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Multi-colour Electron Microscopy: Protein Accumulation and Cellular Activity Surrounding Hydroxyapatite Implants Revealed by Energy Dispersive X-ray Spectrometry

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The interaction of biomaterials and implants with cells and tissues remains a critical issue for patients and researchers involved in the biomedical field. Key research trends include the development of… Click to show full abstract

The interaction of biomaterials and implants with cells and tissues remains a critical issue for patients and researchers involved in the biomedical field. Key research trends include the development of multimodal imaging and contrasting mechanisms to facilitate the visualisation of biological samples and biomaterials/implant materials within the same sample [1]. Energy dispersive x-ray spectrometry (EDS) provides elemental contrast and analytical capabilities within an electron microscope, supplying an opportunity for researchers to analyse ultrastructure and composition within the same session. We used EDS on two research topics investigating tissue interactions with implants and biomaterials. 1. The analysis of wear particles from patient tissue and in vitro studies on cultured macrophages. 2. The degradation of hydroxyapatite implants in vivo. Patient samples were prepared for routine TEM pathology examination. RAW macrophages were seeded onto coverslips in media containing combinations of materials in the form of nanoparticles for 24 hours before being fixed (Karnovsky’s fixation) dehydrated, critical point dried and coated with carbon. Hydroxyapatite implant tissue samples were fixed, embedded in resin and ground to a thickness of approximately 1mm before being stained using uranyl acetate and lead citrate.

Keywords: dispersive ray; microscopy; hydroxyapatite implants; energy dispersive; ray spectrometry

Journal Title: Microscopy and Microanalysis
Year Published: 2020

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