Analysis of proteoglycan expression in human dental pulp

Nahás-Scocate, Ana Carla Raphaelli; Moraes, Gabrielle Ferrante Alves de; Nader, Helena B.; Vicente, Carolina Meloni; Toma, Leny

doi:10.1016/j.archoralbio.2018.03.003

Cited by 7 publications

(3 citation statements)

References 35 publications

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“…15 Throughout the tooth developmental process, the involvement of versican, a large CS-PG; 16 perlecan, a large heparan sulfate (HS)-PG; 17 and syndecans, small cell surface HS-PGs, 18 have been thoroughly investigated. Several studies have also confirmed the involvement of SLRPs in tooth development in humans 13,19,20 and other species. [21][22][23] SLRPs have been proposed to play several roles in tooth development, including cellular differentiation, collagen fibrillogenesis, dental mineralization, and tooth eruption.…”

Section: Introductionmentioning

confidence: 74%

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Expression, localization and synthesis of small leucine-rich proteoglycans in developing mouse molar tooth germ

2020

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The gene expression and protein synthesis of small leucine-rich proteoglycans (SLRPs), including decorin, biglycan, fibromodulin, and lumican, was analyzed in the context of the hypothesis that they are closely related to tooth formation. In situ hybridization, immunohistochemistry, and organ culture with metabolic labeling of [35S] were carried out in mouse first molar tooth germs of different developmental stages using ICR mice at embryonic day (E) 13.5 to postnatal day (P) 7.0. At the bud and cap stage, decorin mRNA was expressed only in the surrounding mesenchyme, but not within the tooth germ. Biglycan mRNA was then expressed in the condensing mesenchyme and the dental papilla of the tooth germ. At the apposition stage (late bell stage), both decorin and biglycan mRNA were expressed in odontoblasts, resulting in a switch of the pattern of expression within the different stages of odontoblast differentiation. Decorin mRNA was expressed earlier in newly differentiating odontoblasts than biglycan. With odontoblast maturation and dentin formation, decorin mRNA expression was diminished and localized to the newly differentiating odontoblasts at the cervical region. Simultaneously, biglycan mRNA took over and extended its expression throughout the new and mature odontoblasts. Both mRNAs were expressed in the dental pulp underlying the respective odontoblasts. At P7.0, both mRNAs were weakly expressed but maintained their spatial expression patterns. Immunostaining showed that biglycan was localized in the dental papillae and pulp. In addition, all four SLRPs showed clear immunostaining in predentin, although the expressions of fibromodulin and lumican mRNAs were not identified in the tooth germs examined. The organ culture data obtained supported the histological findings that biglycan is more predominant than decorin at the apposition stage. These results were used to identify biglycan as the principal molecule among the SLRPs investigated. Our findings indicate that decorin and biglycan show spatial and temporal differential expressions and play their own tissue-specific roles in tooth development.

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Section: Introductionmentioning

confidence: 74%

“…26 Many immunohistochemical studies in tooth germs have also been performed in rodents 21,27 and humans. 13,20 Several in situ hybridization studies in mice have also been carried out. [28][29][30] These studies, however, have not examined the temporal changes in SLRP expression during the process of identical tooth germ formation.…”

Section: Introductionmentioning

confidence: 99%

Expression, localization and synthesis of small leucine-rich proteoglycans in developing mouse molar tooth germ

2020

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“…The 2−ΔΔCt method was used for calculating fold change in gene expression levels versus untreated controls. All the primers were synthesized by Sigma-Aldrich as previously described in Supplemental Table 1 ( 22–27 ).…”

Section: Methodsmentioning

confidence: 99%

Human Milk Oligosaccharides Mediate the Crosstalk Between Intestinal Epithelial Caco-2 Cells and Lactobacillus Plantarum WCFS1in an In Vitro Model with Intestinal Peristaltic Shear Force

Kong

Cheng

Krenning

et al. 2020

The Journal of Nutrition

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Background The intestinal epithelial cells, food molecules, and gut microbiota are continuously exposed to intestinal peristaltic shear force. Shear force may impact the crosstalk of human milk oligosaccharides (hMOs) with commensal bacteria and intestinal epithelial cells. Objectives We investigated how hMOs combined with intestinal peristaltic shear force impact intestinal epithelial cells and crosstalk with a commensal bacterium. Methods We applied the Ibidi system to mimic intestinal peristaltic shear force. Caco-2 cells were exposed to a shear force (5 dynes/cm2) for 3 d, and then stimulated with the hMOs, 2′-fucosyllactose (2′-FL), 3-FL, and lacto-N-triose II (LNT2). In separate experiments, Lactobacillus plantarumWCFS1 adhesion to Caco-2 cells was studied with the same hMOs and shear force. Effects were tested on gene expression of glycocalyx-related molecules (glypican 1 [GPC1], hyaluronan synthase 1 [HAS1], HAS2, HAS3, exostosin glycosyltransferase 1 [EXT1], EXT2), defensin β-1 (DEFB1), and tight junction (tight junction protein 1 [TJP1], claudin 3 [CLDN3]) in Caco-2 cells. Protein expression of tight junctions was also quantified. Results Shear force dramatically decreased gene expression of the main enzymes for making glycosaminoglycan side chains (HAS3 by 43.3% and EXT1 by 68.7%) (P <0.01), but did not affect GPC1 which is the gene responsible for the synthesis of glypican 1 which is a major protein backbone of glycocalyx. Expression of DEFB1, TJP1, and CLDN3 genes was decreased 60.0–94.9% by shear force (P <0.001). The presence of L. plantarumWCFS1 increased GPC1, HAS2, HAS3, and ZO-1 expression by 1.78- to 3.34-fold (P <0.05). Under shear force, all hMOs significantly stimulated DEFB1 and ZO-1, whereas only 3-FL and LNT2 enhanced L. plantarumWCFS1 adhesion by 1.85- to 1.90-fold (P <0.01). Conclusions 3-FL and LNT2 support the crosstalk between the commensal bacterium L. plantarumWCFS1 and Caco-2 intestinal epithelial cells, and shear force can increase the modulating effects of hMOs.

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