Cavalcante,et al.

, J Oral Hyg Health 2015, 3:5

Oral Hygiene & Health http://dx.doi.org/10.4172/2332-0702.1000187

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Neural and Vascular Invasions of Oral Squamous Cell Carcinomas

Cavalcante WS1,2*, Hsieh R1 , Lourenço SV1 , Godoy LM2, De Souza LNG2 , Almeida-Coburn KL2 and Barros LAP2
1
University of São Paulo, Brazil
2
Federal University of Espírito Santo, Brazil

Abstract
Objective: The aim of this retrospective study was to identify perineural and vascular invasions in Oral Squamous
Cell Carcinoma lesions.
Methods: A series of 29 OSSC-diagnosed patients and their clinical and demographic data were collected.
In order to identify invasive process, Hematoxiline and Eosin (HE) staining of all cases were analysed and CD31,
CD34, S100 and PGP9.5 protein expression were performed by immunhistochemistry.
Results: All data were statistically assessed by Kappa test and McNemar’s. We identified vascular and
perineural invasions in 17.2% and 31% of cases, respectively, by HE staining analysis. We found intense peritumoral
microvascular density in 82.8% of cases, by CD31 immunostaining. The CD34 antibody was recorded only 6.9% of
cases with vascular invasion. Perineural invasion was detected in 44.8% of S100 protein immunostained cases, and
58.6% of cases were immunostained by PGP9.5 protein.
Conclusion: According to our H/E and immunohistochemistry analysis, regarding the presence of perineural
invasion, PGP9.5 protein was more effective than the others. It was concluded that the identification of vascular
and perineural invasions is a useful tool for the prognosis of patients with oral squamous cell carcinoma, thus
histopathological and immunohistochemistry methods should be combined for an effective analysis for this tumor.

Keywords: Oral squamous cell carcinoma; Neoplastic invasiveness; has been overcome with the use of immunohistochemistry, which
CD31 antigens; CD34 antigens; S100 proteins; PGP9.5 human protein; support the detection of proteins expression at different locations of a
Human tissue. Basically, its mechanism consists in recognition of antigen by an
antibody associated with tissue epitopes [11].

Introduction CD31, CD34, PGP9.5 and S100 biomarkers have been used in
immunohistochemical studies to identify accurately vessels and
Oral cancer is a public health problem, representing the sixth nerves, supporting the presence of perineural and vascular invasion in
most common malignant neoplasm and more than 90% of cases are different diseases, including the OSCC, moreover it results in deeper
Oral Squamous Cell Carcinoma (OSCC) diagnosis. Due to potential comprehension for morphological analysis related to its spread, and it
aggressiveness and propensity for early and extensive lymphatic contributes to better targeting for treatment and prognosis [9,12-15].
metastasis, this cancer type has been widely investigated in recent
decades [1-3]. Therefore, studies seeking to deepen OSCC clinical and
microscopic evaluation, supported by immunohistochemistry analysis,
The OSCC is defined as invasive epithelial neoplasia with varying could contribute to this tumor pathogenesis understanding, because its
degrees of squamous differentiation and propensity to early and evolution has a direct impact on patient’s quality of life, and also they
extensive lymph node metastasis, it occurs mainly in adults ranging can help specialized services in the prevention and diagnosis of oral
from 50 to 70 years and it could be associated to smoking and alcohol cancer.
consumption [1-3]. This carcinoma can develop mainly in the oral and
oropharynx mucosal membranes [1]. When the primary tumor is associated with the invasions into nerves
and vessels, the OSCC shows capacity to metastasize, increasing high
Commonly OSCC diagnosis is done by histological and recurrence rates and aggressive behavior. So perineural and vascular
morphological analysis. Its histopathological features could define invasion are essential for early diagnosis and invasive conditions,
tumor grading, which is relevant for patient’s prognosis. However, because they will influence directly on the prognosis and survival of
not only tumor grade should be a concern during diagnosis, some the patients [1,4,5]. The invasion of a primary malignant tumor and
process, such as connective tissue invasion, need to be a predictive
factor for low survival of patients. Thus, OSCCs lymphovascular and
perineural invasions have been associated with bad prognosis, as well
*Corresponding author: Wanessa Siqueira Cavalcante, Rua José Alexandre Buaiz,
as its aggressive behavior, increased of recurrence rates, low survival 160, Ed. London Office Tower, sala 101, Enseada do Suá, Brazil, Vitória, ES – CEP:
and metastasis [1,4,5]. Vascular invasion is defined by the presence 29050545, Tel: (55) 27 3315-5742; (55) 27 99892-1221; E-mail: wanessasiqueira@
of tumor cells nests in the vascular space, and perineural invasion is gmail.com.
characterized by the presence of tumor cells nests in the perineural Received: August 13, 2015; Accepted: September 03, 2015; Published: September 07, 2015
space [4-9]. Citation: Cavalcante WS, Hsieh R, Lourenço SV, Godoy LM, De Souza LNG, et
al.(2015) Neural and Vascular Invasions of Oral Squamous Cell Carcinomas. J
Histopathological diagnosis of OSCC has been established by the
Oral Hyg Health 3: 187. doi: 10.4172/2332-0702.1000187
HE staining analysis, and it has been considered as gold standard due to
its low cost, easy to perform and useful in view of various components Copyright: © 2015 Cavalcante WS, et al. This is an open-access article distributed
under the terms of the Creative Commons Attribution License, which permits
of tissues [10]. However, this technique offers little information about unrestricted use, distribution, and reproduction in any medium, provided the
the chemical nature of the components of the tissue, thus this limitation original author and source are credited.

J Oral Hyg Health

ISSN: 2332-0702 JOHH, an open access journal Volume 3 • Issue 5 • 1000187
Citation: Cavalcante WS, Hsieh R, Lourenço SV, Godoy LM, De Souza LNG, et al.(2015) Neural and Vascular Invasions of Oral Squamous Cell Carcinomas. J Oral Hyg
Health 3: 187. doi: 10.4172/2332-0702.1000187

Page 2 of 8

its growth in a distant site occurs from the basement membrane vessels phenotype that are being invaded by the tumor.
degradation, migration of tumor cells through connective tissue, spread
The S100 protein family is a group of low molecular weight acidic
through vascular path, colonization and growth of neoplasms in distant
proteins enriched in neural tissue with two Ca2+ bound regions [29]. It
locations [16].
is present mainly in muscle, glands and epithelial tissue [30,31]. Among
The CD31 protein, also known as PECAM-1, is a glycoprotein of its functions that have been proposed in the literature, including the
molecular weight of 130 kDa, present in endothelial cells and platelets, translation of the Ca2+ signalizing producing a variety of intra and
regarded as a molecule of endothelial cell junction [17-20]. The CD31 extracellular biological effects, which is related to the progression of
molecule is found in continuous endothelium of all types of vessels, the cell cycle, cell differentiation, interactions between cytoskeleton
including arteries, arterioles, venules, and non-sine curves capillaries and membrane, muscle contraction and enzymatic activity. Several
veins, making it a useful immunohistochemistry marker for blood disorders, including neoplastic diseases, are related to increased levels
vessels, it also can be expressed in leukocytes (monocytes, neutrophils of calcium ions [30-33]. The technology of genetic hybridization
and subgroups of T lymphocyte, especially TCD8+ cells), due to the showed that four members of this family (S100A4, S100A6, S100A8
aggregation between the circulating blood cells and the vascular and S100A14) may be involved in the development of OSCC, with their
endothelium [17-19,21]. Additionally, physiological protein roles were down-regulated expressions [34]. Despite this information, further
suggested in the inflammatory process and CD31 on angiogenesis studies are needed to better understand the functional roles of the S100
[18,22]. protein in the progression of this type of cancer.
Using polyclonal and monoclonal antibodies for identification of Researchers [9] conducted an important systematic review of
CD31, some authors [23] have demonstrated involvement in mediating 40 cases diagnosed with SCCs of the oral cavity in order to verify
tumor cell adhesion to vascular endothelium, one of the most important that the incidence of perineural invasion would increase by S100
steps that lead to metastasis. immunohistochemistry detection. The average age of patients diagnosed
was 58 years. Originally, the perineural invasion was found in 30% (12
Another study [14] aimed to analyze the role of angiogenic
cases) tumors, however, after reviewing the HE slides, the number of
and non- angiogenic mechanisms in OSCC evaluating microvessel
cases with perineural invasion increased to 62% (25 cases), later it has
density and intravascular tumor invasion using CD31 and CD34
increased to 82% (33 cases) when S100 maker was used. Comparing
immunohistochemical staining protein in 112 specimens. According to
the original slides, all 45 sections with S100 immunostained, no false-
their results CD31 was detected in peritumoral microvascular density of
positive cases were detected, but 21 false-negative cases. Thus, clearly
all samples, ranging
​​ from 11 to 30 (average medium 20.4) microvessels
the perineural invasion detection accuracy by HE method analysis was
per field in 400X magnification. Regarding the intravascular tumor
significantly lower than immunohistochemical method.
invasion, CD31 showed less sensibility than CD34 .
The PGP 9.5 is a protein first detected in human brain, with a
The CD34 is a surface glycolphosphoprotein that is expressed
molecular weight of 27 kDa. This cytoplasmic protein is specific to
in progenitor, endothelial and hematopoietic stem cells of blood
nerves and is widely distributed in the brains of vertebrates and diffuse
capillaries [24-26]. Although several roles have been assigned to this
neuroendocrine system cells in human [35-38]. Immunolabeling
family of proteins, their functions have not been determined yet.
techniques using PGP 9.5 biomarker are able to demonstrate peripheral
Among the functions described in the literature including the increase
nerve fibers with impressive clarity and sensitivity [39], and it is possible
in proliferation and blockage of the stem cells differentiation, and
to obtain information from nerve endings in human oral mucosa from
promotion of lymphocyte adhesion to specialized vascular endothelium
different intraoral sites, such as vestibule, lip, gingiva, palate, tongue,
in lymphoid tissues [27]. Thus, the protein CD34 has been used in
except the sublingual region [40,41]. Thus, the PGP 9.5 was considered
studies that evaluate the relationship between its immunolabeling and
as the best biomarker for nerves and may demonstrate the nerves and
metastasis of OSCC to lymph nodes [16,28] and blood vessels [16].
their processes in all levels of the nervous system [39].
A group of researchers [28] examined the relationship between
Some authors [42] have been using the PGP9.5 marker to evaluate
the CD34 immunostaining patterns and metastasis of OSCC to lymph
the OSCC innervation pattern in 30 patients, and the aim is to find an
nodes in a series of 40 cases. They identified metastatic lymph nodes in
explanation for the absence of pain symptoms in these cancers. Among
18 cases. They found many microvessels with strong remodeling activity
their series only two (6.6%) nerve fibers were stained by PGP9.5,
as well as non-differentiated endothelial cells and immature endothelial
suggesting that no apparent pattern of innervation within the OSCC
cells in keratinocytes nest areas, and in the infiltrated marginal area
tumor tissue. However, PGP9.5 immunostained nerve fibers in adjacent
of cancer,
​​ which seems to be related to metastasis. The lymph node
tissues to carcinoma in 12 (40%) of 30 cases assessed. It was concluded
metastasis was most common (63%) where the invasion pattern was
that the nerve fibers are surrounding tumor areas.
the penetrating type, it also could be seen microvessels invading tumor
stroma. It was concluded that this penetrating microvascular invasion Another group of researchers [14] compared the expression of
pattern correlates with metastasis, suggesting that metastasis is closely the neuronal marker PGP9.5 to S100 protein expression in 16 cases
associated with the distribution of blood vessels surrounding the nest of of malignant nerve sheath tumors by immunohistochemistry, verified
marginal regions of cancer, with the presence of immature endothelial that the majority of cases (95%) showed positive expression of PGP9.5,
cells. although it is not a specific marker for these kinds of tumor, and the
S100 protein has less sensitivity (38%). These results led the authors to
Other authors [16] found that the immunohistochemical method
conclude that the auxiliary PGP9.5 protein is a useful diagnostic tool
with triple staining using the CD34, cytokeratin and podoplanin in
to confirm the neural origin in sarcoma cell when no expression of the
a single histological section, which was more sensitive in detecting
S100 protein.
invasive events (lymphatic and blood vessel neoplastic invasions) than
HE staining technique. This method allows us to study in more detail Therefore, this present study evaluated perineural and vascular
the vascular invasion process, thus providing better understand of the invasions rates in OSCCs by biomarkers staining, and also the

J Oral Hyg Health

Volume 3 • Issue 5 • 1000187
ISSN: 2332-0702 JOHH, an open access journal
Citation: Cavalcante WS, Hsieh R, Lourenço SV, Godoy LM, De Souza LNG, et al.(2015) Neural and Vascular Invasions of Oral Squamous Cell Carcinomas. J Oral Hyg
Health 3: 187. doi: 10.4172/2332-0702.1000187

Page 3 of 8

correlation of their expression with clinical and histopathological Perineural analysis was considered when presence and/or
features. involvement of isogenous keratinocytes groups, i.e. nests of tumor cells,
in the perineural space [5]. The perineural space consists of connective
Materials and Methods tissue surrounding the nerve [43], with full or partial inclusion of the
Tissue samples identified nerve [9].

This study was approved by the Research Ethics Committee at For S100 and PGP9.5 protein expression analysis, the presence
Federal University of Espirito Santo (UFES) under number 138/10. All of neoplastic perineural invasion, the arrangement of the neoplastic
patient’s sample were collected from Surgical Pathology Department, at cells in nest or isolated compromising the nervous structure and the
Programm for Prevention and Early Diagnosis of Oral Cancer, Dental integrity of nerve morphology were evaluated.
School, UFES, Brazil. Twenty nine formalin-fixed, paraffin-embedded All results from the HE staining and immunohistochemical analysis
OSCC tissues were selected according to the criteria as defined in were tested statistically. The statistical package IBM SPSS Statistics
the World Health Organization Histological Classification of Tumors version 20.0 (IBM) was used.
[1]. The clinicopathological data such as age, sex, race, location of
lesion, smoking, alcoholism, histopathological grading, peritumoral Subsequently, an analysis of agreement between HE histological
inflammatory infitration, intensity of the inflammatory infiltrate, technique and immunohistochemistry methods (S100, PGP9.5) were
neoplastic vascular invasion and perineural invasion were collected and performed using the Kappa coefficient, regarding sensitivity and
are detailed in Table 1. Sections of 4µm thickness of the selected OSCCs specificity parameters. The sensitivity is reflected in the likelihood of
were prepared for HE staining. a method to correctly identify the presence of neoplastic perineural
invasion, while the specificity means the likelihood of a method to be
Immunohistochemistry negative to the neoplastic perineural invasion actually not detected.
Four-micrometer sections from specimens were deparaffinized, The comparison of presence of neoplastic perineural invasion
rehydrated. For S100 and PGP9.5 proteins antigen retrieval, slides percentage between HE, S100 and PGP 9.5 staining was performed by
were heated in citrate buffer at 95oC for 45 min, followed by hydrogen nonparametric McNemar’s.
peroxide incubation for 30 min to block endogenous peroxidase
activity, before primary antibody incubation, on the other hand, for Results
CD31 antigen retrieval, slides were heated in Tris-EDTA buffer at 95oC
for 45 min. Finally, CD34 antigen retrieval was performed by trypsin
Histopathological findings
enzyme incubation during 1 hour at 37oC. The sections were incubated The histopathological analysis for OSCCs classification showed,
with anti-CD31, anti-CD34, anti-S100 and anti-PGP9.5, information of in most cases, well differentiated grading (69%). The peritumoral
these antibodies are described in Table 2. inflammatory infiltrate distribution pattern was most prevalent diffuse
(58.6%) and moderate intensity (58.6%) (Table 1 and Figure 1).
REVEAL SPRING system (Spring Bioscience, Pleasanton, CA)
was used to detect the primary antibodies according to manufacture’s According to HE stained sections analysis, it was possible to observe
instructions. The DAB Substrate System solution - DAB Chromogen the presence of neoplastic invasion of vessels by keratinocytes, arranged
(Spring Bioscience, Pleasanton, CA) were used to show the staining in nest form, with no commitment of vascular morphology in 17.2% of
by brownish color. The specimens were counterstained with Carazzi’s cases. The neoplastic perineural invasion was found in 31% of cases,
hematoxylin, dehydrated and mounted with Permount resin and glass and the morphology of the evaluated nerves was fully preserved (Table
cover slips. 1 and Figure 1).
All morphological, histopathological and immunohistochemical Immunohistochemical findings
analysis were performed, simultaneously, by three blinded researchers
in an optical microscope (Nikon Eclipse E200 Multiple Note - Lot: 851 The immunohistochemical results are summarized in Table 3
979) and the results were registered by a digital camera (AxioCamMRc/ and illustrated in Figures 2 and 3. The analysis of the CD31 protein
Zeiss – Lot: 2 04 07 0902). expression showed intense peritumoral microvascular density in 82.8%
of the cases. Neoplastic invasion from blood vessels was found in 6.9%
Analysis of cases immunostained by CD34. The neoplastic perineural invasion
by nests of keratinocytes was present in 44.8% of cases immunostained
The microscopic criteria used for vascular invasion identification by the S100 protein, moreover in 58.6% of cases immunostained by
was defined by the presence tumor cells in the vascular space, lymph or PGP9.5 protein (Table 3, Figures 2 and Figure 3).
blood vessel space, adhered to the vascular endothelium, maintaining
or not architectural integrity of the invaded vessels [9]. The agreement of the S100 protein expression with the HE method
was strong (kappa = 0.713), however PGP9.5 protein expression with
Semi-quantitative analysis of peritumoral microvascular density the HE method was moderate (kappa = 0.482). It means that the S100
was performed by CD31 immunostaining, it has been considered protein obtained more compatible responses with HE technique, while
intense when found the presence of more than 10 microvessels per PGP9.5 resulted in less compatible responses with the mentioned
examined field in 400x magnification, and it was classified as moderate technique (Table 4).
when up to 10 microvessels were observed. CD34 immunostaining
analysis was evaluated by the presence of neoplastic invasion of blood Finally, the McNemar test was carried out between the three
vessels, the arrangement of the nest or neoplastic cells in isolated methods used in the research in order to compare the percentages of
vascular structure, and compromising the integrity of the vascular presence of neoplastic perineural invasion in the studied cases, showing
morphology. that the PGP9.5 expression was identified in more cases (58.6%) than
the HE method (31%) and S100 (44.8%) immunostaining. Although

J Oral Hyg Health

Volume 3 • Issue 5 • 1000187
ISSN: 2332-0702 JOHH, an open access journal
Citation: Cavalcante WS, Hsieh R, Lourenço SV, Godoy LM, De Souza LNG, et al.(2015) Neural and Vascular Invasions of Oral Squamous Cell Carcinomas. J Oral Hyg
Health 3: 187. doi: 10.4172/2332-0702.1000187

Page 4 of 8

N %
0-20 0 0
21-40 1 3.4
Age (years) 41-60 16 55.2
61-80 11 37.9
>80 1 3.4
Male 24 82.8
Sex
Female 5 17.2
White 12 41.4
Black 4 13.8
Brown 12 41.4
Race/color
Yellow 0 0
Indian 0 0
Not Informed 1 3.4
Mouth floor 7 24.1
Tongue 6 20.7
Lip 1 3.4
Hard palate 1 3.4
Location of the Soft palate 2 6.9
lesions Gingiva 0 0
 2332-0702-3-187_JOHH--15-961CLINICOPATHOLOGICAL Buccal mucosa 1 3.4
INFORMATION Alveolar ridge 5 17.2
Retromolar trigone 0 0
Association of two sites 6 20.7
Yes 24 82.8
Smoking
No 5 17.2
Yes 14 48.3
Alcoholism
No 5 51.7
Well differentiated 20 69
Histopathological
Moderately differentiated 9 31
grading
Little differentiated 0 0
Peritumoral Difuse 17 58.6
inflammatory infiltrate Juxtaposed 12 41.4
Light 3 10.3
Intensity of the
Moderate 17 58.6
inflammatory infiltrate
Intense 9 31.1
Neoplastic vascular Present 5 17.2
invasion Non-detected 24 82.8
Neoplastic perineural Present 9 31
invasion Non-detected 20 69
Total 29 100
Table 1: Clinicopathological data of the OSCCs.

the difference between PGP9.5 and HE methods was statistically sometimes resulting thrombus within these vessels. This result is close
significant, the percentage of cases evaluated by the HE technique did to that of another group [44] who found 21.6% of vascular invasion
not differ significantly from those by S100 immunostaining (Table 5). using the same technique. However, other authors [9] presented higher
values to those already mentioned, having been initially identified 30%
Discussion of vascular invasion in cases already diagnosed with OSCCs, which
One of the main aspects for the prognosis of patients with OSCC is number increased to 35% after the revision of the HE stained slides.
With the same technique of histological identification, the presence of
the identification of neoplastic vascular and perineural invasions, since
perineural invasion by nests of keratinocytes was found in 31% of cases,
they are associated with bad prognosis, as well as the increased rates of
which are in agreement with a study that found 31.1% [44], but other
recurrence, aggressive behavior, low survival and metastases [1,4,5]. In
than that it was reported in other works, 52% [45,46] and 62% [9].
this study, the vascular and perineural invasions rates resulting from
HE and immunohistochemical techniques analysis showed that IHC From the analysis of the expression of CD31 protein, there was
was more efficient in detection of perineural invasion, what is the intense density (> 10 vessels per field examined in 400x magnification)
relevant factor to the prognosis of the patients with OSCCs. microvascular peritumoral. It has been reported that the CD31 protein
has a role in the inflammatory process and angiogenesis [18,19], which
HE staining analysis identified 17.2% of neoplastic vascular
is responsible for the nutrition of tumor cells, thereby contributing
invasion by nests of keratinocytes, either in space or lymphatic vessel,
to tumor progression, at the same time the tumor growth stimulates

J Oral Hyg Health

Volume 3 • Issue 5 • 1000187
ISSN: 2332-0702 JOHH, an open access journal
Citation: Cavalcante WS, Hsieh R, Lourenço SV, Godoy LM, De Souza LNG, et al.(2015) Neural and Vascular Invasions of Oral Squamous Cell Carcinomas. J Oral Hyg
Health 3: 187. doi: 10.4172/2332-0702.1000187

Page 5 of 8

Primary serum Clone Manufacturer Antigen retrieval Titer

Dako (Glostrup, Citrate buffer pH 6.0 for 45 min
S100 Z0311 1:10.000
Denmark) at 95oC / Steamer
Dako (Glostrup, Tris-EDTA pH 9.0 for 45 min at
CD31 M0823 1:2000
Denmark) 95oC/ Steamer
(Wetzlar, Enzyme trypsin pH 7.4 for 20
CD34 QNEnd/10 Leica 1:500
Hesse, Germany) min at room temperature
PGP9.5 ab15503 Abcam (Cambridge,England) Citrate buffer pH 6.0 / Steamer 1:2500
Table 2: Specificity of antibodies.

Figure 1: Distribution pattern of the inflammatory infiltrate and oral SCCs

invasion Distribution of the inflammatory infiltrate (A; B) and aspects of the
vascular (C; D) and perineural (E; F) Invasions in HE. Scale bar = 20 µm.

Figure 3: Histopathological and immunohistochemical aspect of well-

differentiated SSCs, Histopathological (A; B) and immunohistochemistry (C-
F) aspects of the well differentiated OSCCs. Scale bar = 20 µm.

the formation of new microvessels [43]. The analysis of CD34

immunostaining showed low spread of OSCCs by blood vessels (6.9%).
We suggest a more detailed investigation of these invasions with D240
and cytokeratin antibodies.
In histopathological diagnosis, attention should be given to peritumoral
fibrosis, which refers to the presence of concentric rings of fibrous tissue,
when associated to nests of tumor cells, may mimic perineural invasion,
making it difficult to distinguish from nerve tissue without the aid
immunomarkers such as S100 and PGP9.5. A cancer that gains the ability
to invade the perineurium finds a path of low resistance of the protected
host defenses, which facilitates the spread of the cancer along the nerve,
reaching various parts of the body [5]. A more sensitive biomarker to
identify the nerves is the PGP 9.5 than the S100, which demonstrates the
nerve structures at all levels of the nervous system [39]. In addition, isolated
Figure 2: Profile expression of vascular and neural biomarker in oral lesions
SCCs profile of the vascular (A-D) and neural (E-G) biomarkers expression
nerve fibers, nerve bundles, around blood vessels, and they innervate
in OSCC lesions. Scale bar = 20 µm. muscles and glands [40]. PGP 9.5 identifies nerve endings organized in
various regions of the mouth [41].
In addition to the above analysis, HE staining still has been

J Oral Hyg Health

Volume 3 • Issue 5 • 1000187
ISSN: 2332-0702 JOHH, an open access journal
Citation: Cavalcante WS, Hsieh R, Lourenço SV, Godoy LM, De Souza LNG, et al.(2015) Neural and Vascular Invasions of Oral Squamous Cell Carcinomas. J Oral Hyg
Health 3: 187. doi: 10.4172/2332-0702.1000187

Page 6 of 8

Biomarkers Evaluated Criteria Variables Values

      (n) (%)

Moderate 5 17.20%
Peritumoral
CD31
microvascular density
Intense 24 82.80%

Present 2 6.90%
Neoplastic blood vessels
invasion
Non-detected 27 93.10%

CD34 Integrate 1 3.40%

Morphology of the vessel Partially destroyed 1 3.40%

Not evaluated 27 93.10%

Present 13 44.80%
Neoplastic perineural
invasion
Non-detected 16 55.20%

Arrangement of the Isolate 0 0.00%

neoplastic perineural cell
  Nest 13 44.80%
S100  
Not evaluated 16 55.20%

Íntegrate 13 44.80%

Morphology of the nerve Partially destroyed 0 0.00%

Not evaluated 16 55.20%

Present 17 58.60%
Neoplastic perineural
invasion
Non-detected 12 41.40%

Isolate 0 0.00%

Arrangement of the
neoplastic perineural cell Nest 17 58.60%
PGP9.5
Not evaluated 12 41.40%

Íntegrate 17 58.60%

Morphology of the nerve Partially destroyed 0 0.00%

Not evaluated 12 41.40%

    Total 29 100.00%

Table 3: Assessment of vascular and perineural invasion of the osccs subjected to immunohistochemistry technique for CD31, CD34, S100 and PGP9.5.

considered as gold standard than immunohistochemical technique. pathological services. PGP 9.5 was able to mark nerve fibers of the oral
However, it was noted that the data obtained from S100 immunostaining cavity of all sizes with stunning clarity, demonstrating that this antibody
was compatible with those obtained from the technique of HE. On the is the best marker to nerves when compared to S100, corroborating with
other hand, the same was not observed with the expression of PGP 9.5, other results found in the literature [14,39,41].
resulting in less compatible responses between the both techniques.
In summary, the record of the presence of perineural and vascular
However, when the presence of neoplastic perineural invasion was invasion during microscopic analysis of OSCCs can be accomplished
compared among the three methods, the PGP 9.5 identified more cases through conventional HE staining, but vascular and neural structures
than the method HE and the S100, detecting approximately twice as immunostaining extends this analysis to prognosis, and they should also
many neoplastic perineural invasion that the technique used routinely in be performed for a more effective regular pathological diagnosis

J Oral Hyg Health

Volume 3 • Issue 5 • 1000187
ISSN: 2332-0702 JOHH, an open access journal
Citation: Cavalcante WS, Hsieh R, Lourenço SV, Godoy LM, De Souza LNG, et al.(2015) Neural and Vascular Invasions of Oral Squamous Cell Carcinomas. J Oral Hyg
Health 3: 187. doi: 10.4172/2332-0702.1000187

Page 7 of 8

Neoplastic Neoplastic
     
Perineural Invasion Perineural Invasion (HE)

    Present Non-Identified Total Sensitivity Especificity Kappa

Present 9 4 13
S100 100.00% 80.00% 0.713*
Non-Identified 0 16 16

Present 9 8 17
PGP9.5 100.00% 60.00% 0.482*
Non-Identified 0 12 12

Total   9 20 29      

* Statistically significant

Table 4: Analysis of agreement between histopathological and immunohistochemistry methods.

12. Ramos-Vara JA (2005) Technical aspects of immunohistochemistry. Vet Pathol

Categories HE S100 PGP9.5
42: 405-426.
Present 17(58,6%)B
9 (31,0%)A 13(44,8%)A,B
13. Ralhan R, Swain RK, Agarwal S, Kaur J, Nath N, et al. (1999) P-glycoprotein
12(41,4%) is positively correlated with p53 in human oral pre-malignant and malignant
Non-Identified 20 (69,0%) lesions and is associated with poor prognosis. Int J Cancer 84: 80-85.
16(55,2%)
14. Hoang MP, Sinkre P, Albores-Saavedra J (2001) Expression of protein gene
Total 29 (100,0%) 29 (100,0%) 29(100,0%) product 9.5 in epithelioid and conventional malignant peripheral nerve sheath
tumors. Arch Pathol Lab Med 125: 1321-1325.
* Percentages followed by different letters differ significantly by
McNemar test, at a significance level of 5% 15. Shieh YS, Lee HS, Shiah SG, Chu YW, Wu CW, et al. (2004) Role of angiogenic
Table 5: Comparison of the rate of invasion among three methods. and non-angiogenic mechanisms in oral squamous cell carcinoma: correlation
with histologic differentiation and tumor progression. J Oral Pathol Med 33:
601-606.
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ISSN: 2332-0702 JOHH, an open access journal
Citation: Cavalcante WS, Hsieh R, Lourenço SV, Godoy LM, De Souza LNG, et al.(2015) Neural and Vascular Invasions of Oral Squamous Cell Carcinomas. J Oral Hyg
Health 3: 187. doi: 10.4172/2332-0702.1000187

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