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Background <p>Head and neck squamous cell carcinoma (HNSCC) presents significant therapeutic challenges due to its aggressiveness and the morbidity of conventional treatments. Photodynamic therapy (PDT) offers a localized approach, but its efficacy can be limited. This study investigated the potential of nano-silymarin, a bioactive flavonolignan complex with known anticancer properties, in combination with methylene blue-mediated PDT (MB-PDT) in HNSCC cells.</p> Methods <p>The cytotoxic, apoptotic, and anti-migratory effects of various treatments were assessed in HN5 cells. Groups included controls, individual agents (nano-silymarin, MB, laser), dual therapies, and the triple combination (nano-silymarin + MB + laser). Cell viability was measured using the MTT assay, apoptosis by flow cytometry, and migration by the scratch assay. Results were analyzed using one-way ANOVA with Tukey’s post hoc test. Synergy was quantified using the Bliss independence, Loewe additivity, and zero interaction potency models.</p> Results <p>The triple combination (nano-silymarin + MB-PDT) demonstrated the most potent effect, synergistically reducing cell viability to 40.63% (compared with 69.65% for nano-silymarin and 77.32% for MB-PDT). It induced the highest level of total apoptosis (42.93%) and nearly completely inhibited cell migration (97.2% wound openness at 72&#xa0;h). This anti-migratory effect was significantly greater than the sum of the effects of the individual monotherapies, indicating a synergistic blockade of cellular invasion. Synergy analysis confirmed a strong synergistic interaction, with a Combination index of 0.682.</p> Conclusion <p>Nano-silymarin significantly potentiates the anticancer efficacy of MB-PDT against HNSCC cells through strong synergistic interactions, thereby enhancing cytotoxicity, apoptosis, and migration inhibition. These findings support further development of nano-silymarin in combinatorial PDT regimens for HNSCC.</p>

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Nano-silymarin combined with methylene blue photodynamic therapy versus monotherapies of head and neck squamous cell carcinoma: in-vitro study

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年6月29日

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  • Delband Hefzi,
  • Maryam Koopaie,
  • Neda Hakimiha,
  • Shima Younespour

摘要

Background

Head and neck squamous cell carcinoma (HNSCC) presents significant therapeutic challenges due to its aggressiveness and the morbidity of conventional treatments. Photodynamic therapy (PDT) offers a localized approach, but its efficacy can be limited. This study investigated the potential of nano-silymarin, a bioactive flavonolignan complex with known anticancer properties, in combination with methylene blue-mediated PDT (MB-PDT) in HNSCC cells.

Methods

The cytotoxic, apoptotic, and anti-migratory effects of various treatments were assessed in HN5 cells. Groups included controls, individual agents (nano-silymarin, MB, laser), dual therapies, and the triple combination (nano-silymarin + MB + laser). Cell viability was measured using the MTT assay, apoptosis by flow cytometry, and migration by the scratch assay. Results were analyzed using one-way ANOVA with Tukey’s post hoc test. Synergy was quantified using the Bliss independence, Loewe additivity, and zero interaction potency models.

Results

The triple combination (nano-silymarin + MB-PDT) demonstrated the most potent effect, synergistically reducing cell viability to 40.63% (compared with 69.65% for nano-silymarin and 77.32% for MB-PDT). It induced the highest level of total apoptosis (42.93%) and nearly completely inhibited cell migration (97.2% wound openness at 72 h). This anti-migratory effect was significantly greater than the sum of the effects of the individual monotherapies, indicating a synergistic blockade of cellular invasion. Synergy analysis confirmed a strong synergistic interaction, with a Combination index of 0.682.

Conclusion

Nano-silymarin significantly potentiates the anticancer efficacy of MB-PDT against HNSCC cells through strong synergistic interactions, thereby enhancing cytotoxicity, apoptosis, and migration inhibition. These findings support further development of nano-silymarin in combinatorial PDT regimens for HNSCC.