Colloidal DNA carriers for direct localization in cell compartments by pH sensoring.
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.
Reibetanz
Uta
U
School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore 637457, Singapore. ureibetanz@trm.uni-leipzig.de
Chen
Min Hui Averil
MH
Mutukumaraswamy
Shaillender
S
Liaw
Zi Yen
ZY
Oh
Bernice Hui Lin
BH
Venkatraman
Subbu
S
Donath
Edwin
E
Neu
Björn
B
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
D003102:Colloids; D004337:Drug Carriers; D011479:Protamines; D012235:Rhodamines; D004247:DNA; C027755:rhodamine isothiocyanate; D016650:Fluorescein-5-isothiocyanate