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doI 10.11603/IJMMR.2413-6077.2016.2.7028

QUALITATIvE ANd QUANTITATIvE chANgEs IN ThE cELLULAR 
cOmPOsITION Of gUms IN PATIETs WITh PERIOdONTAL dIsEAsEs

N. V. Нasiuk
I. HORBACHEVSKY TERNOPIL STATE MEDICAL UNIVERSITY, TERNOPIL, UKRAINE

Background. The results of a comprehensive study of cytological features of epithelial differentiation 
processes in gums in patients with generalized periodontitis are presented in the article.

Objective. The objects of clinical observations were patients with generalized periodontitis. The disease 
duration was 3–5 years.

Methods. Material for the study was gums epithelium taken from a marginal area of gums in young adults, 
with generalized periodontitis. The epithelium was removed with a spatula, with subsequent transfer to a glass 
plate and drying in the open air for 3–5 minutes. The colouring of the material was done according to Romanovskyy-
Himza, with a next microscopic and morphological analysis, taking into account the percentage of different forms 
of epithelial cells in normal and due to an age aspect. 

Results. One of cytological signs of periodontal pathology in young people is epithelial cells with signs of 
irritation. It should be noted that these cellular elements belong to the intermediate layer of cells that meet 
histological sections and ribbed surface layers are characterized by homogenization and vacuolization of cytoplasm 
as a manifestation of prolonged irritation by bacterial cell aggression. 

Conclusions. Analysis of cytograms proved general periodontitis in the examined patients, in absence of 
adequate therapy as automotive continuous process. its components may regress under the influence of effective 
treatment and re-activate during exacerbation of the inflammatory process under the influence of pathogenic 
factors such as microbial, hygienic and immune.

KEY WORDS: gingiva, epithelium, cells, nucleus, cytoplasm.

Introduction
As a part of the mucous membrane, buccal 

epithelium is active within irritating factors that 
influence it from external and internal environ­
 ment. It allows to use it for studying the phy-
siology and reactivity of mucous membranes, 
and in particular as an indicator of local and 
general disorders of homeostasis [1, 2].

each anatomical area of gums has a zonal 
type of keratinization and on histological 
sections is characterized by basal, ribbed, 
granular and stratum layers; keratinization 
occurs in it due to ortokeratoz — a phased 
process, and as well due to apoptotic changes 
in the upper layer of epithelium [3].

An only exception is gum sulcus epithelium 
that is not keratinized and reduces its barrier 

function and is a target organ at the stage of 
periodontal pathology development [4].

Taken into account the spread of general 
periodontitis among young people [5] and the 
prevailing current points of view about the 
disease pathogenesis [6, 7, 8], the processes of 
differentiation of gums epithelial cells in the 
course of general periodontitis is an urgent 
matter, as well as diagnosis objectification, 
determination of severity degree and invol ve-
ment of adjacent connective tissue to predict 
course of the disease, exacerbation rate and 
relapse [9, 10, 11, 12, 13].

Our study was aimed to determine the 
characteristic features of cellular structure of 
gums cytogram in patients with general perio-
dontitis.

Material and Methods
Material for the study was gums epithelium, 

taken from a marginal area of gums in young 

International Journal of Medicine and Medical Research 
2016, volume 2, Issue 2, p. 26-29
copyright © 2016, TSMU, All Rights Reserved

Corresponding author: Natalya Hasiuk, Department of Thera-
peutic Dentistry, I. Horbachevsky Ternopil State Medical Uni-
versity, 1 Maidan Voli, Ternopil, Ukraine, 46001
Phone number: +380352259802
E-mail: gasyknv@tdmu.edu.ua   

N. V. Нasiuk



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adults, with generalized periodontitis. The 
disease duration was 3–5 years. 

The epithelium was removed with a spatula, 
with subsequent transfer to a glass plate and 
drying in the open air access for 3–5 minutes. 
The colour of the material was done according 
to Romanovskyy-Himza, with a next microscopic 
and morphological analysis, taking into account 
the percentage of different forms of epithelial 
cells in norm and due to an age aspect.

In order to standardize the epithelium layer 
and a deeper studying, in our research we used 
cytological classification; according to it basal, 
parabasal, intermediate, superficial cells were 
in the epithelium of gum in the area of kerati-
nized epithelium of gums, horny scales. 

Statistical analysis of the results was done 
at the Department of Statistical Research of 
I. Horbachevsky Ternopil State Medical Univer-
sity. Parametric methods were used for the 
indi ca tors, their distribution met general requi-
rements. 

To evaluate the nature of distribution the 
coefficients of skewness and kurtosis were 
determined. The test was performed with nor-
mal asymmetry test by Shapiro-Wilkie. Probable 
differences of the results for different groups 
were determined using Student t-test.

The difference was considered likely in 
common in the medical and research error 
probability p<0.05. The probability of error was 
evaluated by the Student t-test tables due to 
the size of the experimental groups. The rate 
of distribution was statistically significantly 
different from the expected normal nonpa ra-
metric criterion (U) Mann-Whitney nonpara-
metric analogue of a Student t-test.

Results
It is established that a major component of 

the cellular composition of the gums’ scraping 
c y t o g ra m  i n  t h e  c o u r s e  o f  g e n e ra l i z e d 
periodontitis are the stratified squamous 

epithelial cells. They are present in cytograms 
under normal conditions and during pathology. 
It should be emphasised that the squamous 
cells are heterogeneous, reflecting the 
heterogeneity of epithelial cells gums’ cover. 
In the case of generalized periodontitis a 
c y t o l o g i c a l  c h a ra c t e r  o f  s c ra p  c h a n g e s 
significantly both in epithelial and connective 
tissue constituents.

Detailed statistical analysis of cytogram, 
made it possible to determine the percentage 
change in gingival epithelial cells of people 
examined due to the different severity of 
generalized periodontitis. 

This ratio is (2,20±0,10) : (9,30±0,16) : 
(41,6±0,36) : (37,0±0,31) : (9,9±0,16) (Table 1). 

Discussion
The qualitative and quantitative cellular 

composition of cytogram in periodontitis 
depends on the clinical course and intensity of 
inflammatory and degenerative processes in 
periodontal tissues. The received data was 
significantly different from the stereotypical 
percentage of epithelial differentiation of 
stratified squamous epithelium of gums [3], 
and ratio that was determined for the examined 
patients and is characterized by violation of 
differentiation: disorder of keratinization as 
evidenced by the presence of all components 
of different epithelial cells. 

This makes it possible to assert that during 
generalised periodontitis the mechanisms of 
epithelial desquamation disruption take place 
causing barrier function of gums at generalized 
periodontitis; that is the outcome of different 
epithelial cells differentiation caused by 
inflammatory infiltrate of lamina propria. 

The presence of parabasal cells in cytograms 
was explained by activation of compensatory 
mechanisms of epithelium, taken into account 
high mitotic activity of gum epithelium and 
increased glycogen-containing epithelial cells.

Table 1. Percentage change in different cells classes of stratified squamous gums epithelium  
in the course of generalized periodontitis

Indicator Cells of smears
Basic Parabasal Intermediate Superficial Horny scales

Norm
(v.l. Bykov)

0 0 60 8 32

Norm for young 
person

0 0 59,20±1,15* 7,60±0,34* 33,2±0,65*

Generalized 
periodontitis

2,20±0,10** 9,30±0,16** 41,60±0,36** 37,0±0,31** 9,90±0,16**

Note: * — p<0,05 compared with the control group; ** — p<0,05 if compared with the experimental group.

N. V. Нasiuk



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At the same time, reflecting the severity, 
course and intensity of general periodontitis 
and intensity of inflammatory and degenerative 
processes in periodontal tissues, cellular 
composition of gums in cytograms was divided 
into four types. The first type is characterized 
by presence of intermediate and parabasal cells 
stained by Romanovskyy-Himza, with smaller 
size compared to intermediate. These epithelial 
layers correspond to deep ribbed cells in 
histological sections. They are larger in size if 
compared to basic cells and have an elongated 
shape. The core is large, well-countered, 
vesicular, with small diffuse located chromatin 
inclusions and relatively wide rim of basophilic 
cytoplasm. The lack of hematogenous cells row 
is noted. It is obvious that this cellular com-
position obtained in cytograms proves evidence 
of proliferation, i.e. reproduction of parabasal 
cells and specific prognostic criterion of gene­
ralized periodontitis of initial stage.

In the second type of cytogram there were 
individual representatives of rod flora with 
advantage of basophilic intermediate (azur 
positive) epithelial cells from cytological smears 
of gums. They have cubic, rectangular or poly-
gonal form; cytoplasm contains azure-positive 
granules. The core is round, sometimes oval. 
The trend towards clusters of cells location of 
this class is noted. Intermediate cells in histo-
logical sections correspond to ribbed parts of 
surface layer. These intermediate cells are found 
in normal and mostly without elements of cyto-
pathology. We argue that this type of cytograms 
proves chronic generalized perio dontitis.

In the third type of cytogram there were 
cytograms of intermediate cells staining by 
Romanovskyy-Himza. They have azure-positive 
cytoplasm nucleus centric located. The cyto-
plasm is elongated. The cells are usually located 
in clusters. The change in the microbial com-
po sition is determined that in this type of 
cytogram is represented mainly by cocci that 
adhere on the surface of epithelial cells.

In addition to the above mentioned epi-
thelial cells in cytogram of the third type of the 
examined people there are segmented leuko-
cytes, which are split into 3–4 segments core 
also there are isolated mononuclear leukocytes 
or young white blood cells it suggests a 
redistribution pool of myeloid cells in response 
to the presence of foci of active inflammatory 
process that causes the process of differentiation 
of leukocytes available. Present cytological 
pattern characterizes pointer of general perio-
dontitis in the examined people.

In the fourth type of cytograms there were 
single basal epithelial cells. During staining by 
Romanovskyy-Himza, if compared with basal 
cells of periodontal pockets they have relatively 
smaller and more rounded shape of cytoplasm. 
The core is round-shaped and has a narrow rim 
of cytoplasm. Basal epithelial cells are sur-
rounded by segmented leukocytes.

The predominant cells in the fourth type of 
cytograms are the cells of intermediate layer, 
their cytological organization corresponds a lot 
to the class affiliation and level of differentiation, 
a pathogenic microflora is evidenced as well as 
the cells of inflammatory response. Superficial 
cells are also verified.

A powerful microbial composition initiates 
further necrobiotic processes in epitheliocytes 
and in segmented leukocytes. Due to phago-
cytosis, the destruction of the cytoplasm of 
segmented leukocytes occur, it is ‘incomplete 
phagocytosis’.

Coccal microflora adheres not only on the 
surface of epithelial cells, but also on the 
surface of segmented leukocytes. Along with 
haematogenous cells in cytograms predo mi-
nantly coccoid flora and single thread of 
pseudomycelia of Candida fungi are evidenced. 

So cytograms of the fourth type present the 
increase of phagocytic reaction of segmented 
leukocytes, and as a result of incomplete 
phagocytosis formation of ‘pus cells’ occurs. 
During phagocytosis of leukocytes specific 
changes in the form of restructuring of its 
nuclear takes place, while in epitheliocytes of 
gums a necrobiotic process occurs primarily in 
the cytoplasm and in the nucleus.

The described type of cytogram corresponds 
to the clinical manifestations of general 
periodontitis with abscess formation.

The emergence of epithelial cells with signs 
of irritation in cytograms is one of cytological 
signs of periodontal pathology in young people. 
These cellular elements belong to the inter-
mediate layer of cells that on histological sec-
tions correspond to ribbed surface layers and 
are characterized by homogenization and 
vacuolization of the cytoplasm as a manifestation 
of prolonged bacterial irritation. The vacuo-
lization of epithelial cells cytoplasm in general 
periodontitis gingival complex proves disorders 
of water and electrolyte and protein metabolism, 
causing the changes in colloid-osmotic pressure 
in the cell. The violation of cell membrane 
permeability, accompanied by their collapse is 
also significant. It leads to lysosomal membranes 
labilization with activation of their hydrolytic 

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enzymes that break intramolecular connection 
with water. In fact, these changes are a mani-
festation of early cell focal necrosis [5].

At the same time the increase in the volume 
of epithelial cells was evidenced, the cytoplasm 
is full of vacuoles, containing a clear liquid. The 
core is moved to the periphery of the cell, 
sometimes are vacuoles or core shrinks. later 
disintegration of ultra-structural elements of 
cells occurs and it overflows with water. These 
changes are the features of the intermediate 
stage of cellular metabolism disorders caused 
by inflammation and preceding cell cyto-
transformation into balloons filled with fluid. 
Such changes characterize balloon cell dege-
neration.

Analysing the cytogram we can reach the 
conclusion that unlike previous smear from 
epithelial gums it has the greater degree of 
destruction accompanied by cariopiknosis and 

kariorexis of nucleus and cytoplasm homo-
genization. Horny scales in cytograms decreased 
in comparison to their quantitative composition 
in cytograms of people with health perio don-
tium. They are mainly eosinophilic, of polygonal 
shape, characterized by no clear contours. 

Conclusions
The changes in the gums in the course of 

generalized periodontitis are subject to rather 
wide limits, which depend on the clinical course 
and exogenous factors. Analysis of cytograms 
gave the opportunity for consideration of 
general periodontitis in the examined patients 
due to the absence of adequate therapy as 
automotive continuous process. Its components 
(inflammatory infiltrative and destructive) may 
decrease after the effective treatment and re-
activate during the exacerbation of inflammatory 
process under the influence of pathogenic 
factors such as microbial, hygienic and immune.

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Received: 2016-12-01

N. V. Нasiuk