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Pre-existing root cementum may promote cementoblast differentiation of human periodontal ligament cells.

ABSTRACT: OBJECTIVES:To observe whether preserved healthy cementum could promote differentiation of human periodontal ligament cells to cementoblasts. MATERIALS AND METHODS:Symmetrical root slices from each healthy premolar were distributed into either the control group (cementum removed) or test group (cementum preserved). After isolation and characterization, human periodontal ligament cells were inoculated onto root slices for 7 days co-culture. Two slices per group were studied for cell morphology by scanning electronic microscopy. Twenty-three slices were detected for expression of cementum attachment protein and cementum protein 23, two putative cementoblast markers, by real-time polymerase chain reaction. Twenty slices were transplanted into nude mice and analysed using histology and immunohistochemistry for osteopontin and bone sialoprotein expression after 8 weeks. RESULTS:Cells of the test group had smoother fibroblast morphology and higher cementum protein 23 and cementum attachment protein expression than those of the control group (P < 0.01). In the test group, 14 root slices revealed cementum-like matrix formation resting on old cementum; no splits were observed between newly formed matrix and old cementum. In the control group, 17 specimens had fibrous tissue formation along the root surface and varying width of splits could be seen between new fibrous tissue and dentine surface. Only three specimens demonstrated presence of newly formed thin cementum-like matrix. Newly formed cementum-like matrix was positive for osteopontin and bone sialoprotein. CONCLUSIONS:The results demonstrate that healthy root cementum may promote differentiation of human periodontal ligament cells towards cementoblasts.

SUBMITTER: Song A

PROVIDER: S-EPMC6495954 | biostudies-literature | 2012 Jun

REPOSITORIES: biostudies-literature

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Pre-existing root cementum may promote cementoblast differentiation of human periodontal ligament cells.

Song A A Cai J J Pan K K Yang P P

Cell proliferation 20120327 3

<h4>Objectives</h4>To observe whether preserved healthy cementum could promote differentiation of human periodontal ligament cells to cementoblasts.<h4>Materials and methods</h4>Symmetrical root slices from each healthy premolar were distributed into either the control group (cementum removed) or test group (cementum preserved). After isolation and characterization, human periodontal ligament cells were inoculated onto root slices for 7 days co-culture. Two slices per group were studied for cell ...[more]

PMID: 22452325

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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:Adaptive 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.Two 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.Reduced 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.Based 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.

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Pre-existing root cementum may promote cementoblast differentiation of human periodontal ligament cells.

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Pre-existing root cementum may promote cementoblast differentiation of human periodontal ligament cells.

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