title: Long chain microRNA conjugates in calcium phosphate nanoparticles for efficient formulation and delivery.
abstract: A long chain microRNA-34a conjugate (lc-miRNA) was prepared by chemical crosslinking in order to improve entrapment efficiency into calcium phosphate nanoparticles (CaPs) and intracellular delivery. Thiol-modified miRNA at both terminal ends was chemically conjugated using crosslinkers to form lc-miRNA which was encapsulated within CaPs by a conventional co-precipitation method. Encapsulation efficiencies, physicochemical properties, and in vitro intracellular delivery efficiencies of the prepared linear polyethyleneimine (LPEI)-coated CaPs (LPEI-CaP) containing common miRNA and lc-miRNA were comparatively evaluated. The prepared lc-miRNA exhibited noticeably enhanced encapsulation efficiency during the CaP formulation process when compared to common miRNA. LPEI-CaP/lc-miRNAs consisted of nano-sized particles with great homogeneity and were observed to be successfully delivered into PC-3 cells. Fabricated LPEI-CaPs with duplex form of lc-miRNA (lc-miRNA-d) suppressed cancer cell proliferation as well as migration much more efficiently than those with duplex form of miRNA (miRNA-d). In addition, LPEI-CaP/lc-miRNA-d conferred negligible cytotoxicity on PC-3 cells. Chemical crosslinking of therapeutic miRNAs via a reducible linkage may allow more efficient encapsulation within CaPs as well as homogeneous particle formulation due to a higher spatial charge density than common miRNAs. The well-formulated LPEI-CaPs with lc-miRNA-d have the potential to provide superior miRNA transfection efficiency and inhibition of cancer proliferation.
authors: Department of Bioscience and Biotechnology, Konkuk University, Seoul, Republic of Korea.
authors: Hyosook
authors: H
authors: Jung
authors: Seung An
authors: SA
authors: Kim
authors: Yong Geun
authors: YG
authors: Yang
authors: Hyundong
authors: H
authors: Yoo
authors: Soo-Jeong
authors: SJ
authors: Lim
authors: Hyejung
authors: H
authors: Mok
mesh_terms: D002130:Calcium Phosphates / Q000008:administration & dosage / Q000737:chemistry*; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002626:Chemistry, Pharmaceutical; D004305:Dose-Response Relationship, Drug; D018014:Gene Transfer Techniques; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000008:administration & dosage / Q000737:chemistry*; D053758:Nanoparticles / Q000008:administration & dosage / Q000737:chemistry*; D014407:Tumor Cells, Cultured
chemical_list: D002130:Calcium Phosphates; C522425:MIRN34 microRNA, human; D035683:MicroRNAs; C020243:calcium phosphate