scholarly journals Effect of Hydroxyapatite Microspheres, Amoxicillin–Hydroxyapatite and Collagen–Hydroxyapatite Composites on Human Dental Pulp-Derived Mesenchymal Stem Cells

Materials ◽  
2021 ◽  
Vol 14 (24) ◽  
pp. 7515
Author(s):  
Yasmine Mendes Pupo ◽  
Lidiane Maria Boldrini Leite ◽  
Alexandra Cristina Senegaglia ◽  
Liziane Antunes ◽  
Jessica Mendes Nadal ◽  
...  
Keyword(s):  
Dental Pulp ◽  
Cell Biology ◽  
Periodontal Regeneration ◽  
Positive Control ◽  
Control Group ◽  
Migration Behavior ◽  
Migration Assay ◽  
Bioactive Scaffolds ◽  
Hydroxyapatite Composite ◽  
Human Dental Pulp

In this study, the preparation and characterization of three hydroxyapatite-based bioactive scaffolds, including hydroxyapatite microspheres (HAps), amoxicillin–hydroxyapatite composite (Amx–HAp), and collagen–hydroxyapatite composite (Col–HAp) were performed. In addition, their behavior in human dental pulp mesenchymal stem cell (hDPSC) culture was investigated. HAps were synthesized through the following methods: microwave hydrothermal, hydrothermal reactor, and precipitation, respectively. hDPSCs were obtained from samples of third molars and characterized by immunophenotypic analysis. Cells were cultured on scaffolds with osteogenic differentiation medium and maintained for 21 days. Cytotoxicity analysis and migration assay of hDPSCs were evaluated. After 21 days of induction, no differences in genes expression were observed. hDPSCs highly expressed the collagen IA and the osteonectin at the mRNA. The cytotoxicity assay using hDPSCs demonstrated that the Col–HAp group presented non-viable cells statistically lower than the control group (p = 0.03). In the migration assay, after 24 h HAps revealed the same migration behavior for hDPSCs observed compared to the positive control. Col–HAp also provided a statistically significant higher migration of hDPSCs than HAps (p = 0.02). Migration results after 48 h for HAps was intermediate from those achieved by the control groups. There was no statistical difference between the positive control and Col–HAp. Specifically, this study demonstrated that hydroxyapatite-based bioactive scaffolds, especially Col-Hap, enhanced the dynamic parameters of cell viability and cell migration capacities for hDPSCs, resulting in suitable adhesion, proliferation, and differentiation of this osteogenic lineage. These data presented are of high clinical importance and hold promise for application in therapeutic areas, because Col–HAp can be used in ridge preservation, minor bone augmentation, and periodontal regeneration. The development of novel hydroxyapatite-based bioactive scaffolds with clinical safety for bone formation from hDPSCs is an important yet challenging task both in biomaterials and cell biology.

scholarly journals Effect of hydroxyapatite microspheres, amoxicillin-hydroxyapatite and collagen-hydroxyapatite composites on human dental pulp-derived mesenchymal stem cells

2021 ◽  
Author(s):  
Yasmine Mendes Pupo ◽  
Lidiane Maria Boldrini Leite ◽  
Alexandra Cristina Senegaglia ◽  
Lisiane Antunes ◽  
Jessica Mendes Nadal ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Dental Pulp ◽  
Positive Control ◽  
Control Group ◽  
Migration Behavior ◽  
Migration Assay ◽  
Hydroxyapatite Composite ◽  
Human Dental Pulp

AbstractThe aim of this study was to evaluate the in vitro behavior of human dental pulp mesenchymal stem cells (hDPSCs) cultured on scaffolds of three hydroxyapatite-based materials: hydroxyapatite microspheres [HAp]; amoxicillin-hydroxyapatite composite [Amx-HAp]; and collagen-hydroxyapatite composite [Col-HAp]. These hydroxyapatites (HAps) were synthesized through three methods: microwave hydrothermal, hydrothermal reactor (teflon pouches), and precipitation, respectively. We performed an in vitro experimental study using dental pulp stem cells obtained from samples of third molars and characterized by immunophenotypic analysis. Cells were cultured on scaffolds with osteogenic differentiation medium and were maintained for 21 days. Cytotoxicity analysis and migration assay of hDPSCs were evaluated. Each experiment was performed in triplicate. Data analysis was performed using Kruskal-Wallis test and Dunn’s post-hoc test. After 21 days of induction, no differences in genes expression were observed. hDPSCs highly expressed the collagen IA and the osteonectin at the mRNA, which indicated these genes plays an important role in odontogenesis regardless of induction stimulus. Cytotoxicity assay using hDPSCs demonstrated that Col-HAp group presented a number of non-viable cells statistically lower than the control group (p=0.03). In the migration assay after 24h, biomaterials HAp, Amx-HAp, and Col-HAp revealed the same migration behavior for hDPSCs observed to the positive control. Col-HAp also provided a statistically significant higher migration of hDPSCs than HAp (p=0.02). The migration results in 48h for HAp, Amx-HAp, and Col-HAp was intermediate from those achieved by control groups. There was no statistical difference between positive control and Col-HAp (p>0.05). In general, Col-HAp scaffold showed better features for these dynamic parameters of cell viability and cell migration capacities for hDPSCs, leading to suitable adhesion, proliferation, and differentiation of this osteogenic lineage. These data present high clinical importance because Col-HAp can be used in a wide variety of therapeutic areas, including ridge preservation, minor bone augmentation, and periodontal regeneration.

scholarly journals Human Dental Pulp Tissue during Orthodontic Tooth Movement: An Immunofluorescence Study

2020 ◽  
Vol 5 (3) ◽  
pp. 65 ◽  
Author(s):  
Giovanna Vermiglio ◽  
Antonio Centofanti ◽  
Giovanni Matarese ◽  
Angela Militi ◽  
Marco Matarese ◽  
...  
Keyword(s):  
Protein Expression ◽  
Dental Pulp ◽  
Tooth Movement ◽  
Orthodontic Tooth Movement ◽  
Control Group ◽  
Pulp Tissue ◽  
Force Application ◽  
Dental Pulp Tissue ◽  
Human Dental Pulp

The orthodontic tooth movement is the last step of several biological processes that take place after the application of external forces. During this process, dental pulp tissue is subjected to structural and protein expression modifications in order to maintain their integrity and functional morphology. The purpose of the present work was to perform an in vivo study, evaluating protein expression modifications in the human dental pulp of patients that have undergone orthodontic tooth movement due to pre-calibrated light force application for 30 days. Dental pulp samples were extracted from molars and premolars of the control group and after 7 and 30 days of treatment; the samples were then processed for immunofluorescence reactions using antibodies against fibronectin, collagen I and vascular endothelial growth factor (VEGF). Our results show that, after 7 days of treatment, all tested proteins change their pattern expression and will reset after 30 days. These data demonstrate that the dental pulp does not involve any irreversible iatrogenic alterations, supporting the efficacy and safety of using pre-calibrated force application to induce orthodontic tooth movement in clinical practice.

scholarly journals miR-6807-5p Inhibited the Odontogenic Differentiation of Human Dental Pulp Stem Cells Through Directly Targeting METTL7A

2021 ◽  
Vol 9 ◽  
Author(s):  
Ning Wang ◽  
Xiao Han ◽  
Haoqing Yang ◽  
Dengsheng Xia ◽  
Zhipeng Fan
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Binding Protein ◽  
Dental Pulp Stem Cells ◽  
Luciferase Reporter ◽  
Control Group ◽  
Alizarin Red ◽  
Odontogenic Differentiation ◽  
Human Dental Pulp ◽  
Tooth Tissue

Background: Tooth tissue regeneration mediated by mesenchymal stem cells (MSCs) has become the most ideal treatment. Although the known regulatory mechanism and some achievements have been discovered, directional differentiation cannot effectively induce regeneration of tooth tissue. In this study, we intended to explore the function and mechanism of miR-6807-5p and its target gene METTL7A in odontogenic differentiation.Methods: In this study, human dental pulp stem cells (DPSCs) were used. Alkaline phosphatase (ALP), Alizarin red staining (ARS), and calcium ion quantification were used to detect the odontogenic differentiation of miR-6807-5p and METTL7A. Real-time RT-PCR, western blot, dual-luciferase reporter assay, and pull-down assay with biotinylated miRNA were used to confirm that METTL7A was the downstream gene of miR-6807-5p. Protein mass spectrometry and co-immunoprecipitation (Co-IP) were used to detect that SNRNP200 was the co-binding protein of METTL7A.Results: After mineralized induction, the odontogenic differentiation was enhanced in the miR-6807-5p-knockdown group and weakened in the miR-6807-5p-overexpressed group compared with the control group. METTL7A was the downstream target of miR-6807-5p. After mineralized induction, the odontogenic differentiation was weakened in the METTL7A-knockdown group and enhanced in the METTL7A-overexpressed group compared with the control group. SNRNP200 was the co-binding protein of METTL7A. The knockdown of SNRNP200 inhibited the odontogenic differentiation of DPSCs.Conclusion: This study verified that miR-6807-5p inhibited the odontogenic differentiation of DPSCs. The binding site of miR-6807-5p was the 3′UTR region of METTL7A, which was silenced by miR-6807-5p. METTL7A promoted the odontogenic differentiation of DPSCs. SNRNP200, a co-binding protein of METTL7A, promoted the odontogenic differentiation of DPSCs.

scholarly journals The Conditioned Medium of Calcined Tooth Powder Promotes the Osteogenic and Odontogenic Differentiation of Human Dental Pulp Stem Cells via MAPK Signaling Pathways

2019 ◽  
Vol 2019 ◽  
pp. 1-13 ◽  
Author(s):  
Jintao Wu ◽  
Na Li ◽  
Yuan Fan ◽  
Yanqiu Wang ◽  
Yongchun Gu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Signaling Pathways ◽  
Conditioned Medium ◽  
Dental Pulp ◽  
Mapk Signaling ◽  
Dental Pulp Stem Cells ◽  
Control Group ◽  
Odontogenic Differentiation ◽  
Human Dental Pulp ◽  
Mapk Signaling Pathways

The calcined tooth powder (CTP), a type of allogeneic biomimetic mineralized material, has been confirmed that can promote new bone formation when obtained at high temperature. The aim of this study was to investigate effects of the conditioned medium of calcined tooth powder (CTP-CM) on the osteogenic and odontogenic differentiation of human dental pulp stem cells (hDPSCs) and the underlying mechanisms involved. First, ALP activity assay determined that 200 μg/mL was the optimal concentration of CTP-CM for the following experiments. CTP-CM had no significant effect on the proliferation of hDPSCs as indicated by CCK-8 and FCM analysis. Both the gene and protein (DSPP/DSPP, RUNX2/RUNX2, OCN/OCN, OSX/OSX, OPN/OPN, ALP/ALP, and COL-1/COL-1) expression levels increased in the CTP-CM-induced hDPSC group as compared with those in the control group at day 3 or 7, showing the positive regulation of CTP-CM on the osteo/odontogenic differentiation of hDPSCs. Mechanistically, MAPK signaling pathways were activated after the CTP-CM treatment, and the inhibitors targeting MAPK were identified which weakened the effects of CTM-CM on the committed differentiation of hDPSCs. These findings could lead to the creation of stem cell therapies for dental regeneration.

scholarly journals TOXICITY ANALYSIS OF RGD-CHITOSAN FROM SHRIMP SHELL SCAFFOLD MEMBRANES TOWARD HUMAN DENTAL PULP CELLS

2017 ◽  
Vol 9 ◽  
pp. 13
Author(s):  
Mauldina Shabrina ◽  
Dewi Fatma Suniarti ◽  
Lisa R Amir ◽  
Erik Idrus
Keyword(s):  
Cell Viability ◽  
Dental Pulp ◽  
Control Group ◽  
Dental Pulp Cells ◽  
Human Dental Pulp Cells ◽  
Membrane Toxicity ◽  
Toxicity Analysis ◽  
Human Dental Pulp ◽  
Pulp Cells ◽  
Group 2

Objective: This study aimed to analyze RGD-Chitosan from Shrimp Shells’ Scaffolds’ (RCSSS) and CSSS membrane toxicity toward human dental pulpcells.Methods: Human dental pulp cells were cultured for 5 days and then exposed to RCSSS or CSSS membranes for 24 hrs. Cell viability was determinedusing an MTT assay method.Results: Cell viability of the RCSSS group and CSSS group was higher than the cell viability of the control group. The cell viability of the RCSSSgroup 2 mg (537.39%) was significantly higher than the CSSS group 2 mg (301.74%).Conclusions: RCSSS membranes were not toxic toward human dental pulp cells and showed better effect toward human dental pulp cells comparedto CSSS membranes.

Human Dental Pulp Stem Cells in Rat Mandibular Bone Defects

2019 ◽  
Vol 207 (3-4) ◽  
pp. 138-148 ◽  
Author(s):  
Rubia Teodoro Stuepp ◽  
Priscilla Barros Delben ◽  
Filipe Modolo ◽  
Andrea Gonçalves Trentin ◽  
Ricardo Castilho Garcez ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Formation ◽  
Dental Pulp ◽  
Bone Defects ◽  
Treated Group ◽  
Dental Pulp Stem Cells ◽  
Control Group ◽  
Mandibular Bone ◽  
Significant Difference ◽  
Human Dental Pulp

This study aimed to evaluate the use of human dental pulp stem cells (hDPSCs) in non-critical-sized mandibular bone defects in rats. hDPSCs from permanent teeth were isolated and engrafted in mandibular bone defects in rats for 7, 14, and 28 days; bone defects without cells formed the control group. Samples were evaluated by scanning electron microscopy (SEM), light microscopy (hematoxylin and eosin staining), and the regeneration area was measured by the Image J program. Before surgery procedures, the human dental pulp cells were characterized as dental pulp stem cells: fusiform morphology, plastic-adherent; expression of CD105, CD73, and CD90; lack of expression of CD45 and CD34, and differentiated into osteoblasts, adipocytes, and chondroblasts. The results indicated that within 7 days the control group presented a pronounced bone formation when compared with the treated group (p < 0.05). After 14 days, the treated group showed an increase in bone formation, but with no statistical difference among the groups (p > 0.05). In the final evaluated period there was no difference between the control group and the treated group (p > 0.05). There was a significant difference between 7 and 14 days (p < 0.05) and between 7 and 28 days (p < 0.05) in the treated group. In conclusion, there is no evidence that the use of hDPSCs in the conditions of this study could improve bone formation in non-critical-sized mandibular bone defects.

scholarly journals Expression of substance P, calcitonin gene-related peptide and vascular endothelial growth factor in human dental pulp under different clinical stimuli

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Javier Caviedes-Bucheli ◽  
Luis Fernando Lopez-Moncayo ◽  
Hernan Dario Muñoz-Alvear ◽  
Jose Francisco Gomez-Sosa ◽  
Luis Eduardo Diaz-Barrera ◽  
...  
Keyword(s):  
Dental Pulp ◽  
Defense Mechanism ◽  
Initial Step ◽  
Control Group ◽  
List Type ◽  
Registration Number ◽  
Human Dental Pulp ◽  
Orthodontic Forces ◽  
Post Hoc ◽  
Endothelial Growth Factor

Abstract Background The aim of this study was to measure the dental pulp inflammatory response through neuropeptides (SP and CGRP) as a response to occlusal trauma, orthodontic movements and a combination of both, as well as the angiogenic defense mechanism through VEGF expression, which could be the initial step to mineralized tissue formation. Methods Forty human dental pulp samples were collected from healthy first premolars with extraction indicated due to orthodontic reasons from a sample of 20 patients. Patients were divided into four groups with 10 premolars each (1 mandibular and 1 maxillary premolar from each patient): healthy pulp control group, occlusal trauma group, moderate orthodontic forces group; and occlusal trauma plus moderate orthodontic forces group. Stimuli were applied for 24 h before tooth extraction in all experimental groups. All samples were processed, and SP, CGRP, and VEGF were measured by radioimmunoassay. The Kruskal–Wallis test was performed to assess significant differences among groups and Mann–Whitney’s U post hoc pairwise comparisons were also performed. Results The highest increase in SP, CGRP, and VEGF expressions was found in the occlusal trauma plus orthodontic forces group, followed by the moderate orthodontic forces, the occlusal trauma and the control groups, with statistically significant differences between all groups for each of the 3 peptides analyzed (Kruskal–Wallis p < 0.001). All possible pairwise post-hoc comparisons were also significant for each peptide analyzed (Mann–Whitney’s U p < 0.001). Conclusion SP, CGRP, and VEGF expressions significantly increase in human dental pulps when stimulated by occlusal trauma combined with moderate orthodontic forces, as compared with these two stimuli applied independently. Name of the registry: Importance of Neurogenic Inflammation in the Angiogenic Response of the Dental Pulp as a Defensive Response. Trial registration number: NCT03804034. Date of registration: 01/15/2019 Retrospectively registered. URL of trial registry record: https://clinicaltrials.gov/ct2/show/NCT03804034?term=NCT03804034&draw=2&rank=1.

scholarly journals Expression of Substance P, Calcitonin Gene-Related Peptide and Vascular Endothelial Growth Factor in Human Dental Pulp Under Different Clinical Stimuli

2020 ◽  
Author(s):  
Javier Caviedes-Bucheli ◽  
Luis Fernando Lopez-Moncayo ◽  
Hernan Dario Munoz-Alvear ◽  
Jose Francisco Gomez-Sosa ◽  
Luis Eduardo Diaz-Barrera ◽  
...  
Keyword(s):  
Dental Pulp ◽  
Control Group ◽  
Vascular Endothelial ◽  
Calcitonin Gene ◽  
Registration Number ◽  
Human Dental Pulp ◽  
Orthodontic Forces ◽  
Post Hoc ◽  
Endothelial Growth Factor ◽  
Occlusal Interferences

Abstract Background To quantify the expression of SP, CGRP, and VEGF in human dental pulps as a response to occlusal interferences, moderate orthodontic forces, and occlusal interferences plus moderate orthodontic forces simultaneously.Methods Forty human dental pulp samples were collected from healthy premolars indicated for extraction for orthodontic reasons. The teeth were divided into four groups with 10 samples each: Healthy pulp control group, occlusal interference group, moderate orthodontic forces group; and occlusal interferences plus moderate orthodontic forces group. Stimuli were applied for 24 h before tooth extraction in all experimental groups. All samples were processed, and SP, CGRP, and VEGF were measured by radioimmunoassay. The ANOVA test was performed to establish significant differences between groups and Tukey’s HSD post hoc comparisons were also performed.Results The highest increase in SP, CGRP, and VEGF expressions was found in the occlusal trauma plus orthodontic forces group, followed by the moderate orthodontic forces, the occlusal interferences and the control groups, with statistically significant differences between all groups (ANOVA p<0.001) and post-hoc comparisons (Tukey HSD p<0.001).Conclusion SP, CGRP, and VEGF expressions significantly increase in human dental pulps when stimulated by occlusal interferences combined with moderate orthodontic forces, as compared with when these two stimuli are applied independently. Peptides expression is directly proportional to the magnitude of the stimulus.Name of the registry: Importance of Neurogenic Inflammation in the Angiogenic Response of the Dental Pulp as a Defensive ResponseTrial registration number: NCT03804034Date of registration: 01/15/2019 Retrospectively registeredURL of trial registry record: https://clinicaltrials.gov/ct2/show/NCT03804034?term=NCT03804034&draw=2&rank=1

scholarly journals Immediate response of human dental pulp capped with mineral trioxide aggregate, Portland cement and biodentin

2019 ◽  
Vol 45 (2) ◽  
pp. 108-116
Author(s):  
AKM Bashar ◽  
A.K.M Nurul Kabir ◽  
Rozina Akhter Rizdina ◽  
Ranjit Ghosh ◽  
Ashis Kumar Biswas ◽  
...  
Keyword(s):  
Portland Cement ◽  
Inflammatory Reaction ◽  
Dental Pulp ◽  
Mineral Trioxide Aggregate ◽  
Tissue Reaction ◽  
Control Group ◽  
Pulp Capping ◽  
Pulp Inflammation ◽  
Significant Difference ◽  
Human Dental Pulp

Background: The initial inflammatory reaction of pulp capping materials on the dental pulp has an intimate relation in promoting the future cellular differentiation and biomaterial mineralisation. So, analysis of immediate pulpal tissue reaction in vivo, is also important for evaluation of ultimate efficacy any pulp capping agent. To observe immediate inflammatory response of Human Dental Pulp capped with Mineral Trioxide Aggregate (MTA), Biodentin and Portland Cement (PC). Methods: A total of 70 permanent premolars teeth planned to be extracted for orthodontic alignment of occlusion were used as study sample. The teeth were divided into 3 experimental groups, MTA (n=20), Biodentin (n=20) Portland cement (n=20) and control group (n=10). After having an occlusal exposure of approximately 1.5 mm in diameter; in group A, pulp of teeth was capped with 2-mm-thick layer of ProRoot White MTA (Dentsply) and in group B, with sterile Biodentin (Septodont) according to the manufacturer’s recommendations. Whereas in group C, pulp of teeth was capped with sterile Portland Cement (PC). After placing the experimental material in each group, all teeth restored with glass i‹xiomer cement. After 24 hours the teeth were extracted, fixed in 10% buffered formalin solution, then decalcified by 10% nitric acid and embedded in paraffin. Finally, sectioned into 2 to 3-micron-thick serial sections in the linguo-buccal plane and stained with hematoxylin-eosin. After then the amount of pulp inflammation (type, intensity, and extension) were determined by using a predetermined evaluation criterion under an optical microscope at 40a magnification. Ten intact teeth, which received no exposure and pulp capping but extracted due to orthodontic purpose were also collected and treated as the control group (group D); undergone same histologic preparation and evaluation. Significantstatistical differences among the experimental groups were to be found (p<0.05). Results: Histologically, all the three tested materials produced immediate pulpal tissue reaction. ‘Biodentin’ found to be most immediate pulpal tissue reactive (reactive in 100% cases) and ‘Portland Cement’ showed least immediate tissue reaction (only in 30.0% cases). whereas, MTA produced immediate tissue reaction only in 50.0% cases. Immediate pulpal inflammatory reaction in response to tested material found to be statistically significant different between ‘Biodentin’ and ‘Portland cmient’ (p=0.01), also between ‘Biodentin’ and ‘MTA’ and (p=0.001); but there was no statistically significant difference between ‘MTA’ and ‘Portland cement’ (p =0.197). Conclusion: Considering the maximum immediate pulpal tissue reaction (Inflammation), Biodentin is expected to produce most favorable ultimate bioactivity (biomaterial mineralization) after pulp capping. Bangladesh Med Res Counc Bull 2019; 45: 108-116

Sign in / Sign up

Export Citation Format

Share Document