Different kinetic pathways of early stage calcium-phosphate cluster aggregation induced by carboxylate-containing polymers.

Acidic proteins are critical to biomineral formation, although their precise mechanistic function remains poorly understood. A number of recent studies have suggested a nonclassical mineralization model that emphasizes the importance of the formation of polymer-stabilized mineral clusters or particles; however, it has been difficult to characterize the precursors experimentally due to their transient nature. Here, we successfully captured stepwise evolution of transient CaP clusters in mineralizing solutions and studied the roles of functional polymers with laser light scattering (LLS) to determine how these polymers influence the stability of nanoclusters. We found that the polymer structure can alter CaP aggregation mechanisms, whereas the polymer concentration strongly influences the rate of CaP aggregation. Our results indicate that the ability of acidic biomolecules to control the formation of relatively stable nanoclusters in the early stages may be critical for intrafibrillar mineralization. More importantly, LLS provided information about the size and the structural evolution of CaP aggregates, which will help define the process of controlled biomineralization.

Cooperative calcium phosphate nucleation within collagen fibrils.

Although "chaperone molecules" rich in negatively charged residues (i.e., glutamic and aspartic acid) are known to play important roles in the biomineralization process, the precise mechanism by which type I collagen acquires intrafibrillar mineral via these chaperone molecules remains unknown. This study demonstrates a mechanism of cooperative nucleation in which three key components (collagen, chaperone molecules, and Ca(2+) and PO(4)(3-)) interact simultaneously. The mineralization of collagen under conditions in which collagen was exposed to pAsp, Ca(2+), and PO(4)(3-) simultaneously or pretreated with the chaperone molecule (in this case, poly(aspartic acid)) before any exposure to the mineralizing solution was compared to deduce the mineralization mechanism. Depending on the exact conditions, intrafibrillar mineral formation could be reduced or even eliminated through pretreatment with the chaperone molecule. Through the use of a fluorescently tagged polymer, it was determined that the adsorption of the chaperone molecule to the collagen surface retarded further adsorption of subsequent molecules, explaining the reduced mineralization rate in pretreated samples. This finding is significant because it indicates that chaperone molecules must interact simultaneously with the ions in solution and collagen for biomimetic mineralization to occur and that the rate of mineralization is highly dependent upon the interaction of collagen with its environment.

Design of stable polyether-magnetite complexes in aqueous media: effects of the anchor group, molecular weight, and chain density.

The colloidal stability of polymer-stabilized nanoparticles is critical for therapeutic use. However, phosphates in physiological media can induce polymer desorption and consequently flocculation. Colloidal characteristics of PEO-magnetite nanoparticles with different anchors for attaching PEO to magnetite were examined in PBS. The effects of the number of anchors, PEO molecular weight, and chain density were examined. It was observed that ammonium phosphonates anchored PEO to magnetite effectively in phosphate-containing solutions because of interactions between the phosphonates and magnetite. Additionally, a method to estimate the magnetite surface coverage was developed and was found to be critical to the prediction of colloidal stability. This is key to understanding how functionalized surfaces interact with their environment.

Synthesis and colloidal properties of polyether-magnetite complexes in water and phosphate-buffered saline.

Biocompatible magnetic nanoparticles show great promise for many biotechnological applications. This paper addresses the synthesis and characterization of magnetite nanoparticles coated with poly(ethylene oxide) (PEO) homopolymers and amphiphilic poly(propylene oxide-b-ethylene oxide) (PPO-b-PEO) copolymers that were anchored through ammonium ions. Predictions and experimental measurements of the colloidal properties of these nanoparticles in water and phosphate-buffered saline (PBS) as functions of the polymer block lengths and polymer loading are reported. The complexes were found to exist as primary particles at high polymer compositions, and most formed small clusters with equilibrium sizes as the polymer loading was reduced. Through implementation of a polymer brush model, the size distributions from dynamic light scattering (DLS) were compared to those from the model. For complexes that did not cluster, the experimental sizes matched the model well. For complexes that clustered, equilibrium diameters were predicted accurately through an empirical fit derived from DLS data and the half-life for doublet formation calculated using the modified Derjaguin-Landau-Verwey-Overbeek (DLVO) theory. Deviation from this empirical fit provided insight into possible additional interparticle hydrophobic interactions for select complexes for which the DLVO theory could not account. While the polymers remained bound to the nanoparticles in water, most of them desorbed slowly in PBS. Desorption was slowed significantly at high polymer chain densities and with hydrophobic PPO anchor blocks. By tailoring the PPO block length and the number of polymer chains on the surface, flocculation of the magnetite complexes in PBS was avoided. This allows for in vitro experiments where appreciable flocculation or sedimentation will not take place within the specified time scale requirements of an experiment.