Dental caries management approaches may influence not only tissue removal but also the microbial composition within the cavity. However, clinical evidence on how different caries removal methods affect the oral microbiome remains limited. This study aims to evaluate the effects of selective and non-selective caries removal methods on the diversity and composition of the oral microbiome using 16S rRNA gene sequencing. The findings are expected to help identify biologically compatible treatment approaches that effectively reduce pathogenic microorganisms.
Dental caries is a highly prevalent chronic disease and a major cause of tooth loss, arising from a disruption in the dynamic balance between demineralization and remineralization processes. Acidogenic microorganisms within the oral biofilm drive mineral loss in enamel and dentin, leading to lesion progression. Beyond mechanical debridement, the method of caries removal plays a critical role in modulating the residual microbial environment and influencing the biological response of the pulp-dentin complex. Conventional (non-selective) caries removal aims to eliminate all infected dentin; however, this approach may result in excessive tissue removal, including dentin with remineralization potential, and may increase the risk of pulpal stress or exposure. In contrast, minimally invasive dentistry emphasizes the preservation of tooth structure and the maintenance of pulp vitality. Selective caries removal has emerged as a biologically oriented strategy in which caries is completely removed at the periphery of the cavity while softened dentin adjacent to the pulp is preserved. This approach is designed to reduce the risk of pulp exposure and to support the reparative and remineralization capacity of the remaining dentin. Despite increasing clinical adoption, the microbiological consequences of different caries removal strategies remain incompletely understood. Traditional culture-based methods provide limited insight into the complexity of the oral microbiota, as a substantial proportion of oral microorganisms cannot be cultivated under standard laboratory conditions. The advent of 16S rRNA gene sequencing has enabled high-resolution, culture-independent characterization of microbial communities, offering a comprehensive view of microbial diversity and composition within carious lesions. This approach provides an opportunity to better understand how clinical interventions shape the oral microbiome. However, clinical evidence evaluating the impact of selective versus non-selective caries removal on the cavity microbiome using 16S rRNA sequencing remains scarce. Given that the quantity and quality of residual dentin may influence both mechanical properties and microbial persistence, elucidating these effects is critical for advancing biologically driven treatment strategies. The aim of this study is to compare the effects of selective and non-selective caries removal methods on the diversity and composition of the oral microbiome using 16S rRNA gene sequencing. The findings are expected to provide insights into the microbiological outcomes of minimally invasive approaches and to inform the development of optimized clinical protocols that balance dentin preservation with effective microbial control.
Study Type
INTERVENTIONAL
Allocation
RANDOMIZED
Purpose
BASIC_SCIENCE
Masking
SINGLE
Enrollment
48
Non-selective caries removal involves the complete excavation of infected dentin using conventional rotary instruments. Both infected and affected dentin are removed until hard dentin is reached, aiming to eliminate all carious tissue.
Selective caries removal involves complete caries removal at the cavity margins while preserving softened dentin near the pulp to reduce the risk of pulp exposure and maintain tooth vitality.
Nuh Naci Yazgan University Faculty of Dentistry
Kayseri, Kocasinan, Turkey (Türkiye)
Microbiome Changes
In all groups, dentin samples will be taken from the affected dentin at the base of the cavity after caries removal using two sterile, size 6 round steel burs moistened with saline solution for microbial analysis. The burs will be placed in 5 ml sterile vials and stored at -80 0C until the analyses are performed. The prepared cavities will be completed with routine and standard treatment protocols. Following the finishing and polishing procedures, occlusion will be checked. The samples collected during the research process will be delivered to the A\&D Genetic Diseases Evaluation Centre (Ankara, Turkey) where DNA isolation and sequence analysis will be performed together with molecular biologists. In the method, following the extraction of total genomic DNA from clinical samples, broad-range 16S rRNA PCR will be performed and MicroSeq 500 16S rRNA Sequencing kit will be used for sequence analysis.
Time frame: 3 months
Relative abundance of specific bacterial taxa
Measured using 16S rRNA sequencing to evaluate changes in key cariogenic and commensal bacteria after caries removal procedures.
Time frame: 3 months
Alpha diversity of the oral microbiome
Assessed using diversity indices (e.g., Shannon index) to evaluate microbial richness and diversity within samples.
Time frame: 3 months
Beta diversity of the oral microbiome
Evaluated to compare differences in microbial community composition between study groups.
Time frame: 3 months
Presence of residual cariogenic bacteria
Assessment of remaining cariogenic microorganisms in the cavity after caries removal.
Time frame: 3 months
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