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INTRODUCTION
Inflammation of dental pulp is similiar to that in other
connective tissue in that it is mediated by cellular and
molecular factors (Fouad, 2002). The inflammatory res-
ponse to dental pulp injury or infection has major clinical
significance. Injury may be caused by dental caries, dental
restorative procedures (iatrogenic), tooth fracture or
attrition (Trowbridge, 2002).
Cytokines are proteins that provide commucation
between cells and play a critical role in a wide veriety of
processes including inflammation. Cytokines released from
cells in an inflammatory process that activate, mediate or
potentiate actions of other cells or tissues. Their actions
may be effected in an autocrine, paracrine or endocrine
manner (Seymour et al., 1995).
Propolis is an adhesive substance produced by
honeybees from the bud and exudates of certain trees and
plants (Bankova et al., 2000). Propolis has a long history as
a general tonic promoting health, due to its several biolog-
ical properties, such as anti-inflamatory (Hu et al., 2005),
antibacterial (Sforcin et al., 2000) and immunomodulatory
(Sforcin, 2007).
Therefore the aim of this report is to review the effect of
propolis on cytokines during dental pulp inflammation, so
could opens a new prespective on the investigation of
propolis mainly with respect to the immune system.
The Effect of Propolis on Cytokines during Dental
Pulp Inflammation
Ardo Sabir*
Department of Conservative Dentistry, Faculty of Dentistry, Hasanuddin University, Makassar 90245, Indonesia
(Received 21 November 2015; Revised 24 April 2016; Accepted 25 April 2016)
97
Propolis has been used empirically for centuries because of its several biological and
pharmacological properties. It was always mentioned as an anti-inflammatory agent. When dental
pulp has inflammation, several mediators will be produced by innate immune cells. Cytokines are
proteins released from cells in inflammatory process. In recent years, in vitro and in vivo assays
provided information concerning about propolis and cytokines, thus a review dealing with the effect of
propolis on cytokines during dental pulp inflammation became imperative. This review complied data
from our study as well as from other researchers, focusing on its chemical composition on
inflammatory process. Its anti-inflammatory property, considering its effect on interleukin, tumour
necrosis factor, transforming growth factor as well as other cytokines during dental pulp inflammation
are discussed. In vitro and in vivo assays demonstrated that propolis could inhibit inflammation
process in dental pulp. Since humans have used propolis for many inflammation diseases and
propolis-containing products have been marketed, the knowledge of its properties with scientific
basis is not only of academic interest but also of those who use propolis as well. This review opens a
new prespective on the investigation of propolis, mainly with respect to the immune system.
Key words: Propolis, Cytokines, Inflammation, Dental pulp.
Abstract |
한국양봉학회지 제25권 제2호 (2010)
Journal of Apiculture 31(1): 97~101 (2016)
*Corresponding author. E-mail: ardo.sabir@yahoo.com
Short
Communication
Ardo Sabir
INFLAMMATION PROCESS
The concept of inflammation has evolved since the
discovery of cells in the 19th century. By this time, infla-
mmation was seen to be preceded by cell and tissue
injuries, and that vascular changes including leukocyte
emigration were secondary events (Trowbridge, 2002).
During the 1920s, the idea that the vascular system
facilitated quick accumulation of great quantities of
phagocytes and antibodies was reviewed. The first phys-
ical-chemical analysis of inflammation, cell stress and local
tissue changes, promoted by an increasing concen-tration
of oxidants and osmotic pressure, were also made at this
time (Mitchell and Cotran, 2003). Therefore, modern
investigators have considered inflammation a primary
event of the host defense system.
Inflammation can be represented by capillary dilatation
with fluid accumulation (oedema) and by phagocyte
emigration and accumulation (neutrophils, monocytes,
macrophages), which also contribute to hyperalgesia
generation and loss of tissue function (Abbas et al., 2007).
Other characteristics, such as erithema and fever, can also
be observed during inflammatory events. The last feature
occurs after cytokine release (Interleukin-1 [IL-1], Tumor
Necrosis Factor-alpha [TNF-α]) by activated macr-
ophages, leading to a vessel dilatation resulting from
smooth muscular relaxation and followed by an increase in
local blood flow (hypothermia) (Fouad, 2002). Inflam-
matory events also involve micro-vascular changes with
increased vascular permeability, flow exudation, including
plasmatic protein and amplification of endogenous chem-
ical mediators (Cirino, 1998).
The acute-phase response involves serous, fibrinous,
supurative or exudative events as well as micro-vascular
and cell events; this response to pathogen occurs within 72
hours. The chronic-phase response includes proliferative
events and histological alterations, different from those in
the acute phase, characterized by cell emigration and inten-
sive mitosis. In addition, inflammation may be physiolog-
ical or pathological, depending mainly on histological
aspects (Mitchell and Cotran, 2003).
PROPOLIS AND ITS CHARACTERISTICS
In a recent years the researchers were searching for
natural products with medicinal properties, particularly
those from plants and bees to found a natural anti-
inflammatory agent. Several plants produce resinous exud-
ates with strong anti-microbial and anti-necrotic properties,
in addition to impermeability provided by populus a
substance from Populus sp. (Greenaway et al., 1990). Bees
collect resin exudates from certain plants and add their
secretion, wood fragments, pollen, and wax; this product
from bees and plants is called propolis. The word propolis
comes from the Greek pro meaning in defense of and polis
city, representing defense of bee cities (or beehives).
Propolis has been used in folk medicine since primeval
times. Nowadays, propolis is still used in home made
remedies and cosmetics. Two characteristics of propolis
are its smell and its various colors from dark green to
brown (Marcucci et al., 1998).
Propolis chemical composition has been correlated with
plant diversity around the beehive (Sforcin et al., 2000). In
general, raw propolis contains about: 50~55% resins and
balsams (phenols, phenolic acids, esters, flavanons
{quercetin, galangin, pinocembrin}, dihydroflavanons,
flavons, flavonols, chalkones, phenolic glycerides, cinn-
amic acid, coumaric acid, prenylated compounds and
artepillin C), 25~30% waxes, 10% volatile oils, 5% pollen
and 5% organic and mineral substances. The components
are rich in vitamins such as B1, B2, B6, C, E and mineral
elements like Mg, Ca, I, K, Na, Cu, Zn, Mn and Fe. It also
contains number of fatty acids and enzymes as succinic
dehydrogenase, glucose-6-phosphatase, adenosine triphos-
phatase and acid phosphatase (Parolia et al., 2000). Con-
sidering that propolis is a complex mixture, synergistic in-
teractions between its compounds must also be considered
as an important factor in its anti-inflammatory activity.
Ethanol, the most commonly used solvent for propolis
preparations, and other solvents such as ethylic ether,
water, methanol, petroleum ether, and chloroform are used
for extracting and identifying many propolis compounds.
Moreover, glycerin, propylene glycol and some solutions
have been used in propolis preparations by the pharmace-
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The Effect of Propolis on Cytokines during Dental Pulp Inflammation
utical and cosmetic industry (Castaldo and Capasso, 2002).
Propolis compounds have recently become the subject of
investigation in order to determine its therapeutic appli-
cation in dentistry (Sabir, 2012); flavonoids are consider as
the most biologically active substance in propolis
(Havsteen et al., 2002). In the last 20 years, there has been
increased commercial interest in propolis use due to its
therapeutic properties to treat many diseases (Parolia et al.,
2010). Nowadays, we can found propolis is commercially
in sprays, ointments, capsules, capillary lotions and tooth-
pastes because of its bacteriostatic activity and pharmac-
ological properties.
EFFECT OF PROPOLIS ON CYTOKINES IN
INFLAMMATION
The knowledge of propolis mechanisms of action on the
immune system has advanced in the last years. However,
only a few reports were published about the effect of
propolis on cytokines during dental pulp inflammation
since it was known that inflammation of dental pulp is
similiar to that in other connective tissue. Many in vitro
and in vivo experiments are performed with Ethanolic
Extracts Propolis (EEP) and Aqueous Extracts Propolis
(AEP) to confirm the anti-inflammatory activity of pro-
polis. EEP anti-inflammatory effects was observed in
inhibit both platelet aggregation and cytokines (Hu et al.,
2005; Missima et al., 2009; Bachiega et al., 2012). Our
previous study using immunohistochemistry method sho-
wed that propolis could delay TNFαexpression on inflam-
med rat dental pulp tissue (Sabir, 2015). This antiinflam-
matory activity of propolis can be explained by the pres-
ence of active flavonoids and cinnamic acid derivatives
(Chirumbolo, 2011). The former includes acacetin,
quercetin, and naringenin (terpenoid constituents may exert
an addictive effect); the latter includes Caffeic Acid Phenyl
Ester (CAPE) (Tolba et al., 2013; Zhang et al., 2014) .
Experiment by Orsi et al. (2000), showed that after
propolis treatment (2.5 and 5mg/kg) of mice for 3 days,
peritoneal macrophages were activated in vitro with
Interferon-gamma (IFN-γ). This fact suggests that propolis
treatment leads macrophages to a higher responsiveness to
stimuli IFN-γ. Another indicative of macrophages
activation is Nitric Oxide (NO). NO is synthesized via L-
arginine oxidation by a family of NO synthases (NOSs)
and several cofactors. Nitric Oxide production by cells in
response to cytokines can destroy host tissue and impair
discrete cellular responses (Clancy et al., 1998). Law et al.
(1999) investigated NO activity in the inflammed pulp of
rat molars. The results showed that there was evidence of a
dramatic increase in NO activity at the site of pulp
irritation. Propolis (50 and 100µg/ml) inhibited NO
generation by peritoneal macrophages (Orsi et al., 2000).
Study by Moriyasu et al. (1994) also observed that propolis
(0.2~1.0mg/ml) inhibited NO production by lipopoly-
saccharides (LPS)-stimulated macrohages. Krol et al.
(1996) stated that this effect to flavonoid. Meanwhile, Hu
et al. (2005) evaluated the action of EEP and WEP in a
murine model of acute inflammation, verifying that both
extracts inhibited NO generation. Transforming Growth
Factors (TGF)-βwas known as an endogenous suppressor
of NOS expression in murine macrophages which destab-
ilized NOS2 mRNA, retarded the synthesis of NOS2
protein and accelerated its degradation (Macmicking et al.,
1997). This was supported by Ansorge et al. (2003), who
found that the concentration of TGF-βwas increased in the
supernatant of T cell or peripheral mononuclear blood cell
culture, after incubation with propolis, and this may be a
possible explanation for propolis inhibit NO production.
These effects mediated by some of propolis constituents
such as CAPE and flavonoids (quercetin and hesperidin).
Study by Bachiega et al. (2012) found that CAPE,
cinnamic acid and coumaric acid in propolis may be
involved in the action of propolis inhibit both IL-6 and IL-
10 but stimulated IL-1βproduction by macrophages. They
were also observed that CAPE and cinnamic acid are
strong Lipoxygenase (LOX) inhibitors, suppressing
leukotriene production by peritoneal macrophages. Their
action on Leukotriene (LTC)-4 was smaller in vivo.
Quercetin inhibits LOX, and at high concentrations blocks
COX. Naringenin only inhibited LTC4 causing weakness.
All these data have demonstrated the strong and different
inhibitory action of several propolis preparations or its
isolated constituents on inflammation events. However, its
anti-inflammatory effects depends mainly on the admini-
99
stration route and its potency (Mirzoeva and Calder, 1996).
CONCLUSION
In vitro and in vivo assays as well as animal model
experiments demonstrated that propolis and its component
has strong anti-inflammatory activity thereby could be
effective to treat dental pulp inflammation. One of the anti-
inflammatory mechanism of propolis is by inhibited cytok-
ines production, although the other mechanisms are not
fully understand. Human have been using propolis for a
long time, so scientific-based information has important
role for further studies on the investigation of propolis to
opening the new prespective, mainly with respect to the
immune system.
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