Zhaozhong Yu, Jun Kuang, Xuan Wan, Chaoyi Liang, Fan Wu, Zhimin Shu, Weidong Liang, Xiaoming Wan, Jianbin Gong
{"title":"The chitosan collagen salvianolic membrane mitigates intervertebral disc degeneration through suppression of the Wnt/β-catenin signaling.","authors":"Zhaozhong Yu, Jun Kuang, Xuan Wan, Chaoyi Liang, Fan Wu, Zhimin Shu, Weidong Liang, Xiaoming Wan, Jianbin Gong","doi":"10.1007/s11626-026-01163-1","DOIUrl":"https://doi.org/10.1007/s11626-026-01163-1","url":null,"abstract":"<p><p>Intervertebral disc degeneration (IVDD), a natural aging process, leads to intervertebral disc pain, with lumbar resection providing pain relief but potential postoperative complications affecting outcomes. The chitosan-collagen-salvianolic (CCS) membrane has shown promise in reducing scar formation after lumbar resection, though the mechanisms remain unclear. A lumbar laminectomy model was used, with materials applied to the surgical site. Tissue samples were collected at 4 and 8 wk post-surgery for histological evaluation to assess pathological changes and apoptosis. Western blot analysis examined protein expression in the Wnt/β-catenin signaling pathway in scar tissue. Rabbit vascular adventitia fibroblast (VAF) cells were isolated, and a Wnt3a overexpression vector was created. Various composite membrane materials and pathway inhibitors were tested, with assessments of cell invasion, apoptosis, viability, cytokine levels, and protein expression. Results showed that lumbar laminectomy activated the Wnt/β-catenin pathway. The CCS membrane facilitated healing by inhibiting this pathway. It enhanced VAF cell activity, reduced apoptosis, and suppressed Wnt3a pathway activity. Additionally, it protected cells from Wnt3a overexpression-induced damage while maintaining normal function. In conclusion, the CCS composite membrane promoted bone tissue repair after lumbar laminectomy by inhibiting Wnt/β-catenin pathway activation and preserving extracellular matrix fibroblast function.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147490993","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}
{"title":"VEGFA is essential for the potentiation of ATRA on BMP9-induced osteogenesis of preadipocytes and osteoporotic fracture healing.","authors":"Hao Wang, Jingjiang Wang, Jiangyu Xiang, Yang Liu","doi":"10.1007/s11626-026-01170-2","DOIUrl":"https://doi.org/10.1007/s11626-026-01170-2","url":null,"abstract":"<p><p>The induction of osteogenic differentiation in preadipocytes may serve as a potential therapeutic approach for treating osteoporosis and osteoporotic fractures. All-trans retinoic acid (ATRA) promotes the bone morphogenetic protein 9 (BMP9)-induced osteogenic differentiation of preadipocytes. The present study further investigated whether vascular endothelial growth factor A (VEGFA) may play a role in this process and the effect of ATRA and BMP9 on osteoporotic fracture healing in rats. The results indicated that ATRA and BMP9 synergistically upregulated VEGFA expression in preadipocytes. Furthermore, knockdown of VEGFA expression abolished the stimulatory effect of ATRA on the BMP9-induced early and late osteogenic differentiation of preadipocytes in vitro, as evidenced by a decrease in alkaline phosphatase (ALP) activity, osteopontin and osteocalcin expression as well as mineralization. The in vivo cell implantation assay showed that ATRA failed to augment BMP9-induced ectopic bone formation in the absence of VEGFA. Subsequently, an osteoporotic femoral fracture rat model was established and micro-CT scans, alongside quantitative analysis, revealed that ATRA effectively promoted BMP9-stimulated callus formation during osteoporotic fracture healing. Moreover, ATRA and BMP9 acted together to significantly elevate VEGFA expression in bone calluses. Mechanistically, ATRA and BMP9 synergistically stimulated the osteogenic transcription factor, runt-related transcription factor 2 (Runx2). Transcriptomic and bioinformatic analyses further revealed that the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway may be crucial for mediating the synergistic effects of ATRA and BMP9, as validated by detecting the phosphorylation of PI3K and Akt. However, the increases in Runx2 expression and Akt phosphorylation induced by the combination of ATRA and BMP9 were inhibited by VEGFA silencing. ATRA also failed to increase the BMP9-induced ALP activity in preadipocytes treated with an Akt inhibitor. These findings suggest that VEGFA may influence the potentiation effect of ATRA on the BMP9-mediated osteogenesis of preadipocytes and in osteoporotic fracture healing through Runx2 and the PI3K/Akt signaling pathway.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-03-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147473629","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}
Zuzana Kiššová, Klaudia Jaszcza, Veronika Vinclérová, Rege Anna Márton, Máté Mackei, Gábor Mátis, Viera Karaffová
{"title":"Dynamics of viability and apoptotic regulation in chicken ileal explant culture.","authors":"Zuzana Kiššová, Klaudia Jaszcza, Veronika Vinclérová, Rege Anna Márton, Máté Mackei, Gábor Mátis, Viera Karaffová","doi":"10.1007/s11626-026-01173-z","DOIUrl":"https://doi.org/10.1007/s11626-026-01173-z","url":null,"abstract":"<p><p>The development of a reliable ex vivo intestinal model is essential for studying gut physiology and host-microbe interactions under controlled conditions. This study evaluated the temporal dynamics of viability, morphology, and apoptotic regulation in chicken ileal explants cultured ex vivo. Ileal tissues from 21-d-old broiler chickens were incubated for 0, 4, and 8 h and analyzed using metabolic assays (MTS, CCK-8), histological staining (hematoxylin and eosin (H&E), Periodic Acid-Schiff (PAS)), and quantitative PCR for apoptosis-related genes (BCL2L11, cytochrome c, caspase-3, caspase-8, and caspase-9). After 4 h, explants retained structural integrity with well-developed villi and active mucus secretion. Metabolic assays confirmed preserved cell viability, while transcriptional analysis revealed moderate upregulation of cytochrome c and downregulation of BCL2L11, suggesting transient adaptation to culture stress. After 8 h, villi appeared shortened with epithelial exfoliation and reduced PAS reactivity, indicating progressive tissue deterioration. Correspondingly, BCL2L11 and cytochrome c were significantly upregulated together with caspase-8, reflecting activation of intrinsic and extrinsic apoptotic pathways. Metabolic activity declined markedly, confirming reduced tissue vitality. These findings demonstrate that chicken ileal explants with a diameter of 4 mm remain viable and functionally stable up to approximately 4 h of ex vivo culture, after which apoptotic processes intensify, compromising tissue integrity. The results define the practical viability window for short-term intestinal explant studies and provide methodological guidance for future investigations on host-pathogen or probiotic interactions in poultry gut models.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147456905","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}
Selinay Şenkal-Turhan, Özüm Begüm Böke, Ezgi Bulut-Okumuş, Hazar Eren Soydan, Neşe Başak Türkmen, Ayça Şenol, Kaan Kadayıfcı, Neslihan Akmaz, Ayşegül Doğan
{"title":"The active plant compounds demonstrated positive activity on mouse intestinal organoids as an inflammation model system.","authors":"Selinay Şenkal-Turhan, Özüm Begüm Böke, Ezgi Bulut-Okumuş, Hazar Eren Soydan, Neşe Başak Türkmen, Ayça Şenol, Kaan Kadayıfcı, Neslihan Akmaz, Ayşegül Doğan","doi":"10.1007/s11626-026-01165-z","DOIUrl":"https://doi.org/10.1007/s11626-026-01165-z","url":null,"abstract":"<p><p>Diet has an important impact on intestinal homeostasis, and the establishment of appropriate experimental models to study the effect of food compounds is of interest. The organoid model can be used to check the positive protective role of active food compounds on intestinal tissue. In the current study, mouse intestinal organoids were used to model air-liquid interface (ALI), lipopolysaccharide (LPS)-induced inflammation, and macrophage co-culture-based inflammation modelling. The activity of hesperidin, capsaicin, allicin, and 18β-glycyrrhetinic acid (18β-GA) was determined in organoid culture. Morphology, crypt number, area, and intensity were analyzed. mRNA expression analysis and immunostaining analysis were performed for inflammation and proliferation markers. The ALI model exerted a suitable organoid culture system to mimic intestinal growth based on our results. Hesperidin, capsaicin, and allicin demonstrated positive effects on LPS-induced inflammation. All of the food compounds showed positive effects in macrophage co-culture for organoid structure and growth but not for macrophage proliferation and viability. All compounds reduced the inflammatory gene expression and increased stem cell marker and proliferation-related gene expression in the ALI model. In addition, capsaicin showed positive effects on organoid growth and maturation. This study generated an experimental model system to test food components and might be used in further research.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-03-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147432556","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}
{"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":"10.1007/s11626-025-01062-x","url":null,"abstract":"<p><p>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.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"389-411"},"PeriodicalIF":1.7,"publicationDate":"2026-03-01","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}
Anita Solhaug, Georgina C Dowd, Vivian R Dayeh, Hilde Sindre, Lucy E J Lee, Niels C Bols
{"title":"Improve your success with fish cell lines-small things that matter.","authors":"Anita Solhaug, Georgina C Dowd, Vivian R Dayeh, Hilde Sindre, Lucy E J Lee, Niels C Bols","doi":"10.1007/s11626-025-01042-1","DOIUrl":"10.1007/s11626-025-01042-1","url":null,"abstract":"<p><p>There is a drive towards reducing animal experiments and developing robust biologically relevant in vitro models based on cell lines, including those derived from fish. At the time of writing, Cellosaurus, the knowledge base of current cell lines used in research, listed more than 900 fish cell lines in its database. One of the key challenges facing fish cell biology is the lack of fundamental technical information regarding the isolation, culture, and application of cell lines. Researchers often work in silos, encountering similar technical challenges, each spending significant time and resources overcoming the same issues for which solutions may not be readily accessible. Here, we share some of the key considerations for the isolation, culture, maintenance, and application of fish cell lines in toxicology, which we have encountered over our collective decades of experience.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"345-363"},"PeriodicalIF":1.7,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13053418/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144008887","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ashaq Sultan Dar, Fayaz Ahmad, Feroz Ahmad Shah, Syed Shariq Nazir Qadiri, Keezia Khurshid
{"title":"Development and characterization of a cell line from the caudal fin of Schizothorax niger (Heckel, 1838) for in vitro toxicity testing.","authors":"Ashaq Sultan Dar, Fayaz Ahmad, Feroz Ahmad Shah, Syed Shariq Nazir Qadiri, Keezia Khurshid","doi":"10.1007/s11626-025-01018-1","DOIUrl":"10.1007/s11626-025-01018-1","url":null,"abstract":"<p><p>Here, we successfully grew the SNCF (Schizothorax niger caudal fin) cell line from the caudal fin explants of S. niger, an important cold-water fish of the Himalayas. The cells were successfully grown up to 22 passages by planting explant tissues in DMEM medium supplemented with FBS. We observed optimal cell growth at a concentration of 18% FBS. We observed the steady generation of cells from explants from days 2 to 5 of seeding, and obtained a complete monolayer at days 7-10. We tested various temperatures, including 10 °C, 13 °C, 16 °C, 19 °C, 22 °C, 25 °C, and 28 °C, and found that 22 °C was the optimal temperature for cell growth. We examined the response to various doses of epidermal growth factor (EGF) and fibroblast growth factor (FGF) (0, 2, 4, 6, 8, and 10 ng/mL) on cell colony growth at an optimal temperature of 22 °C. We characterized the cell line using karyotyping at the 14th and 20th passages. The cell line showed epithelial cell-like growth by morphology, which was confirmed by immunotyping. We further used the cell line to study the impact of three pesticides (chlorpyrifos, dimethoate, and endosulfan), and a fungicide (mancozeb) and bacterial extracellular product (ECP). The DAPI stain assay and MTT assay confirmed the pesticides toxic effects on the cells, revealing disintegration of the cell nuclei by the formation of micronuclei and LC<sub>50</sub> concentrations. ECP treatment showed disruption of the monolayer within 0-36 hrs.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"255-267"},"PeriodicalIF":1.7,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143187760","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}
Gui-Ling Zheng, Yu-Fan Yao, Xiao-Yu Zhang, Qian-Long Yu, Jie Li, Yi-Ping Li, Dong Chu, Chang-You Li
{"title":"Establishment of an embryonic cell line of Grapholita molesta (Lepidoptera: Tortricidae) and in vitro replication of Cydia pomonella granulovirus in it.","authors":"Gui-Ling Zheng, Yu-Fan Yao, Xiao-Yu Zhang, Qian-Long Yu, Jie Li, Yi-Ping Li, Dong Chu, Chang-You Li","doi":"10.1007/s11626-025-01036-z","DOIUrl":"10.1007/s11626-025-01036-z","url":null,"abstract":"<p><p>The oriental fruit moth, Grapholita molesta (Busck) (Lepidoptera: Tortricidae), is a major pest of fruit trees worldwide. In this study, an embryonic cell line QAU-Gm-E-L of the oriental fruit moth was successfully established. The cells grew adherently, round cells and spindle cells accounted for 43.0% and 42.2% of the total population, respectively, and rod-shaped cells accounted for 14.8%. The amplified mitochondrial cytochrome oxidase I subunit (CoI) gene fragment was 651 bp in length, and its similarity with the CoI gene of the oriental fruit moth was 100%. The chromosomes of QAU-Gm-E-L cells were granular or short rod-shaped. Its number varied from 66 to 444, indicating that aneuploidy occurred. The observations were consistent with the chromosome characteristics of lepidopteran insect cell lines. The population doubling time of QAU-Gm-E-L cells was 27.64 h. Real-time fluorescence quantitative polymerase chain reaction (qPCR) confirmed that the number of copies of Cydia pomonella granulovirus (CpGV) gradually increased in QAU-Gm-E-L cells with inoculation time. The electron microscopy observations results showed that occlusion bodies (OBs) of CpGV could be formed in the cells at 4 d post-infection; a large number of OBs were seen in the cells at 8 d post-infection. Hence, the QAU-Gm-E-L cells can support the in vitro replication and proliferation of CpGV, and it will provide an ideal material for the molecular biology research of oriental fruit moth and CpGV.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"329-336"},"PeriodicalIF":1.7,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144012807","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}
{"title":"An in vitro cellular model for measuring the impact of thermal stress on Florida reef sponges.","authors":"Megan Conkling, Tobin Hindle, Zhixiao Xie, Weibo Liu, Timothy Moore, Shirley A Pomponi","doi":"10.1007/s11626-025-01034-1","DOIUrl":"10.1007/s11626-025-01034-1","url":null,"abstract":"<p><p>Coral reefs are threatened by recurrent mortality incidents in their native habitats brought on by natural and anthropogenic stressors. Elevated temperature has been indicated as a major causing factor. Although ongoing research is focused on corals, sponges are an important benthic organism on coral reefs and are often overlooked. An accurate and standardized method is needed to determine the environmental limits and thresholds of sponges commonly found on coral reefs. We established an in vitro sponge cell model and evaluated the effect of elevated temperatures on primary cell cultures of five common Florida reef sponges-Agelas clathrodes, Aplysina fulva, Cliona varians, Geodia neptuni, and Xestospongia muta. Analysis of the results revealed that the impact of increased temperatures had no significant effect at the cellular level, but there are changes at the molecular level. Shifts in the sponges' transcriptomic profiles induced by increased temperatures, trigger processes related to signal transduction, apoptosis, and cell repair pathways. Further elevation of temperature corresponding to local extremes activated the immune response and programmed cell death. The results of the present study are based on both cellular and molecular data obtained from the in vitro cell model which highlight the minimal response of all five species to thermal stress, providing an insight into the mechanisms involved in the adaptive process. Furthermore, they suggest a resilience of these sponges to the current thermal extremes, but a combination of factors could still lead to a loss of sponges on reefs. This study forms the basis for use of in vitro sponge cell models to evaluate other environmental parameters and stressors on additional sponge species.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"313-328"},"PeriodicalIF":1.7,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144127497","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}
{"title":"A comparative genotoxicity study of agrochemicals: nuclear abnormalities, comet assay, and gene expression alterations.","authors":"Ankita Salunke, Parth Pandya, Bhumi Thakkar, Pragna Parikh","doi":"10.1007/s11626-025-01030-5","DOIUrl":"10.1007/s11626-025-01030-5","url":null,"abstract":"<p><p>Agrochemicals (AGs) are known for their ability to have a negative impact on the health of non-target species, despite the fact that they are meant to protect agricultural plants from harmful pests. Catla catla (Hamilton, 1822) gill cells (ICG) were exposed to four AGs: insecticide (Imidacloprid (IMI)), fungicide (Curzate (CZ)), herbicide (pyrazosulfuron ethyl (PE)), and fertilizer micronutrients (MN) with sublethal concentrations 1/20th, 1/10th, and 1/5th of IC<sub>50</sub>, described here as low dose (LD), medium dose (MD), and high dose (HD), respectively. A significant dose-dependent increase in the nuclear abnormalities such as micronuclei formation, bi-nucleated, and lobbed nucleated cells was observed in ICG cells treated with AGs. Of all the AGs, maximum alterations were observed with the HD of IMI followed by CZ, PE, and MN. Concurrently, the genotoxicity was determined by performing comet assays with high dose of all AGs. The gene expression of dnmt and cyp p450 were also studied through q-PCR in ICG cells. The significant increase in expression as well as alteration in cyp p450 and dnmt sequence was reported in ICG cells exposed to HD of IMI. This suggests that IMI has a genotoxic effect and may lead to epigenetic alterations.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"292-302"},"PeriodicalIF":1.7,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144008886","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}