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BMC Oral Health Jan 2023Delphinidin (DP), an anthocyanidin found in blueberries, has antioxidant and anti-inflammatory effects. This study aimed to investigate the efficacy of DP as a storage...
BACKGROUND
Delphinidin (DP), an anthocyanidin found in blueberries, has antioxidant and anti-inflammatory effects. This study aimed to investigate the efficacy of DP as a storage medium for avulsed teeth.
METHODS
Human periodontal ligament cells were cultured and exposed to DP solution (10, 50, and 100 μM), Dulbecco's modified Eagle's medium, Hank's balanced salt solution and tap water. Cell counting kit-8 assays were performed after 0.5, 1, 6, and 24 h to measure the cell viability. Nitric oxide assays and gelatin zymography were performed to evaluate the anti-inflammatory effects of DP. Reverse transcription-polymerase chain reaction was used to determine the expression levels of inflammatory cytokines.
RESULTS
The viability of periodontal ligament cells was greatest at 100 μM DP. At 1 h, 100 μM DP decreased nitric oxide synthesis (p < .0167). Matrix metallopeptidase-9 activity was inhibited by DP in a dose-dependent manner (p < .0167). Moreover, treatment with 100 μM DP decreased the expression levels of tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-8 in periodontal ligament cells (p < .0167).
CONCLUSIONS
Within the limits of this study, DP preserved the viability and suppressed the inflammatory response of periodontal ligament cells. These findings suggest that DP could be promising for preservation of avulsed teeth.
Topics: Humans; Anti-Inflammatory Agents; Cell Survival; Nitric Oxide; Organ Preservation Solutions; Periodontal Ligament; Tooth Avulsion
PubMed: 36641447
DOI: 10.1186/s12903-023-02713-9 -
Journal of Dental Research Feb 2021The most fundamental function of an epithelial tissue is to act as a barrier, regulating interactions between the external environment and the body. This barrier...
The most fundamental function of an epithelial tissue is to act as a barrier, regulating interactions between the external environment and the body. This barrier function typically requires a contiguous cell layer but since teeth penetrate the oral epithelium, a modified barrier has evolved, called the junctional epithelium (JE). In health, the JE attaches to the tooth, sealing the inside of the body against oral micro-organisms. Breakdown of the JE barrier results in periodontal ligament (PDL) disintegration, alveolar bone resorption, and ultimately tooth loss. Using lineage tracing and DNA pulse-chase analyses, we identified an anatomical location in the JE that supported both fast- and slow-cycling Wnt-responsive stem cells that contributed to self-renewal of the tissue. Stem cells produced daughter cells with an extraordinarily high rate of turnover that maintained JE integrity for 1.4 y in mice. Blocking cell proliferation via a chemotherapeutic agent 5-fluorouracil (5-Fu) eliminated fast-cycling stem cells, which caused JE degeneration, PDL destruction, and bone resorption. Upon removal of 5-Fu, slow-cycling stem cells regenerated both the structure and barrier function of the JE. Taken together, our studies identified a stem cell population in the JE and have potential clinical implications for prevention and treatment of periodontitis.
Topics: Animals; Epithelial Attachment; Epithelium; Gingiva; Mice; Periodontal Ligament; Stem Cells; Tooth
PubMed: 32985318
DOI: 10.1177/0022034520960125 -
European Cells & Materials Oct 2021Due to the complexity of the structure of the tooth periodontium, regeneration of the full tooth attachment is not a trivial task. There is also a gap in models that can...
Due to the complexity of the structure of the tooth periodontium, regeneration of the full tooth attachment is not a trivial task. There is also a gap in models that can represent human tooth attachment in vitro and in vivo. The aim of this study was to develop a bilayered in vitro construct that simulated the tooth periodontal ligament and attached alveolar bone, for the purpose of tissue regeneration and investigation of physiological and orthodontic loading. Two types of materials were used to develop this construct: sol-gel 60S10Mg derived scaffold, representing the hard tissue component of the periodontium, and commercially available Geistlich Bio-Gide® collagen membrane, representing the soft tissue component of the tooth attachment. Each scaffold was dynamically seeded with human periodontal ligament cells (HPDLCs). Scaffolds were either cultured separately, or combined in a bilayered construct, for 2 weeks. Characterisation of the individual scaffolds and the bilayered constructs included biological characterisation (cell viability, scanning electron microscopy to confirm cell attachment, gene expression of periodontium regeneration markers), and mechanical characterisation of scaffolds and constructs. HPDLCs enjoyed a biocompatible 3-dimensional environment within the bilayered construct components. There was no drop in cellular gene expression in the bilayered construct, compared to the separate scaffolds.
Topics: Humans; Periodontal Ligament; Periodontium; Tissue Engineering; Tissue Scaffolds; Tooth
PubMed: 34632563
DOI: 10.22203/eCM.v042a17 -
International Journal of Molecular... Jan 2021The aim of the study was to clarify whether orthodontic forces and periodontitis interact with respect to the anti-apoptotic molecules superoxide dismutase 2 (SOD2) and...
The aim of the study was to clarify whether orthodontic forces and periodontitis interact with respect to the anti-apoptotic molecules superoxide dismutase 2 (SOD2) and baculoviral IAP repeat-containing protein 3 (BIRC3). SOD2, BIRC3, and the apoptotic markers caspases 3 (CASP3) and 9 (CASP9) were analyzed in gingiva from periodontally healthy and periodontitis subjects by real-time PCR and immunohistochemistry. SOD2 and BIRC3 were also studied in gingiva from rats with experimental periodontitis and/or orthodontic tooth movement. Additionally, SOD2 and BIRC3 levels were examined in human periodontal fibroblasts incubated with and/or subjected to mechanical forces. Gingiva from periodontitis patients showed significantly higher SOD2, BIRC3, CASP3, and CASP9 levels than periodontally healthy gingiva. SOD2 and BIRC3 expressions were also significantly increased in the gingiva from rats with experimental periodontitis, but the upregulation of both molecules was significantly diminished in the concomitant presence of orthodontic tooth movement. In vitro, SOD2 and BIRC3 levels were significantly increased by , but this stimulatory effect was also significantly inhibited by mechanical forces. Our study suggests that SOD2 and BIRC3 are produced in periodontal infection as a protective mechanism against exaggerated apoptosis. In the concomitant presence of orthodontic forces, this protective anti-apoptotic mechanism may get lost.
Topics: Animals; Apoptosis; Baculoviral IAP Repeat-Containing 3 Protein; Caspase 3; Caspase 9; Cells, Cultured; Fibroblasts; Fusobacterium nucleatum; Gene Expression Regulation; Gingiva; Host-Pathogen Interactions; Humans; Periodontal Ligament; Periodontium; Rats; Superoxide Dismutase
PubMed: 33435582
DOI: 10.3390/ijms22020591 -
Journal of Dental Research Dec 2021The WNT/β-catenin signaling pathway plays a central role in the biology of the periodontium, yet the function of specific extracellular WNT ligands remains poorly...
The WNT/β-catenin signaling pathway plays a central role in the biology of the periodontium, yet the function of specific extracellular WNT ligands remains poorly understood. By using a inducible transgenic mouse model targeting -expressing alveolar osteoblasts, odontoblasts, and cementoblasts, we demonstrate that the WNT ligand WNT1 is a strong promoter of cementum and alveolar bone formation in vivo. We induced expression for 1, 3, or 9 wk in Wnt1Tg mice and analyzed them at the age of 6 wk and 12 wk. Micro-computed tomography (CT) analyses of the mandibles revealed a 1.8-fold increased bone volume after 1 and 3 wk of expression and a 3-fold increased bone volume after 9 wk of expression compared to controls. In addition, the alveolar ridges were higher in Wnt1Tg mice as compared to controls. Nondecalcified histology demonstrated increased acellular cementum thickness and cellular cementum volume after 3 and 9 wk of expression. However, 9 wk of expression was also associated with periodontal breakdown and ectopic mineralization of the pulp. The composition of this ectopic matrix was comparable to those of cellular cementum as demonstrated by quantitative backscattered electron imaging and immunohistochemistry for noncollagenous proteins. Our analyses of 52-wk-old mice after 9 wk of expression revealed that expression affects mandibular bone and growing incisors but not molar teeth, indicating that influences only growing tissues. To further investigate the effect of on cementoblasts, we stably transfected the cementoblast cell line (OCCM-30) with a vector expressing -HA and performed proliferation as well as differentiation experiments. These experiments demonstrated that promotes proliferation but not differentiation of cementoblasts. Taken together, our findings identify, for the first time, as a critical regulator of alveolar bone and cementum formation, as well as provide important insights for harnessing the WNT signal pathway in regenerative dentistry.
Topics: Animals; Cementogenesis; Dental Cementum; Mice; Osteogenesis; Periodontal Ligament; X-Ray Microtomography
PubMed: 34009051
DOI: 10.1177/00220345211012386 -
Mediators of Inflammation 2020Resistin, a proinflammatory adipokine, is elevated in many inflammatory diseases. However, little is known about its performance in periodontitis. The present study is...
Resistin, a proinflammatory adipokine, is elevated in many inflammatory diseases. However, little is known about its performance in periodontitis. The present study is aimed at evaluating resistin expression and synthesis in periodontal cells and tissues under inflammatory/microbial stress in addition to its effects on the periodontium. In vivo, 24 male rats were randomly divided into two groups: control and ligature-induced periodontal disease. After 6 and 12 days, animals were sacrificed to analyze gene expression of adipokines, bone loss, inflammation, and resistin synthesis. In vitro, human periodontal ligament (PDL) fibroblasts were used to evaluate the expression of resistin after inflammatory stimuli. In addition, PDL fibroblasts were exposed to resistin to evaluate its role on soft and hard tissue metabolism markers. The periodontitis group demonstrated significant bone loss, an increase in the number of inflammatory cells and vascular structures, an increase in resistin expression and synthesis, and a decrease in the expression of adiponectin, leptin, and its functional receptor. PDL fibroblasts showed a significant increase in resistin expression and synthesis in response to the inflammatory stimulus by IL-1. Resistin induced an increase in cytokine expression and a decrease in the regulation of some hard tissue and matrix formation genes in PDL fibroblasts. These data indicate that resistin is produced by periodontal cells and tissues, and this effect is enhanced by inflammatory stimuli. Moreover, resistin seems to interfere with soft and hard tissue metabolism during periodontitis by reducing markers related to matrix formation and bone tissue.
Topics: Animals; Bone and Bones; Fibroblasts; Gingiva; Humans; Inflammation; Periodontal Ligament; Periodontitis; Periodontium; Phenotype; Rats; Resistin
PubMed: 32410876
DOI: 10.1155/2020/9817095 -
Odontology Jul 2019The epithelial rests of Malassez (ERMs) might represent a valuable source of oral epithelial cells with stem cell properties. The purpose of this study was to isolate...
The epithelial rests of Malassez (ERMs) might represent a valuable source of oral epithelial cells with stem cell properties. The purpose of this study was to isolate and characterize cells derived from human ERM, and compare them with cells derived from matched normal oral mucosa (NOM). Matched tissue specimens of the periodontal ligament of extracted tooth and NOM were collected. Cells were isolated in culture, then characterized by immunohistochemistry and flow cytometry for expression of pancytokeratin, ESA, PDGFRB, CD31 and CD44. 3D organotypic cultures were constructed by growing epithelial cells on top of fibroblast-populated collagen gels. Both ERM and NOM-isolated cells expressed the markers of epithelial lineage (ESA and pancytokeratin), and to some extent PDGFR, an indicator of a more mesenchymal phenotype, but not the endothelial cell marker CD31. Cells with epithelial morphology were isolated from periodontium of cervical, middle and apical parts of the root, but contained a significantly lower percentage of ESA and pancytokeratin-positive cells than when isolating cells from NOM (p < 0.001). ERM cells expressed a significantly higher percentage of the stem cell-related molecule CD44 (cervical 92.93 ± 0.25%, middle 93.8 ± 0.26%, apical 94.36 ± 0.41%) than cells isolated from NOM (27.8 ± 1.47%, p < 0.001). When grown in 3D organotypic cultures and in collagen gels, ERM cells formed a less differentiated epithelium than NOM cells, but expressing pancytokeratin and vimentin. In conclusion, epithelial cells could be isolated from human periodontium and grown in culture; their in vitro characterization indicates that they have a less differentiated phenotype compared with cells derived from normal oral epithelium.
Topics: Cells, Cultured; Epithelial Cells; Fibroblasts; Humans; Periodontal Ligament; Rest
PubMed: 30478679
DOI: 10.1007/s10266-018-0397-7 -
Life Sciences Sep 2021Stimulation of β-adrenergic receptors (βAR) in osteoblasts by isoproterenol (ISO) was shown to induce Vascular Endothelial Growth Factor (VEGF) and angiogenesis in...
AIMS
Stimulation of β-adrenergic receptors (βAR) in osteoblasts by isoproterenol (ISO) was shown to induce Vascular Endothelial Growth Factor (VEGF) and angiogenesis in long bones. We thus aimed to determine the vascular response of mandibular tissues to βAR stimulation regarding blood vessel formation.
MAIN METHODS
Six-week-old wild-type C57BL6 female mice received daily intraperitoneal injections of ISO or phosphate buffered saline (PBS) for 1 month. Hemimandibles and tibias were collected for immunolocalization of endomucin, tyrosine hydroxylase (TH), neuropeptide Y (NPY) and norepinephrine transporter (NET). Moreover, Vegfa, Il-1 β, Il-6, Adrb2 and Rankl mRNA expression was assessed in mandibles and tibias 2 h after PBS or ISO treatment.
KEY FINDINGS
Despite similar sympathetic innervation and Adrb2 expression between mandibular tissues and tibias, with TH and NPY+ nerve fibers distributed around blood vessels, ISO treatment did not increase endomucin+ vessel area or the total number of endomucin+ vessels in any of the regions investigated (alveolar bone, periodontal ligament, and dental pulp). Consistent with these results, the expression of Vegfα, Il-6, Il-1β, and Rankl in the mandibular molar region did not change following ISO administration. We detected high expression of NET by immunofluorescence in mandible alveolar osteoblasts, osteocytes, and periodontal ligament fibroblasts, in addition to significantly higher Net expression by qPCR compared to the tibia from the same animals.
SIGNIFICANCE
These findings indicate a differential response to βAR agonists between mandibular and tibial tissues, since the angiogenic potential of sympathetic outflow observed in long bones is absent in periodontal tissues.
Topics: Adrenergic beta-Agonists; Animals; Female; Isoproterenol; Mice; Mice, Inbred C57BL; Periodontal Ligament; Receptors, Adrenergic, beta-2; Vascular Endothelial Growth Factor A
PubMed: 34186048
DOI: 10.1016/j.lfs.2021.119776 -
Scientific Reports Apr 2021Periodontal ligament (PDL) possesses a stem/progenitor population to maintain the homeostasis of periodontal tissue. However, transcription factors that regulate this...
Periodontal ligament (PDL) possesses a stem/progenitor population to maintain the homeostasis of periodontal tissue. However, transcription factors that regulate this population have not yet been identified. Thus, we aimed to identify a molecule related to the osteogenic differentiation of PDL progenitors using a single cell-based strategy in this study. We first devised a new protocol to isolate PDL cells from the surface of adult murine molars and established 35 new single cell-derived clones from the PDL explant. Among these clones, six clones with high (high clones, n = 3) and low (low clones, n = 3) osteogenic potential were selected. Despite a clear difference in the osteogenic potential of these clones, no significant differences in their cell morphology, progenitor cell marker expression, alkaline phosphatase activity, proliferation rate, and differentiation-related gene and protein expression were observed. RNA-seq analysis of these clones revealed that Z-DNA binding protein-1 (Zbp1) was significantly expressed in the high osteogenic clones, indicating that Zbp1 could be a possible marker and regulator of the osteogenic differentiation of PDL progenitor cells. Zbp1-positive cells were distributed sparsely throughout the PDL. In vitro Zbp1 expression in the PDL clones remained at a high level during osteogenic differentiation. The CRISPR/Cas9 mediated Zbp1 knockout in the high clones resulted in a delay in cell differentiation. On the other hand, Zbp1 overexpression in the low clones promoted cell differentiation. These findings suggested that Zbp1 marked the PDL progenitors with high osteogenic potential and promoted their osteogenic differentiation. Clarifying the mechanism of differentiation of PDL cells by Zbp1 and other factors in future studies will facilitate a better understanding of periodontal tissue homeostasis and repair, possibly leading to the development of novel therapeutic measures.
Topics: Animals; CRISPR-Cas Systems; Cell Differentiation; Clone Cells; Humans; Mesenchymal Stem Cells; Mice; Osteogenesis; Periodontal Ligament; Periodontium; RNA-Binding Proteins; RNA-Seq; Stem Cells
PubMed: 33824390
DOI: 10.1038/s41598-021-87016-1 -
International Journal of Molecular... Sep 2019Currently, various tissue engineering strategies have been developed for multiple tissue regeneration and integrative structure formations as well as single tissue... (Review)
Review
Currently, various tissue engineering strategies have been developed for multiple tissue regeneration and integrative structure formations as well as single tissue formation in musculoskeletal complexes. In particular, the regeneration of periodontal tissues or tooth-supportive structures is still challenging to spatiotemporally compartmentalize PCL (poly-ε-caprolactone)-cementum constructs with micron-scaled interfaces, integrative tissue (or cementum) formations with optimal dimensions along the tooth-root surfaces, and specific orientations of engineered periodontal ligaments (PDLs). Here, we discuss current advanced approaches to spatiotemporally control PDL orientations with specific angulations and to regenerate cementum layers on the tooth-root surfaces with Sharpey's fiber anchorages for state-of-the-art periodontal tissue engineering.
Topics: Animals; Biocompatible Materials; Dental Cementum; Guided Tissue Regeneration; Humans; Immunohistochemistry; Periodontal Ligament; Printing, Three-Dimensional; Regeneration; Tissue Engineering
PubMed: 31491973
DOI: 10.3390/ijms20184364