Mohamed Hanan R H, Ibrahim Esraa H, Shaheen Shahd E E, Hussein Nesma O E, Diab Ayman, Safwat Gehan
Zoology Department Faculty of Science, Cairo University, Giza, Egypt.
Faculty of Biotechnology, October University for Modern Sciences and Arts, 6th of October, Egypt.
Sci Rep. 2025 Jan 23;15(1):2993. doi: 10.1038/s41598-025-86401-4.
Calcium hydroxide nanoparticles (Ca(OH)NPs) possess potent antimicrobial activities and unique physical and chemical properties, making them valuable across various fields. However, limited information exists regarding their effects on genomic DNA integrity and their potential to induce apoptosis in normal and cancerous human cell lines. This study thus aimed to evaluate the impact of Ca(OH)NPs on cell viability, genomic DNA integrity, and oxidative stress induction in human normal skin fibroblasts (HSF) and cancerous hepatic (HepG2) cells. Cell viability and genomic DNA stability were assessed using the Sulforhodamine B (SRB) assay and alkaline comet assay, respectively. Reactive oxygen species (ROS) levels were measured using 2,7-dichlorofluorescein diacetate, while the expression level of apoptosis-related genes (p53, Bax, and Bcl-2) were quantified using real-time PCR (qRT-PCR). The SRB cytotoxicity assay revealed that a 48-hour exposure to Ca(OH)NPs caused concentration-dependent cell death and proliferation inhibition in both HSF and HepG2 cells, with IC50 values of 271.93 µg/mL for HSF and 291.8 µg/mL for HepG2 cells. Treatment with the IC50 concentration of Ca(OH)NPs selectively induced significant DNA damage, excessive ROS generation, and marked dysregulation of apoptotic (p53 and Bax) and anti-apoptotic (Bcl-2) gene expression in HepG2 cells, triggering apoptosis. In contrast, exposure of HSF cells to the IC50 concentration of Ca(OH)NPs caused no significant changes in genomic DNA integrity, ROS generation, or apoptotic gene expression. These findings indicate that Ca(OH)NPs exhibit concentration-dependent cytotoxicity in both normal HSF and cancerous HepG2 cells. However, exposure to the IC50 concentration was non-genotoxic to normal HSF cells while selectively inducing genotoxicity and apoptosis in HepG2 cancer cells through DNA breaks and ROS-mediated mechanisms. Further studies are required to explore the biological and toxicological properties and therapeutic potential of Ca(OH)NPs in hepatic cancer treatment.
氢氧化钙纳米颗粒（Ca(OH)NPs）具有强大的抗菌活性以及独特的物理和化学性质，这使得它们在各个领域都具有重要价值。然而，关于它们对基因组DNA完整性的影响以及在正常和癌细胞系中诱导细胞凋亡的潜力，目前的信息有限。因此，本研究旨在评估Ca(OH)NPs对人正常皮肤成纤维细胞（HSF）和肝癌细胞（HepG2）的细胞活力、基因组DNA完整性以及氧化应激诱导的影响。分别使用磺酰罗丹明B（SRB）测定法和碱性彗星试验评估细胞活力和基因组DNA稳定性。使用二乙酸2,7-二氯荧光素测量活性氧（ROS）水平，同时使用实时PCR（qRT-PCR）定量凋亡相关基因（p53、Bax和Bcl-2）的表达水平。SRB细胞毒性试验表明，48小时暴露于Ca(OH)NPs会导致HSF和HepG2细胞浓度依赖性的细胞死亡和增殖抑制，HSF细胞的IC50值为271.93μg/mL，HepG2细胞的IC50值为291.8μg/mL。用Ca(OH)NPs的IC50浓度处理选择性地诱导HepG2细胞中显著的DNA损伤、过量的ROS生成以及凋亡（p53和Bax）和抗凋亡（Bcl-2）基因表达的明显失调，从而引发细胞凋亡。相比之下，HSF细胞暴露于Ca(OH)NPs的IC50浓度对基因组DNA完整性、ROS生成或凋亡基因表达没有显著影响。这些发现表明，Ca(OH)NPs在正常HSF细胞和癌细胞HepG2中均表现出浓度依赖性细胞毒性。然而，暴露于IC50浓度对正常HSF细胞无遗传毒性，而通过DNA断裂和ROS介导的机制在HepG2癌细胞中选择性地诱导遗传毒性和细胞凋亡。需要进一步研究以探索Ca(OH)NPs在肝癌治疗中的生物学和毒理学特性以及治疗潜力。
