Mechanistic Approach of Nano Carriers for Targeted in Cancer Chemotherapy: A Newer Strategy for Novel Drug Delivery System

Abstract

The application of nanomedicine represents an innovative approach for the treatment in the modern field of cancer chemotherapy. In the present research work, tamoxifen citrate loaded nanolipid vesicles were prepared conjugated with phosphoethanolamine as the linker molecule, and the specific antibody was tagged with the linker molecule on the bilayer surface of the vesicles. The main objective of this study is to determine the efficacy and biological behavior of antibody conjugated nanoliposome in breast cancer cell lines. Percentage of drug loading and loading efficiency was done and their results were compared to theoretical drug loading. The average diameter of those vesicles was within the 100 nm range, which is revealed in FESEM and TEM images and their lamellarity was observed in cryo-TEM images. The hydrodynamic diameter was done by particle size analysis and the surface charge was determined by the zeta potential parameter. Predominant cellular uptake was observed for antibody conjugated nanolipid vesicles in MCF-7 and MDA-MB-453 human breast cancer cell lines. A cellular apoptosis assay was conducted by flow cytometer (FACS). All experimental data would be more beneficial for the treatment of breast cancer chemotherapy. Further studies are warranted to investigate the efficacy and safety of antibody conjugated nanolipid vesicles in vivo for breast cancer animal model.

Keywords: breast cancer chemotherapy; cellular apoptosis; nanolipid vesicles; specific antibody.

Figure 1

FESEM images of (A) tamoxifen citrate loaded nanolipid vesicles; (B) antibody conjugated tamoxifen citrate-phosphoethanolamine nanolipid vesicles.

Figure 2

TEM images of (A,C) tamoxifen citrate loaded nanolipid vesicles; light peripheral demarcation indicates the phospholipid bilayer (shown by sky blue arrow). (B,D) antibody conjugated tamoxifen citrate-phosphoethanolamine nanolipid vesicles; the presence of antibody with phosphoethanolamine tagging in phospholipid bilayer revealed darker, thicker, polygonal particulate matters (shown by green arrow).

Figure 3

Cryo-TEM images: pink color and sky blue color arrow indicate small unilamellar vesicle (near about 50 nm in diameter) and unilamellar vesicle (near about 100 nm in diameter), respectively. Green color arrow represents two unilamellar vesicles fused with each other.

Figure 4

Fluorescence intensity of TNL-PE nanolipid vesicles (blue color wavelength); fluorescence intensity of TNL-PE-Ab nanolipid vesicles (red color wavelength).

Figure 5

Energy dispersive X-ray (EDX) of (A) TNL-PE and (B) TNL-PE-Ab.

Figure 6

Zeta potential data: (A) tamoxifen citrate loaded nanolipid vesicles; (B) antibody conjugated tamoxifen citrate loaded nano lipid vesicles.

Figure 7

Particle size analysis: (A) tamoxifen citrate loaded nanolipid vesicles, (B) antibody conjugated tamoxifen citrate loaded nanolipid vesicles.

Figure 8

In vitro cellular uptake of antibody conjugated nanolipid vesicles in MCF-7 breast cancer cells. (A) Formulation at concentration 50 µg/mL for 1 h; (B) formulation at concentration 100 µg/mL for 2 h.

Figure 9

(A) Antibody conjugated nanoliposome formulation at concentration 50 µg/mL for 1 h. (B) Antibody conjugated nanoliposome formulation at concentration 100 µg/mL for 1 h. (C) Antibody conjugated nanoliposome formulation at concentration 50 µg/mL for 2 h. (D) Antibody conjugated nanoliposome formulation at concentration 100 µg/mL for 2 h.

Figure 10

Representative FACS analysis of treated cells (A) control cells, (B) free drug solution, (C,D) tamoxifen citrate loaded nanolipid vesicles of 50 and 100 µM, respectively, staining with Annexin V/PI.

Figure 11

(A) Comparison of cell viability percentage between tamoxifen citrate loaded nanolipid vesicles with phosphoethanolamine (TNL-PE), antibody conjugated tamoxifen citrate loaded nanolipid vesicles (TNL-PE-Ab) and free drug solution (TC) on MCF-7 cells for 48 h. (B) Percentage of cell viability by statistical data for free drug (TC), TNL-PE, TNL-PE-Ab and blank nanolipid vesicles (BNL).