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Tissue & Cell Jun 2017Cells sensing changes in their microenvironmental stiffness and composition alter their responses, accordingly. This study determines whether gingival fibroblasts (GFs)...
Cells sensing changes in their microenvironmental stiffness and composition alter their responses, accordingly. This study determines whether gingival fibroblasts (GFs) and dental pulp mesenchymal stem cells (DPMSCs) support the formation of continuous layers in vitro by mimicking the stiffness and protein composition of their native extracellular matrix (ECM). Immortalized cells were incubated with (i) 0-100% Matrigel-ECM (M-ECM) for 7-28d, and with (ii) collagen and fibrin matrices for 14d. Cultures were analyzed by phase-contrast, fluorescence and confocal microscopies. The diameters and surface areas were measured via ImageJ. Self-renewal markers were detected by RT-PCR and immunocytochemistry assays. GFs and DPMSCs developed spheroids interconnected by elongated cell bundles or layers, respectively, expressing the self-renewal markers. Increased matrix stiffness resulted in spheroids replacement by the interconnecting cells/layers. Both cells required 100% M-ECM to reduce their spheroid diameter. However, it reduced the surface area of the interconnecting layers. Those differences led to extended, spindle-shaped GFs vs. compact, ring-shaped DPMSCs constructs. Collagen and fibrin matrices developed continuous layers of tightly connected cells vs. distinctive scattered cell aggregates, respectively. The ability of GFs and DPMSCs to create tissue-like multicellular layers at various matrix conditions may be imprinted by cells' adaptation to mechanical forces and composition in vivo.
Topics: Animals; Cell Line, Transformed; Dental Pulp; Extracellular Matrix; Fibroblasts; Gingiva; Mice
PubMed: 28457531
DOI: 10.1016/j.tice.2017.04.001 -
Journal of Oral Biosciences Jun 2024Regenerative dentistry aims to enhance the structure and function of oral tissues and organs. Modern tissue engineering harnesses cell and gene-based therapies to... (Review)
Review
BACKGROUND
Regenerative dentistry aims to enhance the structure and function of oral tissues and organs. Modern tissue engineering harnesses cell and gene-based therapies to advance traditional treatment approaches. Studies have demonstrated the potential of mesenchymal stem cells (MSCs) in regenerative dentistry, with some progressing to clinical trials. This review comprehensively examines animal studies that have utilized MSCs for various therapeutic applications. Additionally, it seeks to bridge the gap between related findings and the practical implementation of MSC therapies, offering insights into the challenges and translational aspects involved in transitioning from preclinical research to clinical applications.
HIGHLIGHTS
To achieve this objective, we have focused on the protocols and achievements related to pulp-dentin, alveolar bone, and periodontal regeneration using dental-derived MSCs in both animal and clinical studies. Various types of MSCs, including dental-derived cells, bone-marrow stem cells, and umbilical cord stem cells, have been employed in root canals, periodontal defects, socket preservation, and sinus lift procedures. Results of such include significant hard tissue reconstruction, functional pulp regeneration, root elongation, periodontal ligament formation, and cementum deposition. However, cell-based treatments for tooth and periodontium regeneration are still in early stages. The increasing demand for stem cell therapies in personalized medicine underscores the need for scientists and responsible organizations to develop standardized treatment protocols that adhere to good manufacturing practices, ensuring high reproducibility, safety, and cost-efficiency.
CONCLUSION
Cell therapy in regenerative dentistry represents a growing industry with substantial benefits and unique challenges as it strives to establish sustainable, long-term, and effective oral tissue regeneration solutions.
Topics: Humans; Animals; Tissue Engineering; Regeneration; Regenerative Medicine; Mesenchymal Stem Cells; Mesenchymal Stem Cell Transplantation; Dental Pulp; Dentistry
PubMed: 38403241
DOI: 10.1016/j.job.2024.02.006 -
Scientific Reports Dec 2020Implementing the principles of tissue engineering within the clinical management of non-vital immature permanent teeth is of clinical interest. However, the ideal...
Implementing the principles of tissue engineering within the clinical management of non-vital immature permanent teeth is of clinical interest. However, the ideal scaffold remains elusive. The aim of this work was to assess the feasibility of decellularising rat dental pulp tissue and evaluate the ability of such scaffold to support stem cell repopulation. Rat dental pulps were retrieved and divided into control and decellularised groups. The decellularisation protocol incorporated a low detergent concentration and hypotonic buffers. After decellularisation, the scaffolds were characterised histologically, immunohistochemistry and the residual DNA content quantified. Surface topography was also viewed under scanning electron microscopy. Biocompatibility was evaluated using cytotoxicity assays utilising L-929 cell line. Decellularised scaffolds were recellularised with human dental pulp stem cells up to 14 days in vitro. Cellular viability was assessed using LIVE/DEAD stain kit and the recellularised scaffolds were further assessed histologically and immunolabelled using makers for odontoblastic differentiation, cytoskeleton components and growth factors. Analysis of the decellularised scaffolds revealed an acellular matrix with histological preservation of structural components. Decellularised scaffolds were biocompatible and able to support stem cell survival following recellularisation. Immunolabelling of the recellularised scaffolds demonstrated positive cellular expression against the tested markers in culture. This study has demonstrated the feasibility of developing a biocompatible decellularised dental pulp scaffold, which is able to support dental pulp stem cell repopulation. Clinically, decellularised pulp tissue could possibly be a suitable scaffold for use within regenerative (reparative) endodontic techniques.
Topics: Animals; Cell Differentiation; Dental Pulp; Rats; Stem Cells; Tissue Engineering; Tissue Scaffolds
PubMed: 33299073
DOI: 10.1038/s41598-020-78477-x -
Cytokine Feb 2020Pulpitis is known as a typical inflammation of dental pulp tissue, and microorganisms of the oral microbiome are involved in this opportunistic infection. Studies... (Review)
Review
Pulpitis is known as a typical inflammation of dental pulp tissue, and microorganisms of the oral microbiome are involved in this opportunistic infection. Studies indicated that several factors related to host response have a crucial role in pulpitis. Among these factors, inflammatory mediators of the immune system such as cytokines and chemokines contribute to pulpal defense mechanisms. A wide range of cytokines have been observed in dental pulp and these small molecules are able to trigger inflammation and participate in immune cell trafficking, cell proliferation, inflammation, and tissue damage in pulp space. Therefore, the aim of this review was to describe the role of cytokines in the pathogenesis of pulpitis.
Topics: Animals; Bacteria; Cell Proliferation; Cytokines; Dental Pulp; Humans; Inflammation; Inflammation Mediators; Pulpitis
PubMed: 31670007
DOI: 10.1016/j.cyto.2019.154896 -
International Journal of Molecular... Aug 2018Vital pulp therapy (VPT) is to preserve the nerve and maintain healthy dental pulp tissue. Laser irradiation (LI) is beneficial for VPT. Understanding how LI affects...
Vital pulp therapy (VPT) is to preserve the nerve and maintain healthy dental pulp tissue. Laser irradiation (LI) is beneficial for VPT. Understanding how LI affects dental pulp cells and tissues is necessary to elucidate the mechanism of reparative dentin and dentin regeneration. Here, we show how Er:YAG-LI and diode-LI modulated cell proliferation, apoptosis, gene expression, protease activation, and mineralization induction in dental pulp cells and tissues using cell culture, immunohistochemical, genetic, and protein analysis techniques. Both LIs promoted proliferation in porcine dental pulp-derived cell lines (PPU-7), although the cell growth rate between the LIs was different. In addition to proliferation, both LIs also caused apoptosis; however, the apoptotic index for Er:YAG-LI was higher than that for diode-LI. The mRNA level of odontoblastic gene markers-two dentin sialophosphoprotein splicing variants and matrix metalloprotease (MMP)20 were enhanced by diode-LI, whereas MMP2 was increased by Er:YAG-LI. Both LIs enhanced alkaline phosphatase activity, suggesting that they may help induce PPU-7 differentiation into odontoblast-like cells. In terms of mineralization induction, the LIs were not significantly different, although their cell reactivity was likely different. Both LIs activated four MMPs in porcine dental pulp tissues. We helped elucidate how reparative dentin is formed during laser treatments.
Topics: Animals; Apoptosis; Cell Differentiation; Cell Line; Cell Proliferation; Dental Pulp; Extracellular Matrix Proteins; Gene Expression Regulation; Lasers, Semiconductor; Low-Level Light Therapy; Matrix Metalloproteinase 20; Odontoblasts; Phosphoproteins; Sialoglycoproteins; Swine
PubMed: 30126087
DOI: 10.3390/ijms19082429 -
Biochimie Jun 2019Genetic and pharmacological studies provided evidence that serotonin (5-HT) is an important signaling molecule for the development and the maintenance of mineralized... (Review)
Review
Genetic and pharmacological studies provided evidence that serotonin (5-HT) is an important signaling molecule for the development and the maintenance of mineralized tissues. However, how 5-HT takes part to the homeostasis of teeth and bone remains elusive. In the dental field, a major breakthrough comes from the identification of 5-HT but also dopamine (DA) as "damage" signals necessary for stem cell-based tooth repair. Pulpal stem cells express the overall functions of 5-HT and DA neurons including a definite set of functional 5-HT/DA receptors that render cells responsive for circulating bioamines. Upon tooth injury, activated platelets release bulks of 5-HT/DA that mobilize pulpal stem cells for natural dental repair. The contribution of 5-HT to bone metabolism is more documented with description of both anabolic and resorptive effects. By controlling the tissue-non specific alkaline phosphatase (TNAP), 5-HT receptors exert an anabolic function and a pivotal role in mineralization processes. Increasing our understanding of the role of 5-HT receptors in bone metabolism may pave the road for the development of therapeutic strategies towards skeletal-associated pathologies and ectopic calcification.
Topics: Animals; Dental Pulp; Humans; Osteogenesis; Receptors, Serotonin; Serotonin; Stem Cells
PubMed: 30077818
DOI: 10.1016/j.biochi.2018.07.030 -
Dental Traumatology : Official... Dec 2019Dark coronal discoloration is a common outcome of traumatic dental injuries in primary incisors. However, there are only a few research studies on the mechanism that... (Review)
Review
Dark coronal discoloration is a common outcome of traumatic dental injuries in primary incisors. However, there are only a few research studies on the mechanism that causes dark discoloration, the condition of the pulp, and the preferred treatment of such teeth. This article is a summary of the author's research studies and personal experience regarding dark coronal discoloration of traumatized primary incisors.
Topics: Dental Pulp; Dental Pulp Necrosis; Humans; Incisor; Tooth Discoloration; Tooth Injuries; Tooth, Deciduous
PubMed: 31121084
DOI: 10.1111/edt.12483 -
Archives of Oral Biology Jan 2020Fibroblast growth factors (FGFs) are growth factors that play an important role in tooth development, repair, and regeneration. Of the FGF families, basic fibroblast... (Review)
Review
Fibroblast growth factors (FGFs) are growth factors that play an important role in tooth development, repair, and regeneration. Of the FGF families, basic fibroblast growth factor (bFGF) has been the most frequently investigated in dentistry. Numerous studies have reported advantages of bFGF, while others did not find any additional benefit. This review gives a comprehensive summary of the potential role of bFGF in dental pulp wound healing and regeneration in connection with cell proliferation and differentiation, angiogenesis, and neural differentiation from both in vitro and in vivo studies. Furthermore, the possible underlying mechanisms associated with bFGF in promoting dental pulp wound healing are discussed in this review.
Topics: Animals; Cell Differentiation; Cell Proliferation; Dental Pulp; Fibroblast Growth Factor 2; Humans; Neovascularization, Physiologic; Odontogenesis; Regeneration; Wound Healing
PubMed: 31585238
DOI: 10.1016/j.archoralbio.2019.104574 -
Journal of Endodontics May 2018FoxO3a is a member of FoxO transcription factor family that participates in the transcriptional regulation of autophagy. In this study we explored the anti-inflammatory...
INTRODUCTION
FoxO3a is a member of FoxO transcription factor family that participates in the transcriptional regulation of autophagy. In this study we explored the anti-inflammatory function of FoxO3a-regulated autophagy in inflamed human dental pulp and lipopolysaccharide-treated mDPC6T cells.
METHODS
The expression of FoxO3a and autophagy markers in caries and pulpitis from human dental pulp were examined by immunohistochemistry and Western blot. We conducted in vitro studies by treating mDPC6T cells with lipopolysaccharide for various lengths of time. Next, we measured the nuclear translocation of FoxO3a by immunofluorescence and investigated the potential relationship between FoxO3a and autophagy after FoxO3a knockdown using small interfering RNA.
RESULTS
The expression of FoxO3a and autophagy proteins was upregulated in the odontoblasts of human caries and pulpitis samples. In addition, we also observed that the enhanced nuclear translocation of FoxO3a was positively correlated with the progression of inflammation. The results of our in vitro study revealed that 6 hours of lipopolysaccharide treatment increased nuclear translocation of FoxO3a and activated autophagy in mDPC6T cells. We also observed that the knockdown of FoxO3a suppressed autophagy.
CONCLUSIONS
Our data indicate that FoxO3a might play a role in autophagy activation and the maintenance of intracellular homeostasis in inflamed odontoblasts.
Topics: Autophagy; Blotting, Western; Caspase 3; Cells, Cultured; Dental Pulp; Fluorescent Antibody Technique; Forkhead Box Protein O3; Gene Knockdown Techniques; Humans; Inflammation; Odontoblasts
PubMed: 29551204
DOI: 10.1016/j.joen.2017.12.023 -
International Journal of Molecular... Apr 2024Dental tissue stem cells (DTSCs) are well known for their multipotent capacity and regenerative potential. They also play an important role in the immune response of... (Review)
Review
Dental tissue stem cells (DTSCs) are well known for their multipotent capacity and regenerative potential. They also play an important role in the immune response of inflammatory processes derived from caries lesions, periodontitis, and gingivitis. These oral diseases are triggered by toxins known as lipopolysaccharides (LPS) produced by gram-negative bacteria. LPS present molecular patterns associated with pathogens and are recognized by Toll-like receptors (TLRs) in dental stem cells. In this review, we describe the effect of LPS on the biological behavior of DTSCs. We also focus on the molecular sensors, signaling pathways, and emerging players participating in the interaction of DTSCs with lipopolysaccharides. Although the scientific advances generated provide an understanding of the immunomodulatory potential of DTSCs, there are still new reflections to explore with regard to their clinical application in the treatment of oral inflammatory diseases.
Topics: Animals; Humans; Dental Pulp; Lipopolysaccharides; Signal Transduction; Stem Cells; Toll-Like Receptors; Bacterial Infections
PubMed: 38673923
DOI: 10.3390/ijms25084338