scholarly journals ANALYSIS OF LYSATE PLATELET-RICH FIBRIN EFFECTS ON HUMAN DENTAL PULP STEM CELL DIFFERENTIATION THROUGH DENTIN SIALOPHOSPHOPROTEIN EXPRESSION

2020 ◽  
pp. 34-37
Author(s):  
SILVIANA SWASTININGTYAS ◽  
ANGGRAINI MARGONO ◽  
DINI ASRIANTI ◽  
RUNY OKTAYANI ◽  
INDAH YULIANTO
Keyword(s):  
Dental Pulp ◽  
Culture Media ◽  
Stem Cell Differentiation ◽  
Alizarin Red ◽  
Platelet Rich Fibrin ◽  
Dentin Sialophosphoprotein ◽  
Human Dental Pulp ◽  
Stable Growth ◽  
And Control

Objective: In vitro, the culture media in which human dental pulp stem cells (hDPSCs) are grown are supplemented with specific growth factors thatinduce cell cycle entry and differentiation. Lysate platelet-rich fibrin (L-PRF) is a unique and stable growth factor supplement produced from plateletslysed by freezing-thawing. In this study, we aimed to analyze the potential effects of L-PRF on hDPSC differentiation.Methods: We divided hDPSCs isolated from human third molars at the second passage into five culture media groups treated with 1%, 5%, 10%,and 25% L-PRF or 10% fetal bovine serum (control). After 7 days, we evaluated hDPSC differentiation using an enzyme-linked immunosorbent assayspecific for dentin sialophosphoprotein and Alizarin-Red staining.Results: None of our analyses revealed any significant differences between the L-PRF- and control-treated cells.Conclusion: L-PRF could potentially induce the differentiation of hDPSCs in vitro.

scholarly journals TRANSFORMING GROWTH FACTOR-β1 EXPRESSION IN VARIOUS CONCENTRATIONS OF ADVANCED PLATELET-RICH FIBRIN MODULATING HUMAN DENTAL PULP STEM CELL DIFFERENTIATION

2020 ◽  
pp. 38-40
Author(s):  
ANGGRAINI MARGONO ◽  
DINI ASRIANTI ◽  
FRIEDA AYU PRIHADINI ◽  
INDAH JULIANTO
Keyword(s):  
Dental Pulp ◽  
Transforming Growth Factor ◽  
Culture Media ◽  
Stem Cell Differentiation ◽  
Transforming Growth Factor Β1 ◽  
Serum Starvation ◽  
Differentiation Process ◽  
Platelet Rich Fibrin ◽  
Human Dental Pulp ◽  
Tgf Β1

Objective: Modification of the speed and time of centrifugation based on the low-speed centrifugation concept for platelet-rich fibrin (PRF) has resulted in a new type of PRF known as advanced PRF (A-PRF). A-PRF can release several types of growth factors (GFs) that participate in the process of differentiation, such as transforming GF-β1 (TGF-β1). The aim of this study was to analyze TGF-β1 expression in various concentrations of A-PRF in the differentiation process of human dental pulp stem cells (hDPSCs).Methods: hDPSC cultures were obtained from those of previous research (ethical approval form has been attached). These hDPSCs were in the 2nd–3rd passage, and serum starvation was done by reducing fetal bovine serum (FBS) levels in the hDPSC culture media. A-PRF was obtained using 10 ml blood collected from the cubital vein, which was centrifuged at 1500 rpm for 14 min and then divided into four concentration groups. TGF-β1 expression in 1%, 5%, and 25% A-PRF as well as in 10% FBS (control) was analyzed by ELISA on day 7.Results: Although no significant differences were observed in TGF-β1 expression between 1%, 5%, and 25% A-PRF, and 10% FBS, it was observed that the higher the concentration of A-PRF, the greater the TGF-β1 expression.Conclusion: The expression of TGF-β1 was consistent with the increase in A-PRF concentration. The highest TGF-β1 expression was detected in 25% A-PRF among all concentrations in the differentiation process of hDPSCs.

scholarly journals Effect of Vitamins D and E on the Proliferation, Viability, and Differentiation of Human Dental Pulp Stem Cells: An In Vitro Study

2020 ◽  
Vol 2020 ◽  
pp. 1-10
Author(s):  
Lina M. Escobar ◽  
Zita Bendahan ◽  
Andrea Bayona ◽  
Jaime E. Castellanos ◽  
María-Clara González
Keyword(s):  
Stem Cells ◽  
Cell Proliferation ◽  
Dental Pulp ◽  
Morphological Changes ◽  
In Vitro Study ◽  
Osteoblastic Differentiation ◽  
Dental Pulp Stem Cells ◽  
Alizarin Red ◽  
Human Dental Pulp

Introduction. The aim of the present study was to determine the effects of vitamins D and E on the proliferation, morphology, and differentiation of human dental pulp stem cells (hDPSCs). Methods. In this in vitro experimental study, hDPSCs were isolated, characterized, and treated with vitamins D and E, individually and in combination, utilizing different doses and treatment periods. Changes in morphology and cell proliferation were evaluated using light microscopy and the resazurin assay, respectively. Osteoblast differentiation was evaluated with alizarin red S staining and expression of RUNX2, Osterix, and Osteocalcin genes using real-time RT-PCR. Results. Compared with untreated cells, the number of cells significantly reduced following treatment with vitamin D (49%), vitamin E (35%), and vitamins D + E (61%) after 144 h. Compared with cell cultures treated with individual vitamins, cells treated with vitamins D + E demonstrated decreased cell confluence, with more extensive and flatter cytoplasm that initiated the formation of a significantly large number of calcified nodules after 7 days of treatment. After 14 days, treatment with vitamins D, E, and D + E increased the transcription of RUNX2, Osterix, and Osteocalcin genes. Conclusions. Vitamins D and E induced osteoblastic differentiation of hDPSCs, as evidenced by the decrease in cell proliferation, morphological changes, and the formation of calcified nodules, increasing the expression of differentiation genes. Concurrent treatment with vitamins D + E induces a synergistic effect in differentiation toward an osteoblastic lineage.

scholarly journals Synergies of Human Umbilical Vein Endothelial Cell-Laden Calcium Silicate-Activated Gelatin Methacrylate for Accelerating 3D Human Dental Pulp Stem Cell Differentiation for Endodontic Regeneration

Polymers ◽  
2021 ◽  
Vol 13 (19) ◽  
pp. 3301
Author(s):  
Wei-Yun Lai ◽  
Tzu-Hsin Lee ◽  
Jian-Xun Chen ◽  
Hooi-Yee Ng ◽  
Tsui-Hsien Huang ◽  
...  
Keyword(s):  
Dental Pulp ◽  
Vascular Endothelial Cells ◽  
Umbilical Vein ◽  
Stem Cell Differentiation ◽  
Cell Block ◽  
Fish Gelatin ◽  
Alizarin Red ◽  
Covalent Bonds ◽  
Human Dental Pulp ◽  
Dentin Regeneration

According to the Centers for Disease Control and Prevention, tooth caries is a common problem affecting 9 out of every 10 adults worldwide. Dentin regeneration has since become one of the pressing issues in dentistry with tissue engineering emerging as a potential solution for enhancing dentin regeneration. In this study, we fabricated cell blocks with human dental pulp stem cells (hDPSCs)-laden alginate/fish gelatin hydrogels (Alg/FGel) at the center of the cell block and human umbilical vascular endothelial cells (HUVEC)-laden Si ion-infused fish gelatin methacrylate (FGelMa) at the periphery of the cell block. 1H NMR and FTIR results showed the successful fabrication of Alg/FGel and FGelMa. In addition, Si ions in the FGelMa were noted to be bonded via covalent bonds and the increased number of covalent bonds led to an increase in mechanical properties and improved degradation of FGelMa. The Si-containing FGelMa was able to release Si ions, which subsequently significantly not only enhanced the expressions of angiogenic-related protein, but also secreted some cytokines to regulate odontogenesis. Further immunofluorescence results indicated that the cell blocks allowed interactions between the HUVEC and hDPSCs, and taken together, were able to enhance odontogenic-related markers’ expression, such as alkaline phosphatase (ALP), dentin matrix phosphoprotein-1 (DMP-1), and osteocalcin (OC). Subsequent Alizarin Red S stain confirmed the benefits of our cell block and demonstrated that such a novel combination and modification of biomaterials can serve as a platform for future clinical applications and use in dentin regeneration.

Inhibition of Human Dental Pulp Stem Cell Differentiation by Notch Signaling

2008 ◽  
Vol 87 (3) ◽  
pp. 250-255 ◽  
Author(s):  
C. Zhang ◽  
J. Chang ◽  
W. Sonoyama ◽  
S. Shi ◽  
C.-Y. Wang
Keyword(s):  
Stem Cells ◽  
Notch Signaling ◽  
Cell Fate ◽  
Dental Pulp ◽  
Critical Role ◽  
Stem Cell Differentiation ◽  
Over Expression ◽  
Odontoblastic Differentiation ◽  
Human Dental Pulp

Notch signaling plays a critical role in development and cell fate specification. Notch receptors and ligands have been found to be expressed in dental epithelium or mesenchyme in the developing tooth, suggesting that Notch signaling may regulate odontogenesis. Post-natal human dental pulp stem cells (DPSCs) isolated from the dental pulp have characteristics of mesenchymal stem cells and can differentiate into odontoblasts. In this study, we examined whether Notch signaling regulated the odontoblastic differentiation of DPSCs. We found that over-expression of the Notch ligand, Jagged-1, activated the Notch signaling pathway in DPSCs. Jagged-1 inhibited the odontoblastic differentiation of DPSCs in vitro. Jagged-1-expressing DPSCs could not form mineralized tissues in vivo. Moreover, over-expression of the constitutively activated Notch1 intracellular domain (Notch-ICD) also inhibited odontoblastic differentiation of DPSCs. Taken together, our results demonstrate that Notch signaling can inhibit the odontoblastic differentiation of DPSCs.

Biological Effects of Provisional Resin Materials on Human Dental Pulp Stem Cells

2017 ◽  
Vol 42 (2) ◽  
pp. E81-E92 ◽  
Author(s):  
S-K Jun ◽  
C Mahapatra ◽  
H-H Lee ◽  
H-W Kim ◽  
J-H Lee
Keyword(s):  
Stem Cells ◽  
Proinflammatory Cytokines ◽  
Repeated Measures ◽  
Dental Pulp ◽  
Culture Media ◽  
Biological Effects ◽  
Dental Pulp Stem Cells ◽  
Pulp Tissue ◽  
Human Dental Pulp

SUMMARY Objectives: This study investigated the in vitro cytotoxicity as well as the proinflammatory cytokine expression of provisional resin materials on primary cultured human dental pulp stem cells (hDPSCs). Methods: Five commercially available provisional resin materials were chosen (SNAP [SN], Luxatemp [LT], Jet [JE], Revotek LC [RL], and Vipi block [VB]). Eluates that were either polymerizing or already set were added to hDPSCs under serially diluted conditions divided into three different setting times (25% set, 50% set, and 100% set) and incubated for 24 hours with 2× concentrated culture media. Cell cytotoxicity tests were performed by LDH assay and live and dead confocal microscope images. The expression of proinflammatory cytokines in SN and VB was measured using cytokine antibody arrays. Data were analyzed using repeated measures analysis of variance (ANOVA) or ANOVA followed by the Tukey post hoc test at a significance level of p<0.05. Results: Cytotoxicity greater than 30% was observed in the 50% diluted culture in SN, LT, and JE in the already set stage (p<0.05), while it was detected in SN and LT in early or intermediate stage samples. The cytotoxicity of SN, JE, and LT was greater with eluates from the polymerizing phase compared to that from already set samples (p<0.05), as observed by live and dead images. On the other hand, RL and VB did not exhibit cytotoxicity greater than 30%. Proinflammatory cytokines were not detected in 12.5% diluted culture with eluates from VB and early set stage SN. Conclusions: The eluates from chemical-activated provisional resin materials during polymerization (SN, LT, and JE) were cytotoxic to hDPSCs and may adversely affect pulp tissue.

scholarly journals Biocompatibility and Bioactivity of Set Direct Pulp Capping Materials on Human Dental Pulp Stem Cells

Materials ◽  
2020 ◽  
Vol 13 (18) ◽  
pp. 3925
Author(s):  
Yemi Kim ◽  
Donghee Lee ◽  
Dani Song ◽  
Hye-Min Kim ◽  
Sin-Young Kim
Keyword(s):  
Stem Cells ◽  
Cell Migration ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Nodule Formation ◽  
Alizarin Red ◽  
Pulp Capping ◽  
Alp Activity ◽  
Human Dental Pulp ◽  
And Control

In this study, we assessed the biocompatibility and bioactivity of various pulp capping materials—ProRoot MTA (Dentsply Tulsa Dental Specialties), Biodentine (Septodont), TheraCal LC (Bisco), and Dycal (Dentsply Caulk)—on human dental pulp stem cells (hDPSCs). Experimental disks (diameter, 7 mm; height, 4 mm) were stored in a humified incubator at 37 °C for 48 h. Then, the pulp capping materials were tested for cytotoxic effects by methyl-thiazoldiphenyl-tetrazolium and scratch wound healing assays, and for mineralization potential by Alizarin red S (ARS) staining assay and alkaline phosphatase enzyme (ALP) activity. Cell viability and cell migration did not significantly differ between ProRoot MTA, Biodentine, and control (p > 0.05). TheraCal LC exhibited slower cell migration on days 2–4 compared to control (p < 0.05), and Dycal showed no cell migration. ALP activity was highest with Biodentine on days 10 and 14, and was lowered with TheraCal LC and Dycal (p < 0.05). In the ARS assay, hDPSCs grown in ProRoot MTA and TheraCal LC eluates showed significantly increased mineralized nodule formation on day 21 compared to Biodentine, Dycal, and control (p < 0.05). These findings indicate that ProRoot MTA, Biodentine, and TheraCal LC exhibit better biocompatibility and bioactivity than Dycal.

Functionalized self-assembled peptide RAD/Dentonin hydrogel scaffold promotes dental pulp regeneration

2021 ◽  
Author(s):  
Yijuan Liu ◽  
Lina Fan ◽  
Xuemei Lin ◽  
Luning Zou ◽  
Yaoyao Li ◽  
...  
Keyword(s):  
Dental Pulp ◽  
Immunofluorescent Staining ◽  
Hydrogel Scaffold ◽  
Alizarin Red ◽  
Pulp Regeneration ◽  
Odontogenic Differentiation ◽  
Human Dental Pulp ◽  
Self Assembled ◽  
Functional Peptide

Abstract RADA16-Ⅰ is an ion-complementary self-assembled peptide with a regular folded secondary conformation and can be assembled into an ordered nanostructure. Dentonin is an extracellular matrix phosphate glycoprotein functional peptide motif-containing RGD and SGDG motifs. In this experiment, we propose to combine RAD and Dentonin to form a functionalized self-assembled peptide RAD/Dentonin hydrogel scaffold. Furthermore, we expect that the RAD with the addition of functional motif Dentonin can promote pulp regeneration. The study analyzed the physicochemical properties of RAD/Dentonin through Circular dichroism, Morphology scanning, and Rheology. Besides, we examined the scaffold’s biocompatibility by Immunofluorescent staining, CCK-8 method, Live/Dead fluorescent staining, and 3D reconstruction. Finally, we applied ALP activity assay, RT-qPCR, and Alizarin red S staining to detect the effect of RAD/Dentonin on the odontogenic differentiation of human dental pulp stem cells (hDPSCs). The results showed that RAD/Dentonin spontaneously assembles into a hydrogel with a β-sheet-based nanofiber network structure. In vitro, RAD/Dentonin has superior biocompatibility and enhances adhesive proliferation, migration, odontogenic differentiation, and mineralization deposition of hDPSCs. In conclusion, the novel self-assembled peptide RAD/Dentonin is a new scaffold material suitable for cell culture and has promising applications as a scaffold for endodontic tissue engineering.

scholarly journals Increased VEGF-A Expression of Human Dental Pulp Stem Cells (hDPSCs) Cultured with Advanced Platelet Rich Fibrin (A-PRF)

2021 ◽  
Vol 15 (1) ◽  
pp. 569-574
Author(s):  
Dini Asrianti Bagio ◽  
Indah Julianto ◽  
Anggraini Margono ◽  
Endang Suprastiwi
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Control Group ◽  
Platelet Rich Fibrin ◽  
Significant Difference ◽  
Angiogenesis Process ◽  
Human Dental Pulp ◽  
Post Hoc

Background: VEGF-A expression of human dental pulp stem cells (hDPSCs) can induce the angiogenesis process of dental pulp regeneration. This in vitro study aimed to analyze the effect of various concentrations of Advanced Platelet Rich Fibrin (A-PRF) conditioned media (CM) on the increased expression of vascular endothelial growth factor-A (VEGF-A) of hDPSCs. Methods: hDPSCs were collected from ten third molars extracted from nine healthy donors, cultured, and then harvested at the end of the third passage. The hDPSCs were seeded in four different CM (control group: hDPSCs + DMEM; 1% A-PRF CM group: hDPSCs + 1% A-PRF CM; 5% A-PRF CM group: hDPSCs + 5% A-PRF CM; 10% A-PRF CM group: hDPSCs + 10% A-PRF CM). All of the groups were cultured in biological triplicates (Triplo) and observed for 5, 12, and 24 hours. The VEGF-A protein expression of hDPSCs was measured using human VEGF-A ELISA at a wavelength of 405 nm. Data was analyzed with Kruskal Wallis and post hoc Mann Whitney test with p<0.05. Results: The VEGF-A expression rate of hDPSCs among all groups was statistically significantly different at 5, 12 and 24 hours of observations (p<0.05). Post hoc analysis test showed a statistically significant difference of hDPSCs’s VEGF-A expression between 5% A-PRF groups compared to other groups at 5 and 12 hours of observation (p<0.05). However, there were no statistically significant differences observed of hDPSCs’ VEGF-A expression at 24 hours of observation between 1%, 5% and 10% A-PRF groups (p>0.05). Conclusion: 5% A-PRF CM was superior in increasing VEGF-A expression of hDPSCs at 5, 12 and 24 hours of observations.

scholarly journals Human dental pulp stem cell responses to different dental pulp capping materials

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Chawan Manaspon ◽  
Chavin Jongwannasiri ◽  
Sujin Chumprasert ◽  
Noppadol Sa-Ard-Iam ◽  
Rangsini Mahanonda ◽  
...  
Keyword(s):  
Dental Pulp ◽  
Marker Gene ◽  
Serum Free Medium ◽  
Free Medium ◽  
Alizarin Red ◽  
Serum Free ◽  
Pulp Capping ◽  
Dental Pulp Capping ◽  
Human Dental Pulp

Abstract Background Direct pulp capping is a vital pulp therapy for a pin-point dental pulp exposure. Applying a pulp capping material leads to the formation of a dentin bridge and protects pulp vitality. The aim of this study was to compare the effects of four dental materials, DyCal®, ProRoot® MTA, Biodentine™, and TheraCal™ LC in vitro. Methods Human dental pulp stem cells (hDPs) were isolated and characterized. Extraction medium was prepared from the different pulp capping materials. The hDP cytotoxicity, proliferation, and migration were examined. The odonto/osteogenic differentiation was determined by alkaline phosphatase, Von Kossa, and alizarin red s staining. Osteogenic marker gene expression was evaluated using real-time polymerase chain reaction. Results ProRoot® MTA and Biodentine™ generated less cytotoxicity than DyCal® and TheraCal™ LC, which were highly toxic. The hDPs proliferated when cultured with the ProRoot® MTA and Biodentine™ extraction media. The ProRoot® MTA and Biodentine™ extraction medium induced greater cell attachment and spreading. Moreover, the hDPs cultured in the ProRoot® MTA or Biodentine™ extraction medium migrated in a similar manner to those in serum-free medium, while a marked reduction in cell migration was observed in the cells cultured in DyCal® and TheraCal™ LC extraction media. Improved mineralization was detected in hDPs maintained in ProRoot® MTA or Biodentine™ extraction medium compared with those in serum-free medium. Conclusion This study demonstrates the favorable in vitro biocompatibility and bioactive properties of ProRoot® MTA and Biodentine™ on hDPs, suggesting their superior regenerative potential compared with DyCal® and TheraCal™.

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