Effects of nanosilymarin and conditioned medium of adipose-derived mesenchymal stem cells on peripheral blood mononuclear cells co-cultured with HeLa cells

Cervical cancer is the fourth most common malignancy in females worldwide. Silymarin, a flavonoid with established anti-inflammatory properties, shows promise in oncology. Conditioned medium (CM) of Mesenchymal stem cell (MSC) exerts paracrine immunomodulatory effects. This study evaluated the combined impact of CM of adipose tissue-derived MSC (AD-MSC) and nanosilymarin on peripheral blood mononuclear cells (PBMC) from healthy donors co-cultured with HeLa cervical cancer cells. AD-MSCs were isolated from adipose tissue via collagenase I digestion, characterized by immunophenotyping and multilineage differentiation potential. Apoptosis (Annexin V/PI), proliferation (MTT assay), and gene expression of IFN-γ, TNF-α, IL-4, and TGF-β (qRT-PCR) were assessed in PBMC treated with AD-MSC-CM (50%) and/or nanosilymarin (25 µM) for 48 h, in monoculture or HeLa co-culture. In PBMC-HeLa co-culture, nanosilymarin (different concentrations) with 50% MSC-CM significantly reduced PBMC proliferation. Nanosilymarin (25 µM) plus MSC-CM induced marked HeLa apoptosis. Both agents together upregulated IFN-γ expression in PBMC, in monoculture and co-culture, without altering TNF-α, IL-4, or TGF-β. Nanosilymarin combined with AD-MSC-CM enhances PBMC-mediated anti-tumor immunity via IFN-γ upregulation and promotes HeLa apoptosis, suggesting synergistic potential for cervical cancer immunotherapy. Mechanistic studies and dose optimization are warranted. • Nanosilymarin + AD-MSC-CM (50%) reduces PBMC proliferation in HeLa co-culture. • Nanosilymarin (25 µM) + AD-MSC-CM induces HeLa apoptosis in PBMC co-culture. • Nanosilymarin + AD-MSC-CM upregulates IFN-γ expression in PBMC (monoculture/co-culture).