Vacuum-assisted infiltration of chitosan or polycaprolactone as a structural reinforcement for sintered cancellous bovine bone graft.

Sintered cancellous bovine bone (SCBB) offers numerous advantages as a bone graft substitute material; however, its mechanical properties require improvement. In this study, SCBB was infiltrated with ε-polycaprolactone (PCL) or chitosan/monetite to improve mechanical properties while retaining valuable SCBB structure. Organic infiltrating solutions consisted of (i) chitosan and monetite (CaHPO(4)) dissolved in hydrochloric acid; (ii) chitosan, monetite (CaHPO(4)), and genipin dissolved in hydrochloric acid; or (iii) PCL polymer dissolved in tetrahydrofuran (THF). Porous SCBB materials were infiltrated with one of the three solutions using vacuum-assisted infiltration. Chitosan and CaHPO(4) were immobilized in the SCBB structure by reprecipitation following a pH increase in an ammonia atmosphere. Genipin was used in one sample group to immobilize chitosan via crosslinking. PCL was immobilized by evaporating the THF carrier solvent. Mechanical compression testing showed an improvement in ultimate stress for the SCBB with chitosan/CaHPO(4) infiltrates, whereas PCL samples showed an increase in modulus. All SCBB samples were found to demonstrate favorable in vitro biocompatibility when subjected to L929 mouse fibroblast cells but required a vigorous washing regime to eradicate toxic ammonia residue. In conclusion, infiltration of SCBB with a polymeric or organic/mineral composite matrix positively modifies SCBB mechanical properties in favor of bone grafting applications.

Growth of calcium hydroxyapatite (Ca-HAp) on cholesterol and cholestanol crystals from a simulated body fluid: A possible insight into the pathological calcifications associated with atherosclerosis.

An experimental study into calcium phosphate (CP) nucleation and growth on cholesterol and cholestanol surfaces from a supersaturated simulated body fluid (SBF) is presented with the overall aim of gaining some fundamental insights into the pathological calcifications associated with atherosclerosis. Soaking of pressed cholesterol disks at physiological temperature in SBF solutions was found to lead to CP nucleation and growth if the disks were surface roughened and if an SBF with concentrations of the calcium and hydrogen phosphate ions at 2.25x physiological concentrations was used. The CP phase deposited was shown via SEM micrographs to possess a florette type morphology akin to that observed in earlier reported studies. The use of recrystallised cholesterol and cholestanol microcrystals as substrates for soaking in SBF facilitated the observation of CP deposition. In general, cholesterol recrystallised from polar solvents like 95% ethanol as a cholesterol monohydrate phase which was a better substrate for CP growth than cholesterol recrystallised from more non-polar solvents (e.g., benzene) which produced anhydrous cholesterol phases. CP was also observed to form on recrystallised cholestanol microcrystals, a molecule closely related to cholesterol. Inductively coupled plasma optical emission spectrometry (ICP-OES) data gave confirmation that Ca:P mole ratios of the grown CP were 1.3-1.5 suggesting a mixed phase of octacalcium phosphate (OCP) and Ca-deficient HAp and that the CP coating grows (with time of soaking) on the substrates after nucleation in the SBF growth medium. Infrared (IR) spectra of the extracted coatings from the cholesterol substrates confirmed that the CP phase deposited is a semi crystalline HAp with either carbonate substituted into its structure or else co-deposited as calcium carbonate. Soaking experiments involving modified cholesterol substrates in which the OH group in the molecule was replaced with the oleiyl or phosphonate group showed no CP nucleation and growth. This observation illustrates the importance of the known epitaxial relationship between cholesterol and HAp (which theoretically predicts favourable deposition of one phase upon the other) and the consequences of its destruction (by chemical modification of the cholesterol). In the case of the phosphorylated cholesterol, failure of this substrate to nucleate CP phases may have also been caused by the reduction in concentration of free solution Ca2+ in the SBF medium by complexation with the phosphonate groups on the phosphorylated cholesterol. This would have reduced the ion product of Ca2+ and inorganic phosphate and lowered the degree of supersaturation in the SBF medium.