Preparation of Xanthohumol (XN)-incorporated sub-5 ultrafine iron oxide nanoparticles: Investigation of their effects on multiple myeloma cell growth
Location
Suwanee, GA
Start Date
6-5-2025 1:00 PM
End Date
6-5-2025 4:00 PM
Description
Introduction
Multiple myeloma (MM) is a hematological cancer that affects B lymphocytes. MM cells can undergo a homing process within the bone marrow microenvironment (BMM). The BMM helps MM cells undergo proliferation through interactions with bone marrow stromal cells. The proliferation aids in MM’s continued growth and spread throughout the BMM and into neighboring organs by releasing interleukin-6 and vascular endothelial growth factor. Xanthohumol (XN) is categorized as a prenylated flavonoid and has exhibited anticancer effects in in-vitro models. XN has downregulated the NF-kappa B signaling pathway, which is activated by interleukin-6 and aids in MM cell proliferation. Major challenges limiting the use of XN are poor aqueous solubility and lack of targeted drug delivery. To address this, we formulated anti-biofouling polyethylene glycol-block-allyl glycidyl ether (PEG-b-AGE) polymer-coated ultrafine iron oxide nanoparticles (µIONPs) loaded with XN (µIONP/XN) to improve the delivery efficiency to MM cells. An RGD motif of arginine, glycine, and aspartic acid was conjugated to the µIONPs as the targeting ligand for the αvβ3 integrin overexpressed on MM cells. This study compared RGD-µIONP/XN, RGD-µIONP (vehicle control), and µIONP/XN (non-targeted drug control) for cytotoxic effects on MM RPMI 8226 cells in an in-vitro model.
Methods
Throughout this project, several experiments were conducted to gather quantitative data. Presto blue assays were used to analyze cell viability, western blots were used to evaluated the expression of apoptosis-related markers, and a reactive oxidative species assay (ROS) was used to determine the ROS production by the three nanoparticles in RPMI 8226 cells. The release of XN from RGD-µIONP/XN was measured by high-performance liquid chromatography with or without a short-wave radio frequency (RF) stimulation.
Results
XN induced a dose and time-dependent decrease in cellular viability of RPMI 8226 cells with an IC50 value of 12.28 µM at 48 hours and 7.54 µM at 72 hours. The data from the Western blot analysis showed an upregulation of cleaved-caspase 3 with increased XN concentrations. Interestingly, results indicated that the RGD-µIONP (vehicle control) also induced cytotoxic effects on the MM cells in a time dependent manner. This urged us to analyze the generation of ROS by the three nanoparticles. The preliminary ROS results indicated that all the nanoparticles are producing ROS in a time dependent manner. Upon a short-wave RF stimulation for 10 minutes, greater than 30% of XN was released.
Conclusion
The results indicate the potential mechanism of cell death induced by the RGD-µIONP/XN to be at least in part if not all, apoptosis. Additional experimentation will be performed to evaluate the cytotoxic effects of RGD-µIONP, XN-µIONP, and RGD-µIONP/XN on RPMI 8226 cells upon short-wave RF stimulation and the mechanism of another programed cell death, ferroptosis, that these nanoparticles can induce.
Embargo Period
5-19-2025
Preparation of Xanthohumol (XN)-incorporated sub-5 ultrafine iron oxide nanoparticles: Investigation of their effects on multiple myeloma cell growth
Suwanee, GA
Introduction
Multiple myeloma (MM) is a hematological cancer that affects B lymphocytes. MM cells can undergo a homing process within the bone marrow microenvironment (BMM). The BMM helps MM cells undergo proliferation through interactions with bone marrow stromal cells. The proliferation aids in MM’s continued growth and spread throughout the BMM and into neighboring organs by releasing interleukin-6 and vascular endothelial growth factor. Xanthohumol (XN) is categorized as a prenylated flavonoid and has exhibited anticancer effects in in-vitro models. XN has downregulated the NF-kappa B signaling pathway, which is activated by interleukin-6 and aids in MM cell proliferation. Major challenges limiting the use of XN are poor aqueous solubility and lack of targeted drug delivery. To address this, we formulated anti-biofouling polyethylene glycol-block-allyl glycidyl ether (PEG-b-AGE) polymer-coated ultrafine iron oxide nanoparticles (µIONPs) loaded with XN (µIONP/XN) to improve the delivery efficiency to MM cells. An RGD motif of arginine, glycine, and aspartic acid was conjugated to the µIONPs as the targeting ligand for the αvβ3 integrin overexpressed on MM cells. This study compared RGD-µIONP/XN, RGD-µIONP (vehicle control), and µIONP/XN (non-targeted drug control) for cytotoxic effects on MM RPMI 8226 cells in an in-vitro model.
Methods
Throughout this project, several experiments were conducted to gather quantitative data. Presto blue assays were used to analyze cell viability, western blots were used to evaluated the expression of apoptosis-related markers, and a reactive oxidative species assay (ROS) was used to determine the ROS production by the three nanoparticles in RPMI 8226 cells. The release of XN from RGD-µIONP/XN was measured by high-performance liquid chromatography with or without a short-wave radio frequency (RF) stimulation.
Results
XN induced a dose and time-dependent decrease in cellular viability of RPMI 8226 cells with an IC50 value of 12.28 µM at 48 hours and 7.54 µM at 72 hours. The data from the Western blot analysis showed an upregulation of cleaved-caspase 3 with increased XN concentrations. Interestingly, results indicated that the RGD-µIONP (vehicle control) also induced cytotoxic effects on the MM cells in a time dependent manner. This urged us to analyze the generation of ROS by the three nanoparticles. The preliminary ROS results indicated that all the nanoparticles are producing ROS in a time dependent manner. Upon a short-wave RF stimulation for 10 minutes, greater than 30% of XN was released.
Conclusion
The results indicate the potential mechanism of cell death induced by the RGD-µIONP/XN to be at least in part if not all, apoptosis. Additional experimentation will be performed to evaluate the cytotoxic effects of RGD-µIONP, XN-µIONP, and RGD-µIONP/XN on RPMI 8226 cells upon short-wave RF stimulation and the mechanism of another programed cell death, ferroptosis, that these nanoparticles can induce.
Comments
Awarded "Best Original Research: Honorable Mention" at PCOM Georgia Research Day 2025