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The nanostructure of engineered bioscaffolds has a profound impact on cell response, yet its understanding remains incomplete as cells interact with a highly complex interfacial layer rather than the material itself. For bioactive glass scaffolds, this layer comprises of silica gel, hydroxyapatite (HA)/carbonated hydroxyapatite (CHA), and absorbed proteins—all in varying micro/nano structure, composition, and concentration. Here, we examined the response of MC3T3-E1 pre-osteoblast cells to 30 mol% CaO–70 mol% SiO₂porous bioactive glass monoliths that differed only in nanopore size (6–44 nm) yet resulted in the formation of HA/CHA layers with significantly different microstructures. We report that cell response, as quantified by cell attachment and morphology, does not correlate with nanopore size, nor HA/CHO layer micro/nano morphology, or absorbed protein amount (bovine serum albumin, BSA), but with BSA’s secondary conformation as indicated by its β-sheet/α-helix ratio. Our results suggest that the β-sheet structure in BSA interacts electrostatically with the HA/CHA interfacial layer and activates the RGD sequence of absorbed adhesion proteins, such as fibronectin and vitronectin, thus significantly enhancing the attachment of cells. These findings provide new insight into the interaction of cells with the scaffolds’ interfacial layer, which is vital for the continued development of engineered tissue scaffolds.

For hard tissue regeneration, the bioactivity of a material is measured by its ability to induce the formation of hydroxyapatite (HA) under physiological conditions. It depends on the dissolution behavior of the glass, which itself is determined by the composition and structure of glass. The enhanced HA growth on nanoporous than on nonporous glass has been attributed by some to greater specific surface area (SSA), but to nanopore size distribution by others. To decouple the influence of nanopore size and SSA on HA formation, we have successfully fabricated homogeneous 30CaO‐70SiO₂(30C70S) model bioactive glass monoliths with different nanopore sizes, yet similar SSA via a combination of sol–gel, solvent exchange, and sintering processes. After incubation in PBS, HA, and Type‐B carbonated HA (HA/B‐CHA) form on nanoporous monoliths. The XPS, FTIR, and SEM analyses provide the first unambiguous demonstration of the influence of nanopore size alone on the formation pathway, growth rate, and microstructure of HA/CHA. Due to pore‐size limited diffusion of PO₄³⁻, two HA/CHA formation pathways are observed: HA/CHA surface deposition and/or HA/CHA incorporation into nanopores. HA/CHA growth rate on the surface of a nanoporous glass monolith is dominated by the pore‐size limited transport of Ca²⁺ions dissolved from nanoporous glass substrates. Furthermore, with increasing nanopore size, HA/CHA microstructures evolve from needle‐like, plate‐like, to flower‐like appearance. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 886–899, 2019.