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Targeted Delivery of 5-fluorouracil with Monoclonal Antibody Modified Bovine Serum Albumin Nanoparticles.

Fadaeian G, Shojaosadati SA, Kouchakzadeh H, Shokri F, Soleimani M - Iran J Pharm Res (2015)

Bottom Line: No cellular uptake was observed for HER2-negative MCF7 cells.Physicochemical and biological properties of the mAb-modified nanoparticles did not significantly alter after three months of storage at room temperature.This novel system can efficiently be employed for targeted delivery of 5-FU to HER2-positive cancerous cells.

View Article: PubMed Central - PubMed

Affiliation: Biotechnology Group, Chemical Engineering Faculty , Tarbiat Modares University, Tehran, Iran.

ABSTRACT
Herein, 1F2, an anti-HER2 monoclonal antibody (mAb), was covalently coupled to the surface of 5-Fluorouracil (5-FU) loaded bovine serum albumin (BSA) nanoparticles. Concerning two different crosslinkers for conjugation of 1F2, Maleimide-poly (ethylene glycol)-Succinimidyl carbonate (Mal-PEG5000-NHS) was selected due to its higher conjugation efficiency (23 ± 4%) obtained in comparison to N-succinimidyl 3-(2-Pyridyl Dithio) Propionate (SPDP) (8 ± 2%). A slight increase in the average particle size with a negligible prolongation of the 5-FU release was observed after 1F2 coupling. The 1F2-coupled 5-FU-loaded BSA nanoparticles interacted with nearly all HER2 receptors available on the surface of HER2-positive SKBR3 cells. No cellular uptake was observed for HER2-negative MCF7 cells. Physicochemical and biological properties of the mAb-modified nanoparticles did not significantly alter after three months of storage at room temperature. The in-vitro cytotoxicity evaluation by MTT assay, demonstrated lower SKBR3 viability (50.7 ± 9 %) after 5 hours contact with 1F2-coupled 5-FU-loaded BSA nanoparticles in comparison with the other control systems due to their cell attachment and internalization after washing. In addition, no significant toxicity was observed on MCF7 cells. This novel system can efficiently be employed for targeted delivery of 5-FU to HER2-positive cancerous cells.

No MeSH data available.


Related in: MedlinePlus

In-vitro cell viability assay. HER2-positive SKBR3 cells were incubated with six different systems including free 5-FU, BSA nanoparticles, 5-FU-loaded BSA nanoparticles, 5-FU-loaded PEGylated BSA nanoparticles, 1F2-copled BSA nanoparticles and 1F2-coupled 5-FU-loaded BSA nanoparticles for one (□) and 5 hours (■), then washed and incubated for another 72 hours in order to investigate their specificity and toxicity. Cells were assessed by MTT assay (n=3, mean SD).
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Figure 6: In-vitro cell viability assay. HER2-positive SKBR3 cells were incubated with six different systems including free 5-FU, BSA nanoparticles, 5-FU-loaded BSA nanoparticles, 5-FU-loaded PEGylated BSA nanoparticles, 1F2-copled BSA nanoparticles and 1F2-coupled 5-FU-loaded BSA nanoparticles for one (□) and 5 hours (■), then washed and incubated for another 72 hours in order to investigate their specificity and toxicity. Cells were assessed by MTT assay (n=3, mean SD).

Mentions: Cytotoxicity of the BSA nanoparticles, free 5-FU, 5-FU-loaded BSA nanoparticles, 5-FU-loaded PEGylated nanoparticles and 1F2-coupled BSA nanoparticles was investigated in-vitro by MTT assay on SKBR3 cells and compared with the novel produced system. As is shown in Figure 6, the 1F2-coupled 5-FU-loaded BSA nanoparticles showed higher cell cytotoxicity (50.7 ± 9%) in comparison to other systems due to specific cell attachment through 1F2 mAb after washing, leading to accumulation of higher concentrations of 5-FU on targeted cells. Therefore, according to the results, the used concentration of drug (2 mM) is approximately equal to IC50 for 1F2-modified nanoparticles, which demonstrates higher toxicity of drug delivered by this targeted system. Other control systems such as 1F2-coupled nanoparticles did not show significant cytotoxicity, which is in accordance with previous studies (17, 23). Therefore, the most occurred cytotoxicity by drug-loaded modified-nanoparticles is due to their encapsulated 5-FU (37%) rather than 1F2 or nanoparticles. 5-FU toxicity more than 30% is probably due to higher rate of drug internalization by 1F2-modified nanoparticles. The higher 5-FU toxicity obtained by the targeted system in comparison to free drug (11 ± 8 %) after 5 hours contact with the cells continued with another 72 hours incubation, verifies that this system is more efficient.


Targeted Delivery of 5-fluorouracil with Monoclonal Antibody Modified Bovine Serum Albumin Nanoparticles.

Fadaeian G, Shojaosadati SA, Kouchakzadeh H, Shokri F, Soleimani M - Iran J Pharm Res (2015)

In-vitro cell viability assay. HER2-positive SKBR3 cells were incubated with six different systems including free 5-FU, BSA nanoparticles, 5-FU-loaded BSA nanoparticles, 5-FU-loaded PEGylated BSA nanoparticles, 1F2-copled BSA nanoparticles and 1F2-coupled 5-FU-loaded BSA nanoparticles for one (□) and 5 hours (■), then washed and incubated for another 72 hours in order to investigate their specificity and toxicity. Cells were assessed by MTT assay (n=3, mean SD).
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4403055&req=5

Figure 6: In-vitro cell viability assay. HER2-positive SKBR3 cells were incubated with six different systems including free 5-FU, BSA nanoparticles, 5-FU-loaded BSA nanoparticles, 5-FU-loaded PEGylated BSA nanoparticles, 1F2-copled BSA nanoparticles and 1F2-coupled 5-FU-loaded BSA nanoparticles for one (□) and 5 hours (■), then washed and incubated for another 72 hours in order to investigate their specificity and toxicity. Cells were assessed by MTT assay (n=3, mean SD).
Mentions: Cytotoxicity of the BSA nanoparticles, free 5-FU, 5-FU-loaded BSA nanoparticles, 5-FU-loaded PEGylated nanoparticles and 1F2-coupled BSA nanoparticles was investigated in-vitro by MTT assay on SKBR3 cells and compared with the novel produced system. As is shown in Figure 6, the 1F2-coupled 5-FU-loaded BSA nanoparticles showed higher cell cytotoxicity (50.7 ± 9%) in comparison to other systems due to specific cell attachment through 1F2 mAb after washing, leading to accumulation of higher concentrations of 5-FU on targeted cells. Therefore, according to the results, the used concentration of drug (2 mM) is approximately equal to IC50 for 1F2-modified nanoparticles, which demonstrates higher toxicity of drug delivered by this targeted system. Other control systems such as 1F2-coupled nanoparticles did not show significant cytotoxicity, which is in accordance with previous studies (17, 23). Therefore, the most occurred cytotoxicity by drug-loaded modified-nanoparticles is due to their encapsulated 5-FU (37%) rather than 1F2 or nanoparticles. 5-FU toxicity more than 30% is probably due to higher rate of drug internalization by 1F2-modified nanoparticles. The higher 5-FU toxicity obtained by the targeted system in comparison to free drug (11 ± 8 %) after 5 hours contact with the cells continued with another 72 hours incubation, verifies that this system is more efficient.

Bottom Line: No cellular uptake was observed for HER2-negative MCF7 cells.Physicochemical and biological properties of the mAb-modified nanoparticles did not significantly alter after three months of storage at room temperature.This novel system can efficiently be employed for targeted delivery of 5-FU to HER2-positive cancerous cells.

View Article: PubMed Central - PubMed

Affiliation: Biotechnology Group, Chemical Engineering Faculty , Tarbiat Modares University, Tehran, Iran.

ABSTRACT
Herein, 1F2, an anti-HER2 monoclonal antibody (mAb), was covalently coupled to the surface of 5-Fluorouracil (5-FU) loaded bovine serum albumin (BSA) nanoparticles. Concerning two different crosslinkers for conjugation of 1F2, Maleimide-poly (ethylene glycol)-Succinimidyl carbonate (Mal-PEG5000-NHS) was selected due to its higher conjugation efficiency (23 ± 4%) obtained in comparison to N-succinimidyl 3-(2-Pyridyl Dithio) Propionate (SPDP) (8 ± 2%). A slight increase in the average particle size with a negligible prolongation of the 5-FU release was observed after 1F2 coupling. The 1F2-coupled 5-FU-loaded BSA nanoparticles interacted with nearly all HER2 receptors available on the surface of HER2-positive SKBR3 cells. No cellular uptake was observed for HER2-negative MCF7 cells. Physicochemical and biological properties of the mAb-modified nanoparticles did not significantly alter after three months of storage at room temperature. The in-vitro cytotoxicity evaluation by MTT assay, demonstrated lower SKBR3 viability (50.7 ± 9 %) after 5 hours contact with 1F2-coupled 5-FU-loaded BSA nanoparticles in comparison with the other control systems due to their cell attachment and internalization after washing. In addition, no significant toxicity was observed on MCF7 cells. This novel system can efficiently be employed for targeted delivery of 5-FU to HER2-positive cancerous cells.

No MeSH data available.


Related in: MedlinePlus