Because of their well-controlled molecular properties and low toxicity, PAMAM starburstdendrimers have been attractive polymers for potential biomedical applications. In particular,higher generation PAMAM starburst dendrimers have shown extraordinary efficacy as vectors forthe transfection of DNA into mammalian cells.
Some of this efficacy is probably due to theability of the polycationic dendrimer to form a tight, charge-neutralized complex with polyanionicDNA, since neutral molecules are better able to permeate the lipid membranes that surround cells.However, additional factors must be important, since starburst dendrimers are much more effectiveat DNA transfection than linear polymers, such as polylysine, and are more effective thanhyperbranched polyethyleneimine.
Moreover, simply neutralizing the charge on a macromoleculeis not sufficient for membrane permeation, since neutral, hydrophilic polymers such as dextran orpolyethyleneoxide are not membrane permeant.Several unique properties of starburst dendrimers and their complexes with DNA may beimportant for transfection, and these properties need to be elucidated. These include:(1) Packing of DNA. Dendrimers of generation 6 and higher possess sizes as large as theeukaryotic DNA packing proteins, the histones.
It is possible that dendrimers serve as templateswhich condense DNA into a structure that is more readily transported across biologicalmembranes.(2) Membrane binding. As discussed below, the putative pathway for cellular entry of dendrimer-DNA complexes is by entrapment into vesicles that originate as invaginations from thecell surface. However, it has not been definitively established whether the entrapped complexes arefirst bound to the cell surface, or are simply captured in the fluid that is taken into the formingvesicle. The binding of dendrimers and dendrimer-DNA complexes to lipid membranes maydepend on the membrane composition and the stoichiometry of the complexes.(3) Titration properties. DNA transfection by dendrimers is thought to proceed via the so-calledendocytic pathway.
In this process, the cell membrane surface forms an invagination whichpinches off from the extracellular medium, forming a lipid vesicle within the cellular cytoplasm(Figure 2.) This "endosomal vesicle" entraps some of the extracellular fluid, as well as anymembrane-bound molecules; dendrimer-DNA complexes may be in solution or membraneadherent. Once inside the cytoplasm, the endosomal vesicle is actively acidified by proton-pumpingenzymes and anion channels in the endosome membrane. If the pH in the endosome is unusuallywell-buffered, then acidification can result in a large osmotic imbalance (
) caused by the largeinflux of H
and anions. Weak bases, which concentrate in endosomes and act as pH-buffers, cancause endosomal rupture by this mechanism.
Haensler and Szoka
have proposed that dendrimersact similarly, with the physiologically relevant buffering capacity provided by the internal, tertiary