title: Colloidal DNA carriers for direct localization in cell compartments by pH sensoring.
abstract: Multifunctional colloidal microparticles allow the integration of various active agents as well as reporter molecules into one system without interfering combining delivery and sensing functions. In this study, calcium carbonate particles were functionalized with fluorescein isothiocyanate-labeled poly(allylamine hydrochloride) (FITC-PAH) allowing particle localization in cell compartments of different pH. Plasmid DNA (pEGFP-C1 and pDsRed1-N1) as a reporter agent for drug release in the cytoplasm and rhodamine-B-isothiocyanate-labeled protamine (RITC-PRM) were integrated into biocompatible and biodegradable PRM/DXS multilayers. The uptake and processing of the particles by HEK293T/17 cells were investigated via flow cytometry and confocal laser scanning microscopy. The presented data show a clear correlation between the fluorescence intensity of the FITC-labeled core, that is, the particle localization after cellular uptake, and the expression of fluorescent proteins by the cells without further cell staining. In conclusion, this particle design allows the simultaneous study of particle location and processing to monitor the transport and release of active agents and should thus be an invaluable tool for the study and design of nano- and microcarrier systems.
authors.affiliation: School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore 637457, Singapore. ureibetanz@trm.uni-leipzig.de
authors.forename: Uta
authors.initials: U
authors.lastname: Reibetanz
authors.forename: Min Hui Averil
authors.initials: MH
authors.lastname: Chen
authors.forename: Shaillender
authors.initials: S
authors.lastname: Mutukumaraswamy
authors.forename: Zi Yen
authors.initials: ZY
authors.lastname: Liaw
authors.forename: Bernice Hui Lin
authors.initials: BH
authors.lastname: Oh
authors.forename: Subbu
authors.initials: S
authors.lastname: Venkatraman
authors.forename: Edwin
authors.initials: E
authors.lastname: Donath
authors.forename: Björn
authors.initials: B
authors.lastname: Neu
mesh_terms: D002451:Cell Compartmentation; D002460:Cell Line; D003102:Colloids / Q000737:chemistry / Q000493:pharmacokinetics*; D004247:DNA / Q000008:administration & dosage*; D004337:Drug Carriers / Q000737:chemistry / Q000493:pharmacokinetics*; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate; D017930:Genes, Reporter; D006801:Humans; D006863:Hydrogen-Ion Concentration; D018613:Microscopy, Confocal; D010316:Particle Size; D011479:Protamines; D012235:Rhodamines
chemical_list: D003102:Colloids; D004337:Drug Carriers; D011479:Protamines; D012235:Rhodamines; D004247:DNA; C027755:rhodamine isothiocyanate; D016650:Fluorescein-5-isothiocyanate