Mesenchymal stem cells derived from human exfoliated deciduous teeth (SHED) are a promising source for regenerative therapies due to their multipotency, proliferative capacity, and immunomodulatory properties. The present study aimed to isolate and characterize SHED subpopulations based on CD71 and CD146 expression and evaluate their multilineage differentiation potential. SHED were obtained from pediatric donors and separated into CD71⁺, CD71⁻, CD146⁺, and CD146⁻ fractions using magnetic-activated cell sorting (MACS). CD71⁺/CD71⁻ and CD146⁺/CD146⁻ populations were isolated independently; no simultaneous double sorting for both markers was performed. Immunocytochemistry was employed to confirm the expression of surface and intracellular markers, including STRO-1, CD44, nestin, and vimentin. Multilineage differentiation assays toward osteogenic, adipogenic, and chondrogenic lineages revealed that CD71⁺ cells exhibited reduced osteogenic capacity compared to CD71⁻ cells, whereas CD146⁺ cells showed enhanced osteogenic and adipogenic differentiation. Chondrogenic differentiation seemed unaffected by marker expression under the 2D conditions employed. These results highlight functional heterogeneity within SHED populations and indicate that CD71 and CD146 independently influence differentiation outcomes. The selective enrichment of CD146⁺ SHED may enhance osteogenic and adipogenic regenerative applications, while CD71⁺ subsets may serve as a valuable model for studying proliferation and paracrine effects. Limitations include the use of in vitro differentiation assays and the absence of in vivo validation; additionally, combined CD71/CD146 analysis may further clarify the relationship between metabolic activity and stem/progenitor niche characteristics. Overall, marker-based characterization of SHED subpopulations provides insight into their biological properties and potential utility in targeted cell-based therapeutic strategies.

Purpose: The aim of this in vitro study was to evaluate the effect of different irrigation regimens of root dentin microhardness. Materials and methods: Twenty extracted, single-rooted, non-endodontically treated upper incisors were decoronated and further sectioned longitudinally in buccolingual direction. The surfaces of all forty samples were polished, evenly distributed into four groups (n=10) and then immersed in the following solutions: group 1: 2% sodium hypochlorite for 2 minutes followed by 17% EDTA for 2 minutes; group 2: 2% sodium hypochlorite for 2 minutes followed by 17% EDTA for 2 minutes, both ultrasonically activated; group 3: 5.25% sodium hypochlorite for 2 minutes followed by 17% EDTA for 2 minutes; group 4: distilled water (control group). Afterwards, the samples were dried and subjected to microhardness testing by using 300g load with a dwell time of 20 seconds. The averaged values of each sample’s Vicker’s hardness numbers were calculated and statistically analyzed. Results: All irrigation regimens significantly decreased the mean hardness of dentin when compared to the control group 4 (p0.05) regardless of the concentration and the ultrasonic activation of the solutions. Conclusions: The dentin microhardness was decreased regardless of the irrigation protocols used. Additional studies should clarify the clinical relevance of the compromised mechanical properties of dentin tissue and their relationship with the overall treatment success.

Background: Early detection and monitoring of enamel changes during caries lesion formation are essential for preventive management. This study aimed to evaluate time-dependent enamel demineralization using micro-computed tomography (micro-CT) and to compare its diagnostic performance with laser fluorescence and digital colorimetric image analysis. Methods: Twelve sound human permanent teeth were subjected to a gel-based lactic acid demineralization for 14 days. Assessments were performed at baseline and after 3, 7, and 14 days. Enamel mineral density (MD) and demineralization depth (DD) were measured using micro-CT. Laser fluorescence was evaluated using DIAGNOdent, while colorimetric changes were analyzed through standardized digital imaging using the CIE Lab* system, including ΔE and Whiteness Index (WI). Statistical analysis included repeated measures ANOVA and Pearson correlation (p < 0.05). Results: A significant time-dependent progression of enamel demineralization was observed. Demineralization depth increased from 0.0828 mm (3 days) to 0.234 mm (14 days) (p < 0.001), while mineral density decreased significantly over time (p < 0.001). DIAGNOdent values showed significant increases after 7 and 14 days (p = 0.002). Colorimetric analysis revealed early detectable changes, with ΔE exceeding clinically perceptible thresholds as early as day 3. WI values increased progressively, indicating enhanced enamel opacity. A weak but significant negative correlation between MD and DD was found (p = 0.04). Conclusions: Enamel demineralization progresses in a time-dependent manner and can be effectively monitored using micro-CT, laser fluorescence, and colorimetric analysis. Digital colorimetric analysis may serve as a valuable adjunctive tool in caries diagnostics.

Accurate identification of cariogenic bacteria is crucial for understanding caries development in children. Classical culture methods often underestimate microbial diversity, while polymerase chain reaction (PCR) can detect species that are difficult to cultivate. The aim of this study was to compare culture-based and PCR-based methods for detecting key cariogenic microorganisms in the carious dentin of pediatric patients. Thirty dentin samples were collected from the permanent teeth of children aged 8–14 years. Parallel analyses were performed using standard culture techniques and PCR targeting the gtfB gene of S. mutans and the 16S rRNA gene of Lactobacillus spp. Culture results were quantified as colony-forming units, while PCR results were classified as negative, low-positive, or positive. The results show that culture-based methods identified S. mutans in 16.7% of the samples and Lactobacillus spp. in 3.3%, while PCR identified a signal for S. mutans in 43.3% and Lactobacillus spp. in 100% of the samples. PCR-based methods provide higher sensitivity for detecting key cariogenic bacteria, including S. mutans and Lactobacillus spp. However, PCR detects bacterial DNA and does not indicate bacterial viability or activity. Combining molecular and culture-based approaches allows a more comprehensive assessment of the cariogenic microbiota, supporting accurate microbiological evaluation in pediatric caries research.