Supplementary Material for: Deferoxamine-Induced Migration and Odontoblast Differentiation via ROS-Dependent Autophagy in Dental Pulp Stem Cells

<b><i>Background/Aims:</i></b> As a vital degradation and recycling system, autophagy plays an essential role in regulating the differentiation of stem cells. We previously showed that iron chelator deferoxamine (DFO) could promote the repair ability of dental pulp stem cells (DPSCs). Here, we investigated the effect of DFO in autophagy and the role of autophagy in regulating the migration and odontoblast differentiation of DPSCs. <b><i>Methods:</i></b> Transmission electron microscopy, immunofluorescence staining and western blotting were performed to evaluate the autophagic activity of DPSCs. Transmigration assay, alkaline phosphatase staining/activity, alizarin red S staining and quantitative PCR were performed to examine the migration and odontoblast differentiation of DPSCs. Reactive oxygen species (ROS) levels and the effects of ROS scavenger in autophagy induction were also detected. Autophagy inhibitors (3-MA and bafilomycin A1) and lentiviral vectors carrying ATG5 shRNA sequences were used for autophagy inhibition. <b><i>Results:</i></b> Early exposure to DFO promoted the mineralization of DPSCs and increased autophagic activity. Autophagy inhibition suppressed DFO-induced DPSC migration and odontoblast differentiation. Furthermore, DFO treatment could induce autophagy partly through hypoxia-inducible factor 1α/B cell lymphoma 2/adenovirus E1B 19K-interacting protein 3 (HIF-1α/BNIP3) pathway in a ROS-dependent manner. <b><i>Conclusion:</i></b> DFO increased DPSC migration and differentiation, which might be modulated through ROS-induced autophagy

Supplementary Material for: Candida albicans and Early Childhood Caries: A Systematic Review and Meta-Analysis

<p>Oral <i>Candida albicans</i> has been detected in children with early childhood caries (ECC) and has demonstrated cariogenic traits in animal models of the disease. Conversely, other studies found no positive correlation between <i>C. albicans</i> and caries experience in children, while suggesting it may have protective effects as a commensal organism. Thus, this study aimed to examine whether oral <i>C. albicans</i> is associated with ECC. Seven electronic databases were searched. The data from eligible studies were extracted, and the risk of bias was evaluated. A fixed effects model (Mantel-Haenszel estimate) was used for meta-analysis, and the summary effect measure was calculated by odds ratio (OR) and 95% confidence interval (CI). Fifteen cross-sectional studies were included for the qualitative assessment and 9 studies for meta-analysis. Twelve studies revealed higher oral <i>C. albicans</i> prevalence in ECC children than in caries-free children, while 2 studies indicated an equivalent prevalence. A pooled estimate, with OR = 6.51 and 95% CI = 4.94-8.57, indicated a significantly higher ECC experience in children with oral <i>C. albicans</i> than those without <i>C. albicans</i> (<i>p</i> < 0.01). The odds of experiencing ECC in children with <i>C. albicans</i> versus children without <i>C. albicans</i> were 5.26 for salivary, 6.69 for plaque, and 6.3 for oral swab samples. This systematic review indicates that children with oral <i>C. albicans</i> have >5 times higher odds of having ECC compared to those without <i>C. albicans</i>. Further prospective cohort studies are needed to determine whether <i>C. albicans </i>could be a risk factor for ECC, and whether it is dependent on different sample sources (saliva/plaque).</p