To assess the biochemical, mechanical and structural characteristics of retained dentin after applying three novel bromelain-contained chemomechanical caries removal (CMCR) formulations in comparison to the conventional excavation methods (hand and rotary) and a commercial papain-contained gel (Brix 3000). Seventy-two extracted permanent molars with natural occlusal carious lesions (score > 4 following the International Caries Detection and Assessment System (ICDAS-II)) were randomly allocated into six groups (n = 12) according to the excavation methods: hand excavation, rotary excavation, Brix 3000, bromelain-contained gel (F1), bromelain-chloramine-T (F2), and bromelain-chlorhexidine gel (F3). The superficial and deeper layers of residual dentin were examined by Raman microspectroscopy and Vickers microhardness, while the surface morphology was assessed by the scanning electron microscope (SEM). A multivariate analysis of variance followed by Tukey’s test (p > 0.05) was performed for data analysis. The novel formulations showed an ability to preserve the partially demineralized dentin that showed a reduced phosphate content with a higher organic matrix. This was associated with lower Vickers microhardness values in comparison to sound dentin and rotary excavation. The collagen integration ratio in all methods was close to sound dentin (0.9–1.0) at the deeper dentin layer. The bromelain-chloramine-T gel (F2) produced the smoothest smear-free dentin surface with a higher number of opened dentinal tubules. In contrast, dense smearing covering the remaining dentin was observed in the manual and rotary methods with obstructed dentin tubule orifices. The bromelain-contained formulations can be considered a new minimally invasive approach for selectively removing caries in deep cavitated dentin lesions.
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