Mast cells are regarded as complex and multifunctional cells, playing a significant role in immunopathology and a substantial role in tumor angiogenesis. Angiogenesis is a complex process that is tightly regulated by various growth factors in which mast cells act directly by releasing angiogenic factors and henceforth promoting tumor growth and metastasis. The aim of this study is to evaluate the number of mast cells in tissue sections of oral squamous cell carcinoma (OSCC) in comparison with normal mucosa. A total of 40 cases (20 OSCC and 20 normal mucosa) were stained with 1% toluidine blue and the quantitative analysis was done by using light microscope under 400x magnification. A significant increase in the mast cell count was observed in the sections of OSCC when compared to normal mucosa suggesting their contributing role in tumor growth and progression

A Anuradha

B Kiran

G Vijay Srinivas

H K Puneet

Kumar Naik

Ramisetty Sabitha Devi

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Research Article
Incidence of Mast Cells in Oral Squamous
Cell Carcinoma: A Short Study
A. Anuradha,1,2 B. Kiran Kumar Naik,1 G. Vijay Srinivas,1
Ramisetty Sabitha Devi,1 and H. K. Puneet1
1 Department of Oral Pathology, St Joseph Dental College & Hospital, Duggirala, Eluru 534003, India
2 Anuradha ENT Hospital, D/No-10/200, Eluru Road, Gudivada, Krishna District, Andhra Pradesh 521301, India
Correspondence should be addressed to A. Anuradha; anuradhaundavalli@yahoo.com
Received 12 September 2013; Revised 15 November 2013; Accepted 17 November 2013; Published 23 January 2014
Academic Editor: James L. Mulshine
Copyright © 2014 A. Anuradha et al. This is an open access article distributed under the Creative Commons Attribution License,
which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Mast cells are regarded as complex and multifunctional cells, playing a significant role in immunopathology and a substantial role
in tumor angiogenesis. Angiogenesis is a complex process that is tightly regulated by various growth factors in which mast cells act
directly by releasing angiogenic factors and henceforth promoting tumor growth andmetastasis.The aim of this study is to evaluate
the number of mast cells in tissue sections of oral squamous cell carcinoma (OSCC) in comparison with normal mucosa. A total
of 40 cases (20 OSCC and 20 normal mucosa) were stained with 1% toluidine blue and the quantitative analysis was done by using
light microscope under 400x magnification. A significant increase in the mast cell count was observed in the sections of OSCC
when compared to normal mucosa suggesting their contributing role in tumor growth and progression.
1. Introduction
Mast cells are normally present in small numbers in the
connective tissue of all organs and more particularly (around
blood vessels and nerves) in the dermal layer of skin, ranging
from 5 to 15 𝜇m in diameter, and in histologic sections often
appear ovoid, tadpole, or spindle shaped cells with cytoplas-
mic granules of 0.2 to 0.5 cm in size.They exert their influence
locally and systemically by releasing a variety of potent medi-
ators like histamine, leukotrienes, and cytokines through
degranulation and cause neovascularization by producing
angiogenic mediators such as fibroblast growth factor (FGF),
transforming growth factor-𝛽 (TGF), tumor necrosis factor-
𝛼 (TNF), and vascular endothelial growth factor (VEGF).
They occur in various pathological states and also in some
benign and malignant tumors. An attempt has been made
to quantitatively estimate the number of mast cells in OSCC
and to signify their role in tumor growth and progression
[1–3].
2. Materials and Methods
20 paraffin embedded specimens ofOSCCof age groups from
30–80 years with no systemic illness but having the habit
of smoking and alcohol consumption for a period of 5–10
years were retrieved from the archives of the department
of oral pathology, among which seven cases were of well
differentiated squamous cell carcinoma (𝑛 = 7; females = 4,
males = 3) and 13 were moderately differentiated squamous
cell carcinoma (𝑛 = 13; 4 = females, 9 = males). 20
normal oral mucosal biopsies from age groups of 15–20
years with no systemic illness were obtained from 20 adult
patients undergoing extraction for orthodontic treatment.
Serial sections of 5 𝜇m thickness were made from paraffin
embedded tissue blocks using semiautomatic microtome
(Thermo Scientific Microm HM340E). All the sections were
stained with 1% toluidine blue as per Churukian and Schenk
method [4].
Hindawi Publishing Corporation
Journal of Oncology
Volume 2014, Article ID 614291, 3 pages
http://dx.doi.org/10.1155/2014/614291
2 Journal of Oncology
Table 1: Comparison of the average mast cell count in between
OSCC and control group using ANOVA test.
Group Sample size (𝑛) Average number ofmast cells/slide 𝑃 value
OSCC 20 135.90 0.000
Controls 20 42.85
Table 2: Comparison of the average mast cell count within the
OSCC group using ANOVA test.
Group Sample size (𝑛) Average number ofmast cells/slide 𝑃 value
Well dif 7 77.86 0.058
Mod diff 13 167.15
Mast cells were counted under light microscope at a
magnification of 400x in a 𝑍-pattern from left to right and
the data obtained was statistically analyzed using ANOVA.
(Figures 1 and 2).
3. Criteria to Identify the Mast Cells
Mast cell granules are purplish red and the nuclei ofmast cells
appear sky blue in color. Toluidine blue stains the mast cell
granulesmetachromatically due to its reactionwith sulphated
mucopolysaccharides [1].
4. Result
A highly significant (𝑃 = 0.00) threefold increase in the
average number of mast cells/slide was observed in OSCC
when compared to controls (Table 1, Figure 3). No statistical
difference (𝑃 = 0.058) was found in the average number
of mast cells/slide when moderately and well-differentiated
OSCC were compared. (Table 2, Figure 4). Average count of
mast cells/microscopic field under 400x magnifications was
found to be 3 in OSCC and 2 in controls.
5. Discussion
Paul Ehrlich in 1877 discovered a granular cell of loose
connective tissue and named it as “Mastzellen”—a well fed
cell. Studies on mast cells in normal and various pathologic
conditions have shown them to be complex, well-engineered,
andmultifunctional cells playing an essential role in acquired
and innate immunity. They take origin from multipotent
CD 34+ precursor in the bone marrow, later circulate in
the peripheral blood as agranular monocytic cell, and then
migrate into tissues, assuming their typical granular mor-
phology from their immature state. They are normally dis-
tributed throughout the connective tissue, adjacent to blood
or lymphatic vessels, and near or within peripheral nerves.
They are numerous especially beneath the epithelial surfaces
of the skin, in the respiratory system, gastrointestinal and
genitourinary tracts. Many of the mediators are stored within
cytoplasmic granules and these include preformedmediators
Figure 1: Mast cells in controls (40x).
Figure 2: Mast cells in oral squamous cell carcinoma (40x).
like histamine, heparin, and tryptase; lipid derived mediators
like leukotriene’s B4 (LTB4), LTC4, LTD4, and LTE4; proin-
flammatory cytokines like TNF-𝛼, IL-1; mitogenic cytokines
like IL-3, IL-5; and immunomodulatory cytokines like IL-4,
IL-10, and serotonin and other mediators are produced at the
time of mast cell stimulation such as IL-1 [5, 6].
Mast cells have been considered the tissue equivalent of
the circulating basophils though they arise from a common
precursor cell in the bone marrow. There is no evidence that
mature basophils are able to differentiate into mast cells. The
two cell types are readily distinguished by their morphol-
ogy on light microscopy and the presence of chloroacetate
esterase activity in mast cells. They have been studied in
normal gingiva, chronic inflammatory gingivitis, desqua-
mative gingivitis, lichen planus, oral submucous fibrosis
(OSMF), and OSCC. They exhibit phenotypic plasticity and
variation in the mast cell mediators with the change in the
microenvironment. Volumes of literature speak about their
role in the inflammatory reactions and neovascularization. In
somemalignancies, large numbers ofmast cells were detected
before the occurrence of neovascularization [2, 7].
In carcinogenesis, angiogenic regulation is biphasic. In
the premalignant early phase of hyperplasia and dysplasia,
infiltratingmast cells degranulate and activate dermal fibrob-
lasts which intensify angiogenesis. They even activate proge-
latinase B (amember of thematrixmetalloproteinase (MMP)
family) which is involved in both extracellular remodeling [8]
and regulation of angiogenesis [9].Themast cells activate and
progressively intensify angiogenesis by releasing sequestered
Journal of Oncology 3
Co
nt
ro
l
0
100
200
300
400
1 3 5 7 9 11 13 15 17 19Control
Well differentiated
Figure 3: Bar graph showing increase of mast cell count in OSCC
when compared to controls.
W
ell
0
100
200
300
400
1 3 5 7 9 11 13 15 17 19
Well differentiated
Moderately differentiated
di
ffe
re
nt
iat
ed
Figure 4: Bar graph showing mast cell count between well and
moderately differentiated OSCC.
angiogenic activators. As neoplastic progression proceeds,
angiogenic growth factor gene expression is upregulated in
the cancer cells, marking progression to the second cancer
phase, wherein the tumor cells control their angiogenic
phenotype directly instead of depending on themanipulation
of inflammatory cells to indirectly affect neovascularization
[10–14]. This implies that mast cells have significant role in
the early stages of cancer progression and increase in mast
cells is observed in the initial stages whereas, in second cancer
phase, the mast cells decrease because the tumor cells are not
dependent on them anymore for the neovascularization effect
and this could be the reason for the increase in the mast cells
in moderate when compared to well-differentiated OSCC in
our study and also might be due to interobserver mystifica-
tion in classifying thewell- andmoderate differentiated forms
of OSCC when compared to poorly differentiated.
Though many studies have proven the presence and
significance of mast cells, their mediated angiogenesis is
complex and not completely understood. In harmony with
the literature, our study also showed a high mast cell count in
OSCC than in control tissue indicating their supportive role
in tumor progression and metastasis.
6. Conclusion
This study reveals that there is a definitive increase in mast
cells count when compared to normal mucosa substantiating
their contributing role in tumor progression.
Conflict of Interests
The authors declare that they have no conflict of interests
regarding the publication of this paper.
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Incidence of Mast Cells in Oral Squamous Cell Carcinoma: A Short Study

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Incidence of Mast Cells in Oral Squamous Cell Carcinoma: A Short Study

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