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Effects of conservatively treated diseased cementum with or without EMD on in vitro cementoblast differentiation and in vivo cementum-like tissue formation of human periodontal ligament cells.

ABSTRACT: OBJECTIVES:The present study aimed to evaluate the effects of conservatively treated diseased cementum on in vitro cementoblast differentiation and in vivo cementum-like tissue formation of human periodontal ligament cells (hPDLCs), and observe differential effects of enamel matrix derivative (EMD) on in vivo cementum formation by hPDLCs. MATERIALS AND METHODS:Forty-eight cementum slices and 48 dentin slices were prepared from periodontitis compromised teeth, and hPDLCs were inoculated on to all root slices. Twenty-four co-cultured root slices of each group were used for mRNA expression of cementum attachment protein and CEMP1. With application of EMD, 24 co-cultured root slices (divided into groups C, D, C+E, D+E) were transplanted subcutaneously into nude mice. All root fragments were reviewed by histological analysis and immunohistochemical staining for bone sialoprotein. RESULTS:mRNA expressions of cementum attachment protein and cementum protein - 1 from hPDLCs on cementum slices were statistically higher than those of dentin slices. Seven specimens of group C and 10 specimens of group C+E revealed a layer of cementum-like tissue (NFC) on surfaces of pre-existing cementum. NFC was thicker in group C+E than in group C. All NFCs were positively stained for bone sialoprotein, however, there was no NFC formation on dentin slices. CONCLUSION:Conservatively treated diseased cementum promoted in vitro cementoblast differentiation and in vivo cementum-like tissue formation by hPDLCs, and the in vivo effect was enhanced by the presence of EMD.

SUBMITTER: Qi Y

PROVIDER: S-EPMC6496275 | biostudies-literature | 2014 Aug

REPOSITORIES: biostudies-literature

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Effects of conservatively treated diseased cementum with or without EMD on in vitro cementoblast differentiation and in vivo cementum-like tissue formation of human periodontal ligament cells.

Qi Y Y Feng W W Cai J J Sun Q Q Li S S Li M M Song A A Yang P P

Cell proliferation 20140616 4

<h4>Objectives</h4>The present study aimed to evaluate the effects of conservatively treated diseased cementum on in vitro cementoblast differentiation and in vivo cementum-like tissue formation of human periodontal ligament cells (hPDLCs), and observe differential effects of enamel matrix derivative (EMD) on in vivo cementum formation by hPDLCs.<h4>Materials and methods</h4>Forty-eight cementum slices and 48 dentin slices were prepared from periodontitis compromised teeth, and hPDLCs were inocu ...[more]

PMID: 24930868

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Project description:The structural and functional integrity of bone-periodontal ligament (PDL)-cementum complex stems from the load-bearing attachment sites (entheses) between soft (PDL) and hard (bone, cementum) tissues. These attachment sites are responsible for the maintenance of a bone-PDL-cementum complex biomechanical function. The objective was to investigate changes in spatiotemporal expression of key biomolecules in developing and functionally active entheses.Multilabeling technique was performed on hemimandibles of 3 wk and 3 mo-old scleraxis-GFP transgenic mice for CD146, CD31, NG2, osterix and bone sialoprotein. Regions of dominant stretch within the PDL were evaluated by identifying directionality of collagen fibrils, PDL fibroblasts and PDL cell cytoskeleton.CD146+ cells adjacent to CD31+ vasculature were identified at PDL-bone enthesis. NG2+ cells were located at coronal bone-PDL and apical cementum-PDL entheses in the 3-wk-old group, but at 3 mo, NG2 was positive at the entheses of the apical region and alveolar crest. NG2 and osterix were colocalized at the osteoid and cementoid regions of the PDL-bone and PDL-cementum entheses. Bone sialoprotein was prominent at the apical region of 3-wk-old mice. The directionality of collagen fibers, fibroblasts and their cytoskeleton overlapped, except in the apical region of 3 wk.Colocalization of biomolecules at zones of the PDL adjacent to attachment sites may be essential for the formation of precementum and osteoid interfaces at a load-bearing bone-PDL-tooth fibrous joint. Biophysical cues resulting from development and function can regulate recruitment and differentiation of stem cells potentially from a vascular origin toward osteo- and cemento-blastic lineages at the PDL-bone and PDL-cementum entheses. Investigating the coupled effect of biophysical and biochemical stimuli leading to cell differentiation at the functional attachment sites is critical for developing regeneration strategies to enable functional reconstruction of the periodontal complex.

Project description:Background and objectiveAdaptive properties of the bone-periodontal ligament-tooth complex have been identified by changing the magnitude of functional loads using small-scale animal models, such as rodents. Reported adaptive responses as a result of lower loads due to softer diet include decreased muscle development, change in structure-function relationship of the cranium, narrowed periodontal ligament space, and changes in the mineral level of the cortical bone and alveolar jaw bone and in the glycosaminoglycans of the alveolar bone. However, the adaptive role of the dynamic bone-periodontal ligament-cementum complex to prolonged reduced loads has not been fully explained to date, especially with regard to concurrent adaptations of bone, periodontal ligament and cementum. Therefore, in the present study, using a rat model, the temporal effect of reduced functional loads on physical characteristics, such as morphology and mechanical properties and the mineral profiles of the bone-periodontal ligament-cementum complex was investigated.Material and methodsTwo groups of 6-wk-old male Sprague-Dawley rats were fed nutritionally identical food with a stiffness range of 127-158 N/mm for hard pellet or 0.3-0.5 N/mm for soft powder forms. Spatio-temporal adaptation of the bone-periodontal ligament-cementum complex was identified by mapping changes in the following: (i) periodontal ligament collagen orientation and birefringence using polarized light microscopy, bone and cementum adaptation using histochemistry, and bone and cementum morphology using micro-X-ray computed tomography; (ii) mineral profiles of the periodontal ligament-cementum and periodontal ligament-bone interfaces by X-ray attenuation; and (iii) microhardness of bone and cementum by microindentation of specimens at ages 6, 8, 12 and 15 wk.ResultsReduced functional loads over prolonged time resulted in the following adaptations: (i) altered periodontal ligament orientation and decreased periodontal ligament collagen birefringence, indicating decreased periodontal ligament turnover rate and decreased apical cementum resorption; (ii) a gradual increase in X-ray attenuation, owing to mineral differences, at the periodontal ligament-bone and periodontal ligament-cementum interfaces, without significant differences in the gradients for either group; (iii) significantly (p < 0.05) lower microhardness of alveolar bone (0.93 ± 0.16 GPa) and secondary cementum (0.803 ± 0.13 GPa) compared with the higher load group insert bone = (1.10 ± 0.17 and cementum = 0.940 ± 0.15 GPa, respectively) at 15 wk, indicating a temporal effect of loads on the local mineralization of bone and cementum.ConclusionBased on the results from this study, the effect of reduced functional loads for a prolonged time could differentially affect morphology, mechanical properties and mineral variations of the local load-bearing sites in the bone-periodontal ligament-cementum complex. These observed local changes in turn could help to explain the overall biomechanical function and adaptations of the tooth-bone joint. From a clinical translation perspective, our study provides an insight into modulation of load on the complex for improved tooth function during periodontal disease and/or orthodontic and prosthodontic treatments.

Project description:Periodontitis is a common chronic inflammatory oral disease. The objective of periodontal treatment is to control infection whilst regenerating damaged periodontal tissue. The present study aimed to determine the potential effects of pterostilbene (PTE), a representative stilbene compound, on the proliferation and differentiation of human periodontal ligament stem cells (hPDLSCs). Different concentrations (1, 5, 10 and 20 µM) of PTE were applied to hPDLSCs, after which Cell Counting Kit-8 and western blotting assays were performed to examine the protein levels of Ki67, PCNA, p-IκBα, IκBα, Bcl-2, Bax, cleaved caspase3. The effect of PTE on the release of inflammatory factors, including IL-1β, IL-6 and IL-10 was assessed by RT-qPCR. The apoptosis of TNF-α-induced hPDLSCs was evaluated by TUNEL assay and western blotting. Additionally, the role of PTE in hPDLSC mineralization was evaluated using alizarin red staining. The expression levels of mineralization indices, including RUNX2 and ALP were subsequently determined using western blotting. Subsequently, the target of PTE was predicted using TargetNet database and AutoDock v4.2 software and verified using western blotting. The results of the present study revealed that PTE promoted the proliferation of hPDLSCs in a concentration-dependent manner. Furthermore, PTE treatment decreased the release of inflammatory factors and alleviated the apoptosis of TNF-α-induced hPDLSCs. PTE was also demonstrated to promote the formation of mineral nodules in TNF-α-induced hPDLSCs. The Targetnet database, along with molecular docking, indicated that histone deacetylases (HDACs) were the probable targets of PTE upstream of regulating periodontitis. The results of western blotting implied that TNF-α significantly increased expression levels of HDAC2, 4, 6 and 8, whilst PTE treatment markedly decreased HDAC4, 6 and 8 expression in a concentration-dependent manner compared with the TNF-α group, which further confirmed these conclusions. In summary, results of the present study revealed that PTE promoted TNF-α-induced hPDLSC proliferation and differentiation, whilst alleviating inflammation and apoptosis. PTE also inhibited the expression of HDACs, which may be involved in the mechanism of periodontitis.

Dataset Information

Effects of conservatively treated diseased cementum with or without EMD on in vitro cementoblast differentiation and in vivo cementum-like tissue formation of human periodontal ligament cells.

Publications

Effects of conservatively treated diseased cementum with or without EMD on in vitro cementoblast differentiation and in vivo cementum-like tissue formation of human periodontal ligament cells.

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