Enhancing dentin bonding through new adhesives formulations with natural polyphenols, tricalcium phosphate and chitosan

Objectives: The aim of the study was to develop new adhesive formulations that include natural polyphenols extracted from green tea (GTE), tricalcium phosphate (TCP) and chitosan to improve dentin bonding characteristics and cytotoxicity. Methods: Four experimental adhesives were formulated under laboratory conditions. The groups differed in the integration of either GTE and/or TCP + chitosan. The four experimental and one clinically proven reference adhesive underwent shear bond testing after 24 h and 6 months of aging (n = 200) with subsequent fractographic analysis. Bond morphology was analyzed under a scanning electron microscope. The presence of phenolic compounds was validated by high performance liquid chromatography. Cytotoxicity was assessed by the WST-1 colorimetric assay on eluates up to 6 months. Statistical analysis was performed by one- and three-way ANOVA, Games-Howell and Tukey's post-hoc test as well as multiple students t-tests (alpha = 0.05). Weibull analysis was further conducted. Results: The addition of GTE into the bonding agent did show immediate (p = 0.023, p = 0.013) and long-term (p < 0.001) effects on bond strength. After 24 h, GTE doped groups performed equal to the reference (p = 0.501, p = 0.270) and TCP and chitosan displayed improvements in reliability (m=4.0, m=4.3). Bond strength is retained after aging by adding GTE (p = 0.983). The additional presence of TCP and chitosan reduces it (p = 0.026). Excluding cohesive and mixed failures, the reference adhesive performed statistically equal to three of the four experimental groups. No long-term cytotoxic effects were shown. Significance: The integration of GTE can enhance bond strength and a calcium source helps to improve immediate bond reliability.