Triple-Negative Breast Cancer (TNBC) is the most aggressive sub-set of breast cancer, that is also metastatic. TNBC is characterized by its clinical absence of the three most common re ceptors, Estrogen, Progesterone, and Human Epi dermalGrowthFactorReceptor2(Her2)andthera peutic modalities, including the prevalent chemo therapeutic agent Cisplatin. It has the worst sur vival and a lower 5-year survival rate compared to other subtypes, such as luminal A, luminal B, and HER2-positive breast cancers. Thus, there is an unmet needfor alternative, effective and affordable therapies that are gentle on both the body and the purse. Towards this, in this research, we explored the ef ficacy of Resveratrol (Resv), a natural polyphenolic compoundthatiswidelyavailableinredgrapesand other dark colored fruits and vegetables, on MDA MB-231, human, TNBC cell line. For this purpose, a 3D scaffold matrix, composed of hyaluronic acid (HA)andself-assemblingpeptides (SAPs; specific ally, EAbuK), which are further condensed with a Laminin-derived adhesive motif (IKVAV). The major advantage of 3D cell culture is that it mimics more closely the in vivo model, meeting the 3R principles (Replacement, Reduction, andRefinement). Given the known limitations surrounding the bioavailabil ity of Resveratrol, this study integrates Electropor ation (EP) to enhance the uptakeof theintracellular translocation of Resv, employing brief electric fields to transiently permeabilize cellular membranes. MDA-MB-231 human TNBC cell line, derived from a Caucasian woman was seeded within the custom-fabricated HA-EAbuK-IKVAV scaffold that simulates the three-dimensional architecture of bio logical tissue environments. The experimental pro tocol involves the administration of Resv with the application of electric pulses with parameters set at 800–1,000 V/cm and 100µs duration, at a fre quency of 1Hz, followed by a comprehensive as sessment of cell viability and reactive oxygen spe cies (ROS) production. The results of this study are compelling, revealing a cell viability, as low as 24% at 24 hours of treatment with EP and Resv, underscoring the therapeutic potential of this ap proach. Moreover, the concomitant use of Elec troporation and Resveratrol at specified concentra tions significantly augmented ROS generation (4x) comparedtosingular treatments, indicative of a po tent synergistic interaction that exacerbates TNBC cell mortality via enhanced ROS-mediated apop tosis. These robust findings underscore the poten tial utility of Resveratrol as an anti-TNBC agent and highlight the pivotal role of electroporation in aug menting its cellular uptake and therapeutic impact
Electroporation-Enhanced Resveratrol Delivery into 3D-Hyaluronic Acid-Peptide Scaffold Cells for Effective Triple-Negative Breast Cancer Treatments
Monica Dettin;Annj Zamuner;Maria Teresa Conconi;
2024
Abstract
Triple-Negative Breast Cancer (TNBC) is the most aggressive sub-set of breast cancer, that is also metastatic. TNBC is characterized by its clinical absence of the three most common re ceptors, Estrogen, Progesterone, and Human Epi dermalGrowthFactorReceptor2(Her2)andthera peutic modalities, including the prevalent chemo therapeutic agent Cisplatin. It has the worst sur vival and a lower 5-year survival rate compared to other subtypes, such as luminal A, luminal B, and HER2-positive breast cancers. Thus, there is an unmet needfor alternative, effective and affordable therapies that are gentle on both the body and the purse. Towards this, in this research, we explored the ef ficacy of Resveratrol (Resv), a natural polyphenolic compoundthatiswidelyavailableinredgrapesand other dark colored fruits and vegetables, on MDA MB-231, human, TNBC cell line. For this purpose, a 3D scaffold matrix, composed of hyaluronic acid (HA)andself-assemblingpeptides (SAPs; specific ally, EAbuK), which are further condensed with a Laminin-derived adhesive motif (IKVAV). The major advantage of 3D cell culture is that it mimics more closely the in vivo model, meeting the 3R principles (Replacement, Reduction, andRefinement). Given the known limitations surrounding the bioavailabil ity of Resveratrol, this study integrates Electropor ation (EP) to enhance the uptakeof theintracellular translocation of Resv, employing brief electric fields to transiently permeabilize cellular membranes. MDA-MB-231 human TNBC cell line, derived from a Caucasian woman was seeded within the custom-fabricated HA-EAbuK-IKVAV scaffold that simulates the three-dimensional architecture of bio logical tissue environments. The experimental pro tocol involves the administration of Resv with the application of electric pulses with parameters set at 800–1,000 V/cm and 100µs duration, at a fre quency of 1Hz, followed by a comprehensive as sessment of cell viability and reactive oxygen spe cies (ROS) production. The results of this study are compelling, revealing a cell viability, as low as 24% at 24 hours of treatment with EP and Resv, underscoring the therapeutic potential of this ap proach. Moreover, the concomitant use of Elec troporation and Resveratrol at specified concentra tions significantly augmented ROS generation (4x) comparedtosingular treatments, indicative of a po tent synergistic interaction that exacerbates TNBC cell mortality via enhanced ROS-mediated apop tosis. These robust findings underscore the poten tial utility of Resveratrol as an anti-TNBC agent and highlight the pivotal role of electroporation in aug menting its cellular uptake and therapeutic impactPubblicazioni consigliate
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