ORIGINAL RESEARCH PAPER
EFFECT OF HYDROGEN PEROXIDE CONTAINING EXTRACT OF PINEAPPLE &
PAPAYA AS AN ADDITIVE ON HUMAN ENAMEL DURING VITAL BLEACHING USING
REFLECTANCE SPECTROPHOTOMETER: AN IN VITRO STUDY
Post Graduate student, Department of Conservative Dentistry and Endodontics, School
of Dental Sciences, Sharda University, Greater Noida
Dr. G Ratna
Reader, Department of Conservative Dentistry and Endodontics, School of Dental
Sciences, Sharda University, Greater Noida *Corresponding Author
Professor and Head, Department of Conservative Dentistry and Endodontics, School of
Dental Sciences, Sharda University, Greater Noida
Several corrective measures were introduced in the ﬁeld of esthetic
dentistry to treat discolored teeth. Laminate veneers, direct resin
restorations, crowns or ﬁxed prostheses were included in those
corrective measures. A conservative treatment option i.e. dental
bleaching is useful when restorative procedures are performed to
correct color abnormalities
Tobacco, tea, coffee, red wine, medicines etc. can cause extrinsic
staining of teeth. These substances contains chromogens, (e.g.
polyvalent metal salts in iron supplements and cation antiseptics in
chlorhexidine) that may get attached to the tooth, and creates black or
brown characteristic stains.
Prophylactic procedures are successful in removing the extrinsic
stains; but persistent stains requires bleaching procedure.
One of the most effective tooth bleaching agents is the hydrogen
peroxide, whose application in dentistry was described by Harlan as
early as 1884.
Although hydrogen peroxide provides better outstanding results, the
related clinical side effects remains inevitable. The scientiﬁc evidence
of change in surface texture, composition, and micro hardness of
enamel are reported when bleaching is done with hydrogen peroxide.
The enzymes like Proteases could help degrade the stained ﬁlms as
extrinsic stains are incorporated in pellicle. Papaya is a compatible
bleaching agent due to presence of proteolytic enzymes such as papain
and chymopapain Some in vitro study reported that a dentifrice .
containing papain and Bromelain were helpful in removing stains than
the dentifrice without them.
To evaluate bleaching efﬁciency of solutions containing hydrogen
peroxide and extract of Pineapple & Papaya as an additive on human
enamel in two different timings, using reﬂectance spectrophotometer.
MATERIALS & METHODS:
One hundred twenty maxillary central incisors were collected that
were extracted due to periodontal disease. The defective teeth (i.e. the
one with visible cracks, caries defects, and decalciﬁcations) were
discarded. Then, the teeth were cleaned of calculus and the remaining
soft tissue using an ultrasonic scalar (Satelec, India). They were stored
in 0.2% Thymol, and refrigerated at 4°C until use.
Preparation of pineapple extract
Two hundred grams of pineapple were peeled and cut into small pieces.
The pieces were smashed and blended in a blender with 25 ml of
distilled water. The obtained ﬁltrate was further centrifuged at 2000
rpm for 2 min at a temperature of 4°C. The clear liquid was ﬁltered out
and refrigerated at 4°C.
Preparation of Papaya extract
Unripe papaya was washed with distilled water, peeled, deseeded and
the ﬂesh was blended in a blender. Papaya pulp was squeezed through a
mesh cloth to produce the unripe pulp juice (UPJ). The UPJ was
centrifuged at 2000 RPM at 4°C for 15 minutes. Then the supernatant
was ﬁltered using a sterile 0.22 μm syringe ﬁlter. The sterilized UPJ
was used immediately.
Staining of specimen
Deionized water (100 ml) was boiled and a tea bag was dipped into it.
The solution was cooled to room temperature after 5 min. The strained
solution was used for staining. Staining was achieved by immersing
the teeth for 24 h in this solution. The teeth were properly washed and
stored at 37°C, 100% humidity.
Baseline color value
Baseline color values were established by randomly selecting twenty
stained specimens. Specimens were measured over a white background
using a reﬂectance spectrophotometer (X-Rite Gretag Macbeth). This
was helpful in recording the color variables in accordance with
commission Internationale de l'Eclairage (CIE) laboratory system.
The teeth were randomly divided into three groups of 40 teeth each,
based on the concentration of hydrogen peroxide as follows:
Group 1 (n = 40) – Hydrogen peroxide
Group 2 (n = 40) – Hydrogen peroxide with Pineapple extract
Group 3 (n = 40) – Hydrogen peroxide with Papaya extract
They were further subdivided into two subgroups of 20 teeth each
Subgroup A (n = 20) – 20 min
Subgroup B (n = 20) – 10 min
The crown and roots of all teeth were separated by cutting at
cementoenamel junction. Using acrylic resin, they were mounted
INTERNATIONAL JOURNAL OF SCIENTIFIC RESEARCH
International Journal of Scientiﬁc Research
Volume-9 | Issue-4 | April-2020 | PRINT ISSN No. 2277 - 8179 | DOI : 10.36106/ijsr
Aim: To evaluate bleaching efﬁciency of solutions containing hydrogen peroxide and extract of Pineapple & Papaya as an additive on human
enamel in two different timings, using reﬂectance spectrophotometer
Materials and Methods: Baseline color values of 20 randomly selected artiﬁcially stained incisors were obtained. The specimens were divided
into three groups of 40 teeth each: Group 1 – 30% hydrogen peroxide, Group II – Hydrogen peroxide with Pineapple extract, and Group III
–Hydrogen peroxide with Papaya extract. They were further divided into 2 subgroups, according to time of bleaching, i.e. 20 min (Subgroup A) and
10 min (Subgroup B).
Statistical Analysis: The results were statistically analysed using using Kruskal Wallis test
Results: The mean ΔE values of Group 1A (11.49), Group 2A (12.57), and Group 3A (15.82) showed higher values when compared to the mean Δ E
values of Group 1B (10.74), Group 2B (12.21), and Group 3B (15.60).
Conclusion: Natural enzymes enhances the bleaching effect when used along with hydrogen peroxide.
Bleaching, Bromelain, Papain, Spectrophotometer
Volume-9 | Issue-4 | April-2020
International Journal of Scientiﬁc Research
exposing the labial surface.
The bleaching solution contains 1 ml of Pineapple/ Papaya extract + 1
ml of phosphate buffered solution + 28 ml of hydrogen peroxide. The
teeth in group 1 were bleached with hydrogen peroxide, group 2 were
bleached in combination with pineapple extract, and group 3 in
combination with papaya extract. The specimens were immersed in
solutions for 20 min and 10 min from subgroup A and B, respectively.
The teeth were rinsed and stored in artiﬁcial saliva at 37°C for 24 h
The testing was done with reﬂectance spectrophotometer over a white
background, which recorded color variables L*, a*, b* in accordance
to CIEL*a*b color system using a formula:
∆E = ([∆L*] 2+ [∆a*] 2+ [∆b*] 2)½
The results were statistically analysed using using Kruskal Wallis test.
The results of this in vitro study are given in bar graphs 1 and 2.
Bar graph 1. Depicting mean ΔE in subgroup A for all 3 Groups
Bar graph 2. Depicting mean ΔE in subgroup B for all 3 Groups
The mean ΔE values of Group 1A (11.49), Group 2A (12.57), and
Group 3A (15.82) showed higher values when compared to the mean Δ
E values of Group 1B (10.74), Group 2B (12.21), and Group 3B
(15.60) as shown in Table 1.
Table 1. Shows mean ΔE values of all the groups
Managing discolored dentition is a common procedure in day to day
practice. Teeth discoloration can either be intrinsic or extrinsic. The
management protocol basically depends on the etiology starting from
the most conservative procedure like bleaching to extensive full
Commercially available bleaching agents contain a wide spectrum of
the concentration of hydrogen peroxide or its precursor Carbamide
peroxide. Side effects would include the injuries to the hard and soft
tissues of the oral cavity, surface roughness, postoperative sensitivity
and irritation or burns to the soft tissues in the oral cavity.
The use of enzymes has been proposed as a viable alternative for
In this study the vegetative enzyme extracted from Pineapple &
Papaya were used for bleaching. Pineapple contains Bromelain,
catalase, and polyphenol peroxidase. The pH of extract was 3 to 6.5; in
this pH, the components of the extract are stable. At this pH, the extract
causes disruption of adhered proteins causing stains. Papaya contains
enzyme Papain-sulfhydryl protease consisting of single polypeptide
chain. This enzyme is able to hydrolyse the peptide bonds. It can be
used as a non-abrasive whitening agent.
Colorimeter is an alternative mode to evaluate the change in colors. It
is not able to detect color in the curved surface like tooth structure.
Spectrophotometer demonstrates the high level of accuracy and
rep ro ducibil it y. Th e c ol or ch ange value s obtai ne d f rom t he
spectrophotometer were analyzed on the basis of CIE values.
Within the limitation of this study Pineapple & Papaya extract when
used as a bleaching agent resulted in signiﬁcant colour change on
stained enamel. It can be concluded that natural enzymes along with
hydrogen peroxide are more effective when compared to 30%
hydrogen peroxide alone.
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PRINT ISSN No. 2277 - 8179 | DOI : 10.36106/ijsr
20 MINUTES (A)
10 MINUTES (B)