TY - JOUR
T1 - Strategic formulation of apigenin-loaded PLGA nanoparticles for intracellular trafficking, DNA targeting and improved therapeutic effects in skin melanoma in vitro
AU - Das, Sreemanti
AU - Das, Jayeeta
AU - Samadder, Asmita
AU - Paul, Avijit
AU - Khuda-Bukhsh, Anisur Rahman
N1 - Funding Information:
This work was financially supported by a grant sanctioned to Prof. A.R. Khuda-Bukhsh, Department of Zoology, University of Kalyani, Kalyani 741235, India by Boiron Laboratory, Lyon, France . We are also thankful to Dr. Kumaresh Ghosh, Mr. Debasish Kar, Department of Chemistry, Dr. Tarak Das Basu, Ms. Ruchira Chakraborty, Department of Biochemistry, University of Kalyani, India. We are grateful to IACS for AFM, CD spectroscopy, IICB, Kolkata for confocal microscopy and Dr. T.C. Nag and Dr. Sandeep Arya, Department of Anatomy, AIIMS, India for TEM.
PY - 2013/11/25
Y1 - 2013/11/25
N2 - The aim of the present study was the evaluation of anti-proliferative potentials of apigenin (Ap), (a dietary flavonoid) loaded in poly (lactic-co-glycolide) nanoparticles (NAp) in A375 cells in vitro. NAp was characterized for particle size, morphology, zeta potential, drug release and encapsulation. Cellular entry and intracellular localization of NAp were assessed by transmission electron and confocal microscopies. Circular dichroic spectral analysis and stability curve for Gibb's free energy of dsDNA of A375 cells were also analyzed. DNA fragmentation, intracellular ROS accumulation, superoxide-dismutase activity, intracellular glutathione-reductase content and mitochondrial functioning through relevant markers like mitochondrial transmembrane potential, ATPase activity, ATP/ADP ratio, volume changes/swelling, cytochrome-c release, expressions of Apaf-1, bax, bcl-2, caspase-9, 3, and PARP cleavage were analyzed. NAp produced better effects due to their smaller size, faster mobility and site-specific action. Photostability studies revealed that PLGA encapsulations were efficient at preserving apigenin ultraviolet-light mediated photodegradation. NAp readily entered cancer cells, could intercalate with dsDNA, inducing conformational change. Corresponding increase in ROS accumulation and depletion of the antioxidant enzyme activities exacerbated DNA damage, mediating apoptosis through mitochondrial dysfunction. Overall results indicate that therapeutic efficacy of NAp may be enhanced by PLGA nanoparticle formulations to have better ameliorative potentials in combating skin melanoma.
AB - The aim of the present study was the evaluation of anti-proliferative potentials of apigenin (Ap), (a dietary flavonoid) loaded in poly (lactic-co-glycolide) nanoparticles (NAp) in A375 cells in vitro. NAp was characterized for particle size, morphology, zeta potential, drug release and encapsulation. Cellular entry and intracellular localization of NAp were assessed by transmission electron and confocal microscopies. Circular dichroic spectral analysis and stability curve for Gibb's free energy of dsDNA of A375 cells were also analyzed. DNA fragmentation, intracellular ROS accumulation, superoxide-dismutase activity, intracellular glutathione-reductase content and mitochondrial functioning through relevant markers like mitochondrial transmembrane potential, ATPase activity, ATP/ADP ratio, volume changes/swelling, cytochrome-c release, expressions of Apaf-1, bax, bcl-2, caspase-9, 3, and PARP cleavage were analyzed. NAp produced better effects due to their smaller size, faster mobility and site-specific action. Photostability studies revealed that PLGA encapsulations were efficient at preserving apigenin ultraviolet-light mediated photodegradation. NAp readily entered cancer cells, could intercalate with dsDNA, inducing conformational change. Corresponding increase in ROS accumulation and depletion of the antioxidant enzyme activities exacerbated DNA damage, mediating apoptosis through mitochondrial dysfunction. Overall results indicate that therapeutic efficacy of NAp may be enhanced by PLGA nanoparticle formulations to have better ameliorative potentials in combating skin melanoma.
KW - Apigenin
KW - Apoptosis
KW - Drug-DNA interaction
KW - Mitochondrial dysfunction
KW - PLGA-nano-encapsulation
KW - Skin melanoma
UR - http://www.scopus.com/inward/record.url?scp=84886308723&partnerID=8YFLogxK
U2 - 10.1016/j.toxlet.2013.09.012
DO - 10.1016/j.toxlet.2013.09.012
M3 - Article
C2 - 24070738
AN - SCOPUS:84886308723
SN - 0378-4274
VL - 223
SP - 124
EP - 138
JO - Toxicology Letters
JF - Toxicology Letters
IS - 2
ER -
