EFFECT OF AIR FRESHENER EXPOSURE ON THE HISTOLOGY CORNEA OF RATTUS NORVEGICUS
Effect of Air Freshener Exposure on
The Histology Cornea of Rattus Norvegicus
1
2
Yuningtyaswari , Nastiti Chandra Ayu
1
Departement Histology Faculty of Medicine and Health Science Muhammadiyah University of Yogyakarta
2
Student of Faculty of Medicine and Health Science Muhammadiyah University of Yogyakarta
E-mail: yuningtyas_fkumy@yahoo.com
INTRODUCTION
N
CONCLUSION
owadays more and more difficult to obtain clean and quality air. This is
caused by air pollution that occurs everywhere. One attempt to gain air
tasted fresh and fragrant is to use air freshener. In general, people do not
know that by using air freshener can actually add to indoor air pollution. The air
freshener contains several types of chemicals, such as Volatile Organic Compound
(VOC), allergens, benzene, formaldehyde, terpenes, styrene, toluene and diethyl
phtalate (SCHER 2006). Indoor pollution sources that can release gases or particles
into the air is the main cause of indoor air quality problems (USEPA, 2013). Studies
conducted by Cuellar, Lloyd et al. (2003 and 2004) and Burdick, Merrill et al. (2002)
have demonstrated that the solvent component in a product mixture of scents
potentially irritate the eyes. Indoor air quality (Indoor Air Quality) is a problem that
needs attention because it will affect human health. According to a report from the
National Institute of Occupational Safety and Health (NIOSH, 2013) hazardous
chemicals in the air freshener from the results of such research is formaldehyde.
Formaldehyde can cause irritation to skin, eyes, nose, and throat. High levels of
exposure may cause some types of cancer (EPA, 2013). A mixture of formaldehyde
in the solution of their own air freshener specifically damage the cornea and can
cause blindness, California Department of Public Health (2011).
MATERIALS AND METHODS
Exposure
gel air freshener and spray air freshener adversely affect the corneal
histology of Rattus norvegicus which includes the thickness of the cornea, corneal
anterior epithelial thickness, and number of keratocytes.
A
B
C
Figure 1.
A. Corneal Rattus norvegicus control group (K) were cut with paraffin block method, hematoxylin eosin
staining and magnification of 40x10
B. Corneal Rattus norvegicus Gel air freshener group (P1) were cut with paraffin block method,
hematoxylin eosin staining and magnification of 40x10
C. Corneal Rattus norvegicus Spray Air Freshener Group (P2) were cut with paraffin block method,
hematoxylin eosin staining and magnification 40x10
The subjects of this study were 30 male rats Rattus norvegicus (early age 8 days). Air freshener used was produced by one factory in Indonesia, with the gel and spray forms.
Subjects were divided into three groups, namely the control (untreated), the group P1 (given a gel air freshener exposure) and group P2 (given exposure to spray air freshener).
Exposure to the subject of the group P1 and P2 from the age of 8 days up to 67 days, the initial dose of exposure 15 minutes in the morning and afternoon. Subsequent doses
increased by 15 minutes for each week, until the final dose of 4.5 hours. On day 68, subjects were sacrificed to take his eye cornea. Corneal histology preparations made by the
method of paraffin blocks, with staining techniques Hematoxyline Eosine. Histological changes were observed as the overall thickness of the cornea, anterior epithelial
thickness and number of keratinocytes. Histology observations made by light microscopy at a magnification of 10x10 to measure the overall thickness of the cornea and anterior
epithelial thickness. Observations were made on 5 field of vision, with each repetition 5x. Counting the number of keratinocytes done with a light microscope at a magnification of
40x10 calculated at 5 visual field. Corneal thickness data statistically analyzed with Kruskal Wallis test followed by Mann Whitney posthoc. While the data of the anterior
epithelial thickness and number of keratinocytes were analyzed with statistical test One Way Anova followed by Tukey posthoc.
RESEARCH RESULT
R
DISCUSSION
elated to these data results, it is generally seen that exposure gel and spray air freshener causes histologic changes in the cornea of the eye. The results shows that
the two types of air freshener effect on histological changes in the cornea in this case thickness of the cornea. One of the chemicals contained in the two types of
air freshener is formaldehyde. Formaldehyde is a compound that is responsible for the moderate to severe irritation of the cornea. The results showed a white rat
corneal thickness were exposed spray air freshener thicker than the gel air freshener. Big eye irritation potential possessed by liquid air freshener (spray). This is because
the air freshener works by spraying usually also contribute to added pressure gas (propellant) and produces high concentrations of chemical substances. Trimethylamine
(TMA) is one of the substances used as the propellant (EPA, 2010). TMA is very soluble in water and organic material. Trimethylamine has the chemical formula C3H9N.
The effects of exposure to trimetilamnia is eye irritation, eye lesions and respiratory organs associated with it is neurotoxic (AEGLs, 2008). Increased corneal thickness is
caused due to an increase in collagen. Program synthesis of collagen and GAG-collagen interactions change has been linked with corneal dystrophy conditions and wound
healing process (Robert, 2002). Collagen type XII has a role in the development of stromal architecture and maintain the balance of the formation of fibrils. Meanwhile, other
types namely collagen XIV has the role of regulation fibrilogenesis (Young, 2002). Thrombospondin-1 (TSP-1) is a multifunctional protein matrix and usually appears in the
process of wound healing of the cornea. Level TSP-1 on the cornea injured or irritated increased 3 times compared to normal cornea (Koichi, 2004).The results showed no
significant difference between the thickness of the anterior corneal epithelium of the control group to a group that received air freshener exposure. One the mechanism of
wound healing of the cornea.is the mitosis and migration of epithelial cells that cause an increase in thickness of the epithelium especially the anterior epithelium. This
process will run and in a matter of months to years this process will restore the corneal epithelium structure back to normal epithelium as in the eye or eye uninjured normal
(Steven, 2014). The results showed significant differences in the number of keratocytes between the control group (K) and the treatment group (P1 and P2). According to
the Environmental Health Association of Nova Scotia (2008) air freshener contains a number of chemicals including formaldehyde. The content of formaldehyde in the air
freshener used in this research is 0,33ppm. Acetone and formaldehyde is a chemical that can stimulate increased keratocytes in irritation of the cornea (Maurer, 2002). The
reaction of d-limonene and ozone and other substances that are free radicals in the atmosphere is responsible for the irritation (SCHER 2006). Keratoblast proliferate and
synthesize hyaluronan to form the embryo of stromal extracellular matrix. Keratoblas differentiate into keratinocytes that synthesize collagen and proteoglycans which
replaces the keratan sulfate hyaluronan with MES types of solid collagen fibrils that seen in adult cornea transparent. Keratocytes, which is also known as fibroblast
mesenchymal cells was derived from corneal stroma. These cells are normally quiescent, but they can easily respond to repair if there is an injury or irritation. If there is an
infection, keratinocytes play a role also activate the inflammatory cascade by stimulating the secretion of proinflammatory cytokines. The significant difference number of
keratocyte was found between the groups with group spray air freshener and gel air freshener . White rat corneas were exposed to the gel air freshener found the number of
keratinocytes is more than spray air freshener . This is related to the particle size of the gel air freshener. Chemical particle size gel air freshener in its use will be very small
chemical particles are 1μm (Ruzer, 2013). The smaller the particle size of the
gel air freshener better able dispersed into the cornea and activate the dormant keratinocytes corneal irritation not become active, proliferate and synthesis matrix
metalloproteinases in the process of tissue repair irritated (Wilson, 2007).
The Histology Cornea of Rattus Norvegicus
1
2
Yuningtyaswari , Nastiti Chandra Ayu
1
Departement Histology Faculty of Medicine and Health Science Muhammadiyah University of Yogyakarta
2
Student of Faculty of Medicine and Health Science Muhammadiyah University of Yogyakarta
E-mail: yuningtyas_fkumy@yahoo.com
INTRODUCTION
N
CONCLUSION
owadays more and more difficult to obtain clean and quality air. This is
caused by air pollution that occurs everywhere. One attempt to gain air
tasted fresh and fragrant is to use air freshener. In general, people do not
know that by using air freshener can actually add to indoor air pollution. The air
freshener contains several types of chemicals, such as Volatile Organic Compound
(VOC), allergens, benzene, formaldehyde, terpenes, styrene, toluene and diethyl
phtalate (SCHER 2006). Indoor pollution sources that can release gases or particles
into the air is the main cause of indoor air quality problems (USEPA, 2013). Studies
conducted by Cuellar, Lloyd et al. (2003 and 2004) and Burdick, Merrill et al. (2002)
have demonstrated that the solvent component in a product mixture of scents
potentially irritate the eyes. Indoor air quality (Indoor Air Quality) is a problem that
needs attention because it will affect human health. According to a report from the
National Institute of Occupational Safety and Health (NIOSH, 2013) hazardous
chemicals in the air freshener from the results of such research is formaldehyde.
Formaldehyde can cause irritation to skin, eyes, nose, and throat. High levels of
exposure may cause some types of cancer (EPA, 2013). A mixture of formaldehyde
in the solution of their own air freshener specifically damage the cornea and can
cause blindness, California Department of Public Health (2011).
MATERIALS AND METHODS
Exposure
gel air freshener and spray air freshener adversely affect the corneal
histology of Rattus norvegicus which includes the thickness of the cornea, corneal
anterior epithelial thickness, and number of keratocytes.
A
B
C
Figure 1.
A. Corneal Rattus norvegicus control group (K) were cut with paraffin block method, hematoxylin eosin
staining and magnification of 40x10
B. Corneal Rattus norvegicus Gel air freshener group (P1) were cut with paraffin block method,
hematoxylin eosin staining and magnification of 40x10
C. Corneal Rattus norvegicus Spray Air Freshener Group (P2) were cut with paraffin block method,
hematoxylin eosin staining and magnification 40x10
The subjects of this study were 30 male rats Rattus norvegicus (early age 8 days). Air freshener used was produced by one factory in Indonesia, with the gel and spray forms.
Subjects were divided into three groups, namely the control (untreated), the group P1 (given a gel air freshener exposure) and group P2 (given exposure to spray air freshener).
Exposure to the subject of the group P1 and P2 from the age of 8 days up to 67 days, the initial dose of exposure 15 minutes in the morning and afternoon. Subsequent doses
increased by 15 minutes for each week, until the final dose of 4.5 hours. On day 68, subjects were sacrificed to take his eye cornea. Corneal histology preparations made by the
method of paraffin blocks, with staining techniques Hematoxyline Eosine. Histological changes were observed as the overall thickness of the cornea, anterior epithelial
thickness and number of keratinocytes. Histology observations made by light microscopy at a magnification of 10x10 to measure the overall thickness of the cornea and anterior
epithelial thickness. Observations were made on 5 field of vision, with each repetition 5x. Counting the number of keratinocytes done with a light microscope at a magnification of
40x10 calculated at 5 visual field. Corneal thickness data statistically analyzed with Kruskal Wallis test followed by Mann Whitney posthoc. While the data of the anterior
epithelial thickness and number of keratinocytes were analyzed with statistical test One Way Anova followed by Tukey posthoc.
RESEARCH RESULT
R
DISCUSSION
elated to these data results, it is generally seen that exposure gel and spray air freshener causes histologic changes in the cornea of the eye. The results shows that
the two types of air freshener effect on histological changes in the cornea in this case thickness of the cornea. One of the chemicals contained in the two types of
air freshener is formaldehyde. Formaldehyde is a compound that is responsible for the moderate to severe irritation of the cornea. The results showed a white rat
corneal thickness were exposed spray air freshener thicker than the gel air freshener. Big eye irritation potential possessed by liquid air freshener (spray). This is because
the air freshener works by spraying usually also contribute to added pressure gas (propellant) and produces high concentrations of chemical substances. Trimethylamine
(TMA) is one of the substances used as the propellant (EPA, 2010). TMA is very soluble in water and organic material. Trimethylamine has the chemical formula C3H9N.
The effects of exposure to trimetilamnia is eye irritation, eye lesions and respiratory organs associated with it is neurotoxic (AEGLs, 2008). Increased corneal thickness is
caused due to an increase in collagen. Program synthesis of collagen and GAG-collagen interactions change has been linked with corneal dystrophy conditions and wound
healing process (Robert, 2002). Collagen type XII has a role in the development of stromal architecture and maintain the balance of the formation of fibrils. Meanwhile, other
types namely collagen XIV has the role of regulation fibrilogenesis (Young, 2002). Thrombospondin-1 (TSP-1) is a multifunctional protein matrix and usually appears in the
process of wound healing of the cornea. Level TSP-1 on the cornea injured or irritated increased 3 times compared to normal cornea (Koichi, 2004).The results showed no
significant difference between the thickness of the anterior corneal epithelium of the control group to a group that received air freshener exposure. One the mechanism of
wound healing of the cornea.is the mitosis and migration of epithelial cells that cause an increase in thickness of the epithelium especially the anterior epithelium. This
process will run and in a matter of months to years this process will restore the corneal epithelium structure back to normal epithelium as in the eye or eye uninjured normal
(Steven, 2014). The results showed significant differences in the number of keratocytes between the control group (K) and the treatment group (P1 and P2). According to
the Environmental Health Association of Nova Scotia (2008) air freshener contains a number of chemicals including formaldehyde. The content of formaldehyde in the air
freshener used in this research is 0,33ppm. Acetone and formaldehyde is a chemical that can stimulate increased keratocytes in irritation of the cornea (Maurer, 2002). The
reaction of d-limonene and ozone and other substances that are free radicals in the atmosphere is responsible for the irritation (SCHER 2006). Keratoblast proliferate and
synthesize hyaluronan to form the embryo of stromal extracellular matrix. Keratoblas differentiate into keratinocytes that synthesize collagen and proteoglycans which
replaces the keratan sulfate hyaluronan with MES types of solid collagen fibrils that seen in adult cornea transparent. Keratocytes, which is also known as fibroblast
mesenchymal cells was derived from corneal stroma. These cells are normally quiescent, but they can easily respond to repair if there is an injury or irritation. If there is an
infection, keratinocytes play a role also activate the inflammatory cascade by stimulating the secretion of proinflammatory cytokines. The significant difference number of
keratocyte was found between the groups with group spray air freshener and gel air freshener . White rat corneas were exposed to the gel air freshener found the number of
keratinocytes is more than spray air freshener . This is related to the particle size of the gel air freshener. Chemical particle size gel air freshener in its use will be very small
chemical particles are 1μm (Ruzer, 2013). The smaller the particle size of the
gel air freshener better able dispersed into the cornea and activate the dormant keratinocytes corneal irritation not become active, proliferate and synthesis matrix
metalloproteinases in the process of tissue repair irritated (Wilson, 2007).