We are working on an osseointegration project of a new resorbable biomaterial. We implant the biomaterial samples in rabbit condyle and we need to evaluate the local biological effects after implantation following ISO 10993-6 standard (Annex E). We work with undecalcified bone samples embedded in PMMA and cut them with a microtome at 5-10 microns. We would appreciate if you could help us to select the best staining technique/s to identify different cell types/responses: polymorphonuclear cells, lymphocytes, plasma cells, macrophages, giant cells, necrosis, neovascularisation, fibrosis, and fatty infiltrate.
You can do it if you use a tungsten carbide knive and a motorised microtome. We usually obtain 5-7 microns slice. Diamond saw is needed if your samples contain metal implants and, sometimes, when a TCP scaffold is implanted.
It is very important to use an adequate tissue processing procedure to get good PMMA infiltrated samples. In our lab we usually cut bone samples up to 2 cm diameter.
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