Biocompatibility Assays for Titanates

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titanates are insoluble, particulate compounds of titanium and oxygen with crystalline surfaces that bind metal ions. They have been shown to inhibit the growth of several pathogen-causing bacteria and enhance delivery of metal ions into cells. [2] Unfortunately, the mechanisms by which titanates bind metals and suppress cell metabolism are not well understood.

The chemistry of the titanate’s crystalline surface is a primary determinant of its binding properties and biological effects. Initial studies of native titanate (monosodium titanate, MST) and peroxo-titanates showed that a specific domain on the crystalline surface was responsible for binding metals [2].

While this phenomenon seems to work as intended in the context of delivering metals to cells, it poses some problems when measuring the biocompatibility of these materials. The particulate nature of these materials and their’stickiness’ to both cells and culture vessels can confound assays that rely on optical density for measurement.

Specifically, this ‘particulate bias’ can interfere with plate-reader assays that depend on optical density to assess the metabolic state of cells in vitro. Fortunately, several alternatives have been developed to mitigate this problem and allow the cellular effects of titanates to be measured.

These alternatives are based on the fact that MST and APT bind metal ions and inhibit their growth in a concentration-dependent fashion. By using a NanoDrop® spectrophotometer, which requires only 1 mL of sample, and CellTiter-BlueTM (CTB), which reads colorless OD, these assays can be performed with minimal particulate interference.