prof . (dr) mousumi goswamiisppd.org.in/images/pedo_pdf/dr mousumi goswami session 6.pdf · if the...
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Prof . (Dr) Mousumi Goswami
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• No conflict of interest.
• This is an educational presentation with no financial support.
• Compilation of information from various online and offline sources
and are recommended, however others may be referred to.
• Happy to share
DISCLAIMER
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RATIONALE
PROTOCOL
ROLE OF STEM CELLS
RECENT ADVANCES
Definition & Alternate Terminologies
History
Objectives
Stem Cells, Growth Factors Scaffold
Clinical Considerations For Regenerative Endodontic Procedure
Outcomes – Clinical And Histological
Future Prospects
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4
RATIONALE
Pulp of young permanent teeth becomes necrosed due to trauma, caries etc
Tooth becomes non vital and may develop periapical pathosis
It becomes an endodontic challenge. We cannot do regular RCT and resort to
other treatment modalities like apexification or apical plug formation.
Root maturation ceases
Crown- root ratio gets altered
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No potential to restore the vitality of damaged tissue in the
canal space
No potential to promote root maturation (thickening of the
root canal walls and/or apical closure) of immature permanent
teeth with necrotic pulp
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Long Time period required for formation of the calcific bridge
Multiple appointments needed for reapplication
Affects the mechanical properties of the dentin
Increased risk of fracture of tooth
Calcific dentinal bridge formed is of poor quality with pores and tunnels
that often get perforated while obturating
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REGENERATIVE ENDODONTICS
Biologically based procedures designed to physiologically
replace damaged tooth structure, including dentin and root
structures, as well as the PULP-DENTIN COMPLEX
Murray PE, Garcia-Godoy F, Hargreaves KM. Regenerative endodontics: a review of current
status and a call for action. J Endod 2007;33:377–90
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Revascularization : The re-establishment of the vascular
supply to existing pulp in immature permanent teeth
Revitalization :An ingrowth of tissue that may not be
the same as the original lost tissue
Maturogenesis: The continued physiological root
development
Regeneration : pulp revascularization + restoration of
functional odontoblasts and/or nerve fibres
Mao JJ, Kim SG, Zhou J, et al. Regenerative endodontics: Barriers and strategies for clinical translation. Dent Clin North Am.
2012;56(3):639–49.
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Healing of apical periodontitis
Increased thickness of the root canal wall
Continued development of the
root apex
Restoration of pulpal function
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Evaluated the role of the blood clot in healing
Ingrowth of connective tissue into the canal space
Varied levels of mineralized tissue along the canal walls
“Islands” of mineralized tissue embedded within the newly formed tissue
Nygaard-Østby B. The role of the blood clot in endodontic therapy. An experimental histological study. ActaOdontol Scand 1961: 19: 324–353
HISTORY
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Clinically
Iwaya et al. (2001) were the first
group to apply the concept of
revascularization
Antimicrobial agent
metronidazole & ciprofloxacin
was used.
Iwaya SI, Ikawa M, Kubota M. Revascularization of an immature permanent tooth with apical
periodontitis and sinus tract. Dent Traumatol 2001;17:185–7
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LANDMARK ARTICLE
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.
Contemporary
Regenerative
Endodontics
Banchs F, Trope M. Revascularization of immature permanent teeth with apical periodontitis: new treatment
protocol? J Endod 2004: 30: 196–200
Elimination of bacteria
Hermetic coronal
seal
Creation of a
scaffold
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Author Clinical outcome
Nygaard-Ostby, 1961 n=17 .All cases showed ingrowth of connective tissue
The tissue resembles fibrous connective tissue with
numerous capillaries and "undifferentiated
mesenchymal elements" near the capillaries.
Rule and Winter, 1966 n=5 Resolution of signs and symptoms of pathosis,
including periapical radiolucencies. In addition, all
cases showed continued root development and/or
apical closure (6 months–1 year)
Nygaard-Ostby and
Hjortdal, 1971 n=45
Histologic outcome: there was evidence of a
vascularized fibrous connective, with often
deposition of cellular cementum
Iwaya et al, 2001 n=1 Complete resolution of signs and symptoms of
pathosis with complete closure and evidence
of root development at 30-month recall visit.
Banchs and Trope, 2004 n=1 Periradicular lesion healed (6-month recall);
continued root development (12-month recall);
tooth development, apical closure, and cold response
(2-year follow-up)
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STEM CELLS
SCAFFOLD
GROWTH FACTORS
Based on the principles of tissue engineering
Interplay of:--
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STEM CELLS (Short essay)
CONTENTS
Definition
Types
Classification
Dental stem cells
Apical Papilla
Applications
Stem cell therapy
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Stem / Progenitor cells
Self renewal - maintains the stem cell pool
Differentiation – replaces dead or damaged cells throughout life
Embryonic stem (ES) cells from the inner
cell mass of blastocysts
Adult stem cells from adult tissues
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Embryonic stem cells
from inner cell mass
of 3- to 5-day embryo
Totipotent Can give rise to all the cell
types of the body, including
those cells making the
extraembryonic tissues (eg,
placenta)
Unlimited capacity to divide
Each cell can develop into a
new individual
Embryonic stem cells
Induced pluripotent
stem cells
Pluripotent Can form derivatives of all the
embryonic germ layers
(ectoderm, mesoderm, and
endoderm) from a single cell
Adult stem cells
(postnatal)
Multipotent Can differentiate and form a
number of tissue types
Stem Cell Potency
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Hargreaves et al,Treatment Options: Biological Basis of Regenerative Endodontic Procedures ; JOE: 2013
SOURCES OF DENTAL STEM CELLS
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Huang G, Sonoyama W, Shi S.The Hidden Treasure in Apical Papilla: The Potential Role
in Pulp/Dentin Regeneration and BioRoot Engineering J Endod. 2008; 34(6): 645-651.
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Dense reservoir of undifferentiated MSCs
SCAP are regulated by Hertwig’s epithelial root sheath
Readily available rich source of stem cells due to close proximity
W. Sonoyama, Y. Liu, T. Yamaza et al., “Characterization of the apical papilla
and its residing stem cells from human immature permanent teeth: a pilot study,”
Journal of Endodontics, vol. 34, no. 2, pp. 166–171, 2008
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Provide support for cell organization, proliferation,
differentiation and vascularization
Biodegradable
Degradation rate coincide rate of tissue formation
A high porosity and an adequate pore size are necessary to
facilitate cell seeding and diffusion throughout the whole
structure of both cells and nutrients
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Natural polymers
(Collagen, Hyaluronic Acid, Chitosan And Chitin)
Synthetic polymers
(Polylactic Acid, Polyglycolic Acid, Tricalcium Phosphate, Hydroxyapatite)
Hydrogels
Bioceramics
Gotlieb EL, Murray PE, Namerow KN, et al. An ultrastructural investigation oftissue-engineered pulp constructs implanted within endodontically treated teeth.
J Am Dent Assoc 2008;139:457-6
Chandrahasa S, Murray PE, Namerow KN. Proliferation of mature ex vivo human dental pulp using tissue engineering scaffolds. J Endod 2011;37:1236-9
Torabinejad M, Milan M, Shabahang S, et al. Histologic examination of teeth with necrotic pulps and periapical lesions treated with 2 scaffolds: an animal investigation. J Endod 2015; 41: 846–852
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Growth factors are proteins that bind to receptors on the cell
and act as signals to induce cellular proliferation and/or
differentiation
Smith AJ, Scheven BA, Takahashi Y, et al. Dentine as a bioactive extracellular
matrix. Arch Oral Biol 2012;57:109-21
Sun HH, Jin T, Yu Q, et al. Biological approaches toward dental pulp regeneration
by tissue engineering. J Tissue Eng Regen Med 2011;5:e1-16
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Basic fibroblast growth factors (bFGFs) for CHEMOTAXIS AND
ANGIOGENESIS
Vascular endothelial growth factors (VEGFs) for CHEMOTAXIS,
MITOGENESIS, AND ANGIOGENESIS
Platelet-derived growth factors PDGF for ANGIOGENESIS
Nerve growth factors (NGF) for survival and GROWTH OF NERVE
FIBERS
Bone morphogenetic protein-7 (BMP-7) for MINERALIZED TISSUE
FORMATION
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PROTOCOL
CASE SELECTION
INFORMED CONSENT
FIRST APPOINTMENT
SECOND APPOINTMENT
FOLLOW-UP
OUTCOMES
CONTENTS
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Case Selection:
Tooth with necrotic pulp and an immature apex.
Pulp space not needed for post/core, final restoration.
Compliant patient/parent.
Patients not allergic to medicaments and antibiotics necessary to complete procedure (ASA 1 or 2).
Informed Consent : Two (or more) appointments
Use of antimicrobial(s)
Possible adverse effects: staining of crown/root, lack of response to treatment, pain/infection
Alternatives: MTA apexification, no treatment, extraction (when deemed non-salvageable)
Permission to enter information into AAE database (optional).
Clinical Considerations for a Regenerative Procedure
(American Association Of Endodontists )
PROTOCOL
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• Local anesthesia, rubber dam isolation and access opening
• Copious, gentle irrigation with 20ml NaOCl using an irrigation system that
minimizes the possibility of extrusion of irrigants into the periapical space
(e.g., needle with closed end and side-vents, or EndoVac™)
• Lower concentrations of NaOCl are advised
[1.5% NaOCl (20mL/canal,5 min) then irrigated with saline or EDTA (20 mL/canal,5 min),
with irrigating needle positioned about 1 mm from root end, to minimize
cytotoxicity to stem cells in the apical tissues
• Dry canals with paper points.
FIRST APPOINTMENT
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Place calcium hydroxide or low concentration of triple antibiotic paste
If the triple antibiotic paste is used:
1) Consider sealing pulp chamber with a dentin bonding agent
[to minimize risk of staining]
2) Mix 1:1:1 Ciprofloxacin : Metronidazole : Minocycline
to a final concentration of 1 mg/ml
Triple antibiotic paste has been associated with tooth discoloration.
Double antibiotic paste without minocycline or substitution of minocycline
for other antibiotic (e.g., clindamycin; amoxicillin; cefaclor) is another
possible alternative as root canal disinfectant.
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• Deliver into canal system via syringe
• If triple antibiotic is used, ensure that it remains
below CEJ (minimize crown staining)
• Seal with 3-4mm of a temporary restorative material such as Cavit™,
IRM™, glassionomer or another temporary material
Dismiss patient for 1-4 week
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• Assess response to initial treatment. If there are signs/symptoms of
persistent infection, consider additional treatment time with
antimicrobial, or alternative antimicrobial
• Anesthesia with 3% Mepivacaine without vasoconstrictor
• Rubber dam isolation
• Copious, gentle irrigation with 20ml of 17% EDTA
• Dry with paper points
SECOND APPOINTMENT
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It reacts with the mineral portion of dentinal walls
Allow better wettability of the irrigator and removal of the smear layer
Conditions dentin to release transforming growth factor b (TGF-b)
Leads to improved MSC attachment and growth
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Create bleeding into canal system by over-instrumenting
(endo file, endo explorer) (induce by rotating a pre-curved K-file
at 2 mm past the apical foramen with the goal of having the entire
canal filled with blood to the level of the cemento–enamel junction)
An alternative to creating of a blood clot is the use of platelet-rich
plasma (PRP), platelet rich fibrin (PRF) or autologous fibrin matrix
(AFM)
Stop bleeding at a level that allows for 3-4 mm of restorative material
Place a resorbable matrix such as CollaPlug™, Collacote™,
CollaTape™ over the blood clot if necessary and white MTA
as capping material
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It brings along SCAP into root canals.
Showed accumulation of transcripts 400-fold to 600-fold greater for the stem cell markers CD73 and CD105,compared with levels found in the systemic blood
Cells will contribute to the regeneration of
pulp tissue after disinfection of the
root canal space
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Platelet concentrates are comprised of increased concentrations of TGF-β1
and are capable of increasing cell proliferation over time when compared
with a blood clot (Lance et al 2010)
PRP platelet count 1 million/mL
The use of PRP was suggested when it is difficult to produce periapical
bleeding by overextending a file
Zhu X, Zhang C, Huang GT, Cheung GS, Dissanayaka WL, Zhu W. Transplantation of dental pulp stem cells and
plateletrich plasma for pulp regeneration. J Endod. 2012;38(12):1604-9
Torabinejad M, Turman M. Revitalization of tooth with necrotic pulp and open apex by using
platelet-rich plasma: a case report. J Endod. 2011;37(2):265-8
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Choukroun's platelet-rich fibrin (PRF) is defined as an
autologous leukocyte- and platelet-rich fibrin (L-PRF)
Classified as a second generation platelet concentrate as it is prepared as
a natural concentrate without the addition of any anticoagulants
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A 3–4 mm layer of glass ionomer (e.g. Fuji IX™, GC America, Alsip, IL)
is flowed gently over the capping material and light-cured for 40 s.
MTA has been associated with discoloration.
Alternatives to MTA (such as bioceramics or tricalcium silicate cements
[e.g., Biodentine®, Septodont, Lancasted, PA, USA,
EndoSequence® BC RRM-Fast Set Putty, Brasseler, USA])
should be considered in teeth where there is an esthetic concern.
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▪ Anterior and Premolar teeth - Consider use of Collatape/Collaplug and
restoring with 3mm of a nonstaining restorative material followed by bonding
a filled composite to the beveled enamel margin
▪ Molar teeth or teeth with PFM crown - Consider use of
Collatape/Collaplug and restoring with 3mm of MTA, followed by RMGI,
composite or alloy
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No pain, soft tissue swelling or sinus tract
(often observed between first and second appointments)
Resolution of apical radiolucency
(often observed 6-12 months after treatment)
Increased width of root walls
(this is generally observed before apparent increase in root length
and often occurs 12-24 months after treatment)
Increased root length.
Positive Pulp vitality test response
Recommended yearly follow-up after the first 2 years
CBCT is highly recommended for initial evaluation and follow-up visits
Follow-up (6-, 12-, 24-months)Clinical and Radiographic exam
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Primary goal:
The elimination of symptoms and the evidence of bony healing
Secondary goal:
Increased root wall thickness and/or increased root length
(desirable, but perhaps not essential)
Tertiary goal:
Positive response to vitality testing
(which if achieved, could indicate a more organized vital pulp tissue)
OUTCOMES
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Diogenes A,Ruparel NB, Shiloah Y, Hargreaves KM. Regenerative Endodontics
JADA 2016:147(5):372-380
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Elimination of symptoms
Evidence of bony healing
{6-12 months after treatment)
Excellent
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Increased root wall thickness
Increased root length
Occurs 12-24 months after treatment
Inconsistent
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Return of a positive response to pulp sensibility testing
after RET of immature permanent teeth with necrotic pulp
was in 50–60% of published cases
Positive response to vitality testing
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Case report of a 10 year old girl where nerve regeneration was identified and tooth regained sensibility after RET
Histologic and immuohistochemical findings revealed that canal space was filled with fibrous connective tissue, cementum like and bone like tissue together with neurons and nerve fibres
Lishan L,Yuemin C,Ronghui Z,Xiaojing Hand Zhiyu C Histologic And Immuohistochemical Findings Of A Human Immature Permanent Tooth With Apical Periodontitis After Regenerative Endodontic Treatment. J Endod 2015; 41: 1172-79
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Placement of MTA
Coronal edge of MTA should be placed 1-2 mm apical to the CEJ
Use of Collaplug or Collatape over the blood clot before placing MTA
51
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DISCOLORATION
Contact of Minocycline with coronal dentinal walls
MTA might be another source for discoloration
PREVENT DISCOLORATION
Modify TAP replacing minocycline with Cefaclor or Clindamycin
Skip Minocycline / Use DAP
Seal dentinal walls of access cavity with dentin bonding agent and composite resin before placement of triple antibiotic paste inside the canal
Instead of using MTA other alternatives
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Long-term pulp necrosis
Well established biofilms
Long-term periapical infection or dental trauma
Damage SCAP or HERS
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Not requiring laceration of the apical papilla (i.e. no evoked bleeding)
No cell isolation
No immunorejection
No ex vivo cell processing steps
Minimizing the disruption between apical papilla and HERS as a key
determining factor that guides root development
Advantages of chemotactic-based cell homing therapies
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Postnatal Stem Cell Therapy
Pulp Implantation
Murray PE, Garcia-Godoy F, Hargreaves KM. Regenerative endodontics: a review of current
status and a call for action. J Endod 2007;33:377–90
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Scaffold Implantation
Injectable Scaffold Delivery
Murray PE, Garcia-Godoy F, Hargreaves KM. Regenerative endodontics: a review of current status
and a call for action. J Endod 2007;33:377–90
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Three-Dimensional Cell Printing
Gene Therapy
Murray PE, Garcia-Godoy F, Hargreaves KM. Regenerative endodontics: a review of
current status and a call for action. J Endod 2007;33:377–90
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JOE , 2016
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PEDIATRIC DENTISTRY V 3 9 / N O 3 MA Y / JUN 1 7
Two-visit revascularization protocol
Radiographic evidence of complete periradicular healing
and positive response to cold test, and remained symptomless thereafter
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Pulp progenitor cells are defined as resting cells
After infection and injury, these cells require activation
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The biological outcome of the pulp regeneration protocol
may not be predictable and it may not be true pulp
dentin complex regeneration ; but it is Hope for the
hopeless non vital young permanent teeth with immature
apex and apical pathosis
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SUGGESTED READING
1. DCNA JULY 2012 Regenerative Endodontics
2. Kim SG, Malek M, Sigurdsson A, Lin LM, Kahler B. Regenerative endodontics: a
comprehensive review. Int Endod J. 2018;51(12):1367‐1388.
3. Diogenes A, Henry Ma, Fabricio B, Teixeira, Hargreaves KM. An update on clinical
regenerative endodontics .Endodontic Topics 2013, 28, 2–23
4. Sonoyama W, Liu Y, Yamaza T, et al. Characterization of the apical papilla and its residing
stem cells from human immature permanent teeth: a pilot study. J Endod 2008; 34: 166–171
5. Huang GT, Sonoyama W, Liu Y, et al. The hidden treasure in apical papilla: the potential role
in pulp/dentin regeneration and bioroot engineering. J Endod 2008; 34: 645–651
6. Torabinejad M, Nosrat A, Verma P, et al. Regenerative endodontic treatment or mineral
trioxide aggregate apical plug in teeth with necrotic pulps and open apices: a systematic
review and meta-analysis. J Endod 2017; 43:1806–1820.
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7. Banchs F and Trope M. Revascularization of immature permanent teeth with apical
periodontitis: new treatment protocol? J Endod 2004; 30: 196–200.
8. Murray PE, Garcia-Godoy F and Hargreaves KM. Regenerative endodontics: a review
of current status and a call for action. J Endod 2007; 33: 377–390.
9. Ruparel NB, Teixeira FB, Ferraz CC, et al. Direct effect of intracanal medicaments on
survival of stem cells of the apical papilla. J Endod 2012; 38: 1372–1375.
10. American Association of Endodontics. American Association of Endodontics
clinical considerations for a regenerative procedure, https://www.aae.org/specialty
/wp-content/uploads/sites/2/2017/06/currentregenerativeendodonticconsiderations.
11. Hargreaves KM, Diogenes A, Teixeira FB. Treatment options: biological basis of
regenerative endodontic procedures. J Endod. 2013;39(3 Suppl):S30‐S43.
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12. do Couto AM, Espaladori MC, Leite APP, Martins CC, de Aguiar MCF, Abreu LG.
A Systematic Review of Pulp Revascularization Using a Triple Antibiotic Paste.
Pediatr Dent 2019;41(5):341-53
13. Albuquerque MTP, Valera MC, Nakashima M, Nör JE, Bottino MC. Tissue-engineering
-based strategies for Regenerative Endodontics. J Dent Res 2014; 93(12):1222-1231
14. Nazzal H, Duggal MS. Regenerative endodontics: a true paradigm shift or a bandwagon
about to be derailed? Eur Arch Paediatr Dent (2017) 18:3–15
15. H Egusa, W Sonoyama, M Nishimura, Atsuta I, Akiyama K. Stem cells in
dentistry – Part I: Stem cell sources. Journal of Prosthodontic Research (2012) ; 56 151–165