Chemical Fixation Importance of Histologic Samples for Staining of Proliferated Bone During Implant Osseointegration

Abstract

This study aimed to obtain as much and as accurate information as possible about the process of osseointegration of titanium implants, through histological processing. For this purpose, the combination of a mercury chloride fixator (Stieve mixture) and a complex staining method (Goldner’s trichrome) was used, which can highlight the microscopic structures in several colors and shades. The Stieve mixture proved to have distinguished qualities both in the preservation of basophilic and acidophilic structures and especially in the chemical preparation of the substrate for the binding of each structural component with a specific acidophilic dye. On the substrate thus prepared, the acidophilic dyes used in the Goldner’s trichrome staining highlighted the structural components in different colors and shades, offering the possibility of high precision assessment of the newly proliferated bone, in different stages of proliferation, consolidation and reshaping. Complex information was obtained, which allows the appreciation of osseointegration for endosseous implants and the amount of proliferated bone at the bone-implant interface, the stage of proliferation, consolidation and reshaping of the mature bone, the degree of fixation to deep bone structures. The possibility of complex assessment of the osseointegration process for endosseous implants at a given time is of great practical importance for correct assessment of the waiting time required for safe installation of prosthesis. The combination of Stieve fixator mixture with Goldner’s trichrome staining method has proven to be very effective in histology, highlighting on excellent level the existing structural components of each stage, in which the osseointegration process goes through. A very important role in this combination was achieved by mercury chloride mixture, which prepared the substrate for specific reaction with dyes. Regarding the results obtained, we recommend this combination for histological investigations aimed on bone proliferation and repair.