Norman C. Blumenthal

A Raman and infrared spectroscopic investigation of biological hydroxyapatite

Raman spectra were acquired on ox femur samples treated with hydrazine to remove the organic components of bone. A large increase in the signal-noise ratio of the mineral spectrum resulted from the exposure of the mineral surface and the removal of fluorescent components of the organic matrix. The effect of hydrazine treatment of the mineral matrix has been reinvestigated and shown to be slight on the basis of second derivative FTIR data. This is the first time that this high resolution technique has been applied to biological minerals.

Effects of low-intensity AC and/or DC electromagnetic fields on cell attachment and induction of apoptosis

Rat tendon fibroblast (RTF) and rat bone marrow (RBM) osteoprogenitor cells were cultured and exposed to AC and/or DC magnetic fields in a triaxial Helmholtz coil in an incubator for up to 13 days. The AC fields were at 60 and 1000 Hz and up to 0.25 mT peak to peak, and the DC fields were up to 0.25 mT. At various combinations of field strengths and frequencies, AC and/or DC fields resulted in extensive detachment of preattached cells and prevented the normal attachment of cells not previously attached to substrates. In addition, the fields resulted in altered cell morphologies. When RTF and RBM cells were removed from the fields after several days of exposure, they partially reattached and assumed more normal morphologies. An additional set of experiments described in the Appendix corroborates these findings and also shows that low-frequency EMF also initiates apoptosis, i.e., programmed cell death, at the onset of cell detachment. Taken together, these results suggest that the electromagnetic fields result in significant alterations in cell metabolism and cytoskeleton structure. Further work is required to determine the relative effect of the electric and magnetic fields on these phenomena. The research has implications for understanding the role of fields in affecting bone healing in fracture nonunions, in cell detachment in cancer metastasis, and in the effect of EMF on organisms generally.

Evaluation of a low-temperature calcium phosphate particulate implant material: Physical-chemical properties and in vivo bone response

A study was conducted to evaluate the osteoconductive ability of a particulate, low-temperature hydroxylapatite (HA(LT)) material (OsteoGen; Impladent, Holliswood, NY). An implantable chamber model was used to determine the ability of this material to encourage bone ingrowth into channels lined with either rough-surfaced titanium or rough-surfaced plasma-sprayed hydroxylapatite. The HA(LT) material increased bone ingrowth into the titanium-lined channels comparable with that in plasma-sprayed hydroxylapatite-coated channels. It was incorporated into ingrowing bone without intervening soft tissue, with the bone bonding directly to the material surface in much the same fashion as it bonds at the plasma-sprayed hydroxylapatite surface. Mechanical testing of the ingrown bone showed no weakness because particles were incorporated. At 12 weeks, the particles began to show signs of dissolution. It was concluded that the HA(LT) material is a biocompatible, osteoconductive material that conducts bone ingrowth in much the same way as high-temperature particulate hydroxylapatite ceramics. This material has the additional desirable property of being slowly resorbable, a beneficial characteristic for many bone-filling applications.

The effect of cadmium on the formation and properties of hydroxyapatite In vitro and its relation to cadmium toxicity in the skeletal system

In order to understand the biological action of cadmium (Cd) in inducing bone pathologies, the effect of Cd on the formation, structure, and properties of hydroxyapatite (HA) in vitro was investigated using three biologically relevant test systems: (1) direct precipitation of HA with no precursor phase; (2) transformation of amorphous calcium phosphate (ACP) to crystalline HA; and (3) growth of HA seed crystals. Cd-containing HA was prepared by transforming ACP to HA in the presence of Cd at a pH of 10; Cd/Ca ratios of 0.05, 0.10, and 0.20 were obtained. Infrared and x-ray diffraction analyses were performed on the Cd-HA samples, and measurements were made of Cd adsorption on HA and of the dissolution characteristics of Cd-containing HA. Cd incorporation in HA introduced little strain in the lattice but resulted in a decreasing C-axis spacing and a corresponding crystal size decrease in the C-axis direction. Cd incorporation had a nominal effect on HA dissolution. Cd had an inhibitory effect on HA formation kinetics in all three test systems. Infrared spectroscopy of Cd-HA showed a complex series of small changes in the spectra as a function of Cd concentration resulting from some distortion in the crystal perfection and symmetry. The interference of Cd with mineralization can be partially explained by its inhibitory effect on HA nucleation and growth in addition to any cellular involvement. Furthermore, Cd probably has little effect on bone mineral dissolution. Our results explain the Cd incorporation reported in bone.

Molecular structure at the bone-implant interface: A vibrational spectroscopic characterization

SummaryTest implant plates surgically retrieved from distal femurs of dogs were studied by Raman spectroscopy in order to characterize the bone-implant interface. The implant surface consisted of phosphate mineral, plasma sprayed on a titanium substrate. On the basis of its spectroscopic signature, the phosphate mineral of bone and the implant surface formed a mixed phase in the interface.