu, Ting Luo, Yifa Chen, Liqun Zeng, Xiaoyan Li

{"title":"LncRNA TDRG1 facilitates high glucose-induced retinal microvascular endothelial cell injury via miR-7-5p/G3BP2.","authors":"Shuying Xie, Zehong Liu, Ting Luo, Yifa Chen, Liqun Zeng, Xiaoyan Li","doi":"10.1007/s11626-025-01056-9","DOIUrl":"https://doi.org/10.1007/s11626-025-01056-9","url":null,"abstract":"This study explores the mechanism of lncRNA TDRG1 in high glucose (HG)-induced human retinal microvascular endothelial cell (hRMEC) injury. hRMECs were cultured in HG medium, followed by the detection of cell viability, proliferation, migration, and angiogenesis using CCK-8, EdU, Transwell, and tube formation assays. LncRNA TDRG1, miR-7-5p, G3BP2, VEGFA, and CD31 expression in hRMECs was detected by RT-qPCR or western blot. After transfection with lncRNA TDRG1 siRNA or miR-7-5p inhibitor or G3BP2 pcDNA3.1, hRMEC injury induced by HG was evaluated. Dual luciferase, RIP, or RNA pull-down assays were performed to verify the binding of lncRNA TDRG1, miR-7-5p, and G3BP2. HG treatment notably elevated the expressions of lncRNA TDRG1 and G3BP2 in hRMECs but diminished the expression of miR-7-5p. Low expression of lncRNA TDRG1 restrained the proliferation, migration, and angiogenesis of hRMECs while diminishing VEGFA and CD31 expression. Mechanistically, lncRNA TDRG1 upregulated the transcription level of G3BP2 by competitively binding to miR-7-5p. Low expression of miR-7-5p or overexpression of G3BP2 weakened the inhibitory effect of lncRNA TDRG1 silencing on HG-induced hRMEC injury. In conclusion, lncRNA TDRG1 upregulates the transcription level of G3BP2 by competitively binding to miR-7-5p, thus exacerbating HG-induced hRMEC injury.","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144539997","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

MiR-21-5p promotes RPE cell necroptosis by targeting Peli1 in a rat model of AMD. 在AMD大鼠模型中，MiR-21-5p通过靶向Peli1促进RPE细胞坏死。

IF 1.5 4区生物学

In Vitro Cellular & Developmental Biology. Animal Pub Date : 2025-06-30 DOI: 10.1007/s11626-025-01064-9

Yishun Shu, Ziwen Li, Tianyi Zong, Tong Mu, Haoyuan Zhou, Qian Yang, Meili Wu, Yanqiu Liu, Tianhua Xie, Chengye Tan, Miao Zhuang, Xiaolu Wang, Yong Yao

{"title":"MiR-21-5p promotes RPE cell necroptosis by targeting Peli1 in a rat model of AMD.","authors":"Yishun Shu, Ziwen Li, Tianyi Zong, Tong Mu, Haoyuan Zhou, Qian Yang, Meili Wu, Yanqiu Liu, Tianhua Xie, Chengye Tan, Miao Zhuang, Xiaolu Wang, Yong Yao","doi":"10.1007/s11626-025-01064-9","DOIUrl":"https://doi.org/10.1007/s11626-025-01064-9","url":null,"abstract":"Nonexudative age-related macular degeneration (dry AMD) is characterized by the progressive degeneration of retinal pigment epithelial (RPE) cells and photoreceptors, resulting in central vision loss. The disease is primarily marked by the accumulation of drusen and RPE atrophy. Given the emerging role of miR-21-5p in various ocular diseases, including diabetic retinopathy, glaucoma, pterygium, and choroidal neovascularization, we hypothesized that miR-21-5p may also impact RPE cell integrity in AMD. To test this hypothesis, we employed a rat model of dry AMD induced by sodium iodate (NaIO3) and evaluated the effects of miR-21-5p modulation via intravitreal injections of miR-21-5p agomir or antagomir. Comprehensive assessments were performed using optical coherence tomography (OCT), fundus imaging, histopathology, and biochemical markers. Our results demonstrated an upregulation of miR-21-5p in response to NaIO3 treatment. Administration of miR-21-5p agomir exacerbated RPE damage, while pretreatment with miR-21-5p antagomir mitigated these detrimental effects. Furthermore, in vitro experiments revealed that miR-21-5p regulates necroptosis in CoCl2-treated RPE cells by targeting Pellino1 (Peli1) via its 3' untranslated region, thereby inhibiting Peli1 expression. Overexpression of Peli1 effectively counteracted the necroptotic effects induced by CoCl2. These findings highlight the potential of miR-21-5p as a therapeutic target in dry AMD, expanding our understanding of miRNA-mediated regulation of RPE cells and suggesting new avenues for treatment strategies.","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144527752","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

Grb2/Sos1 signaling regulates the number of reserve cells in C2C12 cell culture. Grb2/Sos1信号调控C2C12细胞培养中储备细胞的数量。

IF 1.5 4区生物学

In Vitro Cellular & Developmental Biology. Animal Pub Date : 2025-06-30 DOI: 10.1007/s11626-025-01071-w

Yosuke Nagata, Hiroto Iitsuka, Tomoharu Hagiwara

{"title":"Grb2/Sos1 signaling regulates the number of reserve cells in C2C12 cell culture.","authors":"Yosuke Nagata, Hiroto Iitsuka, Tomoharu Hagiwara","doi":"10.1007/s11626-025-01071-w","DOIUrl":"https://doi.org/10.1007/s11626-025-01071-w","url":null,"abstract":"Skeletal muscle regeneration depends on satellite cells that maintain tissue homeostasis through self-renewal and the production of myoblasts that differentiate into mature myofibers. Dysregulation of these processes can lead to muscle degeneration, highlighting the need to elucidate their molecular mechanisms. In this study, we investigated the role of the Grb2/Sos1 signaling pathway in regulating satellite cell self-renewal and differentiation using C2C12 cells. Knockdown of either Grb2 or Sos1 significantly reduced the formation of Bcl-2-positive reserve cells and increased the proportion of differentiated myotubes. Conversely, forced expression of Grb2 increased the number of reserve cells, whereas the Grb2 P49L mutant, which disrupts its interaction with Sos1, decreased reserve cell formation and resulted in thinner myotubes. Although forced expression of Sos1 alone did not significantly increase reserve cell numbers, the chimeric protein cSos-SH2, which combines elements of Grb2 and Sos1, produced a pronounced increase of reserve cells. These results demonstrate that a precise balance between Grb2 and Sos1, along with their coordinated subcellular localization, is critical for controlling reserve cell populations. Activated by growth factor receptor tyrosine kinases and extracellular matrix/integrin interactions, the Grb2/Sos1 signaling pathway is critical for maintaining the muscle satellite cell pool, thereby playing an essential role in muscle regeneration.","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144527751","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

Efficient cryopreservation of Antillean manatee skin-derived somatic cells via reduced intracellular cryoprotectant concentration. 通过降低细胞内冷冻保护剂浓度高效低温保存安的列斯海牛皮肤来源体细胞。

IF 1.5 4区生物学

In Vitro Cellular & Developmental Biology. Animal Pub Date : 2025-06-27 DOI: 10.1007/s11626-025-01061-y

Luanna Lorenna Vieira Rodrigues, Lhara Ricarliany Medeiros de Oliveira, Yasmin Beatriz França Moura, Yara Letícia Frutuoso E Silva, João Vitor da Silva Viana, Leonardo Vitorino Costa de Aquino, Radan Elvis Matias de Oliveira, Fernanda Loffler Niemeyer Attademo, Fábia Oliveira Luna, Alexsandra Fernandes Pereira

{"title":"Efficient cryopreservation of Antillean manatee skin-derived somatic cells via reduced intracellular cryoprotectant concentration.","authors":"Luanna Lorenna Vieira Rodrigues, Lhara Ricarliany Medeiros de Oliveira, Yasmin Beatriz França Moura, Yara Letícia Frutuoso E Silva, João Vitor da Silva Viana, Leonardo Vitorino Costa de Aquino, Radan Elvis Matias de Oliveira, Fernanda Loffler Niemeyer Attademo, Fábia Oliveira Luna, Alexsandra Fernandes Pereira","doi":"10.1007/s11626-025-01061-y","DOIUrl":"https://doi.org/10.1007/s11626-025-01061-y","url":null,"abstract":"The declining population of the Antillean manatees has prompted interest in developing conservation strategies, including somatic cell cryopreservation. However, the type and concentration of intracellular cryoprotectant agents (CPAs) are limiting factors for its success. Therefore, we evaluated three concentrations (5, 8, 10%) of dimethyl sulfoxide (Me2SO) and ethylene glycol (EG) to assess if reducing CPA concentration is efficient for the cells of these animals. Cells not subjected to cryopreservation were used as a control. All cells were analyzed for morphology, viability, metabolism, proliferative activity (PDT), apoptosis, levels of reactive oxygen species (ROS), and mitochondrial membrane potential (ΔΨm). Regardless of the solution used, the cryopreservation did not change frozen-thawed cells' morphology, metabolism, and apoptosis levels compared to control group cells (p > 0.05). Immediately after thawing, cells derived from the 8% Me2SO group-maintained viability similar to the control; after in vitro culture of thawed cells, this positive response of viability was observed only in cells cryopreserved in solutions containing 5% and 8% CPA, regardless the type of CPA. Interestingly, cells frozen in 8% Me2SO showed a higher PDT value than the other groups (p < 0.05). Cells frozen with 10% EG showed higher ROS than the control group (p < 0.05). Additionally, regardless of the solution used, cryopreservation resulted in a change in ΔΨm. In summary, reducing the concentration of CPAs (5 and 8%) helps with somatic cell quality, regardless of the CPA type used in Antillean manatees.","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144511829","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

Wound healing properties of Biginelli scaffolds in Tilapia gill cell line: an in vitro analysis and computational approaches. 罗非鱼鳃细胞系Biginelli支架的伤口愈合性能：体外分析和计算方法。

IF 1.5 4区生物学

In Vitro Cellular & Developmental Biology. Animal Pub Date : 2025-06-19 DOI: 10.1007/s11626-025-01062-x

Noorudeen Paringamalai, Syed Tajudeen Syed Ameen, Abdul Matheen Ibrahim, Gani Taju, Seepoo Abdul Majeed, Azeez Sait Sahul Hameed, Sivaraj Mithra, Predhanekar Mohamed Imran, Attar Kubaib

{"title":"Wound healing properties of Biginelli scaffolds in Tilapia gill cell line: an in vitro analysis and computational approaches.","authors":"Noorudeen Paringamalai, Syed Tajudeen Syed Ameen, Abdul Matheen Ibrahim, Gani Taju, Seepoo Abdul Majeed, Azeez Sait Sahul Hameed, Sivaraj Mithra, Predhanekar Mohamed Imran, Attar Kubaib","doi":"10.1007/s11626-025-01062-x","DOIUrl":"https://doi.org/10.1007/s11626-025-01062-x","url":null,"abstract":"The skin is a vital organ that regulates the temperature, nutrient absorption, and perception of sensations. Wound healing is a complex biological process in multicellular systems that consists of four key phases: hemostasis, inflammation, proliferation, and remodeling. This study develops a new approach for synthesizing dihydropyrimidinones (DHPM) named Biginelli scaffolds via a simple, rapid, eco-friendly, and cost-effective solvent-free Biginelli reaction for wound healing activities. The synthesis involved a one-pot three-component coupling reaction of β-ketoester derivatives, anisaldehyde, and simple urea in a domestic microwave oven. The synthesized (B1-B4) scaffolds were characterized using melting point, UV-Vis, FT-IR, HRMS, 2D-NMR (NOESY), and proton/carbon NMR spectroscopies. The molecular docking results showed that the synthetic scaffolds (B1-B4) had strong binding abilities, with B3 and B4 having the best interactions in the group, similar to the control compound (curcumin). It exhibited less cytotoxic effects up to 80 µg/mL in Tilapia gill (TG) cells in the MTT assay. The synthesized scaffolds (60 µg/mL) enhanced TG cell growth and had potential applications in wound healing. Biginelli (B1-B4) scaffolds showed good antioxidant properties in the DPPH assay. RT-qPCR analysis indicated that TG cells exposed to different (B1-B4) scaffold concentrations had significantly increased VEGF gene expression. The scaffolds showed no toxic effects on adsorption, distribution, metabolism, excretion, and toxicity (ADMET) analysis, and the structure was optimized using the DFT-B3LYP-6311G-(d,p) hybrid basis set. This method has wide applications in future research and provides insights into tissue engineering and biomedical applications.","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144325493","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

PKCβ expression contributes to M1 macrophage-induced impairment in the osteogenic differentiation of periodontal ligament stem cells. PKCβ表达参与M1巨噬细胞诱导的牙周韧带干细胞成骨分化损伤。

IF 1.5 4区生物学

In Vitro Cellular & Developmental Biology. Animal Pub Date : 2025-06-17 DOI: 10.1007/s11626-025-01032-3

Yang Liu, Zhaocen Liu

{"title":"PKCβ expression contributes to M1 macrophage-induced impairment in the osteogenic differentiation of periodontal ligament stem cells.","authors":"Yang Liu, Zhaocen Liu","doi":"10.1007/s11626-025-01032-3","DOIUrl":"https://doi.org/10.1007/s11626-025-01032-3","url":null,"abstract":"Protein kinase Cβ (PRKCB) is expressed in THP-1 cells and has been found upregulated in periodontitis. Exploring the specific molecular mechanisms that promote the osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) is beneficial to optimizing periodontal regeneration. THP-1 cells were induced to differentiate into macrophages, and the levels of PRKCB in macrophages with different phenotypes were examined, including PKC activity. The effect of pg-LPS induction on osteogenic differentiation of hPDLSCs was measured by measuring alkaline phosphatase, osteocalcin, osteogenic-related proteins, and mineralized nodules. Ruboxistaurin, an inhibitor of PRKCB, was used to treat M1 macrophages to examine its effect on macrophage polarization. Additionally, the cascade effect of ruboxistaurin on osteogenic differentiation was investigated by co-incubating hPDLSCs with medium from macrophages. The results indicated that PRKCB upregulation and increased PKC activity were induced in M1 macrophages upon stimulation with LPS/IFN-γ. pg-LPS resulted in decreased levels of osteogenic-related genes in hPDLSCs, accompanied by a decrease in mineralized nodules. PRKCB inhibitor reduced PKC activity, inhibited macrophage M1 polarization, and reduced M1-related inflammatory cytokine secretion. Exposure of hPDLSCs to M1 macrophage-derived conditioned medium impaired their osteogenic differentiation potentials, which was significantly attenuated by pretreatment of M1 macrophages with ruboxistaurin. Together, inhibition of PRKCB suppressed inflammatory M1 macrophage polarization, thus attenuating M1 macrophage-induced impairment in the osteogenic differentiation of hPDLSCs. These results provide a theoretical and scientific basis for optimizing the potential clinical application of hPDLSC therapy in periodontal regeneration.","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144316827","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

miR- 34c- 5p targets ROCK1 expression to inhibit kidney injury in diabetic nephropathy rats through MAPK/ERK signaling pathway. miR- 34c- 5p通过MAPK/ERK信号通路靶向ROCK1表达，抑制糖尿病肾病大鼠肾损伤。

IF 1.5 4区生物学

In Vitro Cellular & Developmental Biology. Animal Pub Date : 2025-06-12 DOI: 10.1007/s11626-025-01039-w

HuaJuan Wei, Ye Li, HongDe Liu, Li Pan, HuiLing Duo, ShaoYing Dong

{"title":"miR- 34c- 5p targets ROCK1 expression to inhibit kidney injury in diabetic nephropathy rats through MAPK/ERK signaling pathway.","authors":"HuaJuan Wei, Ye Li, HongDe Liu, Li Pan, HuiLing Duo, ShaoYing Dong","doi":"10.1007/s11626-025-01039-w","DOIUrl":"https://doi.org/10.1007/s11626-025-01039-w","url":null,"abstract":"This study was to investigate the mechanism of miR- 34c- 5p in alleviating kidney injury in diabetic nephropathy (DN) rats by targeting ROCK1 and MAPK/ERK signaling pathway. The rat model of DN was established and fasting blood glucose, 24-h proteinuria, blood urea nitrogen, and serum creatinine were measured to quantify kidney injury. Kidney tissue was dissected for H&E and TUNEL staining. Renal injury factor KIM- 1 was measured by Western blot analysis. Retinal Muller cells (RMCs) were treated with high glucose and transfected. Cell viability was detected by CCK- 8 and apoptosis by flow cytometry. Inflammatory factors in DN rats and RMCs were analyzed by ELISA. The targeting effect of miR- 34c- 54p on ROCK1 was demonstrated by RNA pull-down and dual-luciferase reporter gene. Finally, ROCK1, p-MEK1/2, and p-ERK were assessed by Western blot. Elevating miR- 34c- 5p could inhibit DN kidney injury and high glucose-induced cell injury, and reduce inflammation in kidney tissue of DN rats and RMCs. miR- 34c- 5p targeted to regulate ROCK1 expression, and restoring ROCK1 abolished the therapeutic effect of elevating miR- 34c- 5p. Phosphorylated MEK and ERK were increased in DN rats and RMCs induced by high glucose. miR- 34c- 5p can inhibit kidney injury induced by DN by targeting ROCK1, and the MAPK/ERK pathway may represent the pathological mechanism of DN.","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-06-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144274764","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