Solid State Phenomena Submitted: 2021-06-25

ISSN: 1662-9779, Vol. 337, pp 81-86 Revised: 2022-01-30

© 2022 Trans Tech Publications Ltd, Switzerland Accepted: 2022-06-26
Online: 2022-10-14

Carbonate Apatite Crystal Formation in Novel Endodontic Sealer

Elfira Megasari1,a, Hendra Dian Adhita Dharsono1,b,
Myrna Nurlatifah Zakaria4,5,c, Seto Pramudita6,d, Arief Cahyanto2,5,e*
Department of Endodontology and Operative Dentistry, Faculty of Dentistry, Universitas
1

Padjadjaran, Jl. Raya Bandung-Sumedang KM 21, Jatinangor 45363, Indonesia

Department of Dental Materials Science and Technology, Faculty of Dentistry, Universitas
2

Padjadjaran, Jl. Raya Bandung-Sumedang KM 21, Jatinangor 45363, Indonesia

4
Department of Endodontology and Operative Dentistry, Faculty of Dentistry, Universitas Jenderal
Achmad Yani, Jl. Terusan Jenderal Sudirman PO BOX 148, Cimahi, Indonesia
5
Department of Restorative Dentistry, Faculty of Dentistry, University of Malaya, Kuala Lumpur
50603, Federal Territory of Kuala Lumpur, Malaysia
Department of Prosthodontic, Faculty of Dentistry, Universitas Padjadjaran, Jl. Raya Bandung-
6

Sumedang KM 21, Jatinangor 45363, Indonesia

elfira.megasari@unpad.ac.id, , badhita@fkg.unpad.ac.id, cmyrna.nurlatifah@um.edu.my,
a
d
seto.pramudita@unpad.ac.id, e*arief.cahyanto@um.edu.my

Keywords: Carbonate apatite, crystal formation, endodontic sealer, bioceramics

Abstract. Bioceramics play a key role in endodontics, mostly in surgical endodontics and difficult
cases of root canal treatment. Carbonate apatite (CO3Ap) is the natural composition of dental and
bone. The endodontic sealer is used in the obturation phase of endodontic treatment, which requires
sealing ability and bioactivity to promote osseous repair of teeth and alveolar bone. CO3Ap crystal
formation is essential for endodontics sealer as a bioceramic sealer. This study aims to investigate the
crystal characterization of CO3Ap through X-Ray diffraction (XRD) and scanning electron
microscope (SEM). The bioceramics endodontic sealer comprises dicalcium phosphate anhydrous
(DCPA), vaterite, and Ca(OH)2. The powder cement ratio divided into 60% DCPA : 30% vaterite:
10% Ca(OH)2. Powder cement was mixed with 0.2 mol/L Na2HPO4 added by 1% sodium
carboxymethylcellulose and 32μg thymoquinone as an aqueous solution at the liquid to powder ratio
of 0.6 and set at 37°C and 100% of relative humidity for 72h. XRD result showed that all precursor
materials transformed into CO3Ap after 72h treatment. SEM image showed coral-like CO3Ap
morphology that is characteristic of CO3Ap. The initial results of novel CO3Ap endodontic sealers
show that crystal formation occurs and has potential to be used as an endodontic sealer.

Introduction
In particular cases, endodontics, commonly known as root canal treatment involving conventional
treatments (through the crown of the tooth) and surgical intervention, cannot be treated
conventionally. Endodontics is divided into three parts: cleaning and shaping the root canals,
sterilization, and 3-D obturation. To provide a 3-D seal, the root canal filling involves using core
material which is nowadays gutta-percha combined with a root canal sealer. The incidence of voids
in root canal obturation may result in the proliferation of residual bacteria, affecting the structure and
compromising the long-term outcome of root canal treatment. Endodontic sealer plays a critical role
in the obturation phase. It acts as a filler to fill the gap between gutta-percha cones and root canal
dentin surface. Endodontic sealers require sealing ability as ideal root canal sealer properties. As the
root filling material comes into direct contact with the vital tissue at the apical foramen and lateral of
the root through dentinal tubules or indirectly through surface restoration, biocompatibility is also a
crucial requirement of any root canal sealer [1-3].
Only in the last thirty years have bioceramic-based sealers become widely used in endodontics;
this development is directly related to the increased use of bioceramic technology in dentistry,

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82 Solid State Technologies and Modern Production

particularly in relation to dental surgeries. Bioceramic sealer is expected to seal the dentinal tubules
and apical foramen while promoting bone regeneration around the periapical area, especially in cases
of surgical endodontics. There are two major advantages associated with bioceramic materials as root
canal sealers; their biocompatibility in order to adapt to the surrounding tissues. Moreover, Calcium
phosphate is a component of bioceramic materials that improves their setting properties and gives
them a chemical composition and crystalline structure resembling apatite materials found in bone and
tooth. Although the precise process by which bioceramic-based sealers adhere to root dentin is
unknown, the following mechanisms have been proposed: Mechanical interlocking bonds will be
created as a result of the sealer's diffusion into the dentinal tubules. After denaturing the collagen
fibres with a potent alkaline sealer, the mineral content of the sealer diffuses into the intertubular
dentin, creating a mineral infiltration zone. Along the mineral infiltration zone, hydroxyapatite is
formed as a result of the partial reaction of phosphate with calcium hydroxide and calcium silicate
hydrogel, which is produced by the reaction of calcium silicates in the presence of dentin's moisture.
In 2015, research conducted on surface chemistry and topography of materials influenced the
establishment of osseointegration in vivo; while the attachment of cells is the first criteria for success
in vivo, researchers suggest that the presence of CO3Ap is the component necessary to facilitate initial
cell attachment, accelerate cell proliferation based on in vitro cell study. [1, 4-7]
Apatite is one of the main components of a tooth and bone. Calcium phosphate in our body is
performed in the amorphous and crystal phases. Along with amorphous silica and amorphous calcium
phosphate, amorphous calcium carbonate (ACC) is probably the most frequently studied precursor to
crystalline phases in biomineral systems. In nature, particularly in marine organisms like sea urchins,
corals, and crustaceans, CaCO3-based biominerals are used for structural, as seen in coral skeletons
and sea urchin tests, as well as for mechanical and protective purposes. In comparison to their purely
inorganic counterparts, CaCO3-based biominerals have improved properties. For example, calcite
biominerals used for protection are less brittle, and mollusks' nacreous layers of aragonite have been
found to increase the mechanical strength of their shells. The spatially controlled crystallisation of
the biomineral structures gives rise to distinct crystal faces, orientations, and locations, which
necessitates exact control over both the nucleation and growth processes. The characteristics and
capabilities are impressive and can serve as a foundation for the creation of novel synthetic materials
for biomedical, commercial, and technological applications. Examples include bone implants,
nanowires, nanoparticles, crystalline optical materials, and semiconductors, as well as this novel
biomaterial sealer. Structure, organisation, and dynamics are crucial in identifying the functional
responses. The term "amorphous precursor strategy" describes a method by which organisms fill pore
spaces, manage crystallisation, and control the chemistry of the final CaCO3 polymorph by taking
advantage of the flexibility of amorphous phases prior to crystallisation. The composition of natural
waters, their alkalinity, and the solubility of other solid phases are all controlled by the dissolution-
precipitation cycling of CaCO3, which also controls the precipitation of a variety of minerals [4-9].
Carbonate apatite (CO3Ap) constitutes the inorganic components of enamel, dentine, and bone and
has a carbonate concentration of a few percent by weight [10]. The incorporation of carbonate into
apatite structure can influence the structure and the morphology of the apatite and is probably alter
the biological reactivity of the bone mineral, which makes researchs said that CO3Ap is regarded as
better model for bone minerals, it was also stated that CO3Ap thin film coatings are likely to be more
biologically compatible. Studies on fracture resistance of teeth obturated with a combination of gutta-
percha cones as core material and bioceramic root canal sealers have been conducted in recent years.
Increased fracture resistance was observed when using MTA-based bioceramic root canal sealers
compared to conventional root canal sealers, zinc oxide eugenol based in general. The increased
fracture resistance could be due to its chemical bonding through the formation of hydroxyapatite
crystals during the set when the material is in contact with moisture. The researcher designed this
study to observe crystal formation in our novel CO3Ap endodontic sealer [8-11].
 Solid State Phenomena Vol. 337 83

Materials and Methods

Preparation of CO3Ap sealer. The bioceramics endodontics sealer is composed of DCPA (J.T.
Baker Chemical Co., NJ, USA), vaterite (Yabashi Industries Co. Ltd., Ogaki, Gifu, Japan), and
Ca(OH)2 (Merck, Darmstadt, Germany). The powder cement ratio (60% DCPA: 30% vaterite: 10%
Ca(OH)2), the powder was mixed by 0.2mol/L Na2HPO4 added by 1% sodium
carboxymethylcellulose and 32μg thymoquinone (Sigma-Aldrich Inc., St. Louis, USA) as an aqueous
solution at the liquid to powder ratio of 0.6 and set at 37°C and 100% of relative humidity for 72
hours.

Characterization of CO3Ap sealer. The samples were ground into a fine powder and was examined
by X-Ray Diffraction (XRD) (Rigaku SmartLab, Copper cathode, 40kV-30mA, 2theta = 10-60°,
Tokyo, Japan)

Morphological observation. Morphological evaluation on the top surface and the fractured surface
of the sample was evaluated by scanning electron microscope (SEM: JSM-6510A, JEOL Ltd., Tokyo,
Japan) at 10 kV of accelerating voltage after gold sputters coating.

Result and Discussion

Fig. 1 shows XRD of CO3Ap endodontic sealer consisting of 30% Vaterite CaCO3, 60% DCPA,
and 10% Ca(OH)2 mixed with 0.2mol/L Na2HPO4 added by 2% NaCMC and 32µg thymoquinone,
set at 37°C and 100% of relative humidity for 72 h shows that almost all of the precursors have
changed into CO3Ap. CO3Ap was detected by the strong carbonate peaks compared to vaterite and
DCPA peaks. DCPA and CaCO3 vaterite remain in a small amount. There is no evidence of Ca(OH)2
remains, which means in 72 hours, all Ca(OH)2 turned into CO3Ap, possibly also taking a role as a
promotor in CO3Ap nucleation. Ca(OH)2 was initially used as an antibacterial agent for root canal
treatment before obturation. It is known to keep the root canal environment in an alkaline situation.
This characteristic of nucleation shows that heterogeneous nucleation occurs. The distinct crystal
faces, orientations, and locations that result from precisely controlling both the nucleation and growth
processes are what give biominerals their unique structural characteristics. Previous research about
the effect of casein on the nucleation of CO3Ap stated that different substances could affect the
nucleation rate of a solution either as a promotor or inhibitor. Ca(OH)2 as a promotor for crystal
growth is shown by the majority phase of CO3Ap.
84 Solid State Technologies and Modern Production

Figure 1. XRD of CO3Ap endodontic sealer

Fig. 2 shows SEM images of the microstructure surface, and the fractured surface of CO3Ap
endodontic sealer consists of 30% Vaterite, 60% DCPA, and 10% Ca(OH)2 mixed with 0.2mol/L
Na2HPO4 added by 2% NaCMC and 32µg thymoquinone. The SEM micrograph showed
agglomeration of coral-like morphology characteristic of carbonate apatite. Further elaboration of
mean particle size might be needed compared to modification of each precursor to observe which
precursor promotes crystal growth and will appear as an increase in particle size. The mean particle
size of coral-shaped CO3Ap crystal should be around 90-100 nm.[2]
 Solid State Phenomena Vol. 337 85

Figure 2. SEM Images of CO3Ap endodontic sealer (a). before fracture, (b). after fracture.
The sealing ability is related to its solubility and bonding to the dentin and root canal filling cones.
Several studies have evaluated the sealing abilities of different bioceramic-based sealers in vitro.
Bond strength is the force per unit area required to debond the adhesive material from the dentin. The
relationship between leakage and bond strength is indirect, but the "monoblock" concept, in which
the sealer bonds to both the core material and the dentinal wall to form a single unit that improves
sealing and fortifies the root-filled tooth against fracture, has drawn attention to the bond strength
test. This concept increases sealing and increases the tooth's overall resistance to fracture. For the
integrity of the sealer-dentin interface to be preserved during the creation of post-spaces and the
preservation of tooth flexure, a strong bond between root canal sealer and root dentin is required.
Bonds between the dentin and the core filling materials can be formed by bioceramic-based sealers,
which the researcher will investigate further in the future [1].
Result of bond strengths evaluation of several sealers under various moisture conditions in the root
canal from other research present, concluding that a sealer’s bond strength is most excellent in moist
and wet canals, the presence of residual moisture positively affecting the adhesion of the root canal
sealers to root dentin. Bioceramic endodontic sealer (BC sealer) had the highest dislodgment
resistance from the root dentin compared to resin-based endodontic sealer and MTA-based sealer.
The bonding of BC sealer to the root dentin was improved by the prior placement of intracanal
calcium hydroxide dressing as a medication; however, the bonding was less than that of resin sealer
and MTA sealer in the absence of calcium hydroxide. This improvement in bonding might be induced
by the chemical interaction between calcium hydroxide and the BC sealer, increasing the frictional
resistance and micro mechanical retention of the sealer to the root dentine [1]. This CO3Ap endodontic
86 Solid State Technologies and Modern Production

sealer showed a similar BC sealer characteristic, transforming Ca(OH)2 into CO3Ap. Crystal
formation in this CO3Ap endodontic sealer showed promising results as a novel endodontic sealer,
although further studies are needed regarding interaction with the dentinal wall, alkaline phosphatase
reaction, and clinical outcomes associated with the use of this sealer.

Conclusion
The initial results of novel CO3Ap endodontic sealers show that crystal formation occurs and has
the potential to be used as an endodontic sealer. However, further studies of Ca(OH)2 related to the
amount of mass, carbonate concentration, Ca/P molar ratio, and whether it affects the crystal size of
carbonate apatite are still required to clarify the clinical outcomes associated with the use of these
sealers.

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