Additional file 1 of Quercetin-loaded mesoporous nano-delivery system remodels osteoimmune microenvironment to regenerate alveolar bone in periodontitis via the miR-21a-5p/PDCD4/NF-κB pathway
Creators: name: Yang, Shi-Yuan
name: Hu, Yue
name: Zhao, Ran
name: Zhou, Yu-Ning
name: Zhuang, Yu
name: Zhu, Yan
name: Ge, Xiao-Li
name: Lu, Ting-Wei
name: Lin, Kai-Li
name: Xu, Yuan-Jin
Publisher: name: figshare
Year: 2024
Subjects: subject: Biochemistry
subject: Space Science
subject: Medicine
subject: Microbiology
subject: FOS: Biological sciences
subject: Molecular Biology
subject: Pharmacology
subject: Environmental Sciences not elsewhere classified
subject: Chemical Sciences not elsewhere classified
subject: Immunology
subject: FOS: Clinical medicine
subject: Biological Sciences not elsewhere classified
subject: Information Systems not elsewhere classified

description: Additional file 1: Table S1. qRT-PCR primer sequences used in this study. Table S2. qRT-PCR primer sequences used in this study. Fig S1. (A-B) The H &amp; E and Masson staining of samples collected in 8 weeks. Fig S2. (A-B) Proliferation results of PDLSCs incubated with quercetin for 1,4 and 7 days in both physiological and periodontitis environment by using CCK-8 assay. (C) Live/Dead staining of PDLSCs on day 1 under the stimulation of quercetin under periodontitis environment. (#p &lt; 0.05, ##p &lt; 0.01 and ###p &lt; 0.001 compared to Pg.LPS (-) + Quercetin (-); *p &lt; 0.05, **p &lt; 0.01 and ***p &lt; 0.001 compared to Pg.LPS (+) + Quercetin (-); Data re represented as the mean ± SEM, n=3). Fig S3. (A-B) The protein level of PDCD4 in PDLSCs incubated with quercetin for 7 days under periodontitis environment by western blot. (#p &lt; 0.05, ##p &lt; 0.01 and ###p &lt; 0.001 compared to Pg.LPS (-) +Quercetin (-); *p &lt; 0.05, **p &lt; 0.01 and ***p &lt; 0.001 compared to Pg.LPS (+) +Quercetin (-); Data re represented as the mean ± SEM, n=3). Fig S4. (A-C) The expression of inflammation-related genes and protein of PDLSCs under periodontitis microenvironment after incubation with quercetin for 1 day. (#p &lt; 0.05, ##p &lt; 0.01 and ###p &lt; 0.001 compared to Pg.LPS (-) +Quercetin (-); *p &lt; 0.05, **p &lt; 0.01 and ***p &lt; 0.001 compared to Pg.LPS (+) +Quercetin (-); Data re represented as the mean ± SEM, n=3). Fig S5. (A) The Go enrichment analysis of Pg.LPS（+） + Quercetin（-） vs Pg.LPS（+） + Quercetin（+） group. (B)Immunofluorescence images of the p-P65 protein expression in macrophages. (C) The miR-21a-5p expression in RAW264.7 transfected by mir-21a-5p mimic NC, mimic, inhibitor NC and inhibitor. (D-G) qRT-PCR, western blot and immunofluorescence staining of the PDCD4 among Pg.LPS (-) +Quercetin (-), Pg.LPS (+) +Quercetin (-) and Pg.LPS (+) +Quercetin (+) groups. (H-I) The protein level of PDCD4 in siNC, siPDCD4-1-, siPDCD4-2- or siPDCD4-3-transfected macrophages determined by western blot. (In Fig. S5 D-F, #p &lt; 0.05, ##p &lt; 0.01 and ###p &lt; 0.001 compared to Pg.LPS (-) + Quercetin (-) group and *p &lt; 0.05, **p &lt; 0.01 and ***p &lt; 0.001 compared to Pg.LPS (+) + Quercetin (-) group. In Fig. S5 C and Fig. S6 I, *p &lt; 0.05, **p &lt; 0.01 and ***p &lt; 0.001; Data are represented as the mean ± SEM, n=3). Fig S6. (A-B) ALP staining and its quantitative results after incubation of PDLSCs with MBG, Quercetin and Quercetin/MBG for 7 days under periodontitis microenvironment. (C-F) Effect of MBG, Quercetin and Quercetin/MBG on the expression of osteogenic-related genes (OPN and RUNX-2) and angiogenic-related genes (VEGF and bFGF) of PDLSCs under periodontitis microenvironment through qRT-PCR. (G-J) Effect of MBG, Quercetin and Quercetin/MBG on the expression of inflammatory genes (IL-1β, IL-6, iNOS and TNF-α) of RAW264.7 under periodontitis microenvironment through qRT-PCR. (K) Effect of MBG, Quercetin and Quercetin/MBG on the expression of miR-21a-5p of RAW264.7 under periodontitis microenvironment through qRT-PCR. (L) Schematic of Quercetin/MBG on PDLSCs and RAW264.7 under periodontitis microenvironment. (ns, no significant difference; *p &lt; 0.05, **p &lt; 0.01 and ***p &lt; 0.001; Data are represented as the mean ± SEM, n=3).