Intracellular fate of bioresponsive poly(amidoamine)s in vitro and in vivo

Linear poly(amidoamine)s (PAAs) have been designed to exhibit minimal non-specific toxicity, display pH-dependent membrane lysis and deliver genes and toxins in vitro. The aim of this study was to measure PAA cellular uptake using ISA1-OG (and as a reference ISA23-OG) in B16F10 cells in vitro and, by subcellular fractionation, quantitate intracellular trafficking of (125)I-labelled ISA1-tyr in liver cells after intravenous (i.v.) administration to rats. The effect of time after administration (0.5-3h) and ISA1 dose (0.04-100mg/kg) on trafficking, and vesicle permeabilisation (N-acetyl-b-D-glucosaminidase (NAG) release from an isolated vesicular fraction) were also studied. ISA1-OG displayed approximately 60-fold greater B16F10 cell uptake than ISA23-OG. Passage of ISA1 along the liver cell endocytic pathway caused a transient decrease in vesicle buoyant density (also visible by TEM). Increasing ISA1 dose from 10mg/kg to 100mg/kg increased both radioactivity and NAG levels in the cytosolic fraction (5-10 fold) at 1h. Moreover, internalised ISA1 provoked NAG release from an isolated vesicular fraction in a dose-dependent manner. These results provide direct evidence, for the first time, of PAA permeabilisation of endocytic vesicular membranes in vivo, and they have important implications for potential efficacy/toxicity of such polymeric vectors.

Kinetic studies of the effect of varnish on water loss by glass-ionomer cements

OBJECTIVES: This paper reports a study of the water loss behaviour of two commercial glass-ionomer cements coated with varnishes. METHODS: For each cement (Fuji IX Fast or Chemflex), specimens (6mmdiameterx2mm depth) were prepared and cured for 10min at 37 degrees C. They were exposed to a desiccating environment over H(2)SO(4) either uncoated or coated with the appropriate varnish (Fuji Varnish, a solvent-based lacquer, or Fuji Coat, a light-cured varnish). Four specimens were prepared for each material. They were weighed at hourly intervals for 6h, daily for up to 5 days, then weekly thereafter until equilibration. RESULTS: Unlike the uncoated specimens, water loss from varnished cements was not Fickian, but followed the form: mass loss=A/t+B, where t is time, A and B are constants specific to each cement/varnish combination. A varied from 1.22 to 1.30 (mean 1.26, standard deviation 0.04), whereas B varied from 1.54 to 2.09 (mean -1.83, standard deviation 0.29). At equilibrium, varnished specimens lost much less water than unvarnished ones (p>0.01) but there was no significant difference between the solvent-based and the light-cured varnishes. SIGNIFICANCE: Varnishes protect immature glass-ionomer cements from drying out by altering the mechanism of water loss. This slows the rate of drying but does not necessarily change the total amount of water retained. It confirms that, in clinical use, glass-ionomer restoratives should be varnished to allow them to mature satisfactorily.