Novel Gemcitabine Conjugated Albumin Nanoparticles: a Potential Strategy to Enhance Drug Efficacy in Pancreatic Cancer Treatment
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The present study reports a novel conjugate of gemcitabine (GEM) with bovine serum albumin (BSA) and thereof nanoparticles (GEM-BSA NPs) to potentiate the therapeutic efficacy by altering physicochemical properties, improving cellular uptake and stability of GEM.
The synthesized GEM-BSA conjugate was extensively characterized by NMR, FTIR, MALDI-TOF and elemental analysis. Conjugation mediated changes in structural conformation and physicochemical properties were analysed by fluorescence, Raman and CD spectroscopy, DSC and contact angle analysis. Further, BSA nanoparticles were developed from BSA-GEM conjugate and extensively evaluated against in-vitro pancreatic cancer cell lines to explore cellular uptake pathways and therapeutic efficacy.
Various characterization techniques confirmed covalent conjugation of GEM with BSA. GEM-BSA conjugate was then transformed into NPs via high pressure homogenization technique with particle size 147.2 ± 7.3, PDI 0.16 ± 0.06 and ZP -19.2 ± 1.4. The morphological analysis by SEM and AFM revealed the formation of smooth surface spherical nanoparticles. Cellular uptake studies in MIA PaCa-2 (GEM sensitive) and PANC-1 (GEM resistant) pancreatic cell lines confirmed energy dependent clathrin internalization/endocytosis as a primary mechanism of NPs uptake. In-vitro cytotoxicity studies confirmed the hNTs independent transport of GEM in MIA PaCa-2 and PANC-1 cells. Moreover, DNA damage and annexin-V assay revealed significantly higher apoptosis level in case of cells treated with GEM-BSA NPs as compared to free GEM.
GEM-BSA NPs were found to potentiate the therapeutic efficacy by altering physicochemical properties, improving cellular uptake and stability of GEM and thus demonstrated promising therapeutic potential over free drug.
KEY WORDSBSA nanoparticles conjugate gemcitabine MIA PaCa-2 PANC-1
Atomic force microscopy
Annexin V-Cy3.18 conjugate
Bovine serum albumin
Confocal laser microscope
Human concentrative nucleoside transporters
Dulbecco’s Modified Eagle Medium
Differential scanning calorimetry
Ethylene diamine tetra acetic acid
Human equilibrative nucleoside transporters
Fetal bovine serum
Fourier transform infrared
- GEM-BSA NPs
Gemcitabine-Bovine serum albumin nanoparticles
Hank’s Buffered Salt Solution
human nucleoside transporters NMR: Nuclear magnetic resonance
matrix assisted laser desorption ionization time of flight
Powder X-ray diffraction analysis
Sodium dodecyl sulfate polyacrylamide gel electrophoresis
Scanning electron microsc opy
Secreted protein acid rich in cysteine
ACKNOWLEDGMENTS AND DISCLOSURES
The authors are thankful to the Director NIPER, James Graham Brown Cancer Center (University of Louisville, KY, USA) and Strathclyde Institute of Pharmacy & Biomedical Sciences (University of Strathclyde, Glasgow, U.K.) for necessary infrastructure and facilities. Varun Kushwah is also grateful to the Council of Scientific and Industrial Research (CSIR), GOI, New Delhi, United States-India Educational Foundation, New Delhi and Commonwealth commission in the UK for providing research funding and fellowships. He was the 2016–17 Fulbright-Nehru Doctoral and 2015–16 Commonwealth Split-site PhD research fellow at University of Louisville, Louisville, KY and University of Strathclyde, Glasgow, U.K., respectively.
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