Allison A. Campbell

Surface-induced mineralization: a new method for producing calcium phosphate coatings.

Calcium phosphate coatings were nucleated and grown from aqueous solution onto titanium metal substrates via surface-induced mineralization (SIM) processing techniques. This process is based on the observation that in nature organisms use biopolymers to produce ceramic composites, such as teeth, bones, and shells. The SIM process involves modification of a surface to introduce surface functionalization followed by immersion in aqueous supersaturated calcium phosphate solutions. This low-temperature process (< 100 degrees C) has advantages over conventional methods of calcium phosphate deposition in that uniform coatings are produced onto complex-shaped and/or microporous samples. Additionally, because it is a low-temperature process, control of the phase and crystallinity of the deposited material can be maintained.

Antimicrobial efficacy of external fixator pins coated with a lipid stabilized hydroxyapatite/chlorhexidine complex to prevent pin tract infection in a goat model.

BACKGROUND Pin tract infection is a common complication of external fixation. An antiinfective external fixator pin might help to reduce the incidence of pin tract infection and improve pin fixation. METHODS Stainless steel and titanium external fixator pins, with and without a lipid stabilized hydroxyapatite/chlorhexidine coating, were evaluated in a goat model. Two pins contaminated with an identifiable Staphylococcus aureus strain were inserted into each tibia of 12 goats. The pin sites were examined daily. On day 14, the animals were killed, and the pin tips cultured. Insertion and extraction torques were measured. RESULTS Infection developed in 100% of uncoated pins, whereas coated pins demonstrated 4.2% infected, 12.5% colonized, and the remainder, 83.3%, had no growth (p < 0.01). Pin coating decreased the percent loss of fixation torque over uncoated pins (p = 0.04). CONCLUSION These results demonstrate that the lipid stabilized hydroxyapatite/chlorhexidine coating was successful in decreasing infection and improving fixation of external fixator pins.

Oriented growth of calcium oxalate monohydrate crystals beneath phospholipid monolayers

Oriented calcium oxalate crystals have been grown beneath phospholipid monolayers at the air-solution interface from supersaturated calcium oxalate solutions. Mature calcium oxalate crystals grown beneath zwitterionic dipalmitoylphosphatidylcholine (DPPC) monolayers exhibit the characteristic morphology of calcium oxalate monohydrate (COM) crystals with the elongated (101) crystal face preferentially oriented parallel to the plane of the monolayer. Calcium oxalate crystals grown beneath negatively-charged dimyristoylphosphatidylserine (DMPS) monolayers also show a preferential orientation with respect to the monolayer; they do not, however, exhibit the characteristic COM morphology. Raman spectroscopy strongly suggests that the crystals grown beneath either DPPC or DMPS monolayers are the monohydrate phase of calcium oxalate; therefore, differences in crystal morphology are not due to differences in the crystalline phase. Dimyristoylphosphatidylethanolamine (DMPE), dimyristoylphosphatidic acid (DMPA), eicosanoic acid (C20), and eicosanol (C20-OH) monolayers have also been studied to help elucidate the mechanisms of interaction between the lipid monolayers and the calcium oxalate crystals. We discuss the roles of lattice matching, hydrogen bonding, stereochemistry and electrostatics on crystal orientation and morphology.

Protein Electrostatic Surface Distribution Can Determine Whether Calcium Oxalate Crystal Growth is Promoted or Inhibited

Acidic proteins found in mineralized tissues act as natures crystal engineers, where they play a key role in promoting or inhibiting the growth of minerals such as hydroxyapatite and calcium oxalate. Despite their importance in such fundamental physiological processes as bone and tooth formation, however, there is remarkably little known of the protein structure-function relationships that govern crystal recognition. We have taken a model system approach to elucidate some of the relationships between protein surface chemistry and secondary crystal growth of biological minerals. We show here that the distribution of electrostatic surface charge on our model protein, Protein G, determined whether the secondary growth of calcium oxalate, the principal mineral phase of kidney stones, was promoted or inhibited when the proteins were preadsorbed at low and equivalent surface coverages of <10%. The native Protein G, which contains 10 surface carboxylates, increased the rate of calcium oxalate growth from aqueous solution under constant composition conditions up to 97%, whereas a site-directed mutant with six of the surface charges removed inhibited the growth rate by 60%. The adsorption isotherms of both proteins were determined and suggested that the differences in electrostatic surface properties also lead to differences in protein orientation on the crystal surface. These results demonstrate that differences in electrostatic surface potential of proteins can directly determine whether secondary calcium oxalate growth is promoted or inhibited, and a model is proposed that suggests the distribution of carboxylate residues determines the interrelated binding orientation and exposed surface chemistry of the adsorbed Protein G.

Histological and biomechanical evaluation of calcium phosphate coatings applied through surface-induced mineralization to porous titanium implants

The purpose of this pilot study was to evaluate surface-induced mineralization (SIM) as a potential technique to apply ceramic coatings to metal orthopaedic implants. Cylindrical titanium porous-coated implants were either coated by SIM or plasma-spray (PLS) techniques with calcium phosphate, or left uncoated (CTL). The implants were bilaterally implanted into the intramedullary canal of the proximal femur of 24 adult New Zealand white rabbits segregated into the following groups: PLS/CTL, SIM/CTL, and SIM/PLS. After 6 weeks in vivo, biomechanical and histologic evaluations were completed. Biomechanically, SIM had consistently greater mechanical interlock than PLS implants. However, CTL implants had greater mechanical interlock than both PLS and SIM. The small sample size prevented statistical evaluation and definitive biomechanical conclusions. Histologically, SIM and PLS had significantly greater ingrowth than CTL implants (p < 0.05). The SIM coating technique produced similar ingrowth characteristics as standard PLS coatings, yet may prevent osteolysis by providing a stronger, more reliable, covalent bond between the ceramic and metal.

Calcium Oxalate Nucleation and Growth on Oxide Surfaces

The heterogeneous nucleation of calcium oxalate onto colloidal oxides from aqueous solution has been studied as a model system for understanding the role of surface chemistry in biomimetic processes. The Constant Composition technique was used for measuring nucleation induction times. Results show that the dependence of nucleation on supersaturation fit well with classical nucleation theories. Surfaces which are anionic appear to promote calcium oxalate nucleation better that neutral or cationic surfaces. Modifications to positive surfaces via the adsorption of anionic surfactants, lower the effective energy barrier for nucleation, and stimulates the heterogeneous nucleation of calcium oxalate

Heterogeneous nucleation of calcium oxalate on native oxide surfaces

The aqueous deposition of calcium oxalate onto colloidal oxides has been studied as a model system for understanding heterogeneous nucleation processes of importance in biomimetic synthesis of ceramic thin films. Calcium oxalate nucleation has been monitored by measuring induction times for nucleation using Constant Composition techniques and by measuring nucleation densities on extended oxide surfaces using an atomic force microscope. Results show that the dependence of calcium oxalate nucleation on solution supersaturation fits the functional form predicted by classical nucleation theories. Anionic surfaces appear to promote nucleation better than cationic surfaces, lowering the effective energy barrier to heterogeneous nucleation.