An in-vitro evaluation of residual dentin retained after using novel enzymatic-based chemomechanical caries removal agents.

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.

An in vitro assessment of the residual dentin after using three minimally invasive caries removal techniques.

To evaluate the efficiency and effectiveness of three minimally invasive (MI) techniques in removing deep dentin carious lesions. Forty extracted carious molars were treated by conventional rotary excavation (control), chemomechanical caries removal agent (Brix 3000), ultrasonic abrasion (WOODPECKER, GUILIN, China); and Er, Cr: YSGG laser ablation (BIOLASE San Clemente, CA, USA). The assessments include; the excavation time, DIAGNOdent pen, Raman spectroscopy, Vickers microhardness, and scanning electron microscope combined with energy dispersive X-ray spectroscopy (SEM-EDX). The rotary method recorded the shortest excavation time (p < 0.001), Brix 3000 gel was the slowest. DIAGNOdent pen values ranged between 14 and 18 in the remaining dentin and laser-ablated surfaces recorded the lowest reading (p < 0.001). The Ca:P ratios of the remaining dentin were close to sound dentin after all excavation methods; however, it was higher in the ultrasonic technique (p < 0.05). The bur-excavated dentin showed higher phosphate and lower matrix contents with higher tissue hardness that was comparable to sound dentin indicating the non-selectiveness of this technique in removing the potentially repairable dentin tissue. In contrast, the MI techniques exhibited lower phosphate and higher organic contents associated with lower microhardness in the deeper dentin layers. This was associated with smooth residual dentin without smearing and patent dentinal tubules. This study supports the efficiency of using MI methods in caries removal as conservative alternatives to rotary excavation, providing a promising strategy for the clinical dental practice.

The preventive/therapeutic effect of CO2 laser and MI Paste Plus® on intact and demineralized enamel against Streptococcus mutans (In Vitro Study).

Background: To evaluate the preventive and therapeutic effects of CO2 laser and MI paste plus on intact and demineralized enamel surfaces and their impact on bacterial adhesion. Methods: 160 enamel slabs were prepared and randomly allocated into two main groups; sound and demineralized enamel (n = 80 per group), in which specimens were immersed in a demineralizing solution (50 mM acetic acid, pH 4.5) for 72 h at 37 °C. Each group was further divided into four subgroups (n = 20); the control (un treated surfaces), surfaces treated by CO2 laser, MI paste plus (Recaldent™, GC corporation/Germany), and those received a combination of CO2 and MI paste plus. Streptococcus Mutans biofilm was isolated, quantified, and then applied on treated enamel surfaces and incubated anaerobically for 24 h and then quantified by colony-forming unit (CFU). Meanwhile, surface changes were assessed by Vickers microhardness and Scanning Electron Microscope combined with Energy-Dispersive X-Ray Spectroscopy (SEM-EDX). Results: The combined use of CO2 laser followed by MI paste plus significantly (p < 0.000) enhanced surface microhardness of sound and demineralized enamel with a significant reduction in bacterial counts. However, each technique alone was beneficial as they exhibited higher microhardness with lower bacterial viability in comparison to the control. The treatment of demineralized enamel surfaces with MI paste significantly reduced the number of bacterial colonies with the presence of dispersed mineral deposits over the surface. Conclusions: The combined use of CO2 laser and MI paste plus was effective as a preventive and/or therapeutic measures in enhancing surface properties of enamel and reducing the bacterial viability.

The in-vitro development of novel enzyme-based chemo-mechanical caries removal agents.

OBJECTIVES: Bromelain is a potent proteolytic enzyme that has a unique functionality makes it valuable for various therapeutic purposes. This study aimed to develop three novel formulations based on bromelain to be used as chemomechanical caries removal agents. METHODS: The novel agents were prepared using different concentrations of bromelain (10-40 wt. %), with and without 0.1-0.3 wt. % chloramine T or 0.5-1.5 wt. % chlorhexidine (CHX). Based on the enzymatic activity test, three formulations were selected; 30 % bromelain (F1), 30 % bromelain-0.1 % chloramine (F2) and 30 % bromelain-1.5 % CHX (F3). The assessments included molecular docking, Fourier-transform infrared spectroscopy (FTIR), viscosity and pH measurements. The efficiency of caries removal was assessed by DIAGNOdent pen, measuring the excavation time and number of applications, followed by a morphological evaluation of the remaining dentine using scanning electron microscopy (SEM). The results were compared to Brix 3000 as a control. RESULTS: The chloramine and chlorhexidine were chemically compatible with bromelain without compromising the enzyme activity. All experimental formulations showed higher viscosity and pH in comparison to Brix 3000. The DIAGNOdent readings were <20 in all groups, and the lowest readings were observed in F2. The excavation time and number of applications were lowest in F2 and F1. Both F2 and F3 produced smooth dentine surfaces with less tissue debris, but more patent dentine tubules were observed in F1 and F2. CONCLUSIONS: The bromelain-contained formulations showed a potential to be used as chemomechanical caries removal agents in vitro. Further laboratory and clinical studies are needed to validate this claim. CLINICAL SIGNIFICANCE: The bromelain from pineapple stem has broad specificity for cleavage the peptide bonds in denatured protein to facilitate their removal. The study proved the efficiency of this enzyme to remove the dental caries chemomechanically when used alone or conjugated with chloramine and/or chlorhexidine to enhance the disinfecting and cleansing properties.