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<p>Thymoquinone (TQ), a bioactive compound from <i>Nigella sativa</i>, exhibits promising anticancer activity but suffers from poor aqueous solubility, rapid degradation, and low bioavailability, limiting its clinical translation. This study aimed to enhance the therapeutic efficacy of TQ using bovine milk-derived exosomes (Exo-TQ) as a natural nanocarrier. Exosomes were isolated from bovine milk via differential ultracentrifugation and confirmed via immonoblotting, characterized for physicochemical properties, and loaded with TQ using a modified sonication method. The resulting Exo-TQ formulation was evaluated for stability, drug release kinetics, and anticancer activity in MCF-7 breast cancer cells. Cellular assays assessed cytotoxicity, biocompatibility, reactive oxygen species (ROS) generation, apoptosis induction, mitochondrial membrane potential (MMP) disruption, colony formation, and migration inhibition (wound healing and transwell assays). Exo-TQ exhibited a mean particle size of ~ 121&#xa0;nm, high encapsulation efficiency (61.1 ± 3.6%), and maintained structural integrity during storage. Drug release followed a biphasic pattern, with accelerated release at acidic pH. Compared to free TQ, Exo-TQ demonstrated significantly lower IC₅₀ values at all time points, higher ROS production, greater apoptotic induction, and more pronounced MMP loss (<i>p</i> &lt; 0.05). Clonogenic growth was markedly reduced, and migration assays revealed substantial inhibition of wound closure and transwell migration (<i>p</i> &lt; 0.001 and <i>p</i> &lt; 0.05, respectively). Notably, no cytotoxicity was observed in HEK-293 normal cells at concentrations up to 30 µM. Bovine milk-derived exosomes markedly enhance the intracellular delivery and anticancer efficacy of TQ while maintaining biocompatibility in normal cells. The combined effects on proliferation, apoptosis, mitochondrial function, and migration indicate strong therapeutic potential for Exo-TQ as a scalable, natural nanocarrier platform for breast cancer treatment. Further in vivo studies are warranted to confirm these findings and explore translational applications.</p> Graphical Abstract <p></p>

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Bovine Milk-Derived Exosomes as Natural Nanocarriers Enhance the Anticancer Efficacy and Selectivity of Thymoquinone in Breast Cancer Cells

  • 期刊论文

  • 发表日期：

    2026年1月19日

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  • A. Meghana,
  • Ashish Kumar Agrawal

摘要

Thymoquinone (TQ), a bioactive compound from Nigella sativa, exhibits promising anticancer activity but suffers from poor aqueous solubility, rapid degradation, and low bioavailability, limiting its clinical translation. This study aimed to enhance the therapeutic efficacy of TQ using bovine milk-derived exosomes (Exo-TQ) as a natural nanocarrier. Exosomes were isolated from bovine milk via differential ultracentrifugation and confirmed via immonoblotting, characterized for physicochemical properties, and loaded with TQ using a modified sonication method. The resulting Exo-TQ formulation was evaluated for stability, drug release kinetics, and anticancer activity in MCF-7 breast cancer cells. Cellular assays assessed cytotoxicity, biocompatibility, reactive oxygen species (ROS) generation, apoptosis induction, mitochondrial membrane potential (MMP) disruption, colony formation, and migration inhibition (wound healing and transwell assays). Exo-TQ exhibited a mean particle size of ~ 121 nm, high encapsulation efficiency (61.1 ± 3.6%), and maintained structural integrity during storage. Drug release followed a biphasic pattern, with accelerated release at acidic pH. Compared to free TQ, Exo-TQ demonstrated significantly lower IC₅₀ values at all time points, higher ROS production, greater apoptotic induction, and more pronounced MMP loss (p < 0.05). Clonogenic growth was markedly reduced, and migration assays revealed substantial inhibition of wound closure and transwell migration (p < 0.001 and p < 0.05, respectively). Notably, no cytotoxicity was observed in HEK-293 normal cells at concentrations up to 30 µM. Bovine milk-derived exosomes markedly enhance the intracellular delivery and anticancer efficacy of TQ while maintaining biocompatibility in normal cells. The combined effects on proliferation, apoptosis, mitochondrial function, and migration indicate strong therapeutic potential for Exo-TQ as a scalable, natural nanocarrier platform for breast cancer treatment. Further in vivo studies are warranted to confirm these findings and explore translational applications.

Graphical Abstract