Metabolically stabilized long-circulating PEGylated polyacridine peptide polyplexes mediate hydrodynamically stimulated gene expression in liver.

A novel class of PEGylated polyacridine peptides was developed that mediate potent stimulated gene transfer in the liver of mice. Polyacridine peptides, (Acr-X)(n)-Cys-polyethylene glycol (PEG), possessing 2-6 repeats of Lys-acridine (Acr) spaced by either Lys, Arg, Leu or Glu, were Cys derivatized with PEG (PEG(5000 kDa)) and evaluated as in vivo gene transfer agents. An optimal peptide of (Acr-Lys)(6)-Cys-PEG was able to bind to plasmid DNA (pGL3) with high affinity by polyintercalation, stabilize DNA from metabolism by DNAse and extend the pharmacokinetic half-life of DNA in the circulation for up to 2 h. A tail vein dose of PEGylated polyacridine peptide pGL3 polyplexes (1 µg in 50 µl), followed by a stimulatory hydrodynamic dose of normal saline at times ranging from 5 to 60 min post-DNA administration, led to a high level of luciferase expression in the liver, equivalent to levels mediated by direct hydrodynamic dosing of 1 µg of pGL3. The results establish the unique properties of PEGylated polyacridine peptides as a new and promising class of gene delivery peptides that facilitate reversible binding to plasmid DNA, protecting it from DNase in vivo resulting in an extended circulatory half-life, and release of transfection-competent DNA into the liver to mediate a high-level of gene expression upon hydrodynamic boost.

Change in surface hardness of enamel by a cola drink and a CPP-ACP paste.

OBJECTIVES: This in vitro study used surface microhardness to evaluate whether a paste containing casein phosphopeptide amorphous calcium phosphate (CPP-ACP) can reharden tooth enamel softened by a cola drink, and how different saliva-substitute solutions affect the enamel hardness. METHODS: Twenty-four bovine incisors, each tooth consisting of treatment and control halves, were immersed in a cola drink (Coke) for 8 min, then placed under a 0.4 mL/min drip with various saliva-substitute solutions. The saliva-substitute solutions were: saliva-like solution (SLS) with 1 ppm fluoride, SLS without fluoride, and Biotene mouthwash. CPP-ACP paste was applied to the treatment halves for 3 min at 0, 8, 24, and 36 h. Knoop microhardness measurements were performed at baseline, after the cola drink immersion, and after 24 and 48 h contact with saliva-substitute solution. RESULTS: Enamel hardness significantly decreased after immersion in cola drink (ANOVA, p<0.05). After contact with saliva-like solutions for 48 h, those treated with CPP-ACP paste were significantly harder than those untreated regardless of the presence of 1 ppm fluoride in the saliva-like solution (ANOVA, p<0.05). Biotene mouthwash significantly softened the enamel surface (ANOVA, p<0.05). Two-way ANOVA showed significant effects of the CPP-ACP paste application and types of saliva-substitute solutions on the changes in surface hardness of the softened enamel at a significance level of 0.05. CONCLUSION: The application of CPP-ACP paste with continuous replenishment of saliva-like solution for 48 h significantly hardened enamel softened by a cola drink. Biotene mouthwash softened enamel surface after 48 h contact.