AVS 62nd International Symposium & Exhibition | |
Biomaterial Interfaces | Monday Sessions |
Session BI+AS-MoA |
Session: | Characterization of Biological and Biomaterials Surfaces (2) |
Presenter: | Adam Taylor, University of Washington |
Authors: | A.J. Taylor, University of Washington L.J. White, University of Nottingham, UK D.M. Faulk, University of Pittsburgh L.T. Saldin, University of Pittsburgh D.G. Castner, University of Washington S.F. Badylak, University of Pittsburgh B.D. Ratner, University of Washington |
Correspondent: | Click to Email |
Principal components analysis (PCA) revealed spectral differences between treatment groups. High mass peaks associated with specific detergent fragments were observed on the scaffolds exposed to SDS and deoxycholate. Peaks indicative of phospholipid membranes were observed in all samples, but to a greater extent with scaffolds not exposed to detergent. We further probed these data sets to investigate how detergent selection impacts proteinaceous ECM components. Using a reduced peak list of known characteristic amino acid fragments, PCA distinguished native bladder tissue from decellularized UBM and highlighted spectral differences between UBM treated with ionic vs. charge-neutral detergents. Notably, the basement membrane surface of UBM prepared with ionic detergents SDS and deoxycholate yielded less intense characteristic peaks from hydrophobic amino acids than UBM treated with charge neutral detergents CHAPS and Triton X-100. Harsher detergents may denature protein structure and break protein-protein interactions through binding of their hydrophobic tail to hydrophobic amino acid residues. Such damage is hypothesized to cause sub-optimal in vitro and in vivo responses. We further examined cell-matrix interactions of human urothelial cells seeded on the BMC of UBM, investigating how detergent exposure affected cell proliferation and permeability of the cell monolayer. An understanding of the effects of detergent exposure on the structure, composition and surface molecular functionality of decellularized scaffolds will facilitate a rational strategy for successful recellularization and subsequent positive clinical outcomes.