Table 1.
Laminin subunits assessed in the study.
Fig 1.
As expected, Von Kossa assay demonstrated that both, deciduous (DecPDL) and permanent (PermPDL) cells produced mineral nodules in vitro when cultured under osteogenic conditions.
Fig 2.
Summary of secretome analysis of the PDL cells harvested from deciduous and permanent teeth.
(A) Venn diagram of total proteins and their top enriched molecular function (MF) GO terms. The MF GO terms were generated by DAVID software based on biological processes (GO_TERM_MF_2 database). (B) Volcano plot analysis of deciduous and permanent PDL cells secretome. Regulated proteins are painted in blue and orange, for deciduous and permanent teeth, respectively (p<0.05).
Table 2.
List of differentially expressed proteins in permanent and deciduous determined by beta-binomial statistical test (p-value < 0.05).
Fig 3.
Gene Ontology (GO) enrichment analysis of the secretome of deciduous and permanent PDL cells.
GO analysis was generated by the DAVID software based on biological processes (GO_TERM_BP_2 database). (A) and (B) GO biological process (BP) analysis from permanent and deciduous PDL cells secretome, respectively. (C) Barplot of normalized spectral count of differentially expressed proteins in permanent PDL cells with GO BP anatomical structure formation involved in morphogenesis and GO BP regulation of developmental process (mean ± standard deviation and p-value determined by beta-binomial statistical test). (D) Barplot of normalized spectral count of differentially expressed proteins of deciduous PDLs involved with GO BP cell adhesion (mean ± standard deviation and p-value determined by beta-binomial statistical test).
Fig 4.
LAMC1 (laminin subunit gamma 1) protein levels in fresh PDL tissues.
Western blotting analysis (four representative samples for each group) of the protein levels of LAMC1 in fresh PDL tissues harvested from deciduous (DecPDL) and permanent (PermPDL) teeth. Bar graph represents average and standard deviation of six samples per group. * indicates statistical difference by Student’s t-test with α = 5%.
Fig 5.
Gene expression in fresh PDL tissues for laminin chains.
qPCR analysis of mRNA levels for laminin (LAM) chains in fresh PDL tissues harvested from deciduous (DecPDL) and permanent (PermPDL) PDL tissues reveals that deciduous teeth presented higher levels of LAMB1 (laminin subunit beta 1), LAMB3 (laminin subunit beta 3), LAMC1 (laminin subunit gamma 1), and LAMC2 (laminin subunit gamma 2), and lower mRNA levels of LAMB2 (laminin subunit beta 2). * indicates statistical difference by Student’s t-test with α = 5%.
Fig 6.
Histological illustration of deciduous and permanent teeth from mini pigs stained by Mallory’s trichrome.
The general structure of the periodontium and collagen organization is shown in a panoramic view of the lingual face of deciduous (A) and permanent (C) teeth. The areas delimited by yellow squares are shown in columns B and D in higher magnification of the cervical, middle third and apical region of deciduous and permanent teeth, respectively. A and C, bars = 500 μm; B and D, bars = 100 μm. G—gingiva; D—dentin; B—bone; C—cementum; PDL—periodontal ligament.
Fig 7.
Polarizing and bright-field images from picrosirius red stained sections from deciduous and permanent teeth.
Bright field images of the cervical, middle third and apical regions of deciduous (A) and permanent (C) teeth, and their respective birefringence in B and D. The arrow at 45° with the polarizer and analyzer indicates position of maximum birefringence. Magnification = 10x. Bars = 100 μm. D—dentin; B—bone; C—cementum; PDL—periodontal ligament.