A Novel Method For Immunohistochemical Detection Of Osteogenic Markers In Undecalcified Sections Of Bone

摘要

Histologic evaluation of the bone-biomaterial interface requires undecalcified polymethylmethacrylate (PMMA) sections.~(1,2) Ideally, bioactive biomaterials for use in bone regeneration should possess the ability to activate bone formation and, thus, cause the differentiation of osteoprogenitor cells into osteoblasts at their surfaces. Therefore, in order to evaluate the osteogenic potential of novel implant materials it is important to examine the effect of these materials on osteoblastic differentiation. While various assays have been developed which permit studying the effect of biomaterials on the expression of osteogenic markers in vitro, there have been considerable difficulties to visualize the expression of these markers in undecalcified implant material containing sections of bone obtained from in vivo experiments. This is due to the fact that, until recently, it was not possible to perform immunohistochemistry and in situ hybridization on implant materials containing bone specimens embedded in PMMA resin. Only recently have embedding resins and embedding techniques become available that permit immunohistochemical analysis on resin embedded hard tissue sections.~(3,4) In the work presented here, we describe a new technique that facilitates immunohistochemical analysis of osteogenic markers on undecalcified sections of bone.