Pull-out mechanical measurement of tissue-substrate adhesive strength: endothelial cell monolayer sheet formed on a thermoresponsive gelatin layer.

Although adhesive strength of a single cell on substrates has been reported, the adhesive strength at the tissue-substrate interface has not been reported. However, the tissue-substrate adhesive strength must provide important criteria for performance of implant devices. This article deals with the tissue-substrate adhesive strength for fully endothelialized tissue, which was formed on commercial tissue culture dishes with or without a coating layer of thermoresponsive gelatin (poly(N-isopropylacrylamide)-grafted gelatin, which dissolves in water at room temperature but is precipitated at 37 degrees C). To determine tissue-substrate adhesive strength, a pull-out technique using a glue-coated cover glass was used. The adhesive strength of monolayered tissue on a noncoated dish was approximately 560 Pa or 230 nN/cell at 37 degrees C. For dishes coated with thermoresponsive gelatin, the adhesive strengths were 1050 Pa or 584 nN/cell at 37 degrees C, and 26 Pa or 14 nN/cell at room temperature. For noncoated dishes, delamination occurred mostly at the interface between the extracellular matrix (ECM) secreted by the cells and the dish surface; and for coated dishes, it took place fully at the interface between ECM and the dish surface. This technique enables determination of the adhesive strength between a full monolayered tissue and a substrate.