The focus of the present thesis lies on the combination of mass spectrometry imaging (MSI) and state-of-the-art instrumental bioanalytical techniques for the detailed characterization of biomaterials and tissue samples in their biological environment. Method development for MSI applications lead to new findings concerning the impact of physical sample characteristics on reproducibility and accuracy in MSI experiments. Stabilizing additives and fixatives, conventionally used in histology, were investigated regarding their influence on MSI analysis based on results obtained for rat kidney tissue samples. The present thesis focuses on the analysis of ultrahigh molecular weight polyethylene (UHWMPE), a polymer commonly used in acetabular hip joint replacement systems. Material science revealed that material aging, relevant for implant failure, is often correlated to oxidative degradation. The high relevance of the biological environment and its interaction with the polymer could be demonstrated by MSI analysis. In vitro experiments showed the time dependent adsorption of components related to synovial fluid on the polymer. Lipids, identified directly from the surface and associated to joint lubrication, were adsorbed onto and diffused into the polymer samples. Focusing on phospholipids and cholesterol, lipids were extracted and further identified by thin layer chromatography separation followed by MALDI-TOF/RTOF analysis in combination with collision-induced dissociation. Protein analysis revealed high contents of proteins adsorbed on UHMWPE samples, which are either high abundance proteins or associated with lipid transportation and synthesis,. Protein identification was performed by 1-dimensional gel electrophoresis followed by enzymatic digestion and peptide mass fingerprinting and peptide sequencing. The obtained results were confirmed by explanted polymer samples after revision surgery. The established method was further transferred to the analysis of biodegradable graft materials used in artificial vascular prosthetics in mouse models, where biodegradation and associated lipid and protein species were investigated.