Development and In Vitro Evaluation of Kaempferol-Loaded Liposomes in LN-229 Glioblastoma Cells: Potency, Selectivity, and Modulation of ROS-Dependent Cytotoxicity
- 1Florida Gulf Coast University , , , ,
Abstract
Glioblastoma multiforme has a median survival of 12–15 months despite multimodal treatment, due to blood-brain barrier impermeability limiting CNS drug bioavailability. Liposomal nanoformulations circumvent this barrier while offering tunable surface chemistry and amphiphilic drug-loading capacity. Here, we report encapsulation of kaempferol, a dietary flavonoid, within phosphatidylcholine-cholesterol liposomes (Lip-KPF) and evaluate cytotoxic activity against LN-229 glioblastoma cells relative to free kaempferol (KPF) and puromycin aminonucleoside (Puro-A). LCMS confirmed 56% encapsulation efficiency; dynamic light scattering yielded a zeta potential of −15.32 ± 0.83 mV, mean particle size of 200.44 ± 12.22 nm, and PDI of 0.29 ± 0.06, with 31P NMR confirming a multilamellar structure. Lip-KPF potency against LN-229 cells increased progressively across 24–72 h, peaking at 72 h (19.880 μM) with significantly lower IC50 values than free KPF (p < 0.001), while maintaining lower astrocyte toxicity. Selectivity indices for Lip-KPF exceeded 1 at every time point, widening over time. Cell death was confirmed by TUNEL assay and phase-contrast microscopy. RT-qPCR revealed concurrent BAX and TP53 upregulation alongside CASP3/CASP7 suppression in both groups, with no significant difference between KPF and Lip-KPF; ELISA corroborated CASP3 suppression, consistent with a possible caspase-independent death mechanism. In a preliminary antioxidant rescue assay, pretreatment did not significantly alter KPF-induced cytotoxicity but significantly increased it in Lip-KPF-treated cells, indicating a cytotoxic mechanism independent of the ROS pathway contributing to free KPF’s activity. These findings support Lip-KPF as a candidate for continued preclinical investigation in LN-229 glioblastoma cells.
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BibTeX
@article{nwokwu2026development,
title = {{Development and In Vitro Evaluation of Kaempferol-Loaded Liposomes in LN-229 Glioblastoma Cells: Potency, Selectivity, and Modulation of ROS-Dependent Cytotoxicity}},
author = {Chukwumaobim D. O. Nwokwu and Malik H. Walker and Miranda De Andrade and Samantha J. Louis and Austin J. Fearday and Alejandro Bugarin},
journal = {ACS Omega},
year = {2026},
month = nov,
doi = {10.1021/acsomega.6c06597},
url = {https://pubs.acs.org/acsodf/article/doi/10.1021/acsomega.6c06597/5466273/Development-and-In-Vitro-Evaluation-of-Kaempferol},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Development and In Vitro Evaluation of Kaempferol-Loaded Liposomes in LN-229 Glioblastoma Cells: Potency, Selectivity, and Modulation of ROS-Dependent Cytotoxicity
AU - Chukwumaobim D. O. Nwokwu
AU - Malik H. Walker
AU - Miranda De Andrade
AU - Samantha J. Louis
AU - Austin J. Fearday
AU - Alejandro Bugarin
PY - 2026
DA - 2026/11/08
JO - ACS Omega
DO - 10.1021/acsomega.6c06597
UR - https://pubs.acs.org/acsodf/article/doi/10.1021/acsomega.6c06597/5466273/Development-and-In-Vitro-Evaluation-of-Kaempferol
AB - Glioblastoma multiforme has a median survival of 12–15 months despite multimodal treatment, due to blood-brain barrier impermeability limiting CNS drug bioavailability. Liposomal nanoformulations circumvent this barrier while offering tunable surface chemistry and amphiphilic drug-loading capacity. Here, we report encapsulation of kaempferol, a dietary flavonoid, within phosphatidylcholine-cholesterol liposomes (Lip-KPF) and evaluate cytotoxic activity against LN-229 glioblastoma cells relative to free kaempferol (KPF) and puromycin aminonucleoside (Puro-A). LCMS confirmed 56% encapsulation efficiency; dynamic light scattering yielded a zeta potential of −15.32 ± 0.83 mV, mean particle size of 200.44 ± 12.22 nm, and PDI of 0.29 ± 0.06, with 31P NMR confirming a multilamellar structure. Lip-KPF potency against LN-229 cells increased progressively across 24–72 h, peaking at 72 h (19.880 μM) with significantly lower IC50 values than free KPF (p < 0.001), while maintaining lower astrocyte toxicity. Selectivity indices for Lip-KPF exceeded 1 at every time point, widening over time. Cell death was confirmed by TUNEL assay and phase-contrast microscopy. RT-qPCR revealed concurrent BAX and TP53 upregulation alongside CASP3/CASP7 suppression in both groups, with no significant difference between KPF and Lip-KPF; ELISA corroborated CASP3 suppression, consistent with a possible caspase-independent death mechanism. In a preliminary antioxidant rescue assay, pretreatment did not significantly alter KPF-induced cytotoxicity but significantly increased it in Lip-KPF-treated cells, indicating a cytotoxic mechanism independent of the ROS pathway contributing to free KPF’s activity. These findings support Lip-KPF as a candidate for continued preclinical investigation in LN-229 glioblastoma cells.
SN - 2470-1343
ER -