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A Calcium Phosphate Nanoparticle-Based Biocarrier for Efficient Cellular Delivery of Antisense Oligodeoxynucleotides

Hou, Shenglei, Ma, Hongmei, Ji, Yuxuan, Hou, Wenyue, Jia, Nengqin
ACS Applied Materials & Interfaces 2013 v.5 no.3 pp. 1131-1136
Fourier transform infrared spectroscopy, X-ray diffraction, agar gel electrophoresis, apoptosis, calcium phosphates, confocal laser scanning microscopy, flow cytometry, messenger RNA, nanoparticles, neoplasm cells, neoplasms, oligodeoxyribonucleotides, pro-apoptotic proteins, translation (genetics), transmission electron microscopy, ultraviolet-visible spectroscopy
Antisense oligodeoxynucleotides (ASODNs) can bind to some specific RNA of survivin can prevent the mRNA translation at the genetic level, which will inhibit survivin expression and make the cancer cells apoptosis. However, the ASODNs-based therapies are hampered by their instability to cellular nuclease and their weak intracellular penetration. Here we reported a calcium phosphate (CP)-based carrier to achieve efficient delivery of ASODNs into cells. In this study, we used a facile microemulsion approach to prepare spherical and porous ASODNs-CP nanoparticles (ASODNS-CPNPs) with the size of 50-70 nm in diameter, and their structure, morphology and composition were characterized by TEM, XRD, FTIR, ICP and DLS, UV-Vis spectroscopy and agarose gel electrophoresis. The results indicated that the nanoparticles have a high ASODNs loading capacity. Furthermore, cellular uptake and delivery efficiency of the ASODNS-CPNPs, as well as cellular apoptosis induced by the ASODNs doping into the calcium phosphate nanoparticles, were investigated by confocal laser scanning microscopy, biological TEM, flow cytometry, and MTT assay. Efficient intracellular delivery of the nanoparticles was observed. All these results suggested that the prepared calcium phosphate nanoparticles could be used as a promising biocarrier for delivery of ASODNs.