A Transwell Tissue Model for Studying Interactions Between Human Oral Biofilms and Epithelium

Begin with a multi-species bacterial oral biofilm grown on a mineral disc submerged in a buffer.

Sonicate to disrupt the biofilm matrix and generate a uniform single-cell suspension.

Add this bacterial suspension onto a transwell insert containing a human oral tissue model with epithelial cells grown on the membrane.

Place the insert in a fresh multiwell plate with media.

During incubation, the bacteria adhere to the epithelial surface and colonize, forming biofilms that mimic an oral infection.

This triggers the expression of inflammatory signaling molecules in the epithelial cells.

After incubation, remove the insert and discard the residual suspension.

Using a needle, excise the epithelial tissue along with its supporting membrane and transfer it into a tube containing lysis buffer and glass beads.

Homogenize to mechanically lyse the epithelial cells and bacteria, releasing intracellular contents into the buffer.

The lysate is ready for analysis of epithelial responses to oral biofilms.

Meanwhile, to remove the biofilms from the hydroxyapatite discs, use a 19 gauge needle and tweezers to lift the discs from the bottom of the 24 well plate. Transfer the discs into a bijoux containing 1 mil