Dental Stem Cells

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 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

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 Stem Cells
Derived from Dental
Tissues
 Mesenchymal Stem Cells (MSC’s)
 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
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Myogenic
Neurogenic
Tenogenic

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 Dental Tissue MSC’s

 Human Pulp Tissue (DPSC’s, 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

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 Dental Stem Cell
Lineages
 Osteo/Odontogenic

 Adipogenic

 Neurogenic

*Dental Stem cells appear to be more committed to

odontogenic paths than BMMSC’s

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 BMMSC’s

 Colony Forming Unit Fibroblasts (CFU-F’s)

 Self Renewal (like hematopoietic lines)
 30-50 PD’s (population doublings)
 Cell Surface Markers
 Heterogeneity supports stromal hierarchy of
differentiation
 Minor proportion involved with extensive
proliferation
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Dental MSC’s

 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.

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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

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Ectopic Formation of Dentin-Pulp-
like Complex

 Transplanted DPSC’s 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
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SHED (Exfoliated Deciduous
Teeth SC’s)
 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 DPSC’s can differentiate into bone
forming cells.

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SCAP ( Apical Papilla SC’s)

 Odontogenic differentiation
 Adipogenic differentiation

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 DPSC’s vs.
SCAP
 Apical papilla is a precursor to radicular pulp
 Earlier line of stem/progenator cells (SCAP)
 SCAP’s superior source of stem cells

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PDLSC’s (periodontal
ligament sc’s)
 Form cementoblasts and osteoblasts
 Homeostasis and regeneration of perio
tissues
 Cementum-PDL structure unique from
BMMSC’s and DPSC’s

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DFPC’s (Dental Follicle
Precursor Cells)
 Periodontium, cementum, PDL, alveolar bone
precursors
 Source: impacted third molars

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Dental MSC’s vs. BMMSC’s

 Gene Expression: 4000 known human genes

 Cooperative regulation of genes for cell
signaling, cell communication, or metabolism
 BMMSC’s only form bone tissue in mice
 DPSC chondrogenic potential is weak
 BMMSC’s have stronger adipogenic potential
than both DPSC’s and SCAP
 Neurogenicity in dental stem cells more potent
than BMMSC’s (probably due to neural crest
origin)
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 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 sheath
areas

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 MSC Homing

 MSC’s in human blood is low under steady state

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

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Immunomodulation of MSC’s

 Allogenic MSC’s are well tolerated by the

recipient hosts (Xenografts do not take).
 MSC’s have an immunosuppressive effect
 Preliminary study shows interferon may act
to differentiate MSC’s into osteoblasts
 Inflammatory reactions against scaffold
materials and serum components lead to the
production of cytokines

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Dental MSC-Based
Therapy for Regenerative
Medicine
 SCAP and PDLSC’s 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. www.rxdentistry.net
Obstacles to Tooth
Regeneration
 Abnormal (small) tooth size

 Lack of consistent root formation

 Incomplete eruption into functional

occlusion.

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Regeneration of Perio Defects with
PDLSC’s

 PDGF (platelet derived growth factor)

 IGF (insulin derived growth factor)
 PRP (platelet rich plasma)
 Cell based regenerative therapy:
 Ex vivo expanded autologous BMMSC’s 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
 PDLSC’s may be an ideal source to regenerate
PDL (Liu et al., 2008)

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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 DPSC’s SHED

and SCAP, more sophisticated regenerative investigation
has occurred (Huang et al., 2006, 2008; Murray et al.,
2007; Prescott et al., 2008)

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 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 MSC’s residing in

the periapical tissue and BMMSC’s 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)
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Modern Pulp Regeneration
(cont.)
 More realistically: the known characteristics of PDLSC’s,
DPSC’s, and SCAP suggest that it is unlikely that
odontoblasts can be derived from PDL or periapical
bone.

 When BMMSC’s and DPSC’s are transplanted into the

subcutaneous space of immunocompromised mice they
form BM-like and Dentin-pulp like complexes
respectively (Gronthos et al., 2000)

 DPSC’s have shown osteogenic potential but there is no

evidence showing BMMSC’s can give rise to functional
odontoblasts and dentin.

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 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 MSC’s may suppress recipient host short and long
term immunorejection.

 Controlling and preventing ex vivo expanded MSC’s from transformation .

Adipose –derived MSC’s lost genetic stability over time and are prone to
tumor formation (Rubio et al., 2005)

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