Cell migration assays were carried out using transwell inserts with pore sizes of 3 μm (polyester membrane, Sigma-Aldrich, CLS3472) or 8 μm (polycarbonate membrane, Sigma-Aldrich, CLS3422), in 24 well plates, as described by Justus et al. [85 ].
Cells were seeded in the top chamber at a density of 1 × 106 cells/mL and incubated for 4 h, at 37 °C and 5% CO2 in a humidified incubator, to allow cell adhesion and spreading. After attachment, cells were supplemented with new culture media, with or without treatment. Culture media with 10% heat inactivated foetal bovine serum (FBS, Invitrogen, 10,099,141) was added to the bottom well to create a chemotactic gradient. Cells were incubated in a humidified incubator at 37 °C and 5% CO2. The chemotactic gradient was renewed every 24 h via addition of new media in both wells. To assess untreated WK1 cell migration over a period of 72 h, separate transwells were run in parallel and fixed at 24, 48 and 72 h, respectively.
The migration of cells, treated with culture media containing 0.1 mg/mL Tf@pSiNPs, 50 μM NFA (Sigma Aldrich, N0630), or 2 mM orthosilicic acid [33 (link)] was characterised using the cell migration assay described.
Cells were seeded in the top chamber at a density of 1 × 106 cells/mL and incubated for 4 h, at 37 °C and 5% CO2 in a humidified incubator, to allow cell adhesion and spreading. After attachment, cells were supplemented with new culture media, with or without treatment. Culture media with 10% heat inactivated foetal bovine serum (FBS, Invitrogen, 10,099,141) was added to the bottom well to create a chemotactic gradient. Cells were incubated in a humidified incubator at 37 °C and 5% CO2. The chemotactic gradient was renewed every 24 h via addition of new media in both wells. To assess untreated WK1 cell migration over a period of 72 h, separate transwells were run in parallel and fixed at 24, 48 and 72 h, respectively.
The migration of cells, treated with culture media containing 0.1 mg/mL Tf@pSiNPs, 50 μM NFA (Sigma Aldrich, N0630), or 2 mM orthosilicic acid [33 (link)] was characterised using the cell migration assay described.
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