Abstract
The objective of this study is to prepare cationized gelatin nanospheres (cGNS) incorporating a molecular beacon (MB), and visualize cellular apoptosis. Two types of MB to detect the messenger RNA (mRNA) of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (GAP MB), and caspase-3 (casp3 MB) were incorporated in cGNS, respectively. MB incorporated in cGNS showed the DNA sequence specificity in hybridization. The cGNS incorporation enabled MB to enhance the stability against nuclease to a significantly great extent compared with free MB. The cGNS incorporating GAP MB were internalized into the KUM6 of a mouse bone marrow-derived stem cell by an endocytotic pathway. The cGNS were not distributed at the lysosomes. After the incubation with cGNS, the cell apoptosis was induced at different concentrations of camptothecin. No change in the intracellular fluorescence was observed for cGNSGAPMB. On the other hand, for the cGNScasp3MB, the fluorescent intensity significantly enhanced by the apoptosis induction of cells. It is concluded that cGNS incorporating MB is a promising system for the visualization of cellular apoptosis.
Highlights
Cell transplantation is one of the promising therapies in regenerative medicine to induce the regeneration and repairing of damaged tissues and organs[1,2,3]
The objective of this study is the preparation of cationized gelatin nanospheres incorporating Molecular beacon (MB) aiming at the visualization of cell apoptosis
The present study demonstrates that cationized gelatin nanospheres (cGNS) incorporating MB prepared were readily internalized into the cells as expected
Summary
Cell transplantation is one of the promising therapies in regenerative medicine to induce the regeneration and repairing of damaged tissues and organs[1,2,3]. Imaging technologies are effective in non-invasively evaluating the localization and distribution of cells transplanted[4,5,6,7]. It is reported that the majority of cells transplanted undergo apoptosis[11] Under these circumstances, it is of prime importance to non-invasively visualize the apoptosis of cells transplanted which is a key to evaluate the therapeutic efficacy. For the detection of cellular biological functions, activatable imaging probes will be used to visualize the biological function depending on the change of intracellular environment without the cell destruction[12,13,14]. Cationized gelatin nanospheres achieved the intracellular controlled release of pDNA25, and siRNA26. The objective of this study is the preparation of cationized gelatin nanospheres (cGNS) incorporating MB aiming at the visualization of cell apoptosis. We examine the cell apoptosis by the conventional gene expression of caspase-3
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