Collagen stability of dentin matrices after treatment with agents used to inactivate MMPs is critical to the preservation of adhesive interfaces. Objectives: The objective of this study was to evaluate the effect of Gluma dentin desensitizer (composed of 5% glutaraldehyde and 35% HEMA in water) on the mechanical and biochemical stability of dentin matrices. Methods: Human coronal dentin from ten extracted third molars was prepared by removing roots and enamel, and used to obtain twenty dentin beams of 2.0 x 1.0 x 6.0 mm. the beams were completely demineralized by immersion in 10 wt.% liquid phosphoric acid (pH 1.0) for 18h, and then rinsed in deionized water (DW). Initial dry mass was measured after dehydrating the beams. The beams were rehydrated and then equally and randomly divided in two groups to be treated either with Gluma or DW (Control) for 60 seconds. The beams were individually incubated in 1ml of complete media in a water bath at 37°C for up to four weeks. Collagen solubilization from dentin matrices was analyzed by means of hydroxyproline (HYP) release and loss of dry mass after one and four weeks. A two-way ANOVA and Tukey test were used for data analysis at a p<0.05. Results: For both mass loss and HYP release, a significant effect of 'treatment' (p<0.001), but no effect of 'time' or the interaction 'treatment-time' was demonstrated. Differences between the control and Gluma groups were shown in HYP release at four weeks (p=0.001) and percentage of total dry mass loss both at one and four weeks (p<0.05). Conclusion: Gluma treatment for 60 sec. resulted in significantly less dry mass loss and HYP release after four weeks indicating that Gluma ingredients can stabilize dentin matrices.