Stem Cells and Cloning: Advances and Applications Dovepress

open access to scientific and medical research

Open Access Full Text Article

REVIEW

Dental Pulp Stem Cells: Advances to Applications

Stem Cells and Cloning: Advances and Applications downloaded from https://www.dovepress.com/ by 185.50.250.82 on 29-Feb-2020

This article was published in the following Dove Press journal:

Stem Cells and Cloning: Advances and Applications

Takeo W Tsutsui Abstract: Dental pulp stem cells (DPSCs) have a high capacity for differentiation and the
ability to regenerate a dentin/pulp-like complex. Numerous studies have provided evidence
Department of Pharmacology, School of
Life Dentistry at Tokyo, The Nippon of DPSCs’ differentiation capacity, such as in neurogenesis, adipogenesis, osteogenesis,
Dental University, Tokyo, Japan chondrogenesis, angiogenesis, and dentinogenesis. The molecular mechanisms and functions
of DPSCs’ differentiation process are affected by growth factors and scaffolds. For example,
growth factors such as basic fibroblast growth factor (bFGF), transforming growth factor-β
(TGF-β), nerve growth factor (NGF), platelet-derived growth factor (PDGF), and bone
morphogenic proteins (BMPs) influence DPSC fate, including in differentiation, cell prolif-
eration, and wound healing. In addition, several types of scaffolds, such as collagen,
For personal use only.

hydrogel, decellularized bioscaffold, and nanofibrous spongy microspheres, have been used
to characterize DPSC cellular attachment, migration, proliferation, differentiation, and func-
tions. An appropriate combination of growth factors and scaffolds can enhance the differ-
entiation capacity of DPSCs, in terms of optimizing not only dental-related expression but
also dental pulp morphology. For a cell-based clinical approach, focus has been placed on the
tissue engineering triad [cells/bioactive molecules (growth factors)/scaffolds] to characterize
DPSCs. It is clear that a deep understanding of the mechanisms of stem cells, including their
aging, self-renewal, microenvironmental homeostasis, and differentiation correlated with cell
activity, the energy for which is provided from mitochondria, should provide new approaches
for DPSC research and therapeutics. Mitochondrial functions and dynamics are related to the
direction of stem cell differentiation, including glycolysis, oxidative phosphorylation, mito-
chondrial metabolism, mitochondrial transcription factor A (TFAM), mitochondrial elonga-
tion, and mitochondrial fusion and fission proteins. This review summarizes the effects of
major growth factors and scaffolds for regenerating dentin/pulp-like complexes, as well as
elucidating mitochondrial properties of DPSCs for the development of advanced applications
research.
Keywords: dental pulp stem cell, bioactive molecule, growth factor, scaffold, mitochondria,
regenerative therapy

Introduction
Dental pulp stem cells (DPSCs) have great potential for a range of applications in
stem cell research and regenerative medicine. In the life science literature, there are
numerous reports on DPSC properties from in vitro and in vivo studies, such as cell
growth, capacity for differentiation, competence in assays, and potential for pio-
neering stem cell functions. The first report on DPSCs revealed that their stem cell
Correspondence: Takeo W Tsutsui
Department of Pharmacology, The properties are comparable to those of bone marrow stromal cells (BMSCs) in vitro
Nippon Dental University School of Life and in vivo.1
Dentistry at Tokyo, 1-9-20 Fujimi,
Chiyoda-Ku, Tokyo 102-8159, Japan The study of DPSCs by Gronthos’s group1 reported an immunophenotype
Tel +81 3-3261-8311 similar to that of BMSCs, along with the formation of a calcified nodule upon
Fax +81 3-3264-8399
Email [email protected] treatment with differentiation medium in vitro. This group transplanted DPSCs into

submit your manuscript | www.dovepress.com Stem Cells and Cloning: Advances and Applications 2020:13 33–42 33
DovePress © 2020 Tsutsui. This work is published and licensed by Dove Medical Press Limited. The full terms of this license are available at https://www.dovepress.com/terms.php
http://doi.org/10.2147/SCCAA.S166759
and incorporate the Creative Commons Attribution – Non Commercial (unported, v3.0) License (http://creativecommons.org/licenses/by-nc/3.0/). By accessing the work
you hereby accept the Terms. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed. For
permission for commercial use of this work, please see paragraphs 4.2 and 5 of our Terms (https://www.dovepress.com/terms.php).

Powered by TCPDF (www.tcpdf.org)

 Tsutsui Dovepress

the dorsal surface of immunocompromised mice with into immunocompromised mice.1 Other in vivo studies on
hydroxyapatite/tricalcium phosphate (HA/TCP), with the regenerating dentin formation also reported a dentin/pulp-
results showing that DPSCs were able to regenerate like structure.12–15 Moreover, several in vivo studies using
a dentin/pulp-like complex. They also showed DPSC transplantation have shown the capacity for differ-
a difference in the structures formed after transplantation entiation in animal models (eg, osteogenic,16 angiogenic,17
Stem Cells and Cloning: Advances and Applications downloaded from https://www.dovepress.com/ by 185.50.250.82 on 29-Feb-2020

compared with the case for BMSCs. In the literature, they and neurogenic18 functions). For in vivo models, scaffolds
speculated that adult dental pulp tissue might also contain are a key factor for tissue engineering. Several kinds of
a population of stem cells.1 material for scaffolds that influence DPSC properties have
The correlation of the presence of these cells in pulp been reported.19–26 Mitochondria are cytoplasmic organelles
with reparative dentinogenesis has also been explored.2,3 that have critical functions in energy metabolism for the
Reparative dentin is also referred to as tertiary, reactive, or regulation of stem cells. Considering analysis of the energy
irregular secondary dentin. Tertiary dentin is produced in metabolism of stem cells for regenerative research, knowl-
response to various irritants (attrition, caries, or edge of mitochondrial properties is important as it should
a restorative dental procedure) by the stimulus-affected deepen our understanding of the differentiation of these
cells. There are two categories of tertiary dentin: reaction- cells. In one study, the differences in energy metabolism
ary dentin, which is deposited by preexisting odontoblasts; in human MSCs were analyzed over the course of their
and reparative dentin, which is from newly differentiated differentiation.27 Moreover, dysfunction in mitochondrial
odontoblast-like cells.4 The precursors of odontoblasts membrane potential was observed in cells from the decid-
For personal use only.

have been shown to be regulated by growth factors such uous teeth of a Rett syndrome patient, showing the func-
as transforming growth factor-β (TGF-β), basic fibroblast tional importance of stem cells and the value of
growth factor (bFGF), platelet-derived growth factor mitochondrial analysis to explore the effectiveness of cell-
(PDGF), epidermal growth factor (EGF), tumor necrosis based clinical approaches. This review summarizes major
factor-α (TNF-α), and insulin-like growth factors (IGF)I growth factors (bFGF, TGF-β, NDF, BMPs, and PDGF),
and II. PDGF and bFGF were revealed to stimulate [3H] scaffolds, and mitochondrial properties within the research
thymidine incorporation into DNA, while TGF-β, EGF, field of DPSCs.
and TNF-α have less of an effect of this kind.5
DPSCs exhibit greater proliferation than other stem
cells, BMSCs and adipocyte stem cells (ASCs). Analysis Growth Factors
of the cellular growth curve demonstrated that DPSCs Basic Fibroblast Growth Factor (bFGF)
remained in the log phase from 3 to 5 days and that bFGF is a potent modulator of cell proliferation, motility, and
BMSCs had a longer population doubling time (PDT) differentiation.5,28 The bFGF locus is on chromosome429 and
than DPSCs during a 10-day period. A BrdU cell prolif- has been reported to present mRNAs of 4.6 kilobases (kb)
eration assay also showed that DPSCs had higher prolif- and additionally 2.2 kb in hypothalamus.30 The important
erative ability than BMSCs.6 properties of bFGF include a high affinity for heparin/
The immunophenotype of DPSCs has been reported to heparan sulfate (HS) and physiological transfer of glycosa-
feature mesenchymal stem cell markers, for example, minoglycans to the extracellular matrix (ECM). bFGF
CD73,7 CD90,7 and CD105.8 Interestingly, another derived from endothelial cells and bone cells may function
mesenchymal stem cell (MSC) marker, STRO-1, was found in the ECM.31,32 Moreover, this high affinity for heparin/HS
to be coexpressed with CD146 and pericyte antigen 3G5 in influences the maintenance of many different target tissues.
dental pulp, forming a specific niche.9 In addition, DPSCs According to the literature, bFGF has been detected in
express neural lineage markers including those found on DPSCs6 and after the endodontic procedure of irrigation.33
neural stem cells, such as nestin,10,11 musashi-1,10 βIII Basic FGF receptors (FGFR1 and FGFR2) were also
tubulin11 glial fibrillary acidic protein (GFAP),11 and neuro- found to be expressed in human dental pulp cells.34
nal nuclei (NeuN).11 Moreover, treatment of DPSCs with bFGF led to their
DPSCs have the capacity to differentiate into odonto- proliferation and differentiation during neurogenesis35,36
blast-like cells. This differentiation capacity was revealed and osteogenesis.37 Furthermore, Lue’s group36 reported
by the finding that DPSCs mixed with HA/TCP were able functional recovery in a spinal cord injury rat model upon
to regenerate a dentin/pulp-like structure by transplantation the application of heparin–poloxamer hydrogel containing

submit your manuscript | www.dovepress.com Stem Cells and Cloning: Advances and Applications 2020:13
34
DovePress

Powered by TCPDF (www.tcpdf.org)

 Dovepress Tsutsui

DPSCs and bFGF. The duration of treatment with bFGF shown to upregulate alpha-SMA in the differentiation of
was also shown to affect osteogenic differentiation to DPSCs into smooth muscle cells.10 The supplementation of
DPSCs. In the literature, it is demonstrated that 1 week TGF-β1/β3 in culture medium induced DPSCs to undergo
of treatment increased osteogenic differentiation. chondro-differentiation.10 TGF-β treatment induced several
Interestingly, 2 weeks of treatment actually decreased types of differentiation of DPSCs, including calcification,
Stem Cells and Cloning: Advances and Applications downloaded from https://www.dovepress.com/ by 185.50.250.82 on 29-Feb-2020

osteogenic differentiation, with similar results for these SMC-specific gene expression, and chondro-differentiation.
treatment periods being obtained in vitro and in vivo.37
Overall, these findings show that bFGF promotes prolif- Nerve Growth Factor (NGF)
eration and is related to osteogenic and neurogenic NGF is an essential regulator in the development, survival,
differentiation. differentiation, and maintenance of neuronal and non-
neuronal cells. NGF is a member of the neurotrophin
Transforming Growth Factor-β (TGF-β) family, which includes brain-derived growth factor
The TGF-β subfamily is divided into three isoforms, (BDNF), glial-cell-derived neurotrophic factor (GDNF),
TGF-β1, 2, and 3, which are produced as large precursor neurotrophic-3 (NT-3), and neurotrophic-4/5 (NT4/5).
molecules constituting mature TGF-β and the latency- Two NGF receptors have been identified: the trk proto-
associated peptide (LAP).38 LAP is cleaved off by an endo- oncogene product p140trk (trkA) and the p75 neurotrophin
protease and remains noncovalently bound to TGF-β, receptor (p75NTR).49–51 The NGF binding sites of neurons
constituting the small latency complex (SLC).39 The SLC are referred to as high-affinity and low-affinity receptor
For personal use only.

is associated with latent TGF-β binding proteins (LTBPs) binding sites.52 The trkA and p75NTR receptors exhibit
1–4.39,40 Active TGF-β is a potent regulator in biological low-affinity NGF binding.52–54 High-affinity binding sites
processes, including development, carcinogenesis, wound are created when trkA and p75NTR are coexpressed.52 The
healing, hematopoiesis, and immune responses, as well as complex network of signal pathways of trkA includes the
having specific effects on proliferation, differentiation, Ras-MAP kinase cascade.55 TrkA and p75NTR are also
migration, and apoptosis in microenvironments related to expressed in keratinocytes56 and in bone marrow and
particular cell types,41 including stem cells such as bone lymphoid tissues57 for cell proliferation, differentiation,
marrow-derived MSCs (BM-MSCs), adipose tisuue-derived and survival. Mitsiadis’ group reported that NGF, trkA,
MSCs (A-MSCs), and MSCs from dental pulp (DP-MSCs) and p75NTR are expressed in dental tissue and act in cell
producing TGF-β1.42,43 The TGF-β family induces signal- proliferation, differentiation, and odontogenesis, while
ing through transmembrane type I and type II membrane also being expressed in nerve fibers of developing human
binding serine/threonine kinase receptors; there are seven teeth.58
type I receptors [activin receptor like kinase (ALK)] and Moreover, a tiny group of DPSCs was shown to
five type II receptors.44 Upon the binding of a ligand to type express NGF, trkA, and p75NTR, the expression of which
I and type II receptors, type II receptors phosphorylate type was affected by the presence of β-glycerophosphate in
I receptors. This leads to the phosphorylation of R-Smads, culture medium, especially in cells forming mineralized
which induces a downstream signaling pathway.45 nodules.59 DPSCs have the capacity to differentiate into
Several studies have demonstrated that treatment with neurons and to repair injured neural systems. In the case of
recombinant TGF-β1 can enhance BMSC and pulp cell a rat model of spinal cord injury (SCI), recovery of hin-
proliferation.5,46 For example, a three-dimensional (3D) dlimb locomotor functions occurred upon the transplanta-
aggregate of DPSCs cultured with TGF-β3 and BMP-2 in tion of DPSCs with chitosan scaffolds.18 Moreover, in
serum-free medium induced calcification.47 In addition, a comparative study of DPSCs and BMSCs, DPSC secre-
Song’s group showed that TGF-β1 induced DPSCs to dif- tion of NGF, BDNF, and NT-3 was shown to be higher
ferentiate into bladder smooth muscle cells (SMCs).48 than that of BMSCs. Furthermore, DPSC coculture with
Moreover, when DPSCs were exposed to SMC- βIII-tubulin+ retinal cells was associated with a decrease in
conditioned medium with TGF-β1 for 14 days, this led to the number of neurite-bearing cells and the duration of
the increased expression of SMC-specific gene and protein treatment with Trk receptor blockers.60 NGF was found to
markers (alpha-SMA, desmin, and calponin). Furthermore, be expressed in dental tissue undergoing cell proliferation
the mature SMC marker myosin was detected after 11 days and odontogenesis. Furthermore, NGF expression was
of this exposure.48 TGF-β1 in culture medium was also shown to affect the differentiation of DPSCs and their

submit your manuscript | www.dovepress.com

Stem Cells and Cloning: Advances and Applications 2020:13 35
DovePress

Powered by TCPDF (www.tcpdf.org)

 Tsutsui Dovepress

potential to promote recovery from spinal cord injury via Bone Morphogenic Proteins (BMPs)
differentiation into neurons. BMPs, which have been shown to have the ability to
induce bone formation, are important in embryo, heart,
Platelet-Derived Growth Factor (PDGF) neural, cartilage, and tooth development. Many studies
have reported the characterization73 of this protein group,
Stem Cells and Cloning: Advances and Applications downloaded from https://www.dovepress.com/ by 185.50.250.82 on 29-Feb-2020

PDGF was identified in cell-free plasma derived from

serum, a component of whole blood,61 and purified from which belongs to the TGF-β superfamily. In terms of the
human platelets.62,63 In terms of its structure, PDGF con- ligands of BMPs, they bind to type I and type II receptors
sists of two polypeptide A and B chains combined in three that signal through canonical and noncanonical pathways.
disulfide-linked dimers (AA, AB, and BB). PDGF-C64 and Upon ligand binding, the type II receptor activates the
PDGF-D consist of domains: CUB and PDGF/VEGF and type I receptor by phosphorylation and activates smads.
N-linked glycosylation site.65 The gene encoding the This signal plays an important role in early
PDGF-A chain is located on chromosome 7,66 while that odontogenesis74 and tooth development, including tooth
for the PDGF-B chain is located on chromosome 22.66,67 homeostasis,75 number, size, and shape.76
Shi’s group demonstrated BMP signal activation of pre-
The PDGFC gene and PDGFD gene are located on chro-
odontoblasts/odontoblasts, dental pulp, and a small number
mosomes 4 and 11.68
of transit-amplifying cells (TAC) in 1-month-old mice.
PDGF binds two receptor tyrosine kinases, namely, α-
Using 1-month-old Gli1-CreERT2 ;td Tomato mice, Gil1
receptor and β-receptor, which are located on different
+(tdTomato+) cells showed that the progeny of Gil1+
For personal use only.

chromosomes, 4 and 5. The PDGF α-receptor binds the

cells differentiated into odontoblasts and dental pulp cells
PDGF-A chain and the PDGF-B chain with high affinity,
and colocalized with phosphorylated Smad1/5/9 (activated
while the PDGF β-receptor binds the PDGF-B chain with
BMP signaling) after tamoxifen induction in the preodonto-
high affinity. PDGF-C binds to the α-receptor but not the β-
blast region and dental pulp cells in close proximity to this
receptor.64 PDGF-D interacts with the β-receptor, but not
region.75 These findings suggest that BMP signaling main-
the α-receptor.65 PDGF signaling is a key regulator in
tains tooth morphology and homeostasis.
mesenchymal cells. PDGF binding to its receptor induces
BMPs influence DPSCs during the processes of prolif-
dimerization and autophosphorylation, as well as activation
eration and differentiation. BMP2-transfected DPSCs iso-
of a signal transduction molecule containing a cytoplasmic
lated by STRO-1 revealed high levels of alkaline
CH2 domain.69
phosphatase (ALP) activity in vitro and the enhancement
Human DPSCs secrete PDGF-AA and other growth of mineralized tissue upon implantation.77 BMP4 affects
factors, and the titers of NGF, BDNF, and VEGF were the growth of dental pulp cells and enhances the mRNA
revealed by ELISA to be greater than those of human expression levels of ALP, DSPP, and DMP-1.78 BMP7
bone marrow-derived mesenchymal stem cells and human induction resulted in increases in dentin sialophosphoprotein
adipose-derived stem cells.70 The overexpression of (DSPP), osteocalcin (OCN), dentin matrix protein 1 (DMP-
PDGF-BB in human DPSCs increases cell proliferation 1), and runt-related transcription factor 2 (RUNX2) mRNA
and odontoblastic differentiation in particular. In addition, expression levels and the formation of mineralized nodules
the secretion of PDGF-BB by DPSCs can increase the in DPSCs.79 Through the p38 mitogen-activated protein
likelihood of stem cell homing via the PI3K/Akt pathway kinase (MAPK) and WNT canonical pathway, BMP2 was
and improve the DPSC-mediated dentin–pulp complex shown to promote the differentiation and mineralization of
regeneration in vivo.71 Moreover, separated PDGFRβ+ human DPSCs.80 Moreover, the incorporation of BMP2 and
and PDGFRβ+/c-kit+ dental pulp cells show faster prolif- VEGF into a three-dimensional culture model (TDM) using
eration than whole pulp cells and PDGFRβ− cells. human DPSCs enhanced the potency of stem cells to induce
Furthermore, an in vivo study demonstrated that trans- angiogenesis and odontogenesis. Specifically, the human
planted PDGFRβ+/c-kit+ dental pulp cells with hydrogel DPSCs and VEGF were encapsulated in a fibrin gel, and
formed globular dentin and pulp-like tissue in rat inserted into BMP2-coated demineralized dentin discs. The
incisor.72 According to these findings, PDGF enhances qRT-PCR results of this TMD showed higher expression of
DPSC proliferation, odontoblast differentiation, and platelet and endothelial cell adhesion molecule (PECAM),
regeneration of dentin–pulp complex. BSP, DMP-1, OCN, and CBFA1 than in a monolayer control

submit your manuscript | www.dovepress.com Stem Cells and Cloning: Advances and Applications 2020:13
36
DovePress

Powered by TCPDF (www.tcpdf.org)

 Dovepress Tsutsui

group.81 In another study, the autogenous transplantation of also recruited few of them.20 Regarding other cytokines,
BMP2-treated three-dimensional (3D) porcine pulp cell pel- an investigation of STRO-1-sorted cells (human pulp cells;
let culture onto amputated pulp induced reparative dentin immature third molars) treated with growth factors (FGF-2
formation.82 According to these reports, BMPs affect DPSC and TGF-β1) in a biodegradable polymer matrix of lactide
proliferation and differentiation, and increase dentinogen- and glycolide released using a Matrigel-covered dish was
Stem Cells and Cloning: Advances and Applications downloaded from https://www.dovepress.com/ by 185.50.250.82 on 29-Feb-2020

esis-related gene expression; moreover, three-dimensional also reported. FGF-2 increased dental pulp proliferation
culture enhances the properties of DPSCs. and TGF-β1 was observed to exert chemotactic potential.
This Matrigel-covered dish culture showed the controlled
Scaffolds release of growth factors upon investigating the early stage
Many different carriers for cells have been reported of pulp/dentin regeneration.21 In another study, vascular-
(Table 1). Scaffolds support appropriate cellular attach- ized pulp-like tissue and osteodentin were analyzed upon
ment, migration, proliferation, differentiation, and function the transplantation of DPSCs, human umbilical vein
to produce tissue constructs specific to the particular pur- endothelial cells (HUVECs), or co-culture of both types
pose. One such purpose would be to provide support for of cell encapsulated in a three-dimensional (3D)
replacement by transplanted cells, but for scaffolds there is PuraMatrix™ in mice. The results showed that transplan-
concern about the nature of their degradation, cytotoxicity, tation in the co-culture group produced more ECM, vas-
and immune reactions to them by the recipient. cularization, and mineralization than achieved with the
CD105+ DPSCs were transplanted with stromal cell- DPSC monocultures in vivo.22 A further study focused
For personal use only.

derived factor-1 (SDF-1) and collagen into the mature on DPSCs and HUVECs encapsulated in 5% gelatin
teeth of dogs that had undergone pulpectomy. This trans- methacrylate (GelMA) xenogeneic hydrogel and injected
plantation resulted in newly regenerated tissue, which into root segments. This transplantation in mice showed
expressed angiogenic/neurotrophic factors.19 Suzuki’s neovasculature formation.23 Another scaffold type in the
group also reported the migration of dental stem cells form of ECM was supplied by decellularized dental pulp
(DSCs) using collagen gel cylinders. The DSCs were from swine as a bioscaffold for pulp regeneration. The
seeded on the surface of these cylinders and cultured swine pulp was decellularized with a mixed solution of
with stromal-derived factor-1α(SDF-1), bFGF, and 10% sodium dodecyl sulfate and Triton X-100.
BMP7, which induced the recruitment of the cells into Transplantation of human DPSCs with decellularized den-
the cylinders. SDF-1 or bFGF recruited more cells into tal pulp into nude mice demonstrated ECM preservation
collagen gel than the case without cytokines, and BMP7 and a pulp-like tissue structure, as revealed by histological

Table 1 Scaffold, Growth Factors, and Bioactive Molecules

Reference No. Authors Scaffold Growth factors, Bioactive in vitro in vivo
molecule

[19] Iohara et al Collgen type I and type III SDF-1 Dog

[20] Suzuki et al Mixing rat tail collagen type I SDF-1, bFGF, BMP7 3D collagen scaffold Rat
solution and 0.02-N acetic
acid, human teeth
[21] Mathieu et al Matrigel FGF-2 and TGF-β1 (encaspulated FGF-2 and TGF-β1
into a biodegradable polymer of loaded microspher
lactide and glycolide) composed Matrigel
[22] Dissanayaka et al PuraMatrix™, human teeth VEGF (for in vitro) PuraMatrix™ Mouse
[23] Khayat et al GelMA hydrogel GelMA hydrogel Rat
[24] Hu et al. Decellularized dental pulp Mouse
ECM, tooth slice
[25] Zhang et al. Decellularized tooth buds BMP-2 decellularized tooth Mini-pig
buds
[26] Ravindranet al. ECM embedded collagen/ ECM embedded Mouse
chitsosan scaffold collagen/chitsosan
scaffold

submit your manuscript | www.dovepress.com

Stem Cells and Cloning: Advances and Applications 2020:13 37
DovePress

Powered by TCPDF (www.tcpdf.org)

 Tsutsui Dovepress

analysis.24 Decellularized natural porcine tooth bud has signaling in the primary human MSCs.85 Furthermore, dur-
also been shown to be useful as a bioengineered scaffold ing adipocyte differentiation, PPARγ-dependent transcrip-
for tooth regeneration, when transplanted with porcine tion is dependent on mitochondrial complex III-generated
dental epithelial cells, human dental pulp cells, and superoxide.85 Differentiation for adipogenesis and osteogen-
human umbilical vein endothelial cells. The implantation esis has been shown to be correlated with mitochondrial
Stem Cells and Cloning: Advances and Applications downloaded from https://www.dovepress.com/ by 185.50.250.82 on 29-Feb-2020

of samples into the mandibles of mini-pigs revealed den- elongation and increases in Mfn1 and 2 (mitochondrial
tin- and enamel-like tissues.25 In addition, DPSCs were fusion proteins) expression. Forni’s group reported the use
cultured on a 3D scaffold using a decellularized ECM of mouse skin mesenchymal stem cells (msMSCs), which are
embedded in a collagen/chitosan scaffold. The subcuta- CD105+ CD90+ CD73+ CD29+ CD34− mesodermal
neous implantation of the scaffold with DPSCs into nude precursors.86 In addition, chondrogenesis of msMSCs
mice resulted in dental pulp-like tissue and the expression showed increases of Drp1, Fis1, and Fis2 (fission proteins)
of dentin sialoprotein (DSP) and DSPP.26 To obtain expression and mitophagy enhancement.86 The regulation of
a deeper understanding of the stemness of DPSCs, there processes such as fission/fusion, mitochondrial biogenesis,
is a need to analyze their activity including in the presence and oxidative metabolism of mitochondria is thus key for
of scaffolds. A key focus for this analysis should be differentiation and homeostasis in MSCs.
mitochondria, one of the key organelles during the differ- Intriguingly, many studies have reported that mito-
entiation of DPSCs, the energy from which is vital for this chondria are transferred from MSCs to injured cells
process. through tunneling nanotubes. The introduction of MSCs
For personal use only.

into an infarcted heart mouse model resulted in increased

expression of heme oxygenase-1 (HO-1) and peroxisome
Research on Prospective Advanced
proliferator-activator receptor gamma coactivator-1-α
Applications for DPSC (PGC-1-α) genes in MSCs infused in intact myocardium.
Mitochondria This suggested that HO-1 or mitophagy inhibition was
To understand the mechanisms of stem cells including associated with cardiac apoptosis.87 Heart muscle
their aging and self-renewal, the establishment of micro- expresses a high level of the heart muscle protein (HMP)
environmental homeostasis and differentiation should be mitofilin.88 Mitofilin is anchored in the inner mitochon-
developed as a new approach for DPSC research and drial membrane and is a transmembrane protein.89 The
therapeutics. According to the literature, mitochondrial morphology of cristae is maintained by the mitochondrial
functions and dynamics are particularly related to the inner membrane organizing system (MINOS) including
direction of stem cell differentiation, including for DPSCs. mitofilin, which is a core component of it along with
The sequence of human mitochondrial DNA (mtDNA) is Mito10. Mitofilin has been reported to function as
16,569 base pairs long, which includes genes for the 12S and a multifunctional regulator of mitochondrial morphology
16S rRNAs, 22 tRNAs, cytochrome c oxidase subunits I, II, and protein biogenesis.90 In MSCs derived from bone
and III, ATPase subunit 6, cytochrome b, and eight other marrow, mitofilin was shown to regulate their homeostasis
predicted protein-coding genes.83 Mitochondria function in and osteogenesis.91 Earlier induction of osteogenic/denti-
energy metabolism, which regulates the homeostasis of cells nogenic markers in DPSCs was also achieved by the
including stem cells. depletion of mitofilin/3C4 antigens.92 Adipose (AD)-
Undifferentiated stem cells show higher levels of MSCs and bone marrow (BM)-MSCs showed higher mito-
glycolysis compared with stem cells undergoing chondrial transfer than dental pulp (DP)-MSCs and
differentiation.27,84 Differentiation for osteogenesis has Wharton’s jelly (WJ)-MSCs. In addition, DP-MSCs and
been shown to be retarded by exogenous H2O2 and mito- WJ-MSCs had reduced mtROS compared with BM-MSCs
chondrial inhibitors. The transition of mitochondrial energy and AD-MSCs in cardiomyocyte coculture. Moreover,
production from glycolysis to oxidative phosphorylation DP-MSCs and WJ-MSCs revealed higher mitochondrial
induces osteogenesis in human MSCs.27 Adipogenic differ- respiratory abilities.93 The initiation of the differentiation
entiation is inhibited by mitochondrially targeted antioxi- of human DPSCs to odontoblasts was also observed to
dants. During differentiation into adipocytes, there are early involve mitochondrial elongation with developed cristae,
increases in mitochondrial metabolism and reactive oxygen enhancement of the mitochondrial oxygen consumption
species (ROS) generation, which are dependent on mTORC1 rate, increasing mitochondrial ATP production,

submit your manuscript | www.dovepress.com Stem Cells and Cloning: Advances and Applications 2020:13
38
DovePress

Powered by TCPDF (www.tcpdf.org)

 Dovepress Tsutsui

upregulation of mitochondrial glycolytic enzyme activ- expression of major genes and proteins, as an example,
ities, and increased glycolytic capacity and glycolytic dentinogenesis-related genes and proteins were mainly ana-
reserve.94 Disruption of the differentiation of human lyzed. Analysis of DPSC metabolism in the presence of
DPSCs into odontoblasts was also induced by lipopolysac- growth factors and scaffolds should also help us to obtain
charide (LPS), which decreased HO-1 and PGC-1-α a deep understanding of their stemness. Such analysis of
Stem Cells and Cloning: Advances and Applications downloaded from https://www.dovepress.com/ by 185.50.250.82 on 29-Feb-2020

levels.95 LPS simulation is inhibited by Schisandrin metabolism is important because mitochondria are the main
C and activates mitochondrial biogenesis, which increased organelles producing the energy not only for the mainte-
HO-1 and PGC-1-α through the phosphorylated-protein nance of homeostasis, but also during differentiation.
kinase B (p-AKt) and nuclear factor erythroid 2-related DPSCs are a promising cell source in the cutting-edge
factor-2 pathway.96 The above findings show that mito- research field of stem cells and for developing regenerative
chondrial dynamics, metabolism, and function are asso- medicine applications. Experiments should be performed to
ciated with the fate of stem cells including DPSCs. Stem evaluate their clinical application, requiring further explora-
cell differentiation is also related to mitochondrial activity. tion and a deeper understanding of various characteristics of
Dental pulp stem cells from children, another type of stem DPSCs.
cell from human deciduous teeth (SHED), differentiate
into neuronal cells, which was shown to increase mito-
chondrial membrane potential, mitochondrial DNA, and
Acknowledgments
I thank Edanz for editing the English text of a draft of this
elongated mitochondria.97 In patients with Rett syndrome,
For personal use only.

manuscript.
loss-of-function mutations in MECP2 have been identified,
which is a gene encoding methyl-CpG-binding protein
(MeCP2). Using MeCP2-expressing and MeCP2-deficient Disclosure
stem cells from exfoliated deciduous teeth, it was shown The author reports no conflicts of interest in this work.
that differentiating MeCP2-deficient stem cells exhibited
reductions in mitochondrial membrane potential and ATP
production, and restricted mitochondrial distribution in
References
1. Gronthos S, Mankani M, Brahim J, Robey PG, Shi S. Postnatal human
neurites compared with MeCP2-expressing cells. In addi-
dental pulp stem cells (DPSCs) in vitro and in vivo. Proc Natl Acad
tion, central mitochondrial fission factor (dynamin-related Sci U S A. 2000;97(25):13625–13630. doi:10.1073/pnas.24
protein1) showed lower expression in MeCP2-deficient 0309797
2. Smith AJ, Cassidy N, Perry H, Begue-Kirn C, Ruch JV, Lesot H.
cells than in MeCP2-expressing ones.98 These reports Reactionary dentinogenesis. Int J Dev Biol. 1995;39(1):273–280.
suggest the importance of mitochondrial function in stem 3. Kitamura C, Kimura K, Nakayama T, Terashita M. Temporal and
cells. Understanding the molecular profile and morphology spatial expression of c-jun and jun-B proto-oncogenes in pulp cells
involved with reparative dentinogenesis after cavity preparation of rat
of mitochondria is thus important to improve the effective- molars. J Dent Res. 1999;78(2):673–680. doi:10.1177/002203459907
ness of DPSC-based clinical approaches. 80020701
4. Bartold PM, Bianco P, Finkelstein MW, et al. Ten Cate’s Oral
Histology: Development, Structure, and Function. 6. Mosby;2003.
Conclusion and Future Challenges 5. Shiba H, Fujita T, Doi N, et al. Differential effects of various growth
The field of research on DPSCs has great potential because factors and cytokines on the syntheses of DNA, type I collagen,
laminin, fibronectin, osteonectin/secreted protein, acidic and rich in
the cells not only have the characteristics of good differ- cysteine (SPARC), and alkaline phosphatase by human pulp cells in
entiation potential and being easy to culture, but can also be culture. J Cell Physiol. 1998;174(2):194–205. doi:10.1002/(ISSN)
1097-4652
conveniently obtained from extracted teeth, which are 6. Kunimatsu R, Nakajima K, Awada T, et al. Comparative characteriza-
usually discarded. Growth factors and scaffolds strongly tion of stem cells from human exfoliated deciduous teeth, dental pulp,
affect DPSC proliferation and their direction of differentia- and bone marrow-derived mesenchymal stem cells. Biochem Biophys
Res Commun. 2018;501(1):193–198. doi:10.1016/j.bbrc.20
tion. DPSCs can aid the regeneration of dentin/pulp-like 18.04.213
complex or other tissues in the presence of growth factors 7. Huang GT, Yamaza T, Shea LD, et al. Stem/progenitor cell-mediated
de novo regeneration of dental pulp with newly deposited continuous
and scaffolds more efficiently than in their absence.
layer of dentin in an in vivo model. Tissue Eng Part A. 2010;16
Moreover, upon the combination of DPSCs with other (2):605–615. doi:10.1089/ten.tea.2009.0518
cells and bioactive molecules, enhanced DPSC properties 8. Lindroos B, Maenpaa K, Ylikomi T, Oja H, Suuronen R, Miettinen S.
Characterisation of human dental stem cells and buccal mucosa
were obtained in vitro and in vivo. Currently, to demon- fibroblasts. Biochem Biophys Res Commun. 2008;368(2):329–335.
strate the advantage of combining analyses of the doi:10.1016/j.bbrc.2008.01.081

submit your manuscript | www.dovepress.com

Stem Cells and Cloning: Advances and Applications 2020:13 39
DovePress

Powered by TCPDF (www.tcpdf.org)

 Tsutsui Dovepress

9. Shi S, Gronthos S. Perivascular niche of postnatal mesenchymal stem 26. Ravindran S, Zhang Y, Huang CC, George A. Odontogenic induction of
cells in human bone marrow and dental pulp. J Bone Miner Res. dental stem cells by extracellular matrix-inspired three-dimensional
2003;18(4):696–704. doi:10.1359/jbmr.2003.18.4.696 scaffold. Tissue Eng Part A. 2014;20(1–2):92–102. doi:10.1089/ten.
10. Karbanova J, Soukup T, Suchanek J, Pytlik R, Corbeil D, Mokry J. tea.2013.0192
Characterization of dental pulp stem cells from impacted third molars 27. Chen CT, Shih YR, Kuo TK, Lee OK, Wei YH. Coordinated
cultured in low serum-containing medium. Cells Tissues Organs. changes of mitochondrial biogenesis and antioxidant enzymes
Stem Cells and Cloning: Advances and Applications downloaded from https://www.dovepress.com/ by 185.50.250.82 on 29-Feb-2020

2011;193(6):344–365. doi:10.1159/000321160 during osteogenic differentiation of human mesenchymal stem

11. Sakai K, Yamamoto A, Matsubara K, et al. Human dental cells. Stem Cells. 2008;26(4):960–968. doi:10.1634/stem-
pulp-derived stem cells promote locomotor recovery after complete cells.2007-0509
transection of the rat spinal cord by multiple neuro-regenerative 28. Klagsbrun M. The fibroblast growth factor family: structural and
mechanisms. J Clin Invest. 2012;122(1):80–90. doi:10.1172/JCI59 biological properties. Prog Growth Factor Res. 1989;1(4):207–235.
251 doi:10.1016/0955-2235(89)90012-4
12. Batouli S, Miura M, Brahim J, et al. Comparison of stem-cell- 29. Mergia A, Eddy R, Abraham JA, Fiddes JC, Shows TB. The genes
mediated osteogenesis and dentinogenesis. J Dent Res. 2003;82 for basic and acidic fibroblast growth factors are on different human
(12):976–981. doi:10.1177/154405910308201208 chromosomes. Biochem Biophys Res Commun. 1986;138(2):64
13. Tsutsui TW, Inaba T, Fisher LW, Robey PG, Tsutsui T. In vitro 4–651. doi:10.1016/S0006-291X(86)80545-9
chromosome aberration tests using human dental pulp cells to detect 30. Abraham JA, Mergia A, Whang JL, et al. Nucleotide sequence of a bovine
the carcinogenic potential of chemical agents. Odontology. 2006;94 clone encoding the angiogenic protein, basic fibroblast growth factor.
(1):44–50. doi:10.1007/s10266-006-0065-1 Science. 1986;233(4763):545–548. doi:10.1126/science.2425435
14. Matsui M, Kobayashi T, Tsutsui TW. CD146 positive human 31. Vlodavsky I, Folkman J, Sullivan R, et al. Endothelial cell-derived basic
dental pulp stem cells promote regeneration of dentin/pulp-like fibroblast growth factor: synthesis and deposition into subendothelial
structures. Hum Cell. 2018;31(2):127–138. doi:10.1007/s13577- extracellular matrix. Proc Natl Acad Sci U S A. 1987;84(8):2292–2296.
017-0198-2 doi:10.1073/pnas.84.8.2292
15. Prescott RS, Alsanea R, Fayad MI, et al. In vivo generation of dental 32. Globus RK, Plouet J, Gospodarowicz D. Cultured bovine bone cells
synthesize basic fibroblast growth factor and store it in their extra-
For personal use only.

pulp-like tissue by using dental pulp stem cells, a collagen scaffold,

and dentin matrix protein 1 after subcutaneous transplantation in cellular matrix. Endocrinology. 1989;124(3):1539–1547. doi:10.12
mice. J Endod. 2008;34(4):421–426. doi:10.1016/j.joen.2008.02.005 10/endo-124-3-1539
16. Fujii Y, Kawase-Koga Y, Hojo H, et al. Bone regeneration by human 33. Zeng Q, Nguyen S, Zhang H, et al. Release of growth factors into
dental pulp stem cells using a helioxanthin derivative and cell-sheet root canal by irrigations in regenerative endodontics. J Endod.
technology. Stem Cell Res Ther. 2018;9(1):24. doi:10.1186/s13287- 2016;42(12):1760–1766. doi:10.1016/j.joen.2016.04.029
018-0783-7 34. Chang YC, Chang MC, Chen YJ, et al. Basic fibroblast growth factor
17. Angelopoulos I, Brizuela C, Khoury M. Gingival mesenchymal stem regulates gene and protein expression related to proliferation, differentia-
cells outperform haploidentical dental pulp-derived mesenchymal tion, and matrix production of human dental pulp cells. J Endod. 2017;43
stem cells in proliferation rate, migration ability, and angiogenic (6):936–942. doi:10.1016/j.joen.2017.01.024
potential. Cell Transplant. 2018;27(6):967–978. doi:10.1177/096368 35. Zhang J, Lian M, Cao P, et al. Effects of nerve growth factor and
9718759649 basic fibroblast growth factor promote human dental pulp stem cells
18. Zhang J, Lu X, Feng G, et al. Chitosan scaffolds induce human dental to neural differentiation. Neurochem Res. 2017;42(4):1015–1025.
pulp stem cells to neural differentiation: potential roles for spinal doi:10.1007/s11064-016-2134-3
cord injury therapy. Cell Tissue Res. 2016;366(1):129–142. doi:10. 36. Luo L, Albashari AA, Wang X, et al. Effects of transplanted
1007/s00441-016-2402-1 heparin-poloxamer hydrogel combining dental pulp stem cells and
19. Iohara K, Imabayashi K, Ishizaka R, et al. Complete pulp regenera- bFGF on spinal cord injury repair. Stem Cells Int. 2018;2018:
tion after pulpectomy by transplantation of CD105+ stem cells with 2398521. doi:10.1155/2018/2398521
stromal cell-derived factor-1. Tissue Eng Part A. 2011;17(15–- 37. Qian J, Jiayuan W, Wenkai J, et al. Basic fibroblastic growth factor
16):1911–1920. doi:10.1089/ten.tea.2010.0615 affects the osteogenic differentiation of dental pulp stem cells in a
20. Suzuki T, Lee CH, Chen M, et al. Induced migration of dental pulp treatment-dependent manner. Int Endod J. 2015;48(7):690–700.
stem cells for in vivo pulp regeneration. J Dent Res. 2011;90 doi:10.1111/iej.2015.48.issue-7
(8):1013–1018. doi:10.1177/0022034511408426 38. Crane JL, Cao X. Bone marrow mesenchymal stem cells and TGF-beta
21. Mathieu S, Jeanneau C, Sheibat-Othman N, Kalaji N, Fessi H, signaling in bone remodeling. J Clin Invest. 2014;124(2):466–472.
About I. Usefulness of controlled release of growth factors in inves- doi:10.1172/JCI70050
tigating the early events of dentin-pulp regeneration. J Endod. 39. de Araujo Farias V, Carrillo-Galvez AB, Martin F, Anderson P. TGF-
2013;39(2):228–235. doi:10.1016/j.joen.2012.11.007 beta and mesenchymal stromal cells in regenerative medicine, auto-
22. Dissanayaka WL, Hargreaves KM, Jin L, Samaranayake LP, immunity and cancer. Cytokine Growth Factor Rev. 2018;43:25–37.
Zhang C. The interplay of dental pulp stem cells and endothelial doi:10.1016/j.cytogfr.2018.06.002
cells in an injectable peptide hydrogel on angiogenesis and pulp 40. Miyazono K, Olofsson A, Colosetti P, Heldin CH. A role of the
regeneration in vivo. Tissue Eng Part A. 2015;21(3–4):550–563. TGF-binding protein in the assembly and secretion of TGF-beta 1.
doi:10.1089/ten.tea.2014.0154 EMBO J. 1991;10(5):1091–1101. doi:10.1002/j.1460-2075.1991.
23. Khayat A, Monteiro N, Smith EE, et al. GelMA-encapsulated tb08049.x
hDPSCs and HUVECs for dental pulp regeneration. J Dent Res. 41. Blobe GC, Schiemann WP, Lodish HF. Role of transforming growth
2017;96(2):192–199. doi:10.1177/0022034516682005 factor beta in human disease. N Engl J Med. 2000;342
24. Hu L, Gao Z, Xu J, et al. Decellularized swine dental pulp as (18):1350–1358. doi:10.1056/NEJM200005043421807
a bioscaffold for pulp regeneration. Biomed Res Int. 2017;20 42. Heo JS, Choi Y, Kim HS, Kim HO. Comparison of molecular
17:9342714. doi:10.1155/2017/9342714 profiles of human mesenchymal stem cells derived from
25. Zhang W, Vazquez B, Oreadi D, Yelick PC. Decellularized tooth bud bone marrow, umbilical cord blood, placenta and adipose
scaffolds for tooth regeneration. J Dent Res. 2017;96(5):516–523. tissue. Int J Mol Med. 2016;37(1):115–125. doi:10.3892/
doi:10.1177/0022034516689082 ijmm.2015.2413

submit your manuscript | www.dovepress.com Stem Cells and Cloning: Advances and Applications 2020:13
40
DovePress

Powered by TCPDF (www.tcpdf.org)

 Dovepress Tsutsui

43. Tomic S, Djokic J, Vasilijic S, et al. Immunomodulatory properties of 61. Kohler N, Lipton A. Platelets as a source of fibroblast
mesenchymal stem cells derived from dental pulp and dental follicle are growth-promoting activity. Exp Cell Res. 1974;87(2):297–301.
susceptible to activation by toll-like receptor agonists. Stem Cells Dev. doi:10.1016/0014-4827(74)90484-4
2011;20(4):695–708. doi:10.1089/scd.2010.0145 62. Antoniades HN, Scher CD, Stiles CD. Purification of human
44. Schmierer B, Hill CS. TGFbeta-SMAD signal transduction: molecu- platelet-derived growth factor. Proc Natl Acad Sci U S A. 1979;76
lar specificity and functional flexibility. Nat Rev Mol Cell Biol. (4):1809–1813. doi:10.1073/pnas.76.4.1809
Stem Cells and Cloning: Advances and Applications downloaded from https://www.dovepress.com/ by 185.50.250.82 on 29-Feb-2020

2007;8(12):970–982. doi:10.1038/nrm2297 63. Heldin CH, Westermark B, Wasteson A. Platelet-derived growth

45. Oshimori N, Fuchs E. The harmonies played by TGF-beta in stem factor: purification and partial characterization. Proc Natl Acad Sci
cell biology. Cell Stem Cell. 2012;11(6):751–764. doi:10.1016/j.stem. U S A. 1979;76(8):3722–3726. doi:10.1073/pnas.76.8.3722
2012.11.001 64. Li X, Ponten A, Aase K, et al. PDGF-C is a new protease-activated
46. Ng F, Boucher S, Koh S, et al. PDGF, TGF-, and FGF signaling is ligand for the PDGF alpha-receptor. Nat Cell Biol. 2000;2
important for differentiation and growth of mesenchymal stem cells (5):302–309. doi:10.1038/35010579
(MSCs): transcriptional profiling can identify markers and signaling 65. LaRochelle WJ, Jeffers M, McDonald WF, et al. PDGF-D, a new
pathways important in differentiation of MSCs into adipogenic, chon- protease-activated growth factor. Nat Cell Biol. 2001;3(5):517–521.
drogenic, and oste. Blood. 2008;112:295–307. doi:10.1182/blood- doi:10.1038/35074593
2007-07-103697 66. Betsholtz C, Johnsson A, Heldin CH, et al. cDNA sequence and
47. Karbanova J, Soukup T, Suchanek J, Mokry J. Osteogenic differen- chromosomal localization of human platelet-derived growth factor
tiation of human dental pulp-derived stem cells under various ex-vivo A-chain and its expression in tumour cell lines. Nature. 1986;320
culture conditions. Acta Medica (Hradec Kralove). 2010;53 (6064):695–699. doi:10.1038/320695a0
(2):79–84. doi:10.14712/18059694.2016.64 67. Swan DC, McBride OW, Robbins KC, Keithley DA, Reddy EP,
48. Song B, Jiang W, Alraies A, et al. Bladder smooth muscle cells Aaronson SA. Chromosomal mapping of the simian sarcoma virus
differentiation from dental pulp stem cells: future potential for blad- onc gene analogue in human cells. Proc Natl Acad Sci U S A.
der tissue engineering. Stem Cells Int. 2016;2016:6979368. 1982;79(15):4691–4695. doi:10.1073/pnas.79.15.4691
doi:10.1155/2016/6979368 68. Uutela M, Lauren J, Bergsten E, et al. Chromosomal location, exon
49. Johnson D, Lanahan A, Buck CR, et al. Expression and structure of structure, and vascular expression patterns of the human PDGFC and
For personal use only.

the human NGF receptor. Cell. 1986;47(4):545–554. doi:10.1016/ PDGFD genes. Circulation. 2001;103(18):2242–2247. doi:10.1161/
0092-8674(86)90619-7 01.CIR.103.18.2242
50. Kaplan DR, Hempstead BL, Martin-Zanca D, Chao MV, Parada LF. 69. Heldin CH, Ostman A, Ronnstrand L. Signal transduction via
The trk proto-oncogene product: a signal transducing receptor for platelet-derived growth factor receptors. Biochim Biophys Acta.
nerve growth factor. Science. 1991;252(5005):554–558. doi:10.1126/ 1998;1378(1):F79–F113. doi:10.1016/s0304-419x(98)00015-8
science.1850549 70. Mead B, Logan A, Berry M, Leadbeater W, Scheven BA. Paracrine-
51. Hartman DS, McCormack M, Schubenel R, Hertel C. Multiple trkA mediated neuroprotection and neuritogenesis of axotomised retinal gang-
proteins in PC12 cells bind NGF with a slow association rate. J Biol lion cells by human dental pulp stem cells: comparison with human bone
Chem. 1992;267(34):24516–24522. marrow and adipose-derived mesenchymal stem cells. PLoS One. 2014;9
52. Neet KE, Campenot RB. Receptor binding, internalization, and retro- (10):e109305. doi:10.1371/journal.pone.0109305
grade transport of neurotrophic factors. Cell Mol Life Sci. 2001;58 71. Zhang M, Jiang F, Zhang X, et al. The effects of platelet-derived
(8):1021–1035. doi:10.1007/PL00000917 growth factor-bb on human dental pulp stem cells mediated
53. Woo SB, Timm DE, Neet KE. Alteration of NH2-terminal residues of dentin-pulp complex regeneration. Stem Cells Transl Med. 2017;6
nerve growth factor affects activity and Trk binding without affecting (12):2126–2134. doi:10.1002/sctm.17-0033
stability or conformation. J Biol Chem. 1995;270(11):6278–6285. 72. Cai S, Zhang W, Chen W. PDGFRbeta(+)/c-kit(+) pulp cells are
doi:10.1074/jbc.270.11.6278 odontoblastic progenitors capable of producing dentin-like structure
54. Mahadeo D, Kaplan L, Chao MV, Hempstead BL. High affinity nerve in vitro and in vivo. BMC Oral Health. 2016;16(1):113. doi:10.1186/
growth factor binding displays a faster rate of association than p140trk s12903-016-0307-8
binding. Implications for multi-subunit polypeptide receptors. J Biol 73. Wozney JM, Rosen V, Celeste AJ, et al. Novel regulators of bone
Chem. 1994;269(9):6884–6891. formation: molecular clones and activities. Science. 1988;242
55. Sofroniew MV, Howe CL, Mobley WC. Nerve growth factor signal- (4885):1528–1534. doi:10.1126/science.3201241
ing, neuroprotection, and neural repair. Annu Rev Neurosci. 74. Yang G, Yuan G, Ye W, Cho KW, Chen Y. An atypical canonical
2001;24:1217–1281. doi:10.1146/annurev.neuro.24.1.1217 bone morphogenetic protein (BMP) signaling pathway regulates
56. Di Marco E, Mathor M, Bondanza S, et al. Nerve growth factor binds Msh homeobox 1 (Msx1) expression during odontogenesis.
to normal human keratinocytes through high and low affinity recep- J Biol Chem. 2014;289(45):31492–31502. doi:10.1074/jbc.M114.
tors and stimulates their growth by a novel autocrine loop. J Biol 600064
Chem. 1993;268(30):22838–22846. 75. Shi C, Yuan Y, Guo Y, et al. BMP signaling in regulating mesench-
57. Vega JA, Garcia-Suarez O, Hannestad J, Perez-Perez M, Germana A. ymal stem cells in incisor homeostasis. J Dent Res. 2019;98
Neurotrophins and the immune system. J Anat. 2003;203(1):1–19. (8):904–911. doi:10.1177/0022034519850812
doi:10.1046/j.1469-7580.2003.00203.x 76. Plikus MV, Zeichner-David M, Mayer JA, et al. Morphoregulation of
58. Mitsiadis TA, Pagella P. Expression of Nerve Growth Factor (NGF), teeth: modulating the number, size, shape and differentiation by
TrkA, and p75(NTR) in developing human fetal teeth. Front Physiol. tuning Bmp activity. Evol Dev. 2005;7(5):440–457. doi:10.1111/
2016;7:338. doi:10.3389/fphys.2016.00338 j.1525-142X.2005.05048.x
59. Mitsiadis TA, Magloire H, Pagella P. Nerve growth factor signalling 77. Yang X, van der Kraan PM, Bian Z, Fan M, Walboomers XF,
in pathology and regeneration of human teeth. Sci Rep. 2017;7 Jansen JA. Mineralized tissue formation by BMP2-transfected pulp
(1):1327. doi:10.1038/s41598-017-01455-3 stem cells. J Dent Res. 2009;88(11):1020–1025. doi:10.1177/
60. Mead B, Logan A, Berry M, Leadbeater W, Scheven BA. 0022034509346258
Intravitreally transplanted dental pulp stem cells promote neuropro- 78. Sun N, Jiang T, Wu C, Sun H, Zhou Q, Lu L. Expression and
tection and axon regeneration of retinal ganglion cells after optic influence of BMP-4 in human dental pulp cells cultured in vitro.
nerve injury. Invest Ophthalmol Vis Sci. 2013;54(12):7544–7556. Exp Ther Med. 2018;16(6):5112–5116. doi:10.3892/etm.2018.
doi:10.1167/iovs.13-13045 6824

submit your manuscript | www.dovepress.com

Stem Cells and Cloning: Advances and Applications 2020:13 41
DovePress

Powered by TCPDF (www.tcpdf.org)

 Tsutsui Dovepress

79. Zhu L, Na J, Mu R et al. Bone morphogenic protein 7 promotes 90. von der Malsburg K, Muller JM, Bohnert M, et al. Dual role of
odontogenic differentiation of dental pulp stem cells in vitro. Life Sci. mitofilin in mitochondrial membrane organization and protein
2018;202:175–181. biogenesis. Dev Cell. 2011;21(4):694–707. doi:10.1016/j.devcel.20
80. Yang J, Ye L, Hui TQ, et al. Bone morphogenetic protein 2-induced 11.08.026
human dental pulp cell differentiation involves p38 mitogen-activated 91. Lv YJ, Yang Y, Sui BD, et al. Resveratrol counteracts bone loss via
protein kinase-activated canonical WNT pathway. Int J Oral Sci. mitofilin-mediated osteogenic improvement of mesenchymal stem
Stem Cells and Cloning: Advances and Applications downloaded from https://www.dovepress.com/ by 185.50.250.82 on 29-Feb-2020

2015;7(2):95–102. doi:10.1038/ijos.2015.7 cells in senescence-accelerated mice. Theranostics. 2018;8(9):23

81. Aksel H, Ozturk S, Serper A, Ulubayram K. VEGF/BMP-2 loaded 87–2406. doi:10.7150/thno.23620
three-dimensional model for enhanced angiogenic and odontogenic 92. Hwang HI, Lee TH, Jang YJ. Cell proliferation-inducing protein 52/
potential of dental pulp stem cells. Int Endod J. 2018;51(4):420–430. mitofilin is a surface antigen on undifferentiated human dental pulp
doi:10.1111/iej.12869 stem cells. Stem Cells Dev. 2015;24(11):1309–1319. doi:10.1089/
82. Iohara K, Nakashima M, Ito M, Ishikawa M, Nakasima A, scd.2014.0387
Akamine A. Dentin regeneration by dental pulp stem cell therapy 93. Paliwal S, Chaudhuri R, Agrawal A, Mohanty S. Human
with recombinant human bone morphogenetic protein 2. J Dent tissue-specific MSCs demonstrate differential mitochondria transfer
Res. 2004;83(8):590–595. doi:10.1177/154405910408300802 abilities that may determine their regenerative abilities. Stem Cell Res
83. Anderson S, Bankier AT, Barrell BG, et al. Sequence and organization of Ther. 2018;9(1):298. doi:10.1186/s13287-018-1012-0
the human mitochondrial genome. Nature. 1981;290(5806):457–465.
94. Wang L, Cheng L, Wang H, et al. Glycometabolic reprogramming
doi:10.1038/290457a0
associated with the initiation of human dental pulp stem cell
84. Zhang J, Khvorostov I, Hong JS, et al. UCP2 regulates energy metabolism
differentiation. Cell Biol Int. 2016;40(3):308–317. doi:10.1002/cbin.
and differentiation potential of human pluripotent stem cells. EMBO J.
v40.3
2011;30(24):4860–4873. doi:10.1038/emboj.2011.401
95. Takanche JS, Kim JS, Kim JE, Han SH, Yi HK. Schisandrin
85. Tormos KV, Anso E, Hamanaka RB, et al. Mitochondrial complex III
C enhances odontoblastic differentiation through autophagy and
ROS regulate adipocyte differentiation. Cell Metab. 2011;14
mitochondrial biogenesis in human dental pulp cells. Arch Oral
(4):537–544. doi:10.1016/j.cmet.2011.08.007
86. Forni MF, Peloggia J, Trudeau K, Shirihai O, Kowaltowski AJ. Biol. 2018;88:60–66. doi:10.1016/j.archoralbio.2018.01.018
For personal use only.

Murine mesenchymal stem cell commitment to differentiation is 96. Takanche JS, Lee YH, Kim JS, et al. Anti-inflammatory and antiox-
idant properties of Schisandrin C promote mitochondrial biogenesis
regulated by mitochondrial dynamics. Stem Cells. 2016;34(3):74
3–755. doi:10.1002/stem.2248 in human dental pulp cells. Int Endod J. 2018;51(4):438–447.
87. Mahrouf-Yorgov M, Augeul L, Da Silva CC, et al. Mesenchymal doi:10.1111/iej.12861
stem cells sense mitochondria released from damaged cells as 97. Kato H, Thi Mai Pham T, Yamaza H, et al. Mitochondria regulate the
danger signals to activate their rescue properties. Cell Death differentiation of stem cells from human exfoliated deciduous teeth.
Differ. 2017;24(7):1224–1238. doi:10.1038/cdd.2017.51 Cell Struct Funct. 2017;42(2):105–116. doi:10.1247/csf.17012
88. Icho T, Ikeda T, Matsumoto Y, Hanaoka F, Kaji K, Tsuchida N. A novel 98. Hirofuji S, Hirofuji Y, Kato H, et al. Mitochondrial dysfunction in
human gene that is preferentially transcribed in heart muscle. Gene. dopaminergic neurons differentiated from exfoliated deciduous
1994;144(2):301–306. doi:10.1016/0378-1119(94)90394-8 tooth-derived pulp stem cells of a child with Rett syndrome.
89. Gieffers C, Korioth F, Heimann P, Ungermann C, Frey J. Mitofilin is Biochem Biophys Res Commun. 2018;498(4):898–904. doi:10.1016/
a transmembrane protein of the inner mitochondrial membrane j.bbrc.2018.03.077
expressed as two isoforms. Exp Cell Res. 1997;232(2):395–399.
doi:10.1006/excr.1997.3539

Stem Cells and Cloning: Advances and Applications Dovepress

Publish your work in this journal
Stem Cells and Cloning: Advances and Applications is an interna- animal and human therapeutic studies; Philosophical and ethical
tional, peer-reviewed, open access journal. Areas of interest in issues related to stem cell research. This journal is indexed on
established and emerging concepts in stem cell research include: CAS. The manuscript management system is completely online
Embryonic cell stems; Adult stem cells; Blastocysts; Cordblood and includes a very quick and fair peer-review system, which is all
stem cells; Stem cell transformation and culture; Therapeutic easy to use. Visit http://www.dovepress.com/testimonials.php to read
cloning; Umbilical cord blood and bone marrow cells; Laboratory, real quotes from published authors.

Submit your manuscript here: https://www.dovepress.com/stem-cells-and-cloning-advances-and-applications-journal

submit your manuscript | www.dovepress.com Stem Cells and Cloning: Advances and Applications 2020:13
42
DovePress

Powered by TCPDF (www.tcpdf.org)