Cytokine 61 (2013) 892–897

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Cytokine
journal homepage: www.journals.elsevier.com/cytokine

Role of monocyte chemoattractant protein-1 (MCP-1) as an

immune-diagnostic biomarker in the pathogenesis of chronic periodontal disease
Mili Gupta a, Rashi Chaturvedi b,⇑, Ashish Jain b
a
Department of Biochemistry, Dr. Harvansh Singh Judge, Institute of Dental Sciences & Hospital, Panjab University, Chandigarh, India
b
Department of Periodontics, Dr. Harvansh Singh Judge, Institute of Dental Sciences & Hospital, Panjab University, Chandigarh, India

a r t i c l e i n f o a b s t r a c t

Article history: Objective: Monocyte chemoattractant protein-1 (MCP-1) is an important chemokine responsible for the
Received 30 August 2012 initiation, regulation and mobilization of monocytes to the active sites of severe periodontal inflamma-
Received in revised form 6 December 2012 tion. The present study aims at evaluating the levels of MCP-1 in GCF, saliva and serum and to analyze
Accepted 19 December 2012
the changes following phase I periodontal therapy. Assessment of possible correlations between levels
Available online 30 January 2013
of MCP-1 in the three biological fluids was also done.
Methods: Fifteen healthy and 30 patients of severe chronic periodontitis (diseased) participated in the
Keywords:
study. Patients of the diseased group underwent scaling/root planing. Evaluation of PI, GI, PD, CAL and
MCP-1
Chronic periodontitis
collection of samples of GCF, serum and saliva was done at baseline and 6 weeks following periodontal
GCF therapy. MCP-1 levels were quantified in all samples using ELISA.
Saliva Results: Compared to healthy controls, MCP-1 levels were statistically significantly higher in GCF
Serum (p < 0.001), saliva (p = 0.002) and serum (p < 0.001) in subjects with chronic periodontitis. Levels of
MCP-1 in all the three fluids decreased significantly in patients after periodontal therapy (p < 0.001).
There was a significant positive correlation between MCP-1 levels in GCF, saliva and serum in patients
of chronic periodontitis both pre (r > 0.9) and post-treatment (r > 0.6).
Conclusions: The results suggest that levels of MCP-1 in GCF and saliva can be reliable indicators of sever-
ity of periodontal destruction and their serum levels reflect the systemic impact of this local inflamma-
tory disease thereby strengthening the reciprocal oro-systemic association.
Ó 2012 Elsevier Ltd. All rights reserved.

1. Introduction cytokines. It has been postulated that ‘appropriate’ cytokine

Chronic periodontitis is primarily a disease of bacterial etiology,
characterized by the destruction of the supporting periodontal
tissues and alveolar bone. However, the clinical outcome of the
disease is not equivalent in all patients; implicating intrinsic differ-
ences in the host immune responses [1]. The oral cavity provides
an ecological niche to a plethora of micro-organisms which, while
residing as a host associated biofilm serve as an entry portal for a
wide array of antigenic challenges. Components of the biofilm acti-
vate the host defenses and trigger the inflammatory response by
emigration of neutrophils, monocytes and macrophages leading
to progression of periodontal destruction [2]. These cells along
with the local resident cells such as fibroblasts and vascular
endothelial cells, on stimulation synthesize and secrete a broad
spectrum of inflammatory and immune mediator molecules called
⇑ Corresponding author. Address: House No. 1036, Second Floor, Sector-40-B,
Chandigarh, India. Tel.: +91 9815306699.
E-mail addresses: miligupta8@yahoo.com (M. Gupta), rashichaturvedi@yahoo.
co.in (R. Chaturvedi), ashish@justice.com (A. Jain).
production results in protective immunity, while ‘inappropriate’
cytokine production leads to tissue destruction and disease pro-
gression [3]. The appearance of specific type of inflammatory infil-
trate has been found to be propagated through cell specific
chemotactic cytokines called chemokines [4,5].
Chemokines are inducible, pro-inflammatory cytokines which
exert their effects by binding to specific receptors on target cells.
Engagement of chemokine receptors with their respective ligands
affects leukocyte migration by regulation of cyto–skeletal rear-
rangement, integrin–dependant adhesion as well as by the binding
and detachment of cells from their substrates [6,7]. This recruit-
ment of the inflammatory cells to the actual arena of host-bacterial
interactions facilitates the progression of destruction to the deeper
tissue planes. Chemokines are broadly divided into four sub-fami-
lies according to their structure and spacing of cysteine residues
namely CXC, CC, C and CX3C. The CXC chemokines enable recruit-
ment of neutrophils and predominantly include IL-8, whereas the
CC chemokines are critical to the migration of monocytes and T
lymphocytes. Monocyte chemo-attractant protein-1 (MCP-1), also
known as CC Chemokine Ligand 2 (CCL-2) is one of the most potent

1043-4666/$ - see front matter Ó 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.cyto.2012.12.012
 M. Gupta et al. / Cytokine 61 (2013) 892–897
chemoattractant for monocytes [8]. It is produced by a variety of
cell types in response to different signals such as tumour necrosis
factor-alpha (TNF-a), interleukin 1-beta (IL-1b) and interferon-
gamma (IFN-c) [9]. Marked expression of MCP-1 gene has been ob-
served in gingival tissue of adult periodontitis patients, suggesting
its predominant role in monocyte chemotactic activity in the gin-
gival crevicular fluid (GCF) [10–12].
GCF is both a physiological fluid as well as an inflammatory
exudate originating from the plexus of blood vessels in the gingival
corium. The analysis of specific constituents in the GCF has been
known to be accurate predictor of local cellular metabolism that
reflects the patient’s periodontal health status [13]. Emingil et al.
showed statistically significantly higher levels of this marker in
the GCF of patients of generalized aggressive periodontitis [14].
Kurtis et al. in a study showed that the levels MCP-1 in the crevic-
ular fluid was higher in both patients of chronic as well as aggres-
sive periodontitis vis a vis healthy controls, however, no statistical
difference was found between the two types of periodontitis [15].
Pradeep et al. have further reported increase in the levels of MCP-1
in GCF with progression of disease in patients of chronic periodon-
titis [16].
The oral cavity with its multitude of resident microorganisms
and disease associated inflammatory markers is not a localized
closed environment. Bacterial systemic exposure and leakage of
inflammatory markers of periodontal inflammation into the serum
have the propensity to affect various organ systems with serious
implications. Numerous such epidemiological associations be-
tween chronic periodontitis and cardiovascular diseases, stroke,
pulmonary diseases and diabetes have been reported, however,
periodontal disease is still not established as a risk factor for these
inter-relationships [17–19]. Heightened MCP-1 expression has
been observed in several chronic inflammatory diseases such as
atherosclerosis, osteoarthritis, rheumatoid arthritis, tumors and
delayed type hypersensitivity reactions [20–24]. Thus, any increase
in plasma levels of MCP-1 in patients with chronic periodontitis
who are otherwise clinically healthy may be hypothesized to be
a risk indicator for systemic illnesses. Till date, only one study
has assessed serum levels of MCP-1 in patients of chronic peri-
odontitis and found these levels to be significantly elevated. [25]
Thus, research is lacking in this field and further evaluations of ser-
um levels of this marker are imperative.
In periodontitis patients, a GCF marker that is indicative of peri-
odontal breakdown is hypothesized to be significantly elevated in
the whole saliva as well. This formed the premise for our explora-
tion of saliva as a diagnostic fluid for periodontal disease. Analyses
of biomarkers in saliva have been shown to provide an overall
assessment of the disease status. [26]. As yet, to the best of our
knowledge, relationship between salivary levels of MCP-1 and
chronic periodontitis has not been clarified. It is presumed that
the results of this study would fortify the assumption of saliva
being predictable and accurate in establishing evidence of peri-
odontal disease. The correlations between levels of MCP-1 in all
the three fluids have also not been ascertained till date.
Phase I periodontal therapy is a time tested methodology to
effectively minimize the clinical effects of periodontal disease.
[27] Hence, we also explored the role of scaling/root planing in
affecting the levels of MCP-1 in GCF, saliva and serum.
Thus our study consisted of two phases: the first part was a
cross-sectional study that aimed at evaluating and comparing
MCP-1 levels in GCF, saliva and serum of patients with chronic
periodontitis vs healthy controls. The second part was a longitudi-
nal assessment of all clinical indices and MCP-1 levels in the dis-
eased patients, 6 weeks after periodontal therapy. The possible
correlations between the MCP-1 levels in the fluids analyzed
and with the clinical parameters assessed have also been
evaluated.
893
2. Methodology
2.1. Study population
A total of 45 patients; 15 healthy controls (Healthy group) and
30 of severe chronic periodontitis (diseased group) were enrolled
for the study from the Out Patient Department of the Department
of Periodontics at Dr. Harvansh Singh Judge Institute of Dental Sci-
ences and Hospital, Panjab University, Chandigarh in the time per-
iod from May 2009 to December 2011. The study was approved by
the ethical committee of the Panjab University, Chandigarh, India
and informed consent was obtained from each patient after appro-
priately explaining the research protocol to them prior to the
study.
All patients selected exhibited good general health with no evi-
dence of any underlying systemic disease and with the presence of
at least 20 natural teeth excluding third molars. Patients who were
smokers, who had undergone phase I therapy in last 3 months,
consumed antibiotics in last 1 month, with history of allergies, dia-
betes, hypertension, coronary artery disease, any invasive cardiac
treatment within last 6 months or any chronic inflammatory dis-
ease such as idiopathic pulmonary fibrosis, osteoarthritis, rheuma-
toid arthritis, renal diseases or tumors, diabetes, pregnant and
lactating women were all selectively excluded from the study.
The periodontal status of the selected patients was then as-
sessed utilizing plaque index (PI), gingival index (GI), probing
pocket depth (PD) and clinical attachment levels (CAL) using a
Ò
UNC 15 probe (Hu Friedy International ). All the readings were re-
corded by a single examiner, whose reliability was ascertained
using Dahlberg’s formula and was found to be 0.72. Based on these
clinical parameters, the patients were divided into two groups;
Healthy: with GI < 1, PD < 3 mm and CAL = 0 and Diseased: peri-
odontitis patients with two or more inter-proximal sites with
CAL P 6 mm, not on the same tooth, and one or more inter-proxi-
mal sites with PD P 5 mm [28].
The patients of severe chronic periodontitis (diseased) were
administered oral hygiene instructions and underwent scaling
and root planning as part of Phase I periodontal therapy. No
adjunctive antibiotics were administered. The patients were re-
called at 6 weeks and the clinical parameters were evaluated and
the samples collected.
2.2. Sample collection
2.2.1. GCF sampling
From each patient, a standardized volume of 2 ll of GCF was
collected using the white color-coded 1–5 ll calibrated volumetric
Hirschmann’s micro-capillary pipettes (Sigma–Aldrich, St. Louis,
MO, USA) from the most severely affected site; that is the site with
the highest loss of attachment. In case of healthy patients, pooled
samples from various healthy sites were collected to get the requi-
site amount. The sample collection was done on the next day of
periodontal assessment, to avoid contamination of the sample col-
lected. The sites were appropriately dried and isolated using cotton
rolls to prevent salivary contamination, supra-gingival plaque at
the gingival margins was removed using a curette and the GCF
was collected extra-crevicularly by placing the micro-pipette
gently at the gingival margin without inserting it into the crevice.
Any sample that was contaminated with blood or saliva was dis-
carded. The samples collected were transferred to airtight 0.5 ml
eppendorfs using a blast of air through the micro-capillary pipettes
to enable complete removal. These eppendorfs contained 198 ll of
sample diluent provided with the ELISA kit to make up to a volume
of 200 ll. 10 ml of this buffer of 5 concentration was diluted with
40 ml of de-ionized water and kept at 4 °C for intermittent use. The
894 M. Gupta et al. / Cytokine 61 (2013) 892–897
sealed eppendorfs were appropriately labeled, and stored at 70 °C
till the day of the bio-chemical assessment [16].
2.2.2. Serum sampling
5 ml of venous blood was collected from the ante-cubital vein
using a standard veni-puncture method and was immediately
transferred to the laboratory. The blood sample was allowed to clot
at room temperature and, after 1 h, serum was separated by centri-
fugation. The serum was transferred to storage vials and stored at
70 °C till required for biochemical assays. [25].
2.2.3. Saliva sampling
Saliva samples were collected by expectorating 5 ml of un-stim-
ulated whole saliva into sterile plastic containers and the patients
were refrained from eating or drinking at least 2 h prior to sample
collection. To avoid the influence of stress on the secretion rate, all
subjects were asked to rest for at least 10 min before saliva collec-
tion. The whole saliva samples were centrifuged and aliquots of
500 ll were stored at 70 °C until required for the assay. [29].
All the samples that were sent for biochemical assessment were
thawed only once and the MCP-1 levels were analyzed utilizing
ELISA technique.
2.3. MCP-1 assay
MCP-1 levels in GCF, saliva and serum obtained from the study
participants were measured using Human MCP-1 Elisa kit (Cat #
ELH-MCP-1–001, Ray Biotech, Inc.) as per manufacturer’s instruc-
tions. Briefly, all the samples and the standards (recombinant
MCP-1) were incubated in the wells pre-coated with antibody spe-
cific for human MCP-1. MCP-1 present in the standards and sam-
ples was bound to the wells by this immobilized antibody. The
wells were washed and biotinylated anti-human MCP-1antibody
was added. After washing away the unbound biotinylated anti-
body, HRP-conjugated streptavidin was pipetted into the wells.
The wells were again washed; a tetramethyl benzidine substrate
solution was added to the wells. The reaction was stopped using
a stop solution after 30 min which changes color from blue to yel-
low. The intensity of color was measured at 450 nm immediately
on an ELISA reader. The concentration of MCP-1 was obtained
using reference calibrated curve, obtained by plotting the optical
density of standards against their concentration.
3. Statistical analysis
The statistical analysis was carried out using statistical package
for social sciences (SPSS Inc., Chicago, IL, version 15.0 for Win-
dows). In this study for the sample size enrolled, the power of
the correlations came out to be 90% at 95% confidence interval.
All quantitative variables were estimated using measures of central
location (mean, median) and measures of dispersion (standard
deviation). Normality of data was checked by measures of skew-
ness and Kolmogorov Smirnov tests of normality. For the age and
gender, the data was found to be normally distributed and hence
Independent ‘t’ test was applied. In the first part of the study
cross-sectional analysis between various parameters of healthy
and diseased group was done. The pattern of distribution of the
data of variables between these two groups was found to be
skewed for GI and CAL and normal for the rest of the parameters.
For the parametric evaluations, Mann whitney ‘U’ test was applied
for the skewed data and Independent ‘t’ test was applied for the
normally distributed data. In the second part of the study, longitu-
dinal evaluation of the diseased group was conducted 6 weeks
after periodontal therapy. For the comparison of the pre and
post-treatment readings of various parameters of the diseased
group, paired ‘t’ test was applied. All statistical tests were two-
sided and performed at a significance level of a =
05. Karl Pearson’s
correlations were done to assess the correlation between the levels
of MCP-1 in serum, saliva and GCF and the various clinical indices.
4. Results
A total of 45 patients, 15 healthy and 30 with severe periodon-
titis were assessed. The mean age of patients in the healthy group
was 43.93 ± 9.9 years (range 30–61 years). The mean age of pa-
tients in the severe periodontitis group was 41.20 ± 8.775 (range
28–63). In healthy group, 53% were females and 46.7% were males.
In diseased group, 43.3% were females and 56.7% were males. The
two groups were found to be statistically matched for age and gen-
der (p = 0.351 & 0.526 respectively).
Part-1: Cross-sectional analysis: The mean values of various
parameters, i.e. GI, PI, PD, CAL along with GCF, saliva and serum
levels of MCP-1 has been graphically presented Fig. 1. The results
revealed a statistically significant difference in all the readings be-
tween the healthy and diseased. All the clinical parameters in the
diseased group except Plaque Index (PI) correlated significantly
with the levels of MCP-1 in all the three biological fluids (Table 1).
Part-2: Longitudinal analysis: Results of the paired‘t’ test be-
tween pre and post-treatment readings of the diseased group re-
vealed a highly significant decrease in all the parameters
assessed (Fig. 1). The post-treatment MCP-1 levels also showed a
significant positive correlation with all the clinical parameters
(Table 2).
The most important observation of the data was the strong
association between GCF, serum, and salivary levels of MCP-1 in
patients with severe chronic periodontitis both before and after
treatment in the diseased group (Figs. 2 and 3).
5. Discussion
Research has documented elevated levels of MCP-1 in GCF of
patients with chronic as well as aggressive forms of periodontal
disease [14–16]. In the past, immuno-histochemistry studies have
not only demonstrated a high level of MCP-1 in human inflamed
gingival tissues but also a significantly higher MCP-1 gene expres-
sion in patients of chronic periodontitis [10,11]. As documented by
Choi et al., this response was probably implicated due to the
Fig. 1. Mean values of various clinical parameters and levels of MCP-1 in healthy,
diseased (pre-treatment) and diseased (post-treatment) groups. ⁄ indicates p values
60.05 (i.e. significant values) on comparing healthy and diseased (pre-treatment)
and diseased (pre-treatment) vs diseased (post-treatment).

Table 1
Correlations comparing various clinical parameters with levels of MCP-1 in diseased
group (Pre-treatment).
Diseased group (Pre-treatment)
Serum MCP-1 (pg/ml)
GCF MCP-1 (pg/ll)
Saliva MCP-1 (pg/ml)
*
p < 0.05.
**
p < 0.001.
Table 2
Correlations comparing various clinical parameters with levels of MCP-1 in the
diseased group (Post-treatment).
Diseased group (Post-treatment)
Serum MCP-1 (pg/ml)
GCF MCP-1 (pg/ll)
Saliva MCP-1 (pg/ml)
*
p < 0.05.
**
p < 0.001.
Fig. 2. Correlations of MCP-1 levels in GCF, saliva and serum in patients of severe
chronic periodontitis.
Fig. 3. Correlations of MCP-1 levels in GCF, saliva and serum in patients of diseased
group following phase I periodontal therapy.
M. Gupta et al. / Cytokine 61 (2013) 892–897 895
activity of the cell components of various gram negative peri-odon-
topathogens especially Porphyromonas gingivalis which are causa-
tive for severe forms of periodontal disease [30]. In concordance
GI PI PD (mm) CAL (mm) with the above mentioned work, cross-sectional results of our

439*
232
777**
986** study also revealed increased levels of MCP-1 in the GCF of patients

451*
257
730**
969** with severe chronic periodontitis as against healthy controls.

392*
304
779**
953** There are however, differences in the readings of our results as
compared to previously published similar studies. This could be
due to various reasons, primarily; the collection technique [31].
The quantity and quality of GCF samples are highly affected by
the method of collection and analysis. Emingill et al. [14] and Kur-
tis et al. [15] have used perio-paper for GCF collection followed by
the use of Periotron for measuring its volume. This technique
though easy in terms of time and ease of sample collection, is asso-
ciated with certain limitations such as concentration of sample due
GI PI PD (mm) CAL (mm)
to evaporation, non specific attachment of the analyte with filter
paper fibers as well as its inability to measure accurately volumes

408*
395*
475**
636**
of GCF greater than 1 ll. These reasons may contribute to the dis-

444*
428*
636**
736**

340
437*
390*
515** parities in readings of the levels of MCP-1 despite use of similar
technique of GCF sampling.
Another technique of GCF collection is the use of micro-capil-
lary pipettes which entails prolonged and repeated sampling of
the diseased site to obtain an adequate volume. This can affect
the nature of the GCF sample which is likely to change with the
time of collection. Pradeep et al. have used micro-capillary pipettes
to collect 1 ll of GCF samples in a time frame of 10 min per site
[16]. In our study we used the same technique to collect 2 ll of
GCF from each site which took us almost 30 min. This prolonged
sampling could probably have diluted the GCF sample and hence
affected the concentration of MCP-1. Studies have also shown that
total enzyme activity and enzyme concentration are generally
greater in the static and initial GCF samples compared to subse-
quent samples and the fluid volume in the crevice recovers more
rapidly than constituents derived from host cells thereby diluting
the GCF sample on prolonged and repeated sampling [32,33].
Further studies have also shown that the protein concentration
of the initial GCF collected is comparable to the interstitial fluid,
whereas prolonged sampling at the site results in protein concen-
trations approaching those of serum, thereby explaining the dis-
cordances with the readings of previously published data. Thus
different approaches in sampling techniques, sampling times and
data presentation seem to be critical in GCF-profile studies [34].
On analyzing the levels of MCP-1 in serum, the difference was
highly significant between healthy and diseased group. These find-
ings strengthen the hypothesis that periodontal infections due to
their chronic low dose bacteremia and endotoxemia affect sys-
temic vascular physiology which precipitates thrombogenic and/
or atherosclerotic states [17,35,36]. MCP-1 has been identified as
a risk predictor for various cardiac conditions such as atherosclero-
sis (especially Coronary Artery Disease), myocarditis and acute cor-
onary syndromes [37,38]. This indicates that the raised serum
levels of MCP-1 due to a periodontal infection might increase the
susceptibility to develop heart diseases in systemically healthy
individuals and may increase the disease progression in patients
with established CAD. Further research in this area is required to
suggest MCP-1 as one of the markers in understanding the recipro-
cal association between pathogenesis of CAD and periodontal
infection.
Going further, the levels of this marker were analyzed in saliva
as well and the results were found to be statistically significant. A
salivary assessment can be potentially used as a specific non-inva-
sive diagnostic test for periodontitis as it contains locally and sys-
temically derived biomarkers [39]. Sexton et al. had conducted a
case control study on patients of chronic periodontitis and ana-
lyzed numerous pro-inflammatory cytokines as salivary biomark-
ers and their levels in response to treatment and positive
896 M. Gupta et al. / Cytokine 61 (2013) 892–897
indications were seen [40]. Nomura et al. have shown that salivary
biomarkers of various host derived products could be useful for
predicting the progression of chronic periodontitis [41]. Teles
et al. also assessed the levels of numerous cytokines implicated
in the periodontal tissue destruction in saliva of chronic periodon-
titis patients vs controls but, found no statistical difference in the
two groups [42]. There exists, however an imperative need to de-
sign chair side, easy and non-invasive methods for diagnosis and
monitoring the progression of periodontal disease and hence stud-
ies on saliva are currently being majorly focused. Saliva is easy to
obtain especially in outdoor or community settings, thus detecting
infections using saliva samples may be of significant clinical, eco-
nomical and epidemiological importance. Till date, there is no
study that has documented the levels of MCP-1 in saliva of chronic
periodontitis patients and the results of this study can go a long
way in establishing saliva as a simple and quick test in screening
of periodontal diseases.
In diseased patients, the levels of MCP-1 in GCF, saliva and ser-
um correlated positively with the clinical parameters. The ‘r’ value
for most of the parameters was found to be >0.7 proving a strong
correlation. This indicates that MCP-1 levels could be considered
predictive for the progression of periodontal disease severity. Sim-
ilar such correlations in GCF and serum have been seen in studies
by Pradeep et al. [16,25] and Hanazawa et al. [12], but the present
study is the first to correlate salivary levels of MCP-1 with severity
of this disease.
In addition, the present research also reports the positive asso-
ciation amongst the levels of MCP-1 in the three biological fluids in
the same group of patients. There has been no published research
about these correlations till date. Statistically significant correla-
tions between GCF and salivary levels of MCP-1 indicate that sali-
vary levels mirror the localized sub-gingival micro-environment.
Thus saliva may serves as a reliable and accurate diagnostic fluid
particularly for screening large populations especially during sur-
veys. Statistically significant correlations between GCF and salivary
levels of MCP-1 with serum levels in diseased group further indi-
cate that increase in the severity of oral diseases poses a propor-
tionate rise in the potential risk for systemic inflammation. This
observation holds importance due to the fact that more than 50%
of the cases of cardiovascular disease are not associated with clas-
sical risk factors and a chronic inflammatory process like periodon-
tal disease is considered to be an important complimentary factor
in this causal chain [43].
Phase I periodontal therapy was given to the diseased patients
which not only led to an improvement in the clinical parameters
but also significantly decreased the MCP-1 levels in all the three
fluids. In treated cases, majority of the correlations were moderate
with ‘r’ values between 0.3 and 0.7. More importantly, decreased
GCF and salivary MCP-1 levels correlated significantly with de-
creased serum levels. This strongly suggests that any interven-
tional therapy designed to improve the periodontal status, would
not only affect the levels of inflammatory markers in the oral mi-
cro-environment but also may have equally significant suppressive
effects on the systemic plane. Similar findings were recorded by
Freitas et al. in their meta-analysis regarding the hypothesis that
periodontal therapy has a reductive effect on serum C-reactive pro-
tein levels [43]. However, further studies need to be conducted to
confirm these results and improve their external validity.
The present research endorses the importance of exploring
MCP-1 as a risk indicator for severe chronic periodontitis.
Treatment strategies that aim at lowering its levels are being cur-
rently worked on. Parenteral administration of PGE1 appeared to
decrease circulating MCP-1 levels, which was thought to help sup-
press the development of atherosclerotic lesions in patients with
peripheral arterial obstructive disease [44]. Role of administration
of anti-MCP-1 antibody [45], gene therapy with MCP-1 mutant
[46], edaravone [47] and statins [48] in lowering the levels of
MCP-1 have also been favorably explored in animal models as well
as patients of various forms of cardiac diseases. From a futuristic
viewpoint, these agents can be potentially tested in severe peri-
odontitis models to achieve improved adjunctive response to Phase
I periodontal therapy leading to a positive clinical outcome of the
disease not only in the oral cavity but also diminishing risk of sys-
temic illnesses.
Acknowledgement
We would like to sincerely thank Mrs. Kusum Chopra for ana-
lyzing our data and providing us with the results.
References
[1] Fokkema SJ, Loos BG, van der Velden U. Monocyte-derived RANTES is
intrinsically elevated in periodontal disease while MCP-1 levels are related
to inflammation and are inversely correlated with IL-12 levels. Clin Exp
Immunol 2003;131:477–83.
[2] Kornman KS, Page RC, Tonetti MS. The host response to the microbial
challenge in periodontitis: assembling the players. Periodontology
1997;2000(14):33–53.
[3] Gemmell E, Seymour GJ. Immunoregulatory control of Th1/Th2 cytokine
profiles in periodontal disease. Periodontology 2004;2000(35):21–41.
[4] Graves DT. The potential role of chemokines and inflammatory cytokines in
periodontal disease progression. Clin Infect Dis 1999;28:482–90.
[5] Baggiolini M. Chemokines in pathology and medicine. J Intern Med
2001;250:91–104.
[6] Zlotnik A, Yoshie O. Chemokines: a new classification system and their role in
immunity. Immunity 2000;12:121–7.
[7] Silva TA, Garlet GP, Fukada SY, Silva JS, Cunha FQ. Chemokines in oral
inflammatory diseases: apical periodontitis and periodontal disease. J Dent Res
2007;86:306–19.
[8] Rossi D, Zlotnik A. The biology of chemokines and their receptors. Annu Rev
Immunol 2000;18:217–42.
[9] Colotta F, Borre A, Wang JM, Tattanelli M, Maddalena F, Polentarutti N, et al.
Expression of monocyte chemotactic cytokine by human mononuclear
phagocytes. J Immunol 1992;148:760–5.
[10] Yu X, Antoniades HN, Graves DT. Expression of monocyte chemoattractant
protein-1 in human inflamed gingival tissues. Infect Immun 1993;61:4622–8.
[11] Yu X, Graves DT. Fibroblasts, mononuclear phagocytes, and endothelial cells
express monocyte chemoattractant protein-1 (MCP-1) in inflamed human
gingiva. J Periodontol 1995;66:80–8.
[12] Hanazawa S, Kawata Y, Takeshita A, Kumada H, Okithu M, Tanaka S, et al.
Expression of monocyte chemoattractant protein-1 (MCP-1) in adult
periodontal disease: increased monocyte chemotactic activity in crevicular
fluids and induction of MCP-1 expression in gingival tissues. Infect Immun
1993;61:5219–24.
[13] Champagne CM, Buchanan W, Reddy MS, Preisser JS, Beck JD, Offenbacher S.
Potential for gingival crevice fluid measures as predictors of risk for
periodontal diseases. Periodontology 2003;2000(31):167–80.
[14] Emingil G, Atilla G, Hüseyinov A. Gingival crevicular fluid monocyte
chemoattractant protein-1 and RANTES levels in patients with generalized
aggressive periodontitis. J Clin Periodontol 2004;31:829–34.
[15] Kurtisß B, Tüter G, Serdar M, Akdemir P, Uygur C, Firatli E, et al. Gingival
crevicular fluid levels of monocyte chemoattractant protein-1 and tumor
necrosis factor-alpha in patients with chronic and aggressive periodontitis. J
Periodontol 2005;76:1849–55.
[16] Pradeep AR, Daisy H, Hadge P. Gingival crevicular fluid levels of monocyte
chemoattractant protein-1 in periodontal health and disease. Arch Oral Biol
2009;54:503–9.
[17] Paquette DW, Brodala N, Nichols TC. Cardiovascular disease, inflammation,
and periodontal infection. Periodontology 2007;2000(44):113–26.
[18] Scannapieco FA. Role of oral bacteria in respiratory infection. J Periodontol
1999;70:793–802.
[19] Mealey BL, Ocampo GL. Diabetes mellitus and periodontal disease.
Periodontology 2000 2007;44:127–53.
[20] Takeya M, Yoshimura T, Leonard EJ, Takahashi K. Detection of monocyte
chemoattractant protein-1 in human atherosclerotic lesions by an anti-
monocyte chemoattractant protein-1 monoclonal antibody. Hum Pathol
1993;24:534–9.
[21] Antoniades HN, Neville-Golden J, Galanopoulos T, Kradin RL, Valente AJ,
Graves DT. Expression of monocyte chemoattractant protein-1 mRNA in
human idiopathic pulmonary fibrosis. Proc Natl Acad Sci 1992;89:5371–5.
[22] Graves DT, Barnhill R, Galanopoulos T, Antoniades HN. Expression of monocyte
chemotactic protein-1 in human melanoma in vivo. Am J Pathol
1992;140:9–14.
[23] Villiger PM, Terkeltaub R, Lotz M. Production of monocyte chemoattractant
protein-1 by inflamed synovial tissue and cultured synoviocytes. J Immunol
1992;149:722–7.
 M. Gupta et al. / Cytokine 61 (2013) 892–897
[24] Yu XH, Barnhill RL, Graves DT. Expression of monocyte chemoattractant
protein-1 (MCP-1) in delayed type hypersensitivity reactions in the skin. Lab
Invest 1994;71:235–66.
[25] Pradeep AR, Daisy H, Hadge P. Serum levels of monocyte chemoattractant
protein-1 in periodontal health and disease. Cytokine 2009;47:77–81.
[26] Miller CS, King Jr CP, Langub MC, Kryscio RJ, Thomas MV. Salivary biomarkers
of existing periodontal disease: a cross-sectional study. J Am Dent Assoc
2006;137:322–9.
[27] Garrett JS. Effects of non-surgical periodontal therapy on periodontitis in
humans: a review. J Clin Periodontol 1983:510–5.
[28] Page RC, Eke PI. Case definitions for use in population-based surveillance of
periodontitis. J Periodontol 2007;78:1387–99.
[29] Navazesh M. Methods for collecting saliva. Ann NY Acad Sci 1993;694:72–7.
[30] Choi EK, Park SA, Oh WM, Kang HC, Kuramitsu HK, Kim BG, et al. Mechanisms
of porphyromonas gingivalis-induced monocyte chemoattractant protein-1
expression in endothelial cells. FEMS Immunol Med Microbiol 2005;44:51–8.
[31] Griffiths GS. Formation, collection and significance of gingival crevicular fluid.
Periodontology 2003;2000(31):32–42.
[32] Persson GR, Page RC. Effect of sampling time and repetition on gingival
crevicular fluid and aspartate aminotransferase activity. J Periodontal Res
1990;25:236–42.
[33] Lamster IB, Harper D, Goldstein S, Celenti RS, Oshrain RL. The effect of
sequential sampling on crevicular fluid volume and enzyme activity. J Clin
Periodontol 1989;16:252–8.
[34] Curtis MA, Griffiths GS, Price SJ, Coulthurst SK, Johnson NW. The total protein
concentration of gingival crevicular fluid. Variation with sampling time and
gingival inflammation. J Clin Periodontol 1988;15:628–32.
[35] Papapanagiotou D, Nicu EA, Bizzarro S, et al. Periodontitis is associated with
platelet activation. Atherosclerosis 2009;202:605–11.
[36] Herzberg MC, Weyer MW. Dental plaque, platelets and cardiovascular disease.
Ann Periodontol 1998;3:151–60.
[37] Tanaka T, Nakamura Y, Nasuno A, Mezaki T, Higuchi K, Fukunaga H, et al.
Plasma concentrations of monocyte chemoattractant protein-1 (MCP-1) and
neopterin in the coronary circulation of patients with coronary artery disease.
Circ J 2004;68:114–20.
897
[38] Gonzalez-Quesada C, Frangogiannis NG. Monocyte chemoattractant protein
(MCP-1)/CCL2 as a biomarker in acute coronary syndromes. Curr Atheroscler
Rep 2009;11:131–8.
[39] Zhang L, Henson BS, Camargo PM. The clinical value of salivary biomarkers for
periodontal disease. Periodontology 2009;2000(51):25–37.
[40] Sexton WM, Lin Y, Kryscio RJ, Dawson 3rd DR, Ebersole JL, Miller CS. Salivary
biomarkers of periodontal disease in response to treatment. J Clin Periodontol
2011;38:434–41.
[41] Nomura Y, Shimada Y, Hanada N, Numabe Y, Kamoi K, Sato T, et al. Salivary
biomarkers for predicting the progression of chronic periodontitis. Arch Oral
Biol 2012;57:413–20.
[42] Teles RP, Likhari V, Socransky SS, Haffajee AD. Salivary cytokine levels in
chronic periodontitis and periodontally healthy subjects. A cross-sectional
study. J Periodontal Res 2009;44:411–7.
[43] Freitas CO, Gomes-Filho IS, Naves RC, Noqueira Filho Gda R, Cruz SS, Santos CA,
et al. Influence of periodontal therapy on C-reactive protein level: a systematic
review and meta-analysis. J Appl Oral Sci 2012;20:1–8.
[44] Matsui K, Ikeda U, Murakami Y, Yoshioka T, Shimada K. Intravenous
prostaglandin E1 reduces monocyte chemoattractant protein-1 levels in
peripheral arterial obstructive disease. Am Heart J 2003;145:330–3.
[45] Furukawa Y, Matsumori A, Ohashi N, Shioi T, Ono K, Harada A, et al. Anti-
monocyte chemoattractant protein-1/monocyte chemotactic and activating
factor antibody inhibits neointimal hyperplasia in injured rat carotid arteries.
Circ Res 1999;84:306–11.
[46] Yue Y, Gui J, Xu W, Xiong S. Gene therapy with CCL2 (MCP-1) mutant protects
CVB-3 induced myocarditis by compromising Th1 polarisation. Mol Immunol
2011;48:706–13.
[47] Nakamura Y, Yamada Y, Shimomura H, Nagayoshi Y, Tsujita K, Yamashita T,
et al. Effect of edaravone on plasma monocyte chemoattractant protein-1
levels in patients with acute myocardial infarction. J Cardiol 2009;54:416–24.
[48] Yang YP, Dong QL, Zhang XH, Zhang YH, Zhu L, Li SY, et al. Combination of
fluvastatin and losartan relieves atherosclerosis and macrophage infiltration in
atherosclerotic plaques in rabbits. Acta Pharmacol Sin 2011;32:1259–65.