scholarly journals Fibrin scaffolds containing dental pulp stem cells for the repair of periodontal bone defects

2020 ◽  
Vol 7 (1) ◽  
pp. 59-69
Author(s):  
Yu. A. Dombrovskaya ◽  
N. I. Enukashvily ◽  
A. V. Kotova ◽  
S. S. Bilyk ◽  
A. N. Kovalenko ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Tissue ◽  
Fibrin Glue ◽  
Dental Pulp ◽  
Bone Defects ◽  
Dental Pulp Stem Cells ◽  
Osteogenic Potential ◽  
Tooth Pulp ◽  
Alizarin Red ◽  
Periodontal Bone Defects

Background. 3D scaffolds plays an important role in developing new approaches in modern dentistry. They are used to establish optimal conditions for cell differentiation, vascularization and remodeling of regenerating bone tissue.Objective. Evaluation of the possibility of creating scaffolds developed on the basis of 3D modeling of periodontal bone defects and containing tooth pulp stem cells.Materials and Methods. The computer tomography data of the maxillar bone tissue defect were analysed. Anatomical prototype — a mold representing defects of the vestibular and palatal fragments of bone tissue was created by 3D printing. This 3D form was filled with fibrin glue and dental pulp stem cells. The fibrin glue was prepared from autologous blood plasma and mixed with dental pulp stem cells.Results. Fibrin glue prepared from an autologous plasma concentrate (fibrinogen 20 g/l) retains its shape for 4 days. On the day 5, the clot retraction became clearly visible and on the day 7, the clot diameter decreased to 50 % of the original size. The proliferation rate of cells, grown both inside the scaffold and in 2D conditions, did not differ. The immunophenotype of cells of both groups corresponded to the immunophenotype of mesenchy mal stromal cells. The mesenchymal immunophenotype is a feature of dental stem cells. Alizarin red staining of cells both grown on adhesive culture plastic and extracted from glue on day 10 after the induction of osteogenic differentiation did not differ.Conclusion. The fibrin glue is a good material for creation a scaffold with suitable mechanical characteristics. The cells enclosed in the fibrin glue maintain their viability, immunophenotype and osteogenic potential. This technology can be used for bone tissue repair in dentistry and maxillofacial surgery.

scholarly journals Fibrin Glue Implants Seeded with Dental Pulp and Periodontal Ligament Stem Cells for the Repair of Periodontal Bone Defects: A Preclinical Study

2021 ◽  
Vol 8 (6) ◽  
pp. 75
Author(s):  
Natella I. Enukashvily ◽  
Julia A. Dombrovskaya ◽  
Anastasia V. Kotova ◽  
Natalia Semenova ◽  
Irina Karabak ◽  
...  
Keyword(s):  
Stem Cells ◽  
3D Printing ◽  
Fibrin Glue ◽  
Bone Defect ◽  
Dental Pulp ◽  
Alveolar Bone ◽  
Bone Defects ◽  
Osteogenic Potential ◽  
Fibrin Gel ◽  
A Cell

A technology to create a cell-seeded fibrin-based implant matching the size and shape of bone defect is required to create an anatomical implant. The aim of the study was to develop a technology of cell-seeded fibrin gel implant creation that has the same shape and size as the bone defect at the site of implantation. Using computed tomography (CT) images, molds representing bone defects were created by 3D printing. The form was filled with fibrin glue and human dental pulp stem cells (DPSC). The viability, set of surface markers and osteogenic differentiation of DPSC grown in fibrin gel along with the clot retraction time were evaluated. In mice, an alveolar bone defect was created. The defect was filled with fibrin gel seeded with mouse DPSC. After 28 days, the bone repair was analyzed with cone beam CT and by histological examination. The proliferation rate, set of surface antigens and osteogenic potential of cells grown inside the scaffold and in 2D conditions did not differ. In mice, both cell-free and mouse DPSC-seeded implants increased the bone tissue volume and vascularization. In mice with cell-seeded gel implants, the bone remodeling process was more prominent than in animals with a cell-free implant. The technology of 3D-printed forms for molding implants can be used to prepare implants using components that are not suitable for 3D printing.

scholarly journals Enhanced Osteogenesis of Dental Pulp Stem Cells In Vitro Induced by Chitosan–PEG-Incorporated Calcium Phosphate Cement

Polymers ◽  
2021 ◽  
Vol 13 (14) ◽  
pp. 2252
Author(s):  
Jae Eun Kim ◽  
Sangbae Park ◽  
Woong-Sup Lee ◽  
Jinsub Han ◽  
Jae Woon Lim ◽  
...  
Keyword(s):  
Stem Cells ◽  
Calcium Phosphate ◽  
Zeta Potential ◽  
Mechanical Strength ◽  
Calcium Phosphate Cement ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Ion Trapping ◽  
Alizarin Red

The use of bone graft materials is required for the treatment of bone defects damaged beyond the critical defect; therefore, injectable calcium phosphate cement (CPC) is actively used after surgery. The application of various polymers to improve injectability, mechanical strength, and biological function of injection-type CPC is encouraged. We previously developed a chitosan–PEG conjugate (CS/PEG) by a sulfur (VI) fluoride exchange reaction, and the resulting chitosan derivative showed high solubility at a neutral pH. We have demonstrated the CPC incorporated with a poly (ethylene glycol) (PEG)-grafted chitosan (CS/PEG) and developed CS/PEG CPC. The characterization of CS/PEG CPC was conducted using Fourier transform infrared spectroscopy (FT-IR) and X-ray diffraction (XRD). The initial properties of CS/PEG CPCs, such as the pH, porosity, mechanical strength, zeta potential, and in vitro biocompatibility using the WST-1 assay, were also investigated. Moreover, osteocompatibility of CS/PEG CPCs was carried out via Alizarin Red S staining, immunocytochemistry, and Western blot analysis. CS/PEG CPC has enhanced mechanical strength compared to CPC, and the cohesion test also demonstrated in vivo stability. Furthermore, we determined whether CS/PEG CPC is a suitable candidate for promoting the osteogenic ability of Dental Pulp Stem Cells (DPSC). The elution of CS/PEG CPC entraps more calcium ion than CPC, as confirmed through the zeta potential test. Accordingly, the ion trapping effect of CS/PEG is considered to have played a role in promoting osteogenic differentiation of DPSCs. The results strongly suggested that CS/PEG could be used as suitable additives for improving osteogenic induction of bone substitute materials.

scholarly journals Odontoblast-Like Cells Differentiated from Dental Pulp Stem Cells Retain Their Phenotype after Subcultivation

2018 ◽  
Vol 2018 ◽  
pp. 1-12 ◽  
Author(s):  
Paula A. Baldión ◽  
Myriam L. Velandia-Romero ◽  
Jaime E. Castellanos
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Matrix Protein ◽  
Dental Materials ◽  
Dental Pulp Stem Cells ◽  
Cell Physiology ◽  
Pulp Tissue ◽  
Alizarin Red ◽  
Dentin Matrix Protein ◽  
Mineral Deposition

Odontoblasts, the main cell type in teeth pulp tissue, are not cultivable and they are responsible for the first line of response after dental restauration. Studies on dental materials cytotoxicity and odontoblast cells physiology require large quantity of homogenous cells retaining most of the phenotype characteristics. Odontoblast-like cells (OLC) were differentiated from human dental pulp stem cells using differentiation medium (containing TGF-β1), and OLC expanded after trypsinization (EXP-21) were evaluated and compared. Despite a slower cell growth curve, EXP-21 cells express similarly the odontoblast markers dentinal sialophosphoprotein and dentin matrix protein-1 concomitantly with RUNX2 transcripts and low alkaline phosphatase activity as expected. Both OLC and EXP-21 cells showed similar mineral deposition activity evidenced by alizarin red and von Kossa staining. These results pointed out minor changes in phenotype of subcultured EXP-21 regarding the primarily differentiated OLC, making the subcultivation of these cells a useful strategy to obtain odontoblasts for biocompatibility or cell physiology studies in dentistry.

scholarly journals Evaluation of Resin-Based Material Containing Copaiba Oleoresin (Copaifera Reticulata Ducke): Biological Effects on the Human Dental Pulp Stem Cells

Biomolecules ◽  
2020 ◽  
Vol 10 (7) ◽  
pp. 972
Author(s):  
Roberta Souza D’Almeida Couto ◽  
Maria Fernanda Setubal Destro Rodrigues ◽  
Leila Soares Ferreira ◽  
Ivana Márcia Alves Diniz ◽  
Fernando de Sá Silva ◽  
...  
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Biological Effects ◽  
Dental Pulp Stem Cells ◽  
Nodule Formation ◽  
Alizarin Red ◽  
Pulp Capping ◽  
Hsp 27 ◽  
Dental Pulp Capping ◽  
Human Dental Pulp

The purpose of this study was to analyze in vitro the biological effects on human dental pulp stem cells triggered in response to substances leached or dissolved from two experimental cements for dental pulp capping. The experimental materials, based on extracts from Copaifera reticulata Ducke (COP), were compared to calcium hydroxide [Ca(OH)2] and mineral trioxide aggregate (MTA), materials commonly used for direct dental pulp capping in restorative dentistry. For this, human dental pulp stem cells were exposed to COP associated or not with Ca(OH)2 or MTA. Cell cytocompatibility, migration, and differentiation (mineralized nodule formation (Alizarin red assay) and gene expression (RT-qPCR) of OCN, DSPP, and HSP-27 (genes regulated in biomineralization events)) were evaluated. The results showed that the association of COP reduced the cytotoxicity of Ca(OH)2. Upregulations of the OCN, DSPP, and HSP-27 genes were observed in response to the association of COP to MTA, and the DSPP and HSP-27 genes were upregulated in the Ca(OH)2 + COP group. In up to 24 h, cell migration was significantly enhanced in the MTA + COP and Ca(OH)2 + COP groups. In conclusion, the combination of COP with the currently used materials for dental pulp capping [Ca(OH)2 and MTA] improved the cell activities related to pulp repair (i.e., cytocompatibility, differentiation, mineralization, and migration) including a protective effect against the cytotoxicity of Ca(OH)2.

scholarly journals A Comparative In Vitro Analysis of the Osteogenic Potential of Human Dental Pulp Stem Cells Using Various Differentiation Conditions

2020 ◽  
Vol 21 (7) ◽  
pp. 2280 ◽  
Author(s):  
Terezia Okajcekova ◽  
Jan Strnadel ◽  
Michal Pokusa ◽  
Romana Zahumenska ◽  
Maria Janickova ◽  
...  
Keyword(s):  
Stem Cells ◽  
Osteogenic Differentiation ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Osteogenic Potential ◽  
Surface Markers ◽  
Osteogenic Medium ◽  
Differentiated Cells ◽  
Mineralization Potential ◽  
Gene And Protein Expression

Dental pulp stem cells (DPSCs) have excellent proliferative properties, mineralization potential and can be easily obtained from third molar teeth. Recently, many studies have focused on isolation and differentiation of DPSCs. In our study, we focused on biological properties of non-differentiated DPSCs in comparison with osteogenic differentiated cells from DPSCs. We analyzed morphology as well as mineralization potential using three varied osteogenic differentiation media. After fifteen days of differentiation, calcium deposit production was observed in all three osteogenic differentiation media. However, only one osteogenic medium, without animal serum supplement, showed rapid and strong calcification—OsteoMAX-XF™ Differentiation Medium. Therefore, we examined specific surface markers, and gene and protein expression of cells differentiated in this osteogenic medium, and compared them to non-differentiated DPSCs. We proved a decrease in expression of CD9 and CD90 mesenchymal stem cell surface markers, as well as downregulation in the expression of pluripotency genes (NANOG and OCT-4) and increased levels of expression in osteogenic genes (ALP, BSP, OCN and RUNX2). Moreover, osteogenic proteins, such as BSP and OCN, were only produced in differentiated cells. Our findings confirm that carefully selected differentiation conditions for stem cells are essential for their translation into future clinical applications.

Transplantation of Dental Pulp Stem Cells in Experimental Bone Defect

2017 ◽  
Vol 34 ◽  
pp. 94-100 ◽  
Author(s):  
Endang W. Bachtiar ◽  
Fatma S. Dewi ◽  
Ahmad Aulia Yusuf ◽  
Rahmi Ulfiana
Keyword(s):  
Stem Cells ◽  
Animal Model ◽  
Bone Regeneration ◽  
Bone Tissue ◽  
Bone Defect ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Bone Tissue Regeneration ◽  
Histological Evaluation ◽  
Control Rabbit

This is preliminary study in order to investigate the effect of dental pulp stem cells (DPSCs) on bone regeneration in an animal model. New Zealand rabbits were used as animal model. The critical defect was created in femoral bone and transplantation of DPSCs applied into bone defect. A colorimetric assay was used to detect ALP level in rabbit’s serum. Bone tissue regeneration was evaluated by histological analysis. In the 2nd week, the treated rabbit show increasing in the activity of ALP (157,925 μU) compared to control rabbit (155,361 μU). This increasing trend continues significantly in DPSCs rabbit (169.750 μU) compared to control rabbit (160.406) after 4 weeks. Histological evaluation revealed that the amount of bone lamellae and osteocytes were filled the defect area of DPSCs treated rabbit. Conclusions: Transplantation of DPSCs accelerating bone regeneration by raising ALP level and forming new bone tissue.

scholarly journals Isolation and culture of dental pulp stem cells from permanent and deciduous teeth

2019 ◽  
Vol 35 (4) ◽  
Author(s):  
Shagufta Naz ◽  
Farhan Raza Khan ◽  
Raheela Rahmat Zohra ◽  
Sahreena Salim Lakhundi ◽  
Mehwish Sagheer Khan ◽  
...  
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Permanent Teeth ◽  
Deciduous Teeth ◽  
Tooth Pulp ◽  
Human Teeth ◽  
Creative Commons ◽  
Calcified Tissue

Objective: To isolate dental pulp mesenchymal stem cells (MSCs) from non-infected human permanent and deciduous teeth. Methods: It was an in-vitro experimental study. Human teeth were collected from 13 apparently healthy subjects including nine adults and four children. After decoronation dental pulps were extirpated from teeth and cultured via explant method in a stem cell defined media. Data was analyzed by descriptive statistics. Results: As above MSCs emerged exhibiting fibroblast-like morphology. In vitro culture was positive for 100% (9/9) and 75% (3/4) of the permanent and deciduous teeth respectively. First cell appeared from deciduous teeth pulp in 10±6.2 days while permanent teeth pulp took 12.4±3.7 days. Together, 26.6±3.6 and 24.5±3.5 days were required for permanent and deciduous tooth pulp stem cells to be ready for further assays. Conclusions: The protocol we developed is easy and consistent and can be used to generate reliable source of MScs for engineering of calcified and non-calcified tissue for regenerative medicine approaches. doi: https://doi.org/10.12669/pjms.35.4.540 How to cite this:Naz S, Khan FR, Zohra RR, Lakhundi SS, Khan MS, Mohammed N, et al. Isolation and culture of dental pulp stem cells from permanent and deciduous teeth. Pak J Med Sci. 2019;35(4):---------. doi: https://doi.org/10.12669/pjms.35.4.540 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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