Structural and biomechanical changes to dentin extracellular matrix following chemical removal of proteoglycans

Proteoglycans are biomacromolecules with significant biomineralization and structural roles in the dentin extracellular matrix. This study comprehensively assessed the mechanical properties and morphology of the dentin extracellular matrix following chemical removal of proteoglycans to elucidate the structural roles of proteoglycans in dentin. Dentin extracellular matrix was prepared from extracted teeth after complete tissue demineralization. Chemical removal of proteoglycans was carried-out using guanidine hydrochloride for up to 10days. The removal of proteoglycans was determined by dimethylmethylene blue colorimetric assay and histological staining analyses using transmission electron microscopy and optical microscopy. The modulus of elasticity of dentin matrix was determined by a 3-point bending test method. Partial removal of proteoglycans induced significant modifications to the dentin matrix, particularly to type I collagen. Removal of proteoglycans significantly decreased the modulus of elasticity of dentin extracellular matrix (p<0.0001). In conclusion, the subtle disruption of proteoglycans induces pronounced changes to the collagen network packing and the bulk modulus of elasticity of dentin matrix.