Quantitative analysis of early in vivo tissue response to synthetic apatite implants

Studies have shown synthetic calcium phosphates such as hydroxyapatite and beta tricalcium phosphate to be biocompatible in vivo. However, few studies have quantitated histological responses to the implants. The aim of this study was to develop a method for the quantitative assessment of tissue biocompatibility to ceramic materials in vivo and to use this method to compare noncarbonated and carbonated apatite implants. Synthetic sintered apatites of 0, 3, and 6% carbonate by weight were prepared and cut into implants 4 X 4 X 1 mm. These were placed 2 mm into the medial aspect of rat femurs. Following sacrifice at 4 weeks, the femurs were fixed in formalin, demineralized in formic acid, and embedded in glycol methacrylate. Sections were cut on an ultramicrotome set at 1.5 micron and stained with toluidine blue. A point counting technique using standard stereological grids and a low-power microscope was used to measure areas of new bone formation. The width of the connective tissue zone adjacent to muscle was measured using an image analyzer. All implants were well accepted by the host tissues judging from criteria of minimal inflammation and degree of fixation. Results showed an increase in new bone formed in the marrow cavity with increasing carbonate content. This may improve stability of the implant in the host bone, particularly during the initial healing period. A technique which should enable quantitative histological evaluation of different ceramic materials has been developed. The use of this method indicates that further studies are warranted to investigate carbonated apatite as an implant material.