Dental Stem Cells

Reviewed by Terrell F. Pannkuk, D.D.S. M.Sc.D

This presentation is an outline derived from the following article:

Mesenchymal Stem Cells Derived from

Dental Tissues vs. Those from Other
Sources: Their Biology and Role in
Regenerative Medicine
G. T. J. Huang, S. Gronthos, and S. Shi, J Dent Res 88 (9):
792-806, 2009

Stem Cells
Derived from Dental Tissues
Mesenchymal Stem Cells (MSCs)
Sources:
Bone Marrow (Friedenstein et al, 1976; Caplan,
1991; Prockop, 1997; Pittenger et al, 1999; Gronthos
et al, 2003)
Adipose Tissue/Umbilical Cord (Mareschi et al, 2001;
Zuk et al, 2001)

Lineages:

Osteogenic
Chondrogenic
Adipogenic

Other Lineages Possibly

Derived from Bone Marrow
Mesenchymal Stem Cells
Myogenic
Neurogenic
Tenogenic

Dental Tissue MSCs

Human Pulp Tissue (DPSCs, post-natal dental pulp stem

cells)
Gronthos et al, 2000

Exfoliated Deciduous Teeth (SHED)

Miura et al, 2003

Periodontal Ligament (PDLSC)

Seo et al, 2004

Apical Papilla (SCAP)

Sonoyama et al, 2006, 2008

Dental Follicle Precursors (DFPC)

Morsczeck et al, 2005

Dental Stem Cell

Lineages
Osteo/Odontogenic
Adipogenic
Neurogenic

*Dental Stem cells appear to be more committed

to odontogenic paths than BMMSCs

BMMSCs
Colony Forming Unit Fibroblasts (CFU-Fs)
Self Renewal (like hematopoietic lines)
30-50 PDs (population doublings)
Cell Surface Markers
Heterogeneity supports stromal hierarchy of

differentiation
Minor proportion involved with extensive
proliferation

Dental MSCs
Dental tissues are specialized tissues that do

not undergo continuous remodeling as shown

in bony tissues
Dental mesenchyme is termed

ectomesenchyme due to its earlier

interaction with the neural crest.

Isolation of Dental Pulp Stem Cells

Enzymatically isolated and seeded onto

dentin to promote Odontoblast-like cells.

Multilineage differentiation of hDPSC

subpopulations:
Adipogenic
Neurogenic
Osteogenic
Chondrogenic
Myogenic

Ectopic Formation of Dentin-Pulp-like

Complex
Transplanted DPSCs mixed with

hydroxyapatite/tricalcium phosphate (HA/TCP)

forms ectopic pulp-dentin like tissue
complexes in immunocompromised mice.
(Gronthos et al., 2000; Batouli et al., 2003)
Odontoblast-like cells express
sialophosphoprotein (DSPP), producing
dentinal tubules similar to natural dentin

SHED (Exfoliated Deciduous

Teeth SCs)
Fast proliferation
Greater PD (population doubling)
Sphere like cluster formation (cultured

neurogenic medium
Also termed immature stem cells)
Unable to regenerate a complete dentin-pulp
complex in vivo
Unlike DPSCs can differentiate into bone
forming cells.

SCAP ( Apical Papilla

SCs)
Odontogenic differentiation
Adipogenic differentiation

DPSCs vs. SCAP

Apical papilla is a precursor to radicular pulp
Earlier line of stem/progenator cells (SCAP)
SCAPs superior source of stem cells

PDLSCs (periodontal ligament scs)

Form cementoblasts and osteoblasts
Homeostasis and regeneration of perio tissues
Cementum-PDL structure unique from

BMMSCs and DPSCs

DFPCs (Dental Follicle

Precursor Cells)
Periodontium, cementum, PDL, alveolar bone

precursors
Source: impacted third molars

Dental MSCs vs.

BMMSCs
Gene Expression: 4000 known human genes
Cooperative regulation of genes for cell

signaling, cell communication, or metabolism

BMMSCs only form bone tissue in mice
DPSC chondrogenic potential is weak
BMMSCs have stronger adipogenic potential
than both DPSCs and SCAP
Neurogenicity in dental stem cells more
potent than BMMSCs (probably due to neural
crest origin)

MSC Niche
Specialized microenvironment needed to

maintain stem cells in their multipotent state.

(Schofield, 1978)
Considered a fixed compartment:
Regulate proliferation
Control fate of stem cell progeny
Prevent exhaustion and death of stem cells

(Scadden, 2006; Jones and Wagers, 2008)

BMMSC niche-perivascular area of bone

marrow
DPSC niche-perivascular and perineural

MSC
Homing
MSCs in human blood is low under steady state

conditions
Ex Vivo expanded MSCs injected into the blood
stream have a limited capacity to home into
various tissues and organs.
Injected Ex Vivo-expanded BMMSCs through
intravenous infusion lodge mainly in lungs,
smaller amounts in liver, heart, spleen, and
damaged areas of the brain.
No evidence that BMMSCs migrate to orofacial
/dental organs

Immunomodulation of
MSCs
Allogenic MSCs are well tolerated by the

recipient hosts (Xenografts do not take).

MSCs have an immunosuppressive effect
Preliminary study shows interferon may act to
differentiate MSCs into osteoblasts
Inflammatory reactions against scaffold
materials and serum components lead to the
production of cytokines

Dental MSC-Based
Therapy for Regenerative Medicine
SCAP and PDLSCs for Bio-root Engineering
Single cells from dog tooth buds at the bell
stage seeded onto scaffolds and transplanted
back into sockets resulted in some dentin
structure regeneration with no enamel or root
formation (Honda et al., 2006)
Kuo et al., 2007 used pigs, expanded ex vivo
expansion of bud cells from bell stage and
observed some root structures along with
periodontium.

Obstacles to Tooth Regeneration

Abnormal (small) tooth size
Lack of consistent root formation
Incomplete eruption into functional occlusion.

Regeneration of Perio Defects with

PDLSCs
PDGF (platelet derived growth factor)
IGF (insulin derived growth factor)
PRP (platelet rich plasma)
Cell based regenerative therapy:
Ex vivo expanded autologous BMMSCs facilitated

repair of perio defects (Yamada et al., 2006)

PDL regeneration is as important as bone
regeneration otherwise ankylosis ensues
rhBMP-2 therapy does not regenerate PDL
PDLSCs may be an ideal source to regenerate PDL
(Liu et al., 2008)

Pulp Tissue
Engineering/Regeneration
Early attempts (Myers and Fountain, 1974) allowed a

blood clot to form in the canal but only connective

tissue formed.
More recently pulp cells grown on polyglycolic acid

(PGA) formed pulp-like tissue in vitro and in vivo (Gu et

al., 1996; Moony et al., 1996, and Burma et al., 1999)
Since the isolation and characterization of DPSCs

SHED and SCAP, more sophisticated regenerative

investigation has occurred (Huang et al., 2006, 2008;
Murray et al., 2007; Prescott et al., 2008)

Modern Pulp
Regeneration
SHED seeded onto synthetic scaffolds seated

into pulp chamber space formed odontoblast-like

cells that located against the existing dentin
surface. (not orthotopic) (Cordeiro et al., 2008)
Speculation: undifferentiated MSCs residing in

the periapical tissue and BMMSCs in the alveolar

bone of the jaws can be introduced into the root
canal space and via blood clots to allow for pulp
tissue regeneration and formation of
odontoblasts (Myers and Fountain, 1974)

Modern Pulp Regeneration (cont.)

More realistically: the known characteristics of PDLSCs,

DPSCs, and SCAP suggest that it is unlikely that

odontoblasts can be derived from PDL or periapical bone.
When BMMSCs and DPSCs are transplanted into the

subcutaneous space of immunocompromised mice they

form BM-like and Dentin-pulp like complexes respectively
(Gronthos et al., 2000)
DPSCs have shown osteogenic potential but there is no

evidence showing BMMSCs can give rise to functional

odontoblasts and dentin.

The Future:
Need to understand mechanisms of self-renewal and regulate stem

cell growth to generate sufficient numbers

Need to overcome regulation of differentiation into specific tissue

production, specialized extracellular matrices (bone, dentin,

cartilage, and tendon). The production of the extracellular matrix
and its maturation into specialized tissues involves a sequential
activation of cascades of signals.

Need to understand the interactions between stem cells and the

immune system. Allogenic dental MSCs may suppress recipient

host short and long term immunorejection.

Controlling and preventing ex vivo expanded MSCs from

transformation . Adipose derived MSCs lost genetic stability over

time and are prone to tumor formation (Rubio et al., 2005)