scholarly journals Silencing VEGFR-2 Hampers Odontoblastic Differentiation of Dental Pulp Stem Cells

2021 ◽  
Vol 9 ◽  
Author(s):  
Kajohnkiart Janebodin ◽  
Rakchanok Chavanachat ◽  
Aislinn Hays ◽  
Morayma Reyes Gil
Keyword(s):  
Stem Cells ◽  
Blood Vessels ◽  
Dental Pulp ◽  
Alizarin Red ◽  
Odontoblast Differentiation ◽  
Dentin Matrix ◽  
Dentin Regeneration

Dental pulp stem cells (DPSCs) are a source of postnatal stem cells essential for maintenance and regeneration of dentin and pulp tissues. Previous in vivo transplantation studies have shown that DPSCs are able to give rise to odontoblast-like cells, form dentin/pulp-like structures, and induce blood vessel formation. Importantly, dentin formation is closely associated to blood vessels. We have previously demonstrated that DPSC-induced angiogenesis is VEGFR-2-dependent. VEGFR-2 may play an important role in odontoblast differentiation of DPSCs, tooth formation and regeneration. Nevertheless, the role of VEGFR-2 signaling in odontoblast differentiation of DPSCs is still not well understood. Thus, in this study we aimed to determine the role of VEGFR-2 in odontoblast differentiation of DPSCs by knocking down the expression of VEGFR-2 in DPSCs and studying their odontoblast differentiation capacity in vitro and in vivo. Isolation and characterization of murine DPSCs was performed as previously described. DPSCs were induced by VEGFR-2 shRNA viral vectors transfection (MOI = 10:1) to silence the expression of VEGFR-2. The GFP+ expression in CopGFP DPSCs was used as a surrogate to measure the efficiency of transfection and verification that the viral vector does not affect the expression of VEGFR-2. The efficiency of viral transfection was shown by significant reduction in the levels of VEGFR-2 based on the Q-RT-PCR and immunofluorescence in VEGFR-2 knockdown DPSCs, compared to normal DPSCs. VEGFR-2 shRNA DPSCs expressed not only very low level of VEGFR-2, but also that of its ligand, VEGF-A, compared to CopGFP DPSCs in both transcriptional and translational levels. In vitro differentiation of DPSCs in osteo-odontogenic media supplemented with BMP-2 (100 ng/ml) for 21 days demonstrated that CopGFP DPSCs, but not VEGFR-2 shRNA DPSCs, were positive for alkaline phosphatase (ALP) staining and formed mineralized nodules demonstrated by positive Alizarin Red S staining. The expression levels of dentin matrix proteins, dentin matrix protein-1 (Dmp1), dentin sialoprotein (Dspp), and bone sialoprotein (Bsp), were also up-regulated in differentiated CopGFP DPSCs, compared to those in VEGFR-2 shRNA DPSCs, suggesting an impairment of odontoblast differentiation in VEGFR-2 shRNA DPSCs. In vivo subcutaneous transplantation of DPSCs with hydroxyapatite (HAp/TCP) for 5 weeks demonstrated that CopGFP DPSCs were able to differentiate into elongated and polarized odontoblast-like cells forming loose connective tissue resembling pulp-like structures with abundant blood vessels, as demonstrated by H&E, Alizarin Red S, and dentin matrix staining. On the other hand, in VEGFR-2 shRNA DPSC transplants, odontoblast-like cells were not observed. Collagen fibers were seen in replacement of dentin/pulp-like structures. These results indicate that VEGFR-2 may play an important role in dentin regeneration and highlight the potential of VEGFR-2 modulation to enhance dentin regeneration and tissue engineering as a promising clinical application.

scholarly journals Periodontal and Dental Pulp Cell-Derived Small Extracellular Vesicles: A Review of the Current Status

Nanomaterials ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 1858
Author(s):  
Shu Hua ◽  
Peter Mark Bartold ◽  
Karan Gulati ◽  
Corey Stephen Moran ◽  
Sašo Ivanovski ◽  
...  
Keyword(s):  
Stem Cells ◽  
Extracellular Vesicles ◽  
Dental Pulp ◽  
Therapeutic Potential ◽  
Current Status ◽  
Pulp Cell ◽  
Dental Pulp Cell

Extracellular vesicles (EVs) are membrane-bound lipid particles that are secreted by all cell types and function as cell-to-cell communicators through their cargos of protein, nucleic acid, lipids, and metabolites, which are derived from their parent cells. There is limited information on the isolation and the emerging therapeutic role of periodontal and dental pulp cell-derived small EVs (sEVs, <200 nm, or exosome). In this review, we discuss the biogenesis of three EV subtypes (sEVs, microvesicles and apoptotic bodies) and the emerging role of sEVs from periodontal ligament (stem) cells, gingival fibroblasts (or gingival mesenchymal stem cells) and dental pulp cells, and their therapeutic potential in vitro and in vivo. A review of the relevant methodology found that precipitation-based kits and ultracentrifugation are the two most common methods to isolate periodontal (dental pulp) cell sEVs. Periodontal (and pulp) cell sEVs range in size, from 40 nm to 2 μm, due to a lack of standardized isolation protocols. Nevertheless, our review found that these EVs possess anti-inflammatory, osteo/odontogenic, angiogenic and immunomodulatory functions in vitro and in vivo, via reported EV cargos of EV–miRNAs, EV–circRNAs, EV–mRNAs and EV–lncRNAs. This review highlights the considerable therapeutic potential of periodontal and dental pulp cell-derived sEVs in various regenerative applications.

scholarly journals Role of Fzd6 in Regulating the Osteogenic Differentiation of Adipose-derived Stem Cells in Osteoporosis

2021 ◽  
Author(s):  
Tianli Wu ◽  
Zhihao Yao ◽  
Gang Tao ◽  
Fangzhi Lou ◽  
Hui Tang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Osteogenic Differentiation ◽  
Wnt Signaling Pathway ◽  
Osteogenic Potential ◽  
Adipose Derived Stem Cells ◽  
Alizarin Red ◽  
Differentiation Potential

Abstract Objective: Although it has been demonstrated that adipose-derived stem cells (ASCs) from osteoporosis mice (OP-ASCs) exhibit impaired osteogenic differentiation potential, the molecular mechanism has not yet been elucidated. We found that Fzd6 was decreased in OP-ASCs compared with ASCs. This study investigates the effects and underlying mechanisms of Fzd6 in the osteogenic potential of OP-ASCs. Methods: Fzd6 expression in ASCs and OP-ASCs was measured by PCR gene chip. Fzd6 overexpression and silencing lentiviruses were used to evaluate the role of Fzd6 in the osteogenic differentiation of OP-ASCs. Real-time PCR (qPCR) and western blotting (WB) was performed to detect the expression of Fzd6 and bone-related molecules, including runt-related transcription factor 2 (Runx2) and osteopontin (Opn). Alizarin red staining and Alkaline phosphatase (ALP) staining was performed following osteogenic induction. Microscopic CT (Micro-CT), hematoxylin and eosin staining (H&E) staining, and Masson staining were used to assess the role of Fzd6 in osteogenic differentiation of osteoporosis (OP) mice in vivo.Results: Expression of Fzd6 was decreased significantly in OP-ASCs. Fzd6 silencing down-regulated the osteogenic ability of OP-ASCs in vitro. Overexpression of Fzd6 rescued the impaired osteogenic capacity in OP-ASCs in vitro. We obtained similar results in vivo.Conclusions: Fzd6 plays an important role in regulating the osteogenic ability of OP-ASCs both in vivo and in vitro. Overexpression of Fzd6 associated with the Wnt signaling pathway promotes the osteogenic ability of OP-ASCs, which provides new insights for the prevention and treatment of OP.

scholarly journals Dentin Phosphophoryn-Derived Peptide Promotes Odontoblast Differentiation In Vitro and Dentin Regeneration In Vivo

Materials ◽  
2021 ◽  
Vol 14 (4) ◽  
pp. 874
Author(s):  
Bayarchimeg Altankhishig ◽  
Mohammad Ali Akbor Polan ◽  
Youjing Qiu ◽  
Md Riasat Hasan ◽  
Takashi Saito
Keyword(s):  
Calcium Hydroxide ◽  
Alizarin Red ◽  
Odontoblast Differentiation ◽  
Reparative Dentin ◽  
Pulp Inflammation ◽  
Dentin Formation ◽  
Reparative Dentin Formation ◽  
Dentin Regeneration

The purpose of the present study was to investigate the effect of a peptide (i.e., SESDNNSSSRGDASYNSDES) derived from dentin phosphophoryn (DPP) with arginine-glycine-aspartic acid (RGD) motifs on odontoblast differentiation in vitro and to compare it with calcium hydroxide—a material used conventionally for vital pulp therapy—in terms of reparative dentin formation and pulp inflammation in vivo. Alkaline phosphatase activity assay and alizarin red S staining were performed to evaluate odontoblast-differentiation in cell culturing experiments. To observe the reparative dentin formation and pulp inflammation animal experiment was performed and examined by histological methods. The difference between the experimental group and the control group was analyzed statistically using a one-way ANOVA test. The results revealed that the DPP-derived RGD-containing peptide triggered odontoblast differentiation and mineralization in vitro. In rats undergoing direct pulp capping, the DPP-derived RGD-containing peptide was found to induce intensively formed reparative dentin with high compactness at week 4. On histological and morphometrical examinations, a smaller degree of pulpitis was observed in the specimens treated with the peptide than in those treated with calcium hydroxide. This study suggests that the DPP-derived RGD-containing peptide is a biocompatible, biodegradable and bioactive material for dentin regeneration.

Intracellular signaling molecules of nerve tissue progenitors as pharmacological targets for treatment of ethanol-induced neurodegeneration

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Gleb Nikolaevich Zyuz’kov ◽  
Larisa Arkad`evna Miroshnichenko ◽  
Elena Vladislavovna Simanina ◽  
Larisa Alexandrovna Stavrova ◽  
Tatyana Yur`evna Polykova
Keyword(s):  
Stem Cells ◽  
Alcohol Abuse ◽  
Intracellular Signaling ◽  
Signaling Molecules ◽  
Growth Potential ◽  
Nerve Tissue ◽  
Intracellular Signaling Molecules

Abstract Objectives The development of approaches to the treatment of neurodegenerative diseases caused by alcohol abuse by targeted pharmacological regulation of intracellular signaling transduction of progenitor cells of nerve tissue is promising. We studied peculiarities of participation of NF-кB-, сАМР/РКА-, JAKs/STAT3-, ERK1/2-, p38-pathways in the regulation of neural stem cells (NSC) and neuronal-committed progenitors (NCP) in the simulation of ethanol-induced neurodegeneration in vitro and in vivo. Methods In vitro, the role of signaling molecules (NF-кB, сАМР, РКА, JAKs, STAT3, ERK1/2, p38) in realizing the growth potential of neural stem cells (NSC) and neuronal-committed progenitors (NCP) in ethanol-induced neurodegeneration modeled in vitro and in vivo was studied. To do this, the method of the pharmacological blockade with the use of selective inhibitors of individual signaling molecules was used. Results Several of fundamental differences in the role of certain intracellular signaling molecules (SM) in proliferation and specialization of NSC and NCP have been revealed. It has been shown that the effect of ethanol on progenitors is accompanied by the formation of a qualitatively new pattern of signaling pathways. Data have been obtained on the possibility of stimulation of nerve tissue regeneration in ethanol-induced neurodegeneration by NF-кB and STAT3 inhibitors. It has been found that the blockage of these SM stimulates NSC and NCP in conditions of ethanol intoxication and does not have a «negative» effect on the realization of the growth potential of intact progenitors (which will appear de novo during therapy). Conclusions The results may serve as a basis for the development of fundamentally new drugs to the treatment of alcoholic encephalopathy and other diseases of the central nervous system associated with alcohol abuse.

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.

P-EP013. Superior migration propensity of dental pulp stem cells in in vitro and in vivo models of excitotoxic neurodegeneration

2021 ◽  
Vol 132 (8) ◽  
pp. e82-e83
Author(s):  
Sivapriya Senthilkumar ◽  
Chaitra Venugopal ◽  
K. Shobha ◽  
Bindu M. Kutty ◽  
Anandh Dhanushkodi
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
In Vivo Models ◽  
Migration Propensity

scholarly journals Hypoxia-inducible factor 1α induces osteo/odontoblast differentiation of human dental pulp stem cells via Wnt/β-catenin transcriptional cofactor BCL9

2022 ◽  
Vol 12 (1) ◽  
Author(s):  
Shion Orikasa ◽  
Nobuyuki Kawashima ◽  
Kento Tazawa ◽  
Kentaro Hashimoto ◽  
Keisuke Sunada-Nara ◽  
...  
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Target Genes ◽  
Hypoxia Inducible Factor ◽  
Dental Pulp Stem Cells ◽  
Pulp Tissue ◽  
Odontoblast Differentiation ◽  
Hypoxia Inducible Factor 1Α ◽  
Hypoxic Conditions

AbstractAccelerated dental pulp mineralization is a common complication in avulsed/luxated teeth, although the mechanisms underlying this remain unclear. We hypothesized that hypoxia due to vascular severance may induce osteo/odontoblast differentiation of dental pulp stem cells (DPSCs). This study examined the role of B-cell CLL/lymphoma 9 (BCL9), which is downstream of hypoxia-inducible factor 1α (HIF1α) and a Wnt/β-catenin transcriptional cofactor, in the osteo/odontoblastic differentiation of human DPSCs (hDPSCs) under hypoxic conditions. hDPSCs were isolated from extracted healthy wisdom teeth. Hypoxic conditions and HIF1α overexpression induced significant upregulation of mRNAs for osteo/odontoblast markers (RUNX2, ALP, OC), BCL9, and Wnt/β-catenin signaling target genes (AXIN2, TCF1) in hDPSCs. Overexpression and suppression of BCL9 in hDPSCs up- and downregulated, respectively, the mRNAs for AXIN2, TCF1, and the osteo/odontoblast markers. Hypoxic-cultured mouse pulp tissue explants showed the promotion of HIF1α, BCL9, and β-catenin expression and BCL9-β-catenin co-localization. In addition, BCL9 formed a complex with β-catenin in hDPSCs in vitro. This study demonstrated that hypoxia/HIF1α-induced osteo/odontoblast differentiation of hDPSCs was partially dependent on Wnt/β-catenin signaling, where BCL9 acted as a key mediator between HIF1α and Wnt/β-catenin signaling. These findings may reveal part of the mechanisms of dental pulp mineralization after traumatic dental injury.

scholarly journals The role of ProMyelocytic Leukemia (PML) protein in breast cancer and breast cancer stem cells

2018 ◽  
Author(s):  
Νικολέτα-Νίκη Σαχίνη
Keyword(s):  
Breast Cancer ◽  
Stem Cells ◽  
Cancer Stem Cells ◽  
Promyelocytic Leukemia ◽  
Breast Cancer Stem Cells ◽  
Forkhead Box ◽  
Forkhead Box M1

Ο καρκίνος του μαστού αποτελεί την πιο συχνή μορφή καρκίνου στις γυναίκες. Πρόκειται για μια ετερογενή ασθένεια όσον αφορά το φαινότυπό της και το γενετικό της υπόβαθρο. Καρκινικά χαρακτηριστικά, όπως ο ρυθμός πολλαπλασιασμού, η διεισδυτικότητα, η μεταστατικότητα, η ανθεκτικότητα στα φάρμακα και συγκεκριμένες ογκογόνες μεταλλαγές διαφέρουν μεταξύ περιπτώσεων καρκίνου του μαστού. Τέτοιου είδους ετερογένεια (ενδο-ογκική και δι-ογκική) μεταξύ των όγκων προκύπτει από στοχαστικές γενετικές και επιγενετικές αλλαγές οι οποίες προσδίδουν κληρονομήσιμες φαινοτυπικές και λειτουργικές διαφορές μεταξύ των καρκινικών κυττάρων. Παρ’ όλο που υπάρχουν θεραπευτικά σχήματα που αντιμετωπίζουν επιτυχώς την ασθένεια, η ετερογενής της φύση την καθιστά δύσκολο θεραπευτικό στόχο. Επομένως, η κατανόηση των παθογενετικών και μοριακών μηχανισμών οι οποίοι διέπουν τη νόσο και τους διαφορετικούς υποτύπους της είναι απαραίτητη για την ανακάλυψη νέων φαρμακευτικών στόχων και το σχεδιασμό εξατομικευμένης θεραπείας.Η πρωτεΐνη της προμυελοκυτταρικής λευχαιμίας (PML) περιγράφεται συνήθως ως ογκοκατασταλτικός παράγοντας λόγω των προ-αποπτωτικών, ανασταλτικών του κυτταρικού κύκλου και προγηραντικών ιδιοτήτων της. Συνεπώς, η έκφρασή της απουσιάζει από πρωτοπαθή δείγματα διαφόρων νεοπλασιών, συμπεριλαμβανομένου και του καρκίνου του μαστού. Παρ’ όλα αυτά σε πρόσφατες βιβλιογραφικές αναφορές η PML χαρακτηρίζεται ως ογκογόνος παράγοντας. Συγκεκριμένα, στη χρόνια μυελογενή λευχαιμία, σε γλοιοβλαστώματα και σε ορισμένες περιπτώσεις τριπλά αρνητικού καρκίνου του μαστού η PML εκφράζεται σε υψηλά επίπεδα. Οι προ-ογκογόνες ιδιότητες της PML φαίνεται να σχετίζονται με τη ρύθμιση του κυτταρικού κύκλου των φυσιολογικών και καρκινικών βλαστοκυττάρων αλλά και τη διατήρηση της αυτο-ανανέωσης, μέσω μεταβολικών οδών π.χ. οξείδωση των λιπαρών οξέων. Επομένως, προκύπτει ότι υπό συγκεκριμένες συνθήκες η PML έχει διττό ρόλο στην ογκογένεση εκδηλώνοντας ογκοκατασταλτική ή ογκογόνα δράση.Σκοπός της παρούσας διδακτορικής διατριβής ήταν η αποσαφήνιση των μοριακών και επιγενετικών μηχανισμών με τους οποίους η PML ρυθμίζει τον κυτταρικό πολλαπλασιασμό και τα μονοπάτια αυτό-ανανέωσης στον καρκίνο του μαστού. Τα πειραματικά μας δεδομένα υποστηρίζουν ότι η υπερέκφραση (ΥΕ) της PML αναστέλλει τον πολλαπλασιασμό των κυττάρων και μπλοκάρει τον κυτταρικό κύκλο της τριπλά αρνητικά κυτταρικής σειράς καρκίνου του μαστού, MDA-MB-231. Από την ανάλυση του μεταγραφικού προφίλ αυτών των κυττάρων προκύπτει ότι σημαντικά σηματοδοτικά μονοπάτια και βιολογικές λειτουργίες, όπως και οι ρυθμιστές τους, διαταράσσονται μετά από ΥΕ PML. Ενδιαφέρον παρουσιάζει το γεγονός ότι πολλά από τα γονίδια σχετιζόμενα με τον κυτταρικό πολλαπλασιασμό των οποίων η έκφραση μειώνεται μετά από ΥΕ PML είναι και, θετικά, ρυθμιζόμενοι στόχοι του μεταγραφικού παράγοντα FOXM1 (Forkhead Box M1). Η λειτουργική αλληλεπίδραση της PMLIV με την επικράτεια πρόσδεσης του FOXM1 στο DNA, καθώς και η μείωση του FOXM1 σε επίπεδο mRNA και πρωτεΐνης, υποδεικνύουν ότι η PMLIV είναι ένας καταστολέας του FOXM1. Επιπλέον, δείξαμε ότι η PMLIV YE επηρεάζει το μεταγραφικό πρόγραμμα του μεταγραφικού παράγοντα, FOXO3, o οποίος δρα ανοδικά του FOXM1. Συμπερασματικά, προτείνουμε ότι η PML επηρεάζει την ισορροπία των FOXO3 και FOXM1 μεταγραφικών προγραμμάτων στοχεύοντας διακριτά υποσύνολα γονιδίων. Τα αποτελέσματα μας δείχνουν ότι υψηλά επίπεδα PML μπορούν να επηρεάσουν ταυτόχρονα ποικίλα σηματοδοτικά μονοπάτια στοχεύοντας τον άξονα FOXO3-FOXM1, ο οποίος συνδέει τον κυτταρικό πολλαπλασιασμό (FOXM1) με την απόπτωση, διακοπή του κυτταρικού κύκλου και μηχανισμούς επιβίωσης (FOXO3). Βρήκαμε ακόμη ότι εκτός από τους FOXO3 και FOXM1, η ΥΕ PML στοχεύει κι άλλους καίριους μεταγραφικούς παράγοντες (NFYA,E2F,TBP) με γενικό ρόλο στη μεταγραφή και στον κυτταρικό πολλαπλασιασμό. Συνεπώς, θεωρούμε ότι η PMLIV ΥΕ επιδρά σε σημαντικές βιολογικές διεργασίες μέσω ενός σύνθετου δικτύου μεταγραφικών παραγόντων και επιγενετικών αλλαγών. Αρχικά πειράματα PMLIV YE σε κύτταρα διαφορετικού τύπου καρκίνου του μαστού, τα Τ47D, έδειξαν ότι η PMLIV YE οδηγεί σε αναστολή του κυτταρικού πολλαπλασιασμού, αλλά προκαλεί και διαφορετικές αποκρίσεις σε σχέση με τα MDA-MB-231 κύτταρα.Μελετήσαμε ακόμη το ρόλο της PMLIV σε βλαστικά καρκινικά κύτταρα του μαστού. Παρατηρήσαμε ότι η PMLIV YE μειώνει την ικανότητα σχηματισμού ογκοσφαιρών στην καρκινική σειρά MDA-MB-231, υποδηλώνοντας ότι η PMLIV μειώνει την αυτό-ανανέωση των βλαστικών καρκινικών κυττάρων. Επιπλέον, ερευνήσαμε την επίδραση της PMLIV σε απομονωμένους, με βάση την έκφραση των επιφανειακών δεικτών EpCAM/CD24, υποπληθυσμούς καρκινικών κυττάρων και διαπιστώσαμε ότι η PMLIV YE επηρεάζει με διαφορετικό τρόπο την έκφραση γονιδίων που εμπλέκονται στη διαφοροποίηση του επιθηλίου, στην επιθηλιακή προς μεσεγχυματική μετάβαση, στον πολλαπλασιασμό και στη βλαστικότητα του κάθε υποπληθυσμού, καθώς και την ικανότητα αυτο-ανανέωσης η οποία καθορίστηκε in vitro με τη δοκιμασία σφαιρογένεσης και in vivo με ξενομοσχεύματα. Επομένως, τα αρχικά μας δεδομένα, σε αντίθεση με προηγούμενες αναφορές, υποστηρίζουν την άποψη ότι πολύ ογκογόνοι κυτταρικοί υποπληθυσμοί εμπίπτουν σε περισσότερους από έναν EpCAM/CD24 υπότυπους οι οποίοι επηρεάζονται από την PMLIV με διαφορετικό τρόπο.Συμπερασματικά, η ερευνά μας δίνει πληροφορίες για τη ρύθμιση της ανάπτυξης των όγκων από την PML στον καρκίνο του μαστού. Προσδιορίσαμε τους FOXO3 και FOXM1 σαν αλληλεπιδρώντες παράγοντες της PML, οι οποίοι προσδιορίζουν και το λειτουργικό αποτέλεσμα της PML στο κυτταρικό υπόβαθρο των MDA-MB-231 κυττάρων και αναδείξαμε έναν καινούριο, ρυθμιστικό άξονα στον πολλαπλασιασμό του καρκίνου του μαστού, PML/FOXO3/FOXM1, ο οποίος μπορεί να είναι θεραπευτικός στόχος. Επιπλέον, οι αρχικές παρατηρήσεις στα T47D κύτταρα, ενισχύουν την άποψη περί κυτταρικού υποβάθρου εξαρτώμενης δράσης της PML. Η PMLIV YE επηρέασε πολύ λιγότερο τη δημιουργία σφαιρών, ένδειξη αυτο-ανανέωσης, στα T47D σε σχέση με τα MDA-MB-231. Επιπλέον, συγκρίνοντας το μεταγραφικό προφίλ των δύο κυτταρικών σειρών, οι οποίες αντιπροσωπεύουν διαφορετικούς μοριακούς υποτύπους του καρκίνου του μαστού, μετά από PMLIV YE διαπιστώσαμε ότι η PMLIV επηρεάζει τόσο κοινές αλλά και διακριτές ομάδες γονιδίων. Προτείνουμε ότι η PML ευνοεί τόσο την αναστολή της ανάπτυξης του όγκου όσο και μηχανισμούς επιβίωσης σε διαφορετικό βαθμό, ο οποίος καθορίζεται από το εκάστοτε γενετικό και επιγενετικό υπόβαθρο των κυττάρων.

scholarly journals Lithium Chloride Exerts Differential Effects on Dentinogenesis and Osteogenesis in Primary Pulp Cultures

2021 ◽  
Vol 2 ◽  
Author(s):  
Anushree Vijaykumar ◽  
Mina Mina
Keyword(s):  
Dental Pulp ◽  
Osteoblast Differentiation ◽  
Prolonged Treatment ◽  
Odontoblast Differentiation ◽  
Positive Effects ◽  
Reparative Dentin ◽  
Dentin Formation ◽  
Reparative Dentin Formation

Wnt/β-catenin signaling is known to play essential roles in odontoblast differentiation and reparative dentin formation. Various Wnt activators including LiCl have been increasingly studied for their effectiveness to induce repair of the dentin-pulp complex. LiCl is a simple salt thought to activate Wnt/β-catenin signaling by inhibiting GSK3β. Previous in vitro and in vivo studies showed that LiCl increased odontoblast differentiation and enhanced reparative dentin formation. However, the underlying molecular and cellular mechanisms by which LiCl regulates odontoblast and osteoblast differentiation during reparative dentinogenesis are not well-understood. Our in vitro studies show that exposure of early dental pulp progenitors to LiCl increased the survival and the pool of αSMA+ progenitors, leading to enhanced odontoblast and osteoblast differentiation. The positive effects of LiCl in the differentiation of osteoblasts and odontoblasts from αSMA+ progenitors are mediated by Wnt/β-catenin signaling. Our results also showed that continuous and late exposure of dental pulp cells to LiCl increased the expression of odontoblast markers through Wnt/β-catenin signaling, and the number of odontoblasts expressing DMP1-Cherry and DSPP-Cerulean transgenes. However, unlike the early treatment, both continuous and late treatments decreased the expression of Bsp and the expression of BSP-GFPtpz transgene. These observations suggest that prolonged treatment with LiCl in more mature cells of the dental pulp has an inhibitory effect on osteoblast differentiation. The inhibitory effects of LiCl on osteogenesis and Bsp were not mediated through Wnt/β-catenin signaling. These observations suggest that the effects of LiCl, and GSK3β antagonists on reparative dentinogenesis involve multiple pathways and are not specific to Wnt/β-catenin signaling.

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