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Tissue & Cell Feb 2018Dental pulp is a highly vascularized tissue with a high regenerative capacity. This is attributed to its unique blood supply and the presence of progenitor or postnatal...
Dental pulp is a highly vascularized tissue with a high regenerative capacity. This is attributed to its unique blood supply and the presence of progenitor or postnatal dental pulp stem cells. Here we aimed to isolate and compare the angiogenic properties of endothelial cells (EC) prepared from mouse dental pulp and periodontal ligament (PDL). EC were isolated from 4-week-old wild type immorto mice. Mice were sacrificed and after mandible isolation, the molar and incisor teeth and the PDL from molar teeth were dissected. EC were prepared by collagenase digestion of tissues and affinity purification using magnetic beads coated with platelet/endothelial cell adhesion molecule-1 (PECAM-1/CD31) antibody. EC prepared from incisor and molar pulps and PDL were examined for expression of appropriate markers by fluorescence-activated cell sorting (FACS) analysis. The proliferation, migration, and capillary morphogenesis of EC were evaluated. Ex vivo sprouting angiogenesis from various tissues was also compared. Data were analyzed at the level of significance of P<0.05. Pulp EC prepared from incisors proliferated and migrated significantly faster than molar and PDL EC (P<0.05). In addition, molar and PDL EC formed a more extensive capillary network when plated on Matrigel. This is consistent with the lower proliferative and migratory characteristics of these cells compared with incisor EC (P<0.05). However, PDL tissue showed significantly more sprouting area than molar and incisor pulp tissues (P<0.05). Thus, pulp EC from molar and incisor and PDL EC present different proangiogenic properties. Collectively our results suggest that EC from different tooth tissue have unique characteristics related to their target tissue and function.
Topics: Animals; Cell Movement; Cell Proliferation; Dental Pulp; Endothelial Cells; Flow Cytometry; Incisor; Mice; Molar; Neovascularization, Physiologic; Periodontal Ligament; Platelet Endothelial Cell Adhesion Molecule-1
PubMed: 29429515
DOI: 10.1016/j.tice.2017.11.004 -
Cell and Tissue Research Feb 2021The dental pulp, a non-mineralized connective tissue uniquely encased within the cavity of the tooth, provides a niche for diverse arrays of dental mesenchymal stem... (Review)
Review
The dental pulp, a non-mineralized connective tissue uniquely encased within the cavity of the tooth, provides a niche for diverse arrays of dental mesenchymal stem cells. Stem cells in the dental pulp, including dental pulp stem cells (DPSCs), stem cells from human exfoliated deciduous teeth (SHEDs) and stem cells from apical papilla (SCAPs), have been isolated from human tissues with an emphasis on their potential application to regenerative therapies. Recent studies utilizing mouse genetic models shed light on the identities of these mesenchymal progenitor cells derived from neural crest cells (NCCs) in their native conditions, particularly regarding how they contribute to homeostasis and repair of the dental tissue. The current concept is that at least two distinct niches for stem cells exist in the dental pulp, e.g., the perivascular niche and the perineural niche. The precise identities of these stem cells and their niches are now beginning to be unraveled thanks to sophisticated mouse genetic models, which lead to better understanding of the fundamental properties of stem cells in the dental pulp and the apical papilla in humans. The new knowledge will be highly instrumental for developing more effective stem cell-based regenerative therapies to repair teeth in the future.
Topics: Animals; Biomarkers; Dental Papilla; Dental Pulp; Mice; Models, Genetic; Stem Cell Niche; Stem Cells
PubMed: 32803323
DOI: 10.1007/s00441-020-03271-0 -
Journal of Endodontics Jan 2017Endodontics uses cell therapy strategies to treat pulpal and periapical diseases. During these therapies, surgeons aim to reconstruct the natural microenvironments that... (Review)
Review
INTRODUCTION
Endodontics uses cell therapy strategies to treat pulpal and periapical diseases. During these therapies, surgeons aim to reconstruct the natural microenvironments that regulate the activity of dental stem cells.
METHODS
We searched for more than 400 articles in PubMed using key words from regenerative endodontics and dental stem cell biology. In 268 articles, we reviewed what factors may influence histologic results after preclinical dental treatments that use regenerative endodontic procedures after pulpectomy.
RESULTS
Several factors, such as the origin of stem cells, the biomimicry of scaffolds used, and the size of lesions, are considered to influence the histologic appearance of the regenerated pulp-dentin complex after treatments. Information is accumulating on transcription factors that generate the pulp-dentin complex and survival/trophic factors that would benefit niche recovery and histologic results.
CONCLUSIONS
In this article, we discuss the noninterchangeability of stem cells, the influence of dentin-entrapped molecule release on pulp regeneration and survival of stem cells, and the need of positional markers to assess treatments histologically. The ex vivo amplification of appropriate dental stem cells, the search for scaffolds storing the molecular diversity entrapped in the dentin, and the use of positional transcription factors as histologic markers are necessary to improve future preclinical experiments.
Topics: Animals; Dental Pulp; Dentin; Humans; Regeneration; Regenerative Medicine; Root Canal Therapy; Stem Cell Transplantation; Stem Cells
PubMed: 27986102
DOI: 10.1016/j.joen.2016.09.011 -
Australian Dental Journal Mar 2016Trauma to the permanent teeth involves not only the teeth but also the pulp, the periodontal ligament, alveolar bone, gingiva and other associated structures. There are... (Review)
Review
Trauma to the permanent teeth involves not only the teeth but also the pulp, the periodontal ligament, alveolar bone, gingiva and other associated structures. There are many variations in the types of injuries with varying severity and often a tooth may sustain more than one injury at the same time. In more severe trauma cases, there are many different cellular systems of mineralized hard and unmineralized soft tissues involved, each with varying potential for healing. Furthermore, the responses of the different tissues may be interrelated and dependent on each other. Hence, healing subsequent to dental trauma has long been known to be very complex. Because of this complexity, tissue responses and the consequences following dental trauma have been confusing and puzzling for many clinicians. In this review, the tissue responses are described under the tissue compartments typically involved following dental trauma: the pulp, periradicular and associated soft tissues. The factors involved in the mechanisms of trauma are analysed for their effects on the tissue responses. A thorough understanding of the possible tissue responses is imperative for clinicians to overcome the confusion and manage dental trauma adequately and conservatively in order to minimize the consequences following trauma.
Topics: Dental Pulp; Dental Pulp Diseases; Humans; Periapical Tissue; Periodontal Diseases; Periodontium; Tooth Injuries; Tooth Resorption; Wound Healing
PubMed: 26923447
DOI: 10.1111/adj.12397 -
BioMed Research International 2017Endodontic regeneration shows promise in treating dental pulp diseases; however, no suitable scaffolds exist for pulp regeneration. Acellular natural extracellular...
Endodontic regeneration shows promise in treating dental pulp diseases; however, no suitable scaffolds exist for pulp regeneration. Acellular natural extracellular matrix (ECM) is a favorable scaffold for tissue regeneration since the anatomical structure and ECM of the natural tissues or organs are well-preserved. Xenogeneic ECM is superior to autologous or allogeneic ECM in tissue engineering for its unlimited resources. This study investigated the characteristics of decellularized dental pulp ECM from swine and evaluated whether it could mediate pulp regeneration. Dental pulps were acquired from the mandible anterior teeth of swine 12 months of age and decellularized with 10% sodium dodecyl sulfate (SDS) combined with Triton X-100. Pulp regeneration was conducted by seeding human dental pulp stem cells into decellularized pulp and transplanted subcutaneously into nude mice for 8 weeks. The decellularized pulp demonstrated preserved natural shape and structure without any cellular components. Histological analysis showed excellent ECM preservation and pulp-like tissue, and newly formed mineralized tissues were regenerated after being transplanted in vivo. In conclusion, decellularized swine dental pulp maintains ECM components favoring stem cell proliferation and differentiation, thus representing a suitable scaffold for improving clinical outcomes and functions of teeth with dental pulp diseases.
Topics: Animals; Bone Regeneration; Calcification, Physiologic; Dental Pulp; Extracellular Matrix; Humans; Stem Cells; Swine; Tissue Scaffolds
PubMed: 29387727
DOI: 10.1155/2017/9342714 -
BMC Oral Health Dec 2019Helicobacter pylori (H. pylori) colonize the stomach and are considered an etiological agent of gastric cancer. The oral cavity is a transmission route to the stomach,...
BACKGROUND
Helicobacter pylori (H. pylori) colonize the stomach and are considered an etiological agent of gastric cancer. The oral cavity is a transmission route to the stomach, but the exact site of colonization has not yet been explicated. Our study investigated the association between H. pylori infection and presence in oral samples.
METHODS
Dental pulp, supragingival plaque, and saliva from 192 patients visiting the Dentistry's outpatient clinic were collected for testing. The H. pylori ureA gene was identified via Nested PCR. Urine anti-H. pylori antibody test was utilized to detect infection.
RESULTS
Twenty-five subjects were found to be antibody-positive. PCR analysis of dental pulp revealed that 23 subjects possessed the ureA gene. Twenty-one subjects were positive for both antibodies and genes in dental pulp. PCR testing revealed that 2 subjects were positive in dental plaque but negative for saliva. The subjects positive for H. pylori in dental pulp expressed clinical signs of severe dental caries.
CONCLUSIONS
H. pylori infected subjects expressed H. pylori in samples from the oral cavity. The main reservoir for infection within the oral cavity was determined to be dental pulp. Moreover, H. pylori are likely transmitted from dental caries to the root canal.
Topics: Adult; Dental Caries; Dental Pulp; Female; Helicobacter Infections; Helicobacter pylori; Humans; Japan; Male; Saliva
PubMed: 31791309
DOI: 10.1186/s12903-019-0967-2 -
Biomedical Papers of the Medical... Nov 2021Teeth extracted are usually disposed as bio-waste whereas they could serve as an autologous tissue for culturing multipotent dental pulp cells which have application...
BACKGROUND AND AIMS
Teeth extracted are usually disposed as bio-waste whereas they could serve as an autologous tissue for culturing multipotent dental pulp cells which have application potential in regenerative medicine. This study aimed to examine the feasibility of cryopreserving dental pulp tissue at teeth extraction for later culturing of cells.
METHODS
The pulp tissue from each of a total of 10 teeth was cut into small fragments which were then divided into two portions. One portion was directly used for culturing pulp cells using the explant method. The other portion was cryopreserved with 10% DMSO in liquid nitrogen for at least one month and then thawed for culturing pulp cells.
RESULTS
Vital cells were obtained from all the 10 pulp fragment suspensions which went through cryopreservation. The cell outgrowth from the explants of cryopreserved pulp fragments was two days later than that of corresponding fresh pulp tissue. Otherwise, no difference was observed in proliferation, expression of stem cell markers and differentiation into adipose cells and osteoblasts between the two groups of cells cultured from the fresh or the cryopreserved pulp fragments.
CONCLUSIONS
Cryopreserving fragmented dental pulp tissue provides a feasible option for saving pulp tissues as autologous cell sources for possible later application.
Topics: Cell Culture Techniques; Cell Differentiation; Cell Proliferation; Cells, Cultured; Cryopreservation; Dental Pulp; Humans; Stem Cells
PubMed: 33325456
DOI: 10.5507/bp.2020.061 -
STAR Protocols Dec 2021Teeth and the surrounding periodontal tissues are affected by many pathologies that compromise their integrity and significantly affect life quality. The study of the...
Teeth and the surrounding periodontal tissues are affected by many pathologies that compromise their integrity and significantly affect life quality. The study of the main dental tissues, the dental pulp and periodontium, is made arduous by their close association with highly mineralized tissues (dentin, cementum, and alveolar bone). Here we describe a protocol to isolate all cells composing human dental pulp and periodontium for single-cell RNA sequencing analysis. For complete details on the use and execution of this protocol, please refer to Pagella et al. (2021).
Topics: Cell Culture Techniques; Cell Separation; Cells, Cultured; Dental Pulp; Humans; Periodontium; Sequence Analysis, RNA; Single-Cell Analysis; Tooth
PubMed: 34825216
DOI: 10.1016/j.xpro.2021.100953 -
Bioscience Reports Jun 2020Dental pulp stem cells (DPSCs) regenerate injured/diseased pulp tissue and deposit tertiary dentin. DPSCs stress response can be activated by exposing cells to the...
Dental pulp stem cells (DPSCs) regenerate injured/diseased pulp tissue and deposit tertiary dentin. DPSCs stress response can be activated by exposing cells to the monomer triethyleneglycol dimethacrylate (TEGDMA) and inducing the DNA-damage inducible transcript 4 (DDIT4) protein expression. The goal of the present study was to determine the impact of TEGDMA on the ability of DPSCs to maintain their self-renewal capabilities, develop and preserve their 3D structures and deposit the mineral. Human primary and immortalized DPSCs were cultured in extracellular matrix/basement membrane (ECM/BM) to support stemness and to create multicellular interacting layers (microtissues). The microtissues were exposed to the toxic concentrations of TEGDMA (0.5 and 1.5 mmol/l). The DPSCs spatial architecture was assessed by confocal microscopy. Mineral deposition was detected by alizarin red staining and visualized by stereoscopy. Cellular self-renewal transcription factor SOX2 was determined by immunocytochemistry. The microtissue thicknesses/vertical growth, surface area of the mineralizing microtissues, the percentage of area covered by the deposited mineral, and the fluorescence intensity of the immunostained cells were quantified ImageJ. DDIT4 expression was determined by a single molecule RNA-FISH technique and the cell phenotype was determined morphologically. DDIT4 expression was correlated with the cytotoxic phenotype. TEGDMA affected the structures of developing and mature microtissues. It inhibited the deposition of the mineral in the matrix while not affecting the SOX2 expression. Our data demonstrate that DPSCs retained their self-renewal capacity although their other functions were impeded. Since the DPSCs pool remained preserved, properties effected by the irritant should be restored by a proper rescue therapy.
Topics: Adult; Cell Line; Cell Self Renewal; Composite Resins; Dental Pulp; Dentin; Dentinogenesis; Humans; Phenotype; Polyethylene Glycols; Polymethacrylic Acids; Primary Cell Culture; SOXB1 Transcription Factors; Signal Transduction; Stem Cells; Transcription Factors; Young Adult
PubMed: 32495822
DOI: 10.1042/BSR20200210 -
Frontiers in Cellular and Infection... 2022The prevalence of dental caries in the Mexican adult population aged 20 to 85 years is around 93.3%, and 50% in Mexican children and adolescents. Worldwide, it is the...
The prevalence of dental caries in the Mexican adult population aged 20 to 85 years is around 93.3%, and 50% in Mexican children and adolescents. Worldwide, it is the most common non-communicable disease. One of the main etiological factors for dental caries is the oral microbiome and changes in its structure and function, with an expansion of pathogenic bacteria like . The exposed dental pulp tissue triggers an innate immune response to counteract this bacterial invasion. The relation between oral dysbiosis and innate immune responses remains unclear. We aimed to understand the relationship between innate immune response and the oral microbiota by quantifying the expression of Toll-like receptors (TLRs) and proinflammatory markers (cytokines and a chemokine) in dental pulp tissue, either exposed or not to carious dentin, and to correlate this information with the oral microbiome found in healthy teeth and those with moderate caries. RNA was purified from pulp tissue, subjected to RT-qPCR and analysed with the method. Supragingival dental plaque of non-carious teeth and dentin of carious teeth were subjected to 16S targeted sequencing. Principal coordinate analysis, permutational multivariate ANOVA, and linear discriminant analysis were used to assess differences between non-carious and carious teeth. Correlations were assessed with Spearman´s test and corrected for multiple comparisons using the FDR method. The relative abundance (RA) of , and was increased in carious teeth; while the RA of and decreased. and were only detected in carious teeth. Significant overexpression of interleukin 1 beta (IL1 β), IL6, and CXCL8 was detected in pulp tissue exposed to carious dentin. IL1β correlated positively with TLR2 and ; yet negatively with These findings suggest that immune response of pulp tissue chronically exposed to cariogenic microbiome is triggered by proinflammatory cytokines IL1β and IL6 and the chemokine CXCL8.
Topics: Adolescent; Adult; Child; Humans; Actinobacteria; Actinomyces; Cytokines; Dental Caries; Dental Pulp; Dentin; Interleukin-6; Microbiota; Streptococcus mutans
PubMed: 36569197
DOI: 10.3389/fcimb.2022.958722