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VOLUME 1 ISSUE 2, APRIL- JUNE 2013

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INTERNATIONAL JOURNAL OF ORAL HEALTH SCIENCES AND ADVANCES

EVALUATION OF HISTOLOGICAL RESPONSE OF DENTAL PULP TO
BIOGRAFT HA A RESEARCH STUDY
Capt. Tapan Singh* Maj. Gen. Jatinder Pal Singh** Maj. Ronauk Singh*** Dr. Ripin Garewal****
*Command Military Dental Centre, Kolkata **Commandant, Command Military Dental Centre, Kolkata ***Corps Dental Unit,
Siliguri, ****Sr. Lecturer Department of Pedodontics and Preventive Dentistry, National Dental College & Hospital, Punjab.













INTRODUCTION
The pulp in primary teeth has a high potential for
repair because of a high degree of cellularity and
vascularity in this tissue
1
. The rationale for the
pulpotomy procedure is that the radicular pulp tissue
is healthy and capable of healing after amputation of
the coronal pulp. Thus pulpotomy helps to maintain
the primary dentition in an intact state until normal
exfoliation occurs.
There is ample information about pulpotomy in
deciduous molars using formocresol, glutaraldehyde,
electrosurgery, ferric sulphate, calcium hydroxide,
etc. The vital pulpotomy process using formocresol
has been widely accepted in primary tooth pulp
therapy because of its simplicity and good prognosis
2
.
However, much concern has arisen over the
mutagenic and carcinogenic potential of
formaldehyde containing products, the cytotoxic
effects of formocresol and the possible diffusion into

the surrounding and systemic tissues
3
. In order to
avoid the possible harmful effects of formocresol and
other pulpotomy agents, an ideal agent for vital
pulpotomy procedure is being sought. Synthetic
hydroxyapatite (HA) is promising biomaterials
specifically in the orthopaedic and dental fields, as
their chemical composition is similar to that of bone.
Due to the need for safer bone graft applications,
these bone graft substitutes are gradually gaining
increased acceptability. In our endeavour to find an
ideal pulpotomy agent, the use of hydroxyapatite
crystals was assessed for its regenerative potential.
Hydroxyapatite has been shown to be an extremely
biocompatible material for soft tissues and bone
4
. It
has been reported to be effective in alveolar ridge
augmentation
4
, healing of periodontal bone defects
5
,
osseointegration of titanium implants
6
and direct pulp
capping
7
. Hydroxyapatite, which is the main
constituent of dental hard tissues, may immediately
ABSTRACT
Purpose: To evaluate Histological assessment of dental pulp response to BioGraft HA
Method: fifteen deciduous molars were treated with pulpotomy using BioGraft HA.
Results: histological evaluation was observed at the end of 4 weeks. On light microscopic observations, minimal
inflammatory reaction, hyperemia and formation of dentinal bridge was observed. None of the samples showed Pulpal
necrosis
Clinical significance: The results of this study revealed that BioGraft HA is a potential pulpotomy agent for
deciduous molars.

KEYWORDS: Pulpotomy, Bioceramics, Deciduous molars, Pulpal response, Formocresol, Hydroxyapatite.







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INTERNATIONAL JOURNAL OF ORAL HEALTH SCIENCES AND ADVANCES

provide an artificial barrier. Despite the putative
abilities of hydroxyapatite to be osteoconductive,
osteogenic and dentinogenic
7
, little research has been
done with this material as a pulp healing agent. The
present study was undertaken to evaluate
histologically the response of pulp to hydroxyapatite
crystals (BioGraft HA).
MATERIALS AND METHOD
This study was carried out on fifteen patients, with
deep carious primary mandibular teeth indicated for
pulpotomy deemed for serial extraction. All
experimental procedures were carried out according
to the protocol approved by the Ethical Committee.
The criteria for tooth selection in this study were:
1- Primary dentition with vital carious pulp exposures
that bleed upon entering the pulp chambers.
2- No clinical symptoms or evidence of pulp
degeneration, such as pain on percussion, history of
swelling, or sinus tracts.
3- No radiographic signs of internal or external
resorption and no periapical or furcation
radiolucency.
4- Teeth would be restorable with posterior stainless
steel crowns.
Following profound local anaesthesia and rubber dam
application, a sterile No 1 round high-speed bur was
used to remove the roof of the coronal pulp chamber.
The coronal pulp tissue was amputated using a sterile
sharp spoon excavator. Pulpal hemorrhage was
controlled by moist cotton pellet. Amputated pulpal
stumps were covered with BioGraft HA

. In all
treated teeth, Cavit (3M ESPE, Germany) base
material was placed over the treated pulps, followed
by a layer of light cured glass ionomer cement (GC
Gold Label

). Teeth were extracted after 4 weeks to

assess the response of the pulp. After extraction, teeth
were fixed in 4% neutral buffered formaline, and then
apical foramina were occluded with composite.
Demineralization was performed in 5% trichloro-
aceticacid for 10 days. Following decalcification, the
samples were processed for embedding in paraffin.
Longitudinal serial sections of (5m) were prepared,
processed and stained with Haematoxylin & Eosin
stain. The specimens were examined under light
microscope to assess the histological response of the
treated pulpal tissue. Each specimen was observed
for pulp vitality, pulp inflammation, dentin bridge
formation and calcific deposits. Series of sections
containing pulp tissue were observed under light
microscope equipped with a digital camera (Olympus
Micro-Image, Maryland, and U.S.A.)
RESULTS
Based on light microscopic observations, minimal
inflammatory reaction was seen in 60 Percent of
samples. Hyperemia was observed in 53.3 percent of
BioGraft HA

samples, Pulpal necrosis was

observed in none of the samples. The formation of
dentinal bridge was observed in 80 percent of
BioGraft HA

. (Figure 1, 2). Few blood vessels

were also appreciated in 72.3% of sample.
DISCUSSION
Potential risk of viral hepatitis, acquired immune
deficiency syndrome in the process to harvest
autograft and allograft, forced us to consider using a
substitute, thereby to avoid the use of autogenous
graft also. Bioceramics are nowadays used in
numerous medical and/or dental applications
including in repairing bone fractures, in attaching
bone plates and other prostheses, in bridging
communicated fractures, and in filling or aligning
dental cavities. Result of this study showed mild to
moderate inflammatory changes this may be an
expected protective response from pulp to any
foreign materials and an alternative reason can also






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be because the sample collected were aged near there
exfoliation of primary tooth. Inflammatory responses
in the pulp tissue caused by HA can be attributed to
particle size and the shape of the particles in this
study
8
. Irregularly shaped, sharp-edged particles have
been shown to prompt more inflammatory response
than the rounded particles of the same size
8
. In the
present study, hydroxyapatite with particle size
ranging from 0.4-0.9 mm (400-900 m) was used. In
addition, little particle size of HA material has been
shown to cause severe inflammatory response
compared with calcium hydroxide in the pulp tissue
9
.
However, Jaber et al
10
.Have reported that granular
free-flowing resorbable form of the HA, which allow
for better adaptation to the dental pulp, caused
scattered dystrophic calcification in the dental pulp.
Jaber et al.
10
stated that intense and scattered
inflammatory reaction of HA-treated pulps of rat
teeth might be due to the pH of the HA in their study.
In our study, although HA- treated cases showed
inflammatory response, we did not relate this with the
alkalinity of the HA material. The pH of the HA
material we used was 12.1 which is very close to
calcium hydroxide. It is an interesting observation
however the alkalinity of HA used in the study did
not induce a focal necrosis when it contact with the
pulp tissue. Higashi and Okamato
11
have reported that
they were observed osteodentin-like tissue
organization around the HA. Added advantage of HA
was reported by Tin-Oo MM
12
in his study that HA
has evinced antibacterial activity, against S. mutans a
cariogenic organism, by HA, it validates its use as a
potential candidate for dental applications such as
pulp capping or as a base liner material.
In conclusion, we showed in this study that pure HA
materials did allow the pulp tissue to regenerate
dentine
ACKNOWLEDGMENTS
I would like to thank my teacher and guide Dr
Suleman Abbas for his support. The above study is
self funded; no consultants' fees or other financial
arrangements were paid to investigators. All co-
authors confirm that this study is free from conflict of
interest.
BIBLIOGRAPHY
1. Camp JH. Pediatric endodontic treatment.
In: Cohen S, Burns RC (Eds): Pathways of
the Pulp, ed. 7. St. Louis, CV Mosby, 1998.
2. Rivera N, Reyes E, Mazzaoui S, Moron A.
Pulpal therapy for primary teeth:
Formocresol vs electrosurgery: A clinical
study. J Dent Child (Chic) 2003 Jan-
Apr;70(1):71-3.
3. Hill SD, Berry CW, Seale NS, Kaga M.
Comparison of antimicrobial and cytotoxic
effects of glutaraldehyde and formocresol.
Oral Surg Oral Med Oral Pathol 1991
Jan;71(1): 89-95.
4. Frame JW. Hydroxyapatite as a biomaterial
for alveolar ridge augmentation. Int J Oral
Maxillofac Surg. 1987 Dec;16(6): 642-55.
5. De Lange GL. Repair of periodontal bone
defect with hydroxylapatite implants. Int J
Oral Implantol. 1990;7(1):54-8.
6. De Lange GL, Donath K. Interface between
bone tissue and implants of solid
hydroxyapatite or hydroxyapatite coated
titanium implants. Biomaterials 1989
Mar;10(2): 121-5.
7. Jaber L, Mascres C, Donohue WB. Reaction
of the dental pulp to hydroxyapatite. Oral
Surg Oral Med Oral Pathol 1992
Jan;73(1):92-8.






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8. Ogilvie A, Frank RM, Benqu EP, Gineste
M, Heughebaert M, Hemmerle J. The
biocompatibility of hydroxyapatite
implanted in the human periodontium. J
Periodontal Res. 1987 Jul;22(4):270-83.
9. Subay R.K., A S. Human pulpal response
to hydroxyapatite and a calcium hydroxide
material as direct capping agents. Oral Surg
Oral Med Oral Pathol. 1993 Oct;76(4):485-
92.
10. Jaber L, Mascrs C, Donohue WB.Reaction
of the dental pulp to hydroxyapatite. Oral
Surg Oral Med Oral Pathol. 1992
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11. Higashi T., Okamato H. Characteristics and
effects of calcified degenerative zones on the
formation of hard tissue barriers in
amputated canine dental pulp. J Endod.
1996 Apr;22(4):168-72.
12. Tin-Oo MM, Gopalakrishnan V, Samsuddin
AR, Al Salihi KA, Shamsuria O.
Antibacterial property of locally produced
hydroxyapatite. Archives of Orofacial
Sciences (2007) 2, 41-44















Figure1-A= blood vessels
B= small spicule of dentine
C= soft tissue showing inflammatory cells

Figure 2-Soft tissue showing inflammatory cells
and Blood vessel

ADDRESS FOR CORRESPONDENCE:
Capt. Tapan Singh
Command Military Dental Centre, Kolkata
Cell: +91-8697212213
Email: tapandr@in.com

SOURCE OF SUPPORT-NIL
CONFLICT OF INTEREST NONE DECLARED
LEGENDS

A
B
C