Stem Cells-based and Molecular-based Approaches in Regenerative Dentistry: A Topical Review

Background:: Regenerative medicine is a growing branch of medicine aimed to treat damaged or lost tissues, to promote tissue formation and to restore both aesthetics and function. In the last years, several scientific articles have been focused on the regenerative procedures aimed to increase the survival rate of compromised teeth; the most effective approaches have been based on molecularbased and on cellular-based protocols; however, to date, both these techniques have not been carefully analysed and discussed, to know in details the advantages of each of them. Methods:: A literature search was undertaken on three electronic scientific databases: Medline via PubMed, EMBASE and Google Scholar. Authors aimed to select such articles published in the time span from January 1961 until December 2017. The authors screened the titles and the abstracts including the following keywords combinations: “Pulp AND Therapy”, “Regenerative AND Endodontic”, and “Endodontics AND Tissue engineering”. After the exclusion of any not related articles, the full text of such papers related to the topics was included in this review. Results:: Following the removal of duplicate articles and of other types of publications (such as erratum and corrigendum), 621 articles were selected to be included and analysed in our topical review. The articles were analysed into the following sections: cellular-based approaches for dental regeneration, molecular-based and combined cellular/molecular-based approaches for dental regeneration, and translational applications of regenerative dentistry. Conclusion:: This topical review has been focused on the main, the most promising and the most innovative strategies for achieving the regeneration of dental pulp or dental tissues. The main and surprising “take-home message” is related to the great interest towards the dental-derived stem cells, characterized by a high angiogenic and neurogenic commitment. Future challenges will be focused on the development of biological-friendly regenerative strategies: the new approaches should overcome the current biological limitations, to promote the combined cellular and molecular-based treatments, able to ensure predictable clinical evidence, with the achievement of the regeneration/repairing of the compromised dental pulp and of the entire tooth structure.

Download Full-text

Novel Molecule Nell-1 Promotes the Angiogenic Differentiation of Dental Pulp Stem Cells

Frontiers in Physiology ◽

10.3389/fphys.2021.703593 ◽

2021 ◽

Vol 12 ◽

Author(s):

Mengyue Li ◽

Qiang Wang ◽

Qi Han ◽

Jiameng Wu ◽

Hongfan Zhu ◽

...

Keyword(s):

Stem Cells ◽

Dental Pulp ◽

Umbilical Vein ◽

Dental Pulp Stem Cells ◽

Exposure Model ◽

Dental Tissues ◽

Normal Rat ◽

Pulp Exposure ◽

Angiogenic Markers

IntroductionThis work aimed to reveal the crucial role of Nell-1 in the angiogenic differentiation of human dental pulp stem cells (DPSCs) alone or co-cultured with human umbilical vein endothelial cell (HUVECs) in vitro and whether this molecule is involved in the pulp exposure model in vivo.MethodsImmunofluorescence was conducted to ascertain the location of Nell-1 on DPSCs, HUVECs, and normal rat dental tissues. RT-PCR, Western blot, and ELISA were performed to observe the expression levels of angiogenic markers and determine the angiogenic differentiation of Nell-1 on DPSCs alone or co-cultured with HUVECs, as well as in vitro tube formation assay. Blood vessel number for all groups was observed and compared using immunohistochemistry by establishing a rat pulp exposure model.ResultsNell-1 is highly expressed in the nucleus of DPSCs and HUVECs and is co-expressed with angiogenic markers in normal rat pulp tissues. Hence, Nell-1 can promote the angiogenic marker expression in DPSCs alone and co-cultured with other cells and can enhance angiogenesis in vitro as well as in the pulp exposure model.ConclusionNell-1 may play a positive role in the angiogenic differentiation of DPSCs.

Download Full-text

Tooth Formation: Are the Hardest Tissues of Human Body Hard to Regenerate?

International Journal of Molecular Sciences ◽

10.3390/ijms21114031 ◽

2020 ◽

Vol 21 (11) ◽

pp. 4031 ◽

Cited By ~ 1

Author(s):

Juliana Baranova ◽

Dominik Büchner ◽

Werner Götz ◽

Margit Schulze ◽

Edda Tobiasch

Keyword(s):

Stem Cells ◽

Drug Release ◽

Adult Stem Cells ◽

Complex Structure ◽

Tooth Germ ◽

Tooth Regeneration ◽

Dental Tissue ◽

Dental Tissues ◽

Regenerative Dentistry

With increasing life expectancy, demands for dental tissue and whole-tooth regeneration are becoming more significant. Despite great progress in medicine, including regenerative therapies, the complex structure of dental tissues introduces several challenges to the field of regenerative dentistry. Interdisciplinary efforts from cellular biologists, material scientists, and clinical odontologists are being made to establish strategies and find the solutions for dental tissue regeneration and/or whole-tooth regeneration. In recent years, many significant discoveries were done regarding signaling pathways and factors shaping calcified tissue genesis, including those of tooth. Novel biocompatible scaffolds and polymer-based drug release systems are under development and may soon result in clinically applicable biomaterials with the potential to modulate signaling cascades involved in dental tissue genesis and regeneration. Approaches for whole-tooth regeneration utilizing adult stem cells, induced pluripotent stem cells, or tooth germ cells transplantation are emerging as promising alternatives to overcome existing in vitro tissue generation hurdles. In this interdisciplinary review, most recent advances in cellular signaling guiding dental tissue genesis, novel functionalized scaffolds and drug release material, various odontogenic cell sources, and methods for tooth regeneration are discussed thus providing a multi-faceted, up-to-date, and illustrative overview on the tooth regeneration matter, alongside hints for future directions in the challenging field of regenerative dentistry.

Download Full-text

Cell-Based Transplantation versus Cell Homing Approaches for Pulp-Dentin Complex Regeneration

Stem Cells International ◽

10.1155/2021/8483668 ◽

2021 ◽

Vol 2021 ◽

pp. 1-23

Author(s):

Geraldine M. Ahmed ◽

Eman A. Abouauf ◽

Nermeen AbuBakr ◽

Asmaa M. Fouad ◽

Christof E. Dörfer ◽

...

Keyword(s):

Stem Cells ◽

Causative Factor ◽

Major Health ◽

Cell Homing ◽

New Era ◽

Dental Tissues ◽

Regenerative Dentistry ◽

Pulp Vitality ◽

Regenerative Therapies

Regenerative dentistry has paved the way for a new era for the replacement of damaged dental tissues. Whether the causative factor is dental caries, trauma, or chemical insult, the loss of the pulp vitality constitutes one of the major health problems worldwide. Two regenerative therapies were introduced for a fully functional pulp-dentin complex regeneration, namely, cell-based (cell transplantation) and cell homing (through revascularization or homing by injection of stem cells in situ or intravenously) therapies, with each demonstrating advantages as well as drawback, especially in clinical application. The present review is aimed at elaborating on these two techniques in the treatment of irreversibly inflamed or necrotic pulp, which is aimed at regenerating a fully functional pulp-dentin complex.

Download Full-text

Neurogenic Differentiation of Human Dental Pulp Stem Cells on Graphene-Polycaprolactone Hybrid Nanofibers

Nanomaterials ◽

10.3390/nano8070554 ◽

2018 ◽

Vol 8 (7) ◽

pp. 554 ◽

Cited By ~ 8

Author(s):

Hoon Seonwoo ◽

Kyung-Je Jang ◽

Dohyeon Lee ◽

Sunho Park ◽

Myungchul Lee ◽

...

Keyword(s):

Stem Cells ◽

Dental Pulp ◽

Morphological Characteristics ◽

Electrospun Nanofibers ◽

Dental Pulp Stem Cells ◽

Neurogenic Differentiation ◽

Dental Tissues ◽

Differentiated Cells ◽

Human Dental Pulp

Stem cells derived from dental tissues—dental stem cells—are favored due to their easy acquisition. Among them, dental pulp stem cells (DPSCs) extracted from the dental pulp have many advantages, such as high proliferation and a highly purified population. Although their ability for neurogenic differentiation has been highlighted and neurogenic differentiation using electrospun nanofibers (NFs) has been performed, graphene-incorporated NFs have never been applied for DPSC neurogenic differentiation. Here, reduced graphene oxide (RGO)-polycaprolactone (PCL) hybrid electrospun NFs were developed and applied for enhanced neurogenesis of DPSCs. First, RGO-PCL NFs were fabricated by electrospinning with incorporation of RGO and alignments, and their chemical and morphological characteristics were evaluated. Furthermore, in vitro NF properties, such as influence on the cellular alignments and cell viability of DPSCs, were also analyzed. The influences of NFs on DPSCs neurogenesis were also analyzed. The results confirmed that an appropriate concentration of RGO promoted better DPSC neurogenesis. Furthermore, the use of random NFs facilitated contiguous junctions of differentiated cells, whereas the use of aligned NFs facilitated an aligned junction of differentiated cells along the direction of NF alignments. Our findings showed that RGO-PCL NFs can be a useful tool for DPSC neurogenesis, which will help regeneration in neurodegenerative and neurodefective diseases.

Download Full-text

Analysis of growth kinetics and colony-forming unit efficiency of human dental pulp stem cells cultured in different media and supplements

Research Journal of Biotechnology ◽

10.25303/167rjbt20321 ◽

2021 ◽

Vol 16 (7) ◽

pp. 203-210

Author(s):

Kumar Chethan ◽

Shishir Shetty ◽

Basan Gowda Kurkalli ◽

Veena Shetty ◽

Kumar Basavarajappa Mohana

Keyword(s):

Stem Cells ◽

Growth Kinetics ◽

Dental Pulp ◽

Culture Conditions ◽

Dental Pulp Stem Cells ◽

Population Doubling ◽

Animal Origin ◽

Dental Tissues ◽

Colony Forming Unit ◽

Human Dental Pulp

Dental tissues are considered as ideal autologous sources of multipotent stem cells. Presently, human dental pulp stem cells (DPSCs) are largely being isolated and expanded in media containing fetal bovine serum (FBS). However, the use of FBS has limitations due to its animal origin. Therefore, the present study evaluated the morphology, proliferation rate, population doubling time (PDT) and colony-forming unit fibroblast (CFU-F) efficiency of DPSCs cultured in animal serum-containing medium (SCM) and serumfree medium (SFM) in addition to serum-free culture conditions by supplementing human blood-derivatives such as platelet lysate (PL), fresh frozen plasma (FFP) and umbilical cord blood serum (UCS) at 2.5%, 5% and 7.5% concentrations. Established DPSCs had spindle-shape during primary culture but acquired characteristic fibroblast-like features when cultured in PL, FFP and UCS. DPSCs in SCM, SFM and PL had significantly (P<0.05) higher proliferative potential than those in UCS and FFP and these observations were supported by PDT values. The CFU efficiency of DPSCs was confirmed in all culture conditions with a slightly varied clonogenic potential in blood-derived components. Based on the growth kinetics and CFU ability, it is concluded that PL could be considered as a suitable alternative to FBS for the ex vivo expansion of DPSCs.

Download Full-text

Neural Crest Stem Cells from Dental Tissues: A New Hope for Dental and Neural Regeneration

Stem Cells International ◽

10.1155/2012/103503 ◽

2012 ◽

Vol 2012 ◽

pp. 1-12 ◽

Cited By ~ 39

Author(s):

Gaskon Ibarretxe ◽

Olatz Crende ◽

Maitane Aurrekoetxea ◽

Victoria García-Murga ◽

Javier Etxaniz ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Cell Therapy ◽

Neural Crest ◽

Neural Regeneration ◽

Dental Tissues ◽

Adult Human ◽

Regenerative Dentistry ◽

Cord Injury

Several stem cell sources persist in the adult human body, which opens the doors to both allogeneic and autologous cell therapies. Tooth tissues have proven to be a surprisingly rich and accessible source of neural crest-derived ectomesenchymal stem cells (EMSCs), which may be employed to repair disease-affected oral tissues in advanced regenerative dentistry. Additionally, one area of medicine that demands intensive research on new sources of stem cells is nervous system regeneration, since this constitutes a therapeutic hope for patients affected by highly invalidating conditions such as spinal cord injury, stroke, or neurodegenerative diseases. However, endogenous adult sources of neural stem cells present major drawbacks, such as their scarcity and complicated obtention. In this context, EMSCs from dental tissues emerge as good alternative candidates, since they are preserved in adult human individuals, and retain both high proliferation ability and a neural-like phenotypein vitro. In this paper, we discuss some important aspects of tissue regeneration by cell therapy and point out some advantages that EMSCs provide for dental and neural regeneration. We will finally review some of the latest research featuring experimental approaches and benefits of dental stem cell therapy.

Download Full-text

Extracellular IL-37 Enhances Osteogenic and Odontogenic Differentiation of Human Dental Pulp Stem Cells via Autophagy Pathway

10.21203/rs.3.rs-291517/v1 ◽

2021 ◽

Author(s):

Na Li ◽

Yan Chen ◽

Ming Yan ◽

Yanqiu Wang ◽

Jintao Wu ◽

...

Keyword(s):

Stem Cells ◽

Dental Pulp ◽

Dental Pulp Stem Cells ◽

Inflammatory Factor ◽

Alizarin Red ◽

Dental Tissues ◽

Alp Activity ◽

Odontogenic Differentiation ◽

Marrow Mesenchymal Stem Cells

Abstract BackgroundThe osteogenic and odontogenic differentiation of dental pulp stem cells (DPSCs) contributes to the restoration and regeneration of dental tissues. Previous study indicated that IL-37 has often been identified as an anti-inflammatory factor that affects other pro-inflammatory signals. It is known to be a factor capable of inducing in vitro osteogenic differentiation of bone marrow mesenchymal stem cells (BMMSCs). The aims of this study were to explore the effects of IL-37 on the differentiation of DPSCs.MethodsDPSCs were cultured in growth medium with different concentration of IL-37, ALP activity was done to detect the optimal concentration for the following experiments. CCK-8 were conducted to assess the effect of IL-37 on proliferation of DPSCs. To assess differentiation, alkaline phosphatase activity, ALP staining, alizarin red S staining and real‐time RT‐PCR of DSPP, Runx2, ALP, and OSX were measured. Western blot was conducted to examine the levels of autophagy related markers (Beclin1, P62, LC3). ResultsCells cultured with 1 ng/mL IL-37 owned the highest ALP activity. IL-37 enhanced the osteogenic and odontogenic differentiation of DPSCs following upregulated the expression of Beclin1, downregulated the expression of P62, and reduced the ratio of LC3II/I, whereas depletion of autophagy suppressed DPSCs osteogenic and odontogenic differentiation. ConclusionIL-37 increased osteogenic and odontogenic differentiation via autophagy.

Download Full-text

UNDERSTANDING THE ROLE OF DYSFUNCTIONAL AND HEALTHY MITOCHONDRIA IN STROKE PATHOLOGY AND ITS TREATMENT

10.20944/preprints201806.0302.v1 ◽

2018 ◽

Author(s):

Hung Nguyen ◽

Sydney Zarriello ◽

Mira Rajani ◽

Julian Tuazon ◽

Eleonora Napoli ◽

...

Keyword(s):

Stem Cells ◽

Neurovascular Unit ◽

The United States ◽

Stroke Therapy ◽

Innovative Strategies ◽

Mitochondria Transfer ◽

Stem Cells Transplantation ◽

And Function ◽

Research Endeavor

Stroke remains a major cause of death and disability in the United States and around the world. Solid safety and efficacy profiles of novel stroke therapeutics have been generated in the laboratory, but most failed in clinical trials. Investigations into the pathology and treatment of the disease remain a key research endeavor in advancing scientific understanding and clinical applications. In particular, cell-based regenerative medicine, specifically stem cells transplantation, may hold promise as stroke therapy because grafted cells and their components may recapitulate the growth and function of the neurovascular unit, which arguably represents the alpha and omega of stroke brain pathology and recovery. Recent evidence has implicated mitochondria, organelles with a central role in energy metabolism and stress response, in stroke progression. Recognizing that stem cells offer a source of healthy mitochondria, potentially transferrable into ischemic cells, may provide a new therapeutic tool. To this end, deciphering cellular and molecular processes underlying dysfunctional mitochondria may reveal innovative strategies for stroke therapy. Here, we review recent studies capturing the intimate participation of mitochondrial impairment in stroke pathology, and showcase promising methods of healthy mitochondria transfer into ischemic cells, to critically evaluate the potential of mitochondria-based stem cell therapy for stroke.

Download Full-text

Dental Pulp and Periodontal Ligament Cells Support Osteoclastic Differentiation

Journal of Dental Research ◽

10.1177/0022034509340008 ◽

2009 ◽

Vol 88 (7) ◽

pp. 609-614 ◽

Cited By ~ 38

Author(s):

M. Uchiyama ◽

Y. Nakamichi ◽

M. Nakamura ◽

S. Kinugawa ◽

H. Yamada ◽

...

Keyword(s):

Dental Pulp ◽

Periodontal Ligament ◽

Human Peripheral Blood ◽

Cell Types ◽

Periodontal Ligament Cells ◽

Dental Tissues ◽

Tooth Roots ◽

Human Dental Pulp ◽

Osteoclastic Differentiation ◽

And Function

Odontoclasts and cementoclasts are considered to play major roles in the internal resorption of dentin and the external resorption of tooth roots. In this study, we evaluated the osteoclast-inducing ability of human dental pulp and periodontal ligament cells, which are mesenchymal cells in dental tissues. These cells expressed RANKL and OPG mRNA constitutively. As osteoclast precursors, CD14+ monocytes derived from human peripheral blood were isolated, and incubated together with human dental pulp or periodontal ligament cells. Both cell types spontaneously induced the differentiation of CD14+ monocytes into osteoclasts without osteotropic factors. These results suggest that dental pulp and periodontal ligament cells are involved in regulating the differentiation and function of osteoclasts.

Download Full-text