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Aptamers are DNA or RNA based oligonucleotides sequences that possess secondary and tertiary structures that have affinity to their targets such as cell surface receptors. Recently, aptamers have attracted much attention in widespread range of investigation and emerge as a potentially powerful molecules that can be used in NDDSs as the targeting ligand. The overexpression of EpCAM has been reported in CSC of various solid tumors. CSC or TIC are cells which have self-renewal, the ability to produce more cells of the same types, that have key roles in tumor development and metastasis. ĮpCAM is demonstrated as cancer stem cell (CSC) or tumor initiating cell (TIC) marker, which its expression in cancer is related to the poor prognosis. This differential expression makes EpCAM as a very interesting ligand for drug delivery which could improve the therapeutic index of drug. During the cancer development the expression pattern of EpCAM change from basal and basolateral membrane in normal epithelial to the apical surface in tumor epithelial cells. Recent findings indicated that the EpCAM has normal low expression healthy epithelial cells, while in cancer cells its expression become in higher levels (up to 1000-fold). The epithelial cell adhesion molecule (EpCAM) is a transmembrane glycoprotein that considered as a candidate ligand for active targeting.
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An ideal targeting agent for targeted delivery is a molecule that have affinity to the cell surface proteins or receptors upregulated by particular cells or tissue components. Īctive targeted delivery is important approach that helps NDDSs to deliver therapeutic agents more efficient to the tumors and minimizing the exposure to non-target tissues. However in case of inorganic nanomaterials attention must be paid to their toxicities and environmental safeties. There are many different inorganic nanomaterials which have the ability to use as anticancer agents. These NDDSs could response to the exogenous stimuli such as light and have the capacity to use in tumor imaging. Different strategies have been used to improve tumor accumulation of NDDSs using endogenous and exogenous stimuli. Lack of the ability to penetrate in the dense extra cellular matrix (ECM) of tumor, return of the released drug to circulation and heterogeneity of tumors are the reasons responsible for this failure. However, recent data indicated that only less than 1% of administrated drug could reach to the tumor site. This occurs through the loose endothelial lining and weak lymphatic drainage. Nano drug delivery systems (NDDSs) with the size of 100–200 nm are passively accumulated in the tumor microenvironment via enhanced permeability and retention (EPR) effect. These results suggest that the functionalization of Caelyx® with anti-EpCAM aptamer is promising in cancer treatment and merits further investigation. In addition, ED-lip effectively improved the tumor accumulation of DOX and promoted survival of animals compared to Caelyx®. The pharmacokinetic and tissue biodistribution of formulations in mice bearing C26 tumors demonstrated that ED-lip did not affect the distribution profile of DOX compared to Caelyx® in animal model. The ED-lip also had more cytotoxic effects than Caelyx® which indicated the efficacy of anti-EpCAM aptamer as targeting ligand.
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The results of flow cytometry and fluorescent microscopy have shown that ED-lip enhanced the rate of cell uptake on C26 cell line compared to Caelyx®. The characterization of the ED-lip demonstrated the slightly increase in size and PDI along with the decrease in zeta potential which indicated that post-insertion efficiently done. The size, charge, release profile, and cytotoxicity and cellular uptake of formulation were determined. In this study, we have surface-functionalized PEGylated-nanoliposomal doxorubicin (DOX) with anti-EpCAM (epithelial cell adhesion molecule) aptamer via post-insertion of anti-EpCAM aptamer-conjugated DSPE-mPEG 2000 into Caelyx® (ED-lip).