Comparative Analysis of Stillman’s and Modified Bass Brushing Method Using Carabao
Mango (Mangifera indica) Twigs as Chewing Stick Against Bacterial Growth
A Research Paper Presented to the
Faculty of Science, Technology, Engineering, and Mathematics
FEU High School
In Partial Fulfillment of the Requirements for the Course:
Practical Research 2
Alvarez, Zimon Gabriel A.
Barrozo, John Emmanuel S.
Carlos, Rafielle Raine O.
Diaz, Zylem Mitzwilroz Q.
Estanislao, Kevin Roy C.
Payot, Precie Jane E.
Sison, Yshiniah Ellora Rogue G.
Usita, Lark Marione P.
Lorane G. Padilla
Research Adviser
January 2023
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CHAPTER 1
THE PROBLEM AND ITS BACKGROUND
This chapter serves as the study's introduction, providing fresh insights into the research
that will be expanded upon in subsequent chapters. This chapter will have the following sections:
Background of the study, conceptual framework, statement of the problem, research hypothesis,
significance of the study, scope and delimitations, definition of terms, respectively.
Background of the Study
With the present focus on COVID-19, many had forgotten about other prominent diseases
that are now widespread. Among them is oral disease, with dental caries (tooth decay) being more
widespread than other dental issues. Oral disorders, as mentioned by Dr. Peres (2019), are among
the most widespread diseases worldwide and impose significant health and economic
consequences, significantly diminishing the quality of life of people affected. According to Deo
and Deshmukh (2019), the oral cavity contains about 700 species of bacteria, the second biggest
and most varied microbiota after the gut. It provides a habitat for countless species, including
bacteria, fungus, viruses, and protozoa. Oral bacteria can cause bacterial infections, dental caries,
periodontal disease, and other common mouth diseases in children and adults. People who have
dental diseases may have a difficult time because of its impact on human oral hygiene; however,
this issue could be resolved by inhibiting bacteria through brushing or cleaning one's teeth.
Dental diseases can be avoided, and regular brushing prevents tooth decay and gum disease
by reducing the quantity of bacteria and plaque on and around the teeth and gums. Keeping the
teeth clean by brushing twice a day is the primary line of protection; but, if it is not executed
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properly, dangerous bacterial plaque can slip in and cause tooth decay and gum disease. There are
a variety of brushing methods that can effectively prevent oral diseases and two of these are the
Modified Bass brushing method and the Stillman’s method. The Modified Bass method emphasizes
plaque removal from the region slightly above and below the gingival edge. According to Baruah
et al. (2017), this method is beneficial for those with periodontitis. On the other hand, the Stillman’s
method is recommended to be applied on gingival inflammatory areas with a broad distribution
(Valiathan & Suhasini.S, 2020). These brushing methods can enhance the efficiency and
effectiveness of the oral health environment.
Even if toothbrushes and toothpaste are highly available on the market, the impoverished
and socially excluded groups are not prioritizing buying them (Saini, 2018). The first toothbrushes
that were originally made from natural materials have now been replaced by nylons or plastics, and
with no other choice but to maintain having clean teeth, people can only purchase them at varying
prices (Borunda, 2019). But people from underdeveloped nations who have poor living conditions
have limited to no access to dental supplies, and because of that, they are at risk of developing
tooth decay, gum disease, bad breath, and other serious health issues (Peres et al., 2019; Geisinger,
2017).
Nirupad et al. (2017) did a study in which they found that extracts from Mangifera indica
and eucalyptus branches were particularly good at killing bacteria. They found that the extract from
Mangifera indica twigs was more effective against germs and had a larger zone of inhibition than
the extract from eucalyptus twigs. They also said that "the astringent tannins, bitter gum, and resins
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in Mangifera indica twigs protect the enamel and form a layer over the mucous membrane." Based
on the findings in the research made by Chandana et al. (2016), Mangifera indica has antibacterial
properties and was effective against both Streptococcus mutans and Lactobacillus acidophilus. It
is an efficient chewing stick that could inhibit bacterial growth inside the mouth.
This study uses a quantitative research approach, and the design is experimental, as it
compares the efficiency of the different brushing methods that can inhibit the bacterial growth
inside the human mouth. The researchers seek to determine which brushing method may be used
to combat bacterial growth in a person's mouth while using a Carabao Mangifera indica twig as a
chewing stick. The research’s target population are individuals ages 18 and above and should be
able to at least know how to hold a toothbrush. This target audience is chosen simply because they
will have better understanding of the topic and may assist the study's aim and the results based on
their opinions and experiences regarding the matter and the factors that led to each of their own
inferences.
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Conceptual Framework
Figure 1. The Difference between the Stillman’s Brushing Method and Modified Bass Brushing Method
Using Carabao mango (Mangifera indica) Twigs as Chewing Stick Against Bacterial Growth
Inside Homo sapiens sapiens' Mouth
The conceptual structure of the study is shown in Figure 1 named "The Difference between
the Stillman’s Brushing Method and Modified Bass Brushing Method Using Carabao Mangifera indica
Twigs as Chewing Stick Against Bacterial Growth Inside Homo sapiens sapiens' Mouth" This research
employs the independent variable-dependent variable model (IV-DV). It includes two independent
variables: (1) Carabao Mangifera indica twigs as Chewing sticks using Stillman's Brushing Method
and (2) Carabao Mangifera indica twigs as Chewing sticks using the Modified Bass Brushing
Method. The dependent variable in this study is the bacterial growth inside a Homo sapiens sapiens'
mouth. The other variables included are the control variables which may have varying effects on
the efficiency of Mangifera indica twigs as a chewing stick against bacterial growth inside Homo
sapiens sapiens' mouth using the Stillman’s brushing method and the Modified Bass brushing method.
First, there is the bristle length, which is a respective 1.00 inch in length. Second, the ideal amount
of time spent brushing teeth in order to reduce the number of bacteria that can grow in the human
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mouth is two minutes. The oral features of the participants are the last control variable, and to
remove any possibility of bias, the participants must have healthy oral features with no records of
any dental issues.
Statement of the Problem
This study aims to determine the most effective brushing technique for cleaning one's teeth
to inhibit the growth of bacteria in the mouth while using a Carabao Mangifera indica twig as a
chewing stick. Specifically, the researchers of the study seek to answer the following questions:
1. What is the bacterial count in colony forming units after the application of Mangifera
indica twigs as chewing sticks with the following brushing method:
1.1. Stillman's Brushing Method
1.2. Modified Bass Brushing Method
2. Is there a significant difference between the bacterial count before and after the
application of Stillman’s and Modified Bass brushing method?
3. Is there a significant difference in the bacterial count between the two brushing methods?
4. Which is the most efficient brushing method for inhibiting bacterial growth in the human
mouth in terms of using Carabao Mangifera indica twigs as chewing sticks?
Specific Objectives:
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1. To determine the bacterial count in colony forming units after the application of Carabao
Mangifera indica twigs as chewing sticks with the following brushing method:
(1) Stillman's Brushing Method
(2) Modified Bass Brushing Method
2. To find out the significant difference between the bacterial count before and after the
application of Stillman’s and Modified Bass brushing method.
3. To discover if there is a significant difference in the bacterial count between the two
brushing methods.
4. To determine which brushing method is the most efficient for inhibiting bacterial growth
in the human mouth while using Carabao Mangifera indica twigs as chewing sticks.
Research Hypotheses
The researchers intend to investigate the difference of Carabao Mangifera indica twigs’
suppression of bacterial development in the mouth when used with two different brushing
strategies. According to Dubuqui (2019), the Modified Bass brushing method and Stillman's
brushing method are highly comparable and significant in terms of how they clean the teeth and
gums but differ mainly in how they are executed. Using that knowledge, the researchers offer two
hypotheses that seek to explain the anticipated result of the study:
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H0 : There is no discernible difference between the two brushing methods while using a
Carabao Mangifera indica twig to reduce oral bacterial growth.
H1 : Inhibiting oral bacterial growth has shown to be significantly different between the two
brushing methods with the Carabao Mangifera indica twig.
Significance of the Study
Oral disorders have been prominent for a very long time, and according to the World Health
Organization (2022), it is estimated that about 3.5 billion individuals suffer from oral diseases. This
research compares the effectiveness of Stillman’s brushing method and the Modified Bass brushing
method in inhibiting bacterial growth in the human mouth while utilizing Carabao Mangifera
indica twigs as chewing sticks. Moreover, the following will benefit from the study's findings:
The researchers will benefit from this study considering that they are Science,
Technology, Engineering, and Mathematics (STEM) students specializing in Biomedical Sciences.
This research will pique the interest of aspiring medical professionals, such as the researchers, in
Biomedical Problems that the world is now encountering. It will be used by the researchers to gain
understanding and enhance learning about the subject matter. Furthermore, their inquisitiveness
and curiosity will be ignited to the extent that they will want to consider pursuing a career in dental
medicine.
Students, With the aid of the study, students will be more knowledgeable of the
significance of brushing their teeth and the significance of brush it the correct way. This study will
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further inform them on the many ways to clean their teeth, as well as the potential consequences of
not brushing every day.
Parents, The findings of this study may help parents realize how vital it is to teach their
children to brush their teeth every day with the proper brushing method and that there are natural
alternatives to plastic toothbrushes that are also effective and better for the environment.
Medical Institutions, The findings of this study may be advantageous for medical
institutions since they will have another alternative for treating oral diseases. Aside from that it
also gives knowledge to them by being introduced to a new alternative.
The researchers will benefit from this study considering that they are Science,
Technology, Engineering, and Mathematics (STEM) students specializing in Biomedical Sciences.
This research will pique the interest of aspiring medical professionals, such as the researchers, in
Biomedical Problems that the world is now encountering. It will be used by the researchers to gain
understanding and enhance learning about the subject matter. Furthermore, their inquisitiveness
and curiosity will be ignited to the extent that they will want to consider pursuing a career in dental
medicine.
Future Researchers, Future researchers can benefit from this study for the data that they
will be able to withdraw and will be used in their own study. Having the study’s data and their
own, the information of the researchers will improve not only in quality but also in reliability
addition to that is the greater depth that the study will be when finished.
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Scope and Delimitations
This study aims to determine the significant difference between two distinct brushing
techniques using a Carabao Mangifera indica twig chewing stick in preventing oral bacterial
growth. In addition, the experiment will be undertaken at Neomedica Ambulatory Care Clinic. The
research will be conducted in vivo, which means the experiments will be conducted inside a living
organism, in this case the human mouth (Dornell, 2021). Following collection, the participant's
mouth swab will be examined in a laboratory. The research will be experimental in nature and will
employ a method of purposive sampling. The researchers will modify the independent variable
alongside its control variables in order to determine its effect on the dependent variable. The period
permitted for the experiment, which includes the time required to prepare supplies and carry out
the method, will be at least seven days. Due to the need for precise results, there will only be six
research participants, each of whom will get two (2) days to prepare for the experiment. Certain
intervening variables may have a substantial impact on the outcomes, such as the food consumed,
which may contain substances that promote or prevent the growth of oral bacteria. The individual
may have consumed acidic foods, which inhibit bacterial development, or protein-rich foods, which
encourage bacterial growth. The individual's mood can also influence the outcomes since there are
instances in which they are so exhausted that they forget to brush their teeth. This may boost
bacterial growth and the potential outcome of the investigation. Other intervening variables may
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include the individual's brushing habits, such as whether they wash their teeth daily and how often
they do so. Moreover, a budget of roughly 15,000 pesos would be allocated for the study's
materials. The budget will be utilized for expenses such as the materials that will be used to create
the product, the equipment required to execute the experiment, and other essentials required for the
study.
Commented [ZA1]: Paki-finish na ito please, andami pa
ring n.d.
Definition of Terms
The following terms are lexically defined for a better perspective and understanding.
Bacterium - Single-celled organisms that exist in millions, both inside and outside of other
organisms (Brazier, 2019).
Modified Bass Brushing Method - Brush each tooth with a gentle circular motion while holding
the toothbrush at a 45-degree angle to the gum line. Once you've thoroughly cleaned each tooth,
flick the toothbrush along the tooth and away from the gum line to cover all of the tooth's surfaces,
including the top, inner, and exterior surfaces. Lastly, continue to brush your tongue in a circular
motion. (TLC Dental, 2018)
Candida albicans – A fungus that lives on your body in small amounts, located in your mouth,
skin and intestines (Cleveland Clinic, 2022).
Carabao Mangifera indica - The Carabao Mangifera indica is the most prevalent and most
exported mango variety in the Philippines due to its exotic taste and sweetness (Ardepolla et al.,
2019).
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Cariogenic Bacteria - In which sugars are metabolized to create acid, demineralizing the tooth
structure over time (Rathee & Sapra, 2020).
Decay - The damage to the surface of a tooth, also known as enamel. It occurs when bacteria in
your mouth produce acids that destroy your tooth enamel. Cavities (dental caries) are holes in your
teeth caused by tooth decay (Tooth Decay, 2019).
Dental Carries - A process of demineralization of tooth enamel that results in enamel and dentin
destruction and tooth cavitation (Abou Neel et al., 2016).
Enamel - The strongest and most mineralized tissue in extinct and extant vertebrate species, and it
provides maximal resilience, allowing teeth to function as weapons, tools, and food processors.
(Lacruz et al., 2017).
Gingival - An inflammatory disorder of the gingival tissue caused primarily by bacterial infection
(Rathee & Jain, 2022).
Gingival margin - Follows the cementoenamel junction, and the tooth has a straight or slightly
concave surface as it exits soft tissue and continues into the mouth (ScienceDirect Topics, 2017).
Gingival recession - Happens when the gingival tissue's edge (called the gingival margin) slips
away from the tooth's crown (Gingival Recession, 2015).
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Gram-positive organisms - Exhibit a wide range of growth and resistance patterns (Sizar &
Unakal, 2022).
Inhibit - Repression or restriction of a physical or chemical action. “To inhibit” means to “hold in
or back” (Biology Online Dictionary, 2021).
Kasturi Mangifera indica - One of the vulnerable germplasms in South Kalimantan. It has the
shape of a little mango, a very sweet taste, is sticky, and has a distinct odor (Darmawan, 2015).
Lactobacillus acidophilus - A bacterium found in the mouth, intestine and vagina, is used as a
probiotic (Mayo Clinic, 2017).
Lysis – The breakdown of a cell caused by damage to its plasma (outer) membrane (National
Cancer Institute, 2019).
Miswak - An oral hygiene aid with widespread use among Muslim population due to religious
norms (Saha et al., 2012).
Mucous Membrane – A layer of moist skin that covers cavities that are open to the outside or
ducts of the body (Nutrition, 2022).
Peptidoglycan (murein) sacculus – A distinct and necessary structural component of most
bacteria's cell wall. The sacculus is a closed, bag-shaped structure made of glycan strands crosslinked by short peptides that surrounds the cytoplasmic membrane (Vollmer et al., 2008).
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Permeability – The volume of a fluid of unit viscosity moving through a unit cross section of a
medium in unit time under the influence of a unit pressure gradient (ScienceDirect Topics, 2016).
Plaque - A sticky film that forms on the teeth when bacteria in the mouth mix with sugary or
starchy foods (Cleveland Clinic, 2020).
Staphylococcus aureus - A gram-positive bacteria that causes a wide variety of clinical diseases
(Taylor & Unakal, 2018).
Stillman’s Brushing Method - The bristles are held at a 45° angle toward the gum line. Half of
the bristles should be covering the gums, and the other half of the bristles should be on the tooth
surface (Toothbrushing Techniques, 2015).
Streptococcus mitis – A viridans streptococcus that is a normal oropharyngeal commensal. It can
escape from this niche and cause a wide range of infectious consequences including infective
endocarditis, bacteraemia, and septicaemia (Mitchell, 2011).
Streptococcus mutans – A bacterial species linked to the development of dental caries in humans
and some other animals, as well as subacute endocarditis (Nomura et al., 2020).
Tannins - Secondary plant metabolites originally recognized because they interact strongly with
collagen (Mbaiogaou et al., 2022).
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Viscosity - The resistance to gradual deformation of a medium under shear stress (Hack, 2018).
CHAPTER 2
REVIEW OF RELATED LITERATURE AND STUDIES
This chapter presents the related literature and studies derived from global and local
contexts. The related literature is synthesized to form different themes which supports the
framework of the study. Furthermore, the related studies are categorized into foreign and local
studies to clearly discuss the problems and issues concerning the study.
Availability of Carabao mango twigs
The Philippines is considered a third-world or developing nation with a high infant
mortality rate, restricted healthcare options, poor gross domestic product (GDP) per capita, and
citizens gaining lower-middle incomes (Kovachek, 2022). According to Probst et. al (2019), people
with low income and poor level of education are particularly at risk for untreated dental caries
because they lack access to dental care services and good oral hygiene habits. As per the University
of Illinois Chicago (2019), having financial limitations usually prevent people from receiving
appropriate dental care, and due to having the inability to afford one, they are often prone to oral
health diseases. In addition to that, some people cannot afford pricey modern toothbrushes due to
financial restrictions.
According to the World Health Organization (2022), approximately 88% of people in
underdeveloped countries rely on traditional medicine as their major source of health care, with
several using chewing sticks to prevent dental caries. It has been claimed that traditional chewing
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sticks contain antibacterial, antioxidant, anti-inflammatory, and antiseptic properties. They also
contribute significantly to maintaining proper dental hygiene because of their mechanical action in
removing caries (decays), plaque, and others (Beshah, 2021). To this day, people all over the world
utilize chewing sticks for their many benefits, including their ability to clean teeth, its low cost, the
variety of therapeutic qualities it offers, and the ease with which they can be found in rural areas
of developing countries.
In the Philippines, 87.4% of Filipinos have dental cavities, while 48.3% were recorded to
have periodontal disease. More than 90% of dental caries go untreated in most low-income
developing nations, which is why the Philippines has difficulties fighting against oral health
diseases (Alfaro, 2017). With the problems brought about by poor dental care, natural chewing
sticks such as mango twigs are significant because they serve as a cheaper alternative to modern
toothbrushes and can help prevent pollution by reducing nylon and plastics.
According to Agri Farming (2022), about 1 million metric tons of Mangifera indica, or
3.5% of the world's production, are produced in the Philippines each year; 95% are produced for
domestic consumption. Moreover, Carabao Mangifera indica trees can be seen in the backyards of
most Filipino homes (Mendoza, 2021). This increased the amount of Carabao Mangifera indica
available, giving Filipinos the chance to produce their natural toothbrush with no needed cost by
only using the Mangifera indica twigs. Due to the Mangifera indica resilience to heat, it is not just
available in the Philippines but also other tropical developing nations and countries with higher
temperatures.
Efficiency of Mangifera indica Twigs
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Antimicrobial Properties
As dental caries continues to be prevalent in this society, with 2 billion adults and 520
million children suffering from permanent tooth caries in 2019, the Global Burden of Disease has
confirmed that this condition remains the most common diagnosis worldwide (World Health
Organization, 2022). This illustrates that dental caries is a serious condition that must be discussed
due to the large number of people it endangers. However, despite its prevalence, dental caries is
curable. Brushing one's teeth helps stop dental caries by preventing the formation of bacteria such
as S. Mutans and Lactobacillus in the mouth. As per Dr. Frissbee (2021) of WebMD, tooth decay
can be prevented by brushing the teeth at least twice a day, after each meal, and before bed. Despite
all this information about preventing such conditions, tooth decay remains prevalent due primarily
to frequent snacking, drinking sugary beverages, and poor dental hygiene of the masses (World
Health Organization, 2015). To address this problem, the researchers propose a toothbrush with
antimicrobial properties and a natural composition that is accessible to the general public.
Mangifera indica are abundant in the Philippines as to why it is also considered its national
fruit. In addition, it is said that all its parts also contain medicinal properties that are beneficial to
the body. According to Snyder (2021), Mangifera indica are abundant in vitamins, minerals, and
antioxidants and have been linked to numerous health benefits, such as potential anticancer effects,
as well as improved immunity, digestive, and eye health. In addition, studies also suggest that
extracts from Mangifera indica are good in isolating organisms causing dental caries. Mangifera
indica have multiple beneficial properties for oral health. It has unique antimicrobial properties
that allow for the penetration of the thick peptidoglycan layers of gram-positive bacteria (Kumar
et al., 2016). In the study conducted by Suhendar al. (2019), it was discovered that Kasturi
Mangifera indica extracts possess antibacterial properties that inhibit the development of dental
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caries. With a 25% extract concentration causing cell lysis or shrinkage and a 50% extract
concentration causing necrosis or cell death. The study proves that Mangifera indica do contain
antimicrobial properties that are effective against bacteria in the mouth. With that in mind,
Mangifera indica twigs are the primary research instrument of this study.
In 2012, The Indian Journal of Dental Research published a study on the antimicrobial
properties of four distinct chewing sticks, including Mangifera indica twigs. The researchers had
to develop aqueous extracts of all chewing sticks by combining 100 ml of deionized distilled water
with 100 mg of chewing stick powder to do the experimental research. The above procedure was
repeated until 5%, 10%, and 50% concentrations of each type of chewing stick were obtained. By
the end of the experiment, the research concluded that Mangifera indica twigs, in high
concentration, are effective against dental caries-causing bacteria such as S. Mutans and L.
acidophilus. Tannins, bitter gum, and resins are the reason for their antimicrobial activity against
a few microorganisms (Elangovan et al., 2012).
Resins are natural or synthetic organic compounds that consist of a non-crystalline or
viscous liquid substance in Mangifera indica twigs (Britannica, 2019). Yellow and viscous, the
natural resin contains antibiotic components that can double the lifespan of tiny consumers such as
ants. This sap is produced by plant secretions and is soluble in a variety of organic liquids but not
water. Tannins, on the other hand, are a category of polyphenolic substances that are abundant in
plant regions and contain a variety of biological properties, such as antibacterial, antiparasitic,
antiviral, antioxidant, anti-inflammatory, immunomodulatory, and many more. (Huang et al.,
2017). It is a type of plant polyphenol, which is a natural chemical found in fruits and responsible
for their astringent or bitter flavor. In addition to diminishing the nutritional value of foods, these
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chemicals bind to and precipitate proteins. Tannins are abundant in the seed, pulp, flesh, and
branches of Mangifera indica. In contrast to the two, bitter gum is a unique resin derived from trees
that have been utilized to improve digestion, oral health, and liver health. It also reportedly contains
antioxidants that support its therapeutic properties (Cronkleton, 2018).
Furthermore, Mangifera indica also has the following antibacterial properties; tannic acids,
saponins, and flavonoids. According to Kaczmarek (2020), tannic acids have antibacterial
properties that deactivate microbial cell adhesion, enzymes, and polypeptide cell wall. The
presence of tannin on the cell wall kills bacteria by liquefying its cell wall. On the other hand,
saponins are detergent-like compounds with antibacterial and anticancer properties (Arabski et al.,
2012). Saponin disrupts the stability of bacterial cell membranes, resulting in the rupture of
bacterial cells and damage to the cell membranes, as well as the release of proteins, nucleic acid,
and nucleotides from the bacterial cell, thereby preventing bacterial cell growth and development.
Flavonoids are well-known antibacterial agents that are effective against a wide variety of
pathogenic microorganisms (Xie et al., 2015). Flavonoids can deactivate proteins so that they
interfere with bacterial cell metabolism, and the interactions between flavonoid and bacterial DNA
(Deoxyribonucleic acid) alter the permeability of the cell wall, microsome, and lysosome.
As stated by Chandana (2016), good dental health and good teeth contribute to our body's
general health and well-being, which is why healthy oral well-being must be preserved. However,
for a very long time, it has been responsible for the world's most widespread condition, and it
remains prevalent. In response, researchers would conduct a study regarding Carabao Mangifera
indica twigs as chewing sticks against bacterial growth. The tannin, resin, and bitter gum-rich
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extracts, as well as tannic acids, saponins, and flavonoids, are the antimicrobial properties of
Mangifera indica that can inhibit bacterial growth.
Inhibition Efficiency Against Bacterial Growth
Poor oral hygiene, which includes bad breath, dry mouth, gum diseases, and others, has
been a problem for many. Among these side effects, tooth decay is one of the most prevalent.
According to Colgate (2020), entire colonies of bacteria reside in human mouths. Although the
average person only has 34 to 72 distinct types of bacteria in their mouths, over 700 different strains
have been found there. As far as human health is concerned, most of these bacterial species seem
to be unharmful. However, certain bacteria are bad for people since it leads to gum disease and
tooth decay.
Different microorganisms thrive in various environments. Others adhere to the tongue
while some attach to the teeth. These microorganisms include bacteria, fungi, and other pathogens.
Sugary meals and beverages provide sustenance for some microbes, which in turn helps them
proliferate and spread throughout the mouth. Some of the sugar-loving microorganisms can convert
sugar to acid and matrix. The enamel of a person's teeth is destroyed by acid (News in Health,
2019). Nevertheless, Mangifera indica twigs have properties that can protect the enamel. Nirupad
et al. (2017) quoted that "the astringent tannins, bitter gum, and resins in Mangifera indica twigs
protect the enamel and form a layer over the mucous membrane."
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Mangifera indica twigs have barks surrounding them that when extracted, increases the
inhibition of bacterial growth. At extracts of 0.15 ml and 0.10 ml, the mean of minimum inhibitory
concentration stays the same with a mean of 3.90 while for the 0.5 ml extract, the mean increased
to 4.17 total (Eziamaka et. al, 2022). In addition to that, according to a study conducted by Sari et
al. (2020), they used the minimum bactericidal concentration of mango as an extract requiring no
more than 30 mg/ml. At this concentration, the bacteria have already been killed thus showing
evidence of antimicrobial properties present in its chemical composition. Once extracted and
exposed to bacteria, it reacts with certain parts of the cell which causes bacterial growth inhibition
or termination.
In a study conducted by Anand et. al (2015), Mangifera indica twigs were compared to
cashew extracts to investigate which of the two can be efficient mitigators of periodontal and
gingival diseases. Among the compared extracts, Mangifera indica twigs had the lowest amount
of concentration which induces bacterial growth inhibition (Minimum Inhibitory Concentration)
and antimicrobial properties (MBC). A lower amount of concentration for bacteria is better since
that means less of the extract is needed to inhibit the growth of bacteria.
Another reason Mangifera indica twigs are efficient is the capability to reduce the amount
of bacterial growth in the mouth thus improving overall gingival health and reducing plaque
formation as well as increasing salivary pH levels. In a study conducted by Dandekar & Winnier
(2020) in which the research utilized the one-way analysis of variance (ANOVA), wherein the
results from experimentation of Mangifera indica extract from baseline to the 21st day were
compared. In all the tests, there were significant differences between the means of the baseline, the
7th day and the 21st day. Which means the salivary pH level was increased for every time the
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experimentation occurred. The results were a ph level of 5.93, 6.40, 7.10 on the baseline, 7th day,
and 21st day respectively. a pH level close to, equal, or lower than 5.5 promotes bacterial growth
within the mouth which causes tooth decay (Does the PH of Your Mouth Matter, 2019.).
Furthermore, throughout the experimentation, the gingival score has greatly decreased using
Mangifera indica extracts. The results were a gingival score of 1.240, 0.960, 0.597 on the baseline,
7th day, and 21st day respectively. A gingival score of close to or equal to 1 means there is mild
inflammation while a score of 0 indicates no inflammation in the gums. Inflammation in the gums
is caused by bacteria from poor oral hygiene. (Periobasics, 2020). Lastly, the plaque score was
greatly reduced by the Mangifera indica extract. The results were a plaque score of 0.550, 0.450,
0.237, on the baseline, 7th day, and 21st day respectively. A lower plaque score suggests a lower
number of bacteria present in the mouth.
Ultimately, Mangifera indica twigs can inhibit bacterial growth in the mouth due to
chemicals present inside the twig itself which has antibacterial properties lethal to gram positive
bacteria. Furthermore, when compared to other well-known plant species, Mangifera indica twigs
are the most potent in terms of its minimum inhibitory concentration (MIC). It also increases the
salivary ph levels and lower gingival and plaque scores.
Significance of Brushing Methods
Stillman’s Brushing Method
Having poor dental conditions required great attention in order to not worsen the situation.
Brushing one's teeth can help prevent tooth decay which can lead to dental cavities, and aids in the
prevention of foul breath and gum disease. Brushing removes germs, which are the major cause of
oral health issues, and prevents the production of plaque. Refusing to brush one's teeth for multiple
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days might result in the development of gum and dental diseases (Community Health of Central
Washington, n.d.). Although there are many ways to clean or brush teeth, some might harm the
tooth and gum tissues. With the appropriate brushing method, keeping the teeth clean and mouth
healthy is easy. According to Christian Johnson D.M.D. (2017), without the proper method, one
may be missing crucial regions of germs and food accumulation that will not be removed by
flossing or washing. Therefore, numerous brushing techniques have been advocated by dental
specialists to treat various oral health concerns. These brushing techniques certainly improved gum
and tooth health without causing harm. According to Colgate (2022), oral hygiene is essential for
a healthy smile and general wellbeing, and brushing under and around the gum line, where bacteria
and plaque gather, can prevent, and treat gum illnesses.
One of the efficient brushing techniques recommended by dental specialists is Stillman's
brushing method, which was introduced to the globe in 1932 and developed as gingival massage
and soft tissue simulation (Sang Mi Jeon et al., 2015). Here, the bristles should be oriented 45
degrees toward the gum line, with half of the bristles on the gingival margin and the other half on
the tooth surface (Davis Dental Care, 2015). The bristles of the brush are partially on the cervical
third of the teeth and partly on the gingival border at an oblique angle to the tooth's long axis (Dr.
Himanshu Ajwani, 2017). Then, occlusal movement followed by brief horizontal back-and-forth
strokes or movements on the linked gingiva, gingival margin, and tooth (Dentistry zone, 2022).
Such technique requires brushes with soft to medium bristles. Otherwise, bristles may cause
gingival injury if they are too abrasive. As stated by Westermeier Martin Dental Care (2019),
Stillman’s brushing method is effective in cleaning more debris from between the teeth, cleaning
the gingival recession, and root exposure to prevent tissue destruction which made the method
recommended for people who have gingivitis.
23
Having a brushing technique can inhibit the spread or growth of any bacteria or plaque in
the human mouth. To maintain this, according to Larissa Hirsch (2021), one must brush their teeth
twice a day to keep them healthy and prevent arising of bacterial growth. Stillman’s method
involves toothbrush bristles placed initially on the gum, (Karen Kaiser, 2013) applying gentle
pressure, vibrating and rolling the bristles with a slow and controlled motion interproximal and
over the tissues. In short, this method requires a soft tufted brush and little pressure. According to
Lisa Haney (2022), brushing too hard can harm one’s teeth and gums, causing enamel erosion and
receding gums, which can contribute to dental sensitivity. Brushing including the upper and lower
teeth gently, going tooth by tooth might assist in slowing down and not missing any spots.
Overbrushing is possible but brushing too hard can cause abrasion, gum recession, and discomfort
(Colgate, 2022). Stillman’s method required light pressure; hence possible to have a high
possibility of less discomfort and effectivity of inhibition of bacterial growth.
Stillman's approach gives good gingival massage and inter-proximal cleaning, which makes
it effective and one of the ways recommended by professionals for dental safety and cleansing.
Modified Bass brushing method
Oral health is the condition of a person's mouth; poor oral health can lead to oral ailments
such as cavities, tooth decay, foul breath, and others. According to Dr. Suhasini (2020), these
disorders are caused by improper plaque removal from the teeth. Because it removes plaque and
limits its growth, cleans the teeth, stimulates the gingival tissues, and applies dentifrice with the
necessary characteristics to prevent diseases, brushing teeth is crucial to an individual's oral health
(Baruah 2017). Therefore, it is essential to know the right procedures and methods for preventing
24
bacterial growth inside the mouth. A review article written by Janakiram et al. (2018) shows that
between the five studies that measured the plaque scores of each participant, The Modified Bass
approach was found to be more effective at removing plaque than the standard toothbrushing
techniques in a meta-analysis of the studies by Poyato et al. and Harnacke D. et al. (2018).
Additionally, another study showed that the Modified Bass was superior to Stillman's technique in
terms of removing plaque. Lastly, a meta-analysis was also conducted between the Modified Bass
technique with Routine technique. The quantitative analysis had a 95% significance rate with a 1.22-confidence interval (-1.62, -0.81). with a 30.30, p<0.00001 and I2 =97% Chi-square value.
Revealing that the Modified Bass technique is efficient in terms of plaque removal.
Commented [ZA2]: paki-elaborate toh like ano ung
isinagawa niyang experiment to prove the conclusion na
nakuha niya
Commented [KE3R2]: ok na ba to
According to Kumar's (2018) research, brushing practices significantly inhibit
bacterial growth and preserve the integrity of the tooth and its surrounding periodontium.
Moreover, with the addition of brushing techniques the efficiency of removing bacteria in the
mouth improves significantly and this is why having techniques such as modified brushing
technique is important. As per Dr. Suhasini (2020), the Modified Bass technique has the advantages
of providing good gingival stimulation, being flexible for open interproximal and cervical areas,
and being effective in plaque removal beneath the gingival margin. It is time-consuming and can
result
in
gingival
damage.
As stated by Sensodyne (2015), the Modified Bass technique is the technique of brushing
that is most extensively used and most effective for adults. In terms of its step, it requires the
toothbrush to be held at a 45-degree angle towards the tooth surface and the bristles should contact
both lines of the gums and tooth. Next, move the brush with a circular motion and clean the surface
of the back teeth and up and down strokes for the inside surface of the teeth. And lastly, for the
25
inside surface of the teeth tilt the brush vertically with a motion of up and down strokes and the
biting
surfaces
of
the
teeth
with
a
back-and-forth
motion
In conclusion, the various toothbrushing techniques, such as the Fones, Leonard,
Stillman’s, Charters, Bass (Sulcular), rolling stroke (press roll), Smith-Bell, and horizontal scrub
techniques, are proven to show great applicability in terms of cleaning the facial, lingual, and some
occlusal surfaces of the teeth. However, when it comes to cleaning the sulcus and interproximal
areas, only the Modified Bass technique is proven to be effective. It is also the first technique to
concentrate on cleaning the gingival sulcus of plaque and debris (Baruah, 2017). Therefore, proving
the
effectiveness
of
utilizing
the
Modified
Bass
brushing
technique.
26
Commented [KE4]: pa chek mga idolo
CHAPTER 3
METHODOLOGY
This chapter introduces the methods and the techniques that the researchers used to gather
their respondents and what the researchers used in order to produce and analyze the data from the
respondents. This chapter also identifies the research design in which the variables in the study are
introduced.
Research Design
This research employs a quantitative research approach using an experimental research
design. According to Voxco (2022), the procedure of gathering and interpreting numerical data and
using it to discover trends, averages, forecasts, as well as cause-and-effect connections between
the variables being investigated is referred to as quantitative research. An experimental method of
research is utilized in this study. As per Pubrica-academy (2022), experimental research designs
necessitate the collection and statistical analysis of quantitative data during the project. These
designs are most often associated with laboratory testing procedures. The concept of an
experimental research design is a research strategy intended to explore the interconnection between
independent and dependent variables in order to discover a cause-and-effect relation (Siago, 2022).
The influence of independent factors on dependent variables is usually observed and documented
over time to assist researchers in coming to a reasonable conclusion regarding the relationship
between these two types of variables (Pubrica-academy, 2022). In this study, the brushing method
using Carabao Mangifera indica twigs as chewing stick is the independent variable, while the
bacterial growth inside Homo sapiens sapiens’ mouth is the dependent variable. This study
evaluated the interaction between the aforementioned independent and dependent variables, which
27
explains the difference in bacterial growth between Modified Bass and Stillman’s as brushing
methods when utilizing Carabao Mangifera indica twigs as chewing sticks.
Research Locale and Sampling Technique
The research was carried out in The NeoMedica Ambulatory Care Clinic, Quezon City. It
was chosen for its ideal location and the availability of tests required for the experiment. A letter
of intent was delivered to the qualifying respondent as well as the research adviser prior to the
commencement of the data collection process. In addition to this, the researchers made it a point
to clarify and expound on the purpose of the study as much as possible in order to answer any
questions or concerns raised by the participants. The researcher sought the advice and direction of
professionals with extensive knowledge in bacteriology and microbiology. After that, the
respondent signed a letter of consent as a formal acceptance to participate in the study. This served
as an indication that the participant was willing to take part in the research. In addition, the
researchers used a method called purposive sampling to enroll participants who were suitable for
the experiment the study. This sampling technique is chosen to select participants with specific
characteristics that help answer the research question (Nikoloupuoluo, 2022). In this research, the
chosen participants had braceless teeth and the same oral features. This was done to lessen the
variables that may affect the results of this study.
Materials and Instruments
The critical data collectors were the researchers. The study's general technique was the
suppression of bacteria by using a bacteria culture test. During this investigation, six individuals
28
were selected to have swabs on their teeth and gums. The participants were required to wash their
teeth before beginning the experiment. The participants are then asked to consume food and liquids
before being swabbed. The participants were then instructed to brush their teeth and consume food
for a second time. After an hour, they rewashed their teeth, this time with a Carabao Mangifera
indica branch, and later swabbed their teeth. The swabbed bacteria sample was then transported to
test tubes for analysis in a laboratory testing facility. The sample in the test tubes was then subjected
to a culture and sensitivity test. The researchers ensured that the samples were delivered carefully
to the testing facility. Researchers assured that selected participants were physically and orally
healthy for the process. Participants in the study were instructed to use a branch as a toothbrush
and either Stillman's or the Modified Brass Brushing Method. Three responders were instructed to
perform the first option, while three others were advised to complete the second. Two minutes were
spent using the Mangifera indica twig toothbrush with one-inch-long bristles.
Mangifera indica twigs: Mangifera indica twigs were the primary material used because of the
research surrounding their efficiency. These were collected in San Fabian, Pangasinan, during field
visits.
Test tubes: Test tubes were used in the study as a vessel for the samples that will be tested and
observed. These test tubes were brought carefully to a testing center.
Sterilized swab: These swabs were used to collect bacterial colonies, move microorganisms, and
put samples on culture media plates.
29
Data Collection Procedure
1. Mangifera indica twigs collection and identification
Materials required to execute experimentation were collected. One week was given to the
researchers to provide ample time to gather the materials for the experiment. The materials needed
were the Mangifera indica twigs, agar plates, and inoculated loop required to acquire the results.
During field visits in San Fabian Pangasinan, a Mangifera indica expert and a guide helped
researchers identify Mangifera indica bushes and collect the twigs needed for the study. Because
Ilocos is a significant Mangifera indica producer, the researchers chose San Fabian Pangasinan for
data collection. Based on the Philippine Statistics Authority (PSA) data, as of 2021, Ilocos Region
produced 124.68 thousand metric tons of Mangifera indica, 22.4 percent of the national total,
followed by SOCCSKSARGEN and the Zamboanga Peninsula. Other mentioned materials, such
as test tubes and sterilized swabs, were all provided by the chosen testing center.
2. Gathering of research participants
There was a total of six research participants gathered. To complete the number of
participants required, this research used a purposive sampling technique, a non-probability
sampling technique wherein the participants were chosen due to specific characteristics that they
possessed that were suitable for the experiment.
30
3. Selection of Testing Center
Once the six research participants were gathered, the selection of a testing center where the
population of the bacteria was counted was selected. The testing center was the NeoMedica
Ambulatory Care Clinic in Quezon City. In this institute, the utilized test was a culture and
sensitivity test, costing about 1000 pesos per sample. The culture test was used to determine the
number of infectious bacteria within the sample while the Sensitivity test was utilized to determine
the efficiency of the Mangifera indica twigs against the cultivated bacteria (Culture and Sensitivity,
2021). Due to this, the researchers prepared a budget of 15 000 pesos, including the cost of the test
for all the samples collected and others needed for this research.
4. Cultivation of Bacteria
To cultivate bacteria, the six research participants waited an hour after initially brushing
their teeth using a plastic toothbrush. Tooth decay develops after one whole week of not brushing
the teeth (Belmont dental, 2021). Whereas plaque forms after two hours, resulting to irreversible
bacterial colonization, gingivitis, and other oral illnesses over extended periods (Pandula, 2012).
In addition, it takes bacteria 20 minutes to complete binary fission (Binary fission, n.d.); therefore,
an hour was plenty. Making an hour the optimal time for cultivating the needed bacteria since
bacteria multiply throughout the mouth while avoiding plaque formation and tooth decay.
5. Preparation of Mangifera indica Twigs
31
Twelve Mangifera indica twigs were collected, 6 of which were processed into a
toothbrush. At the same time, the remaining six were set aside and only used as a contingency in
case of errors that may occur during experimentation. The collected twigs were girdled 1 inch from
one end. Girdling is a process wherein a chosen species' twig’s outer layer is removed, exposing
the inner ground tissue of the plant (Beaulieu, 2022). This is primarily done to kill a plant or tree;
however, its purpose for this research was to expose the inner layer, which became the bristles of
the brush. Then the twigs were placed underwater, making the girdled part soft. Afterward, the
research participants will chew the exposed part making bristles. The saliva further softens the
girdled part, and the crushing motion of the chewing separates the fibers of the twig forming the
bristles of the brush.
6. Brushing of teeth using the Carabao Mangifera indica twigs
Once the Carabao Mangifera indica twigs had been prepared, the research participants who
did not brushed their teeth for one hour used the twig as their toothbrush using two different
brushing methods at two different stages of the experiment. These two brushing methods are well
known for being more efficient than other methods. These were the Stillman's Brushing Method
and Modified Bass Brushing Method. Both methods include a 45-degree angle to remove plaque
above the gumline. The only difference is that the Modified Bass method must be gentler, with
slight vibratory motions instead of long horizontal linear motions, since its primary purpose is to
aid patients with periodontitis (Davis Dental Carte, 2015). The experiment was divided into two
phases with the before and after results.
32
1st phase
In this phase, the six research participants brushed their teeth using a regular
plastic toothbrush, after which they were all given the same meals ideal for
cultivating bacteria within the mouth. This meal consisted of 2 slices of bread, dried
fruits, and a carbonated soft drink. Once these foods were chewed, it was placed
between the participants' teeth as complex and simple sugars such as scratches,
which makes this ideal for cultivating bacteria in the mouth (Kubala, 2021). The
participants then waited an hour to cultivate another batch of bacteria. Afterward,
the same participants were all swabbed with a sterilized swab containing the
samples placed within test tubes. These test samples were rushed to the chosen
testing center within an hour, where the estimated population of their bacteria was
counted. As specified by the testing center, culture and sensitivity tests were to
calculate the number of bacteria. The purpose of his phase was to gather the number
of bacteria living within the mouths of the participants before brushing using
Carabao Mangifera indica twig chewing stick, the results of which were later
compared to the results of the second experiment.
2nd phase
In this phase, the six research participants waited for another hour after the
initial phase to cultivate another batch of bacteria in their mouths. The six
33
participants were divided into two groups that consisted of three members each. The
first group used the prepared Mangifera indica twig toothbrush with a 1-inch bristle
length using Stillman’s brushing method, which lasted for 2 minutes. The second
group used a Mangifera indica twig toothbrush with the same bristle length using
the Modified Bass method, which lasted for 2 minutes. The participants were all
swabbed using a sterilized swab with the same procedure using a sterilized swab.
The samples were all placed within a test tube and rushed to the chosen testing
center within an hour where the estimated population of bacteria were counted.
A total of 12 samples were collected, 6 of which were collected from the first phase, while
the remaining six were collected from the second phase. The estimated population of bacteria from
the chosen testing center were recorded as the data that was later compared to determine which
brushing technique was best used with the Mangifera indica twig toothbrush.
Figure 2. Data collection procedure for removal of bacteria From Human Mouth Using Carabao mango
Twig Toothbrush
34
35
Data Analysis Procedure
The data was analyzed through statistical analysis, particularly descriptive and inferential
analysis. As per Bhandari (2020), inferential analysis is the use of measures from the experiment's
sample of individuals to compare treatment groups and generate conclusions about the greater
population of subjects. Whereas descriptive analysis is a type of data analysis that assists in
describing, displaying, or summarizing data points in a constructive manner so that patterns may
form that satisfy every condition of the data (Rawat, 2021). Among the several statistical tests, the
researchers determined that the experiment would be best examined using the t test, a statistical
test used to compare the means of two groups (Bevans, 2020). Given that the experiment only
required one sample involved in different variables, the researchers determined that a pairedsamples t test would be utilized. The experiment required six participants that underwent the
experiment with different brushing methods and was tested before and after the experiment, which
is why the paired sample t test best fitted the experiment.
The experiment yielded to the bacterial count of all six participants before and after the
experiment leading to 12 samples overall. These samples were divided into the bacterial count
before the experiment and the bacterial count after undergoing the experiment; both were averaged
in order to get the average number of bacteria before the experiment and the average number of
bacteria after brushing their teeth using the Stillman’s and Modified Bass brushing methods. After
which the numbers were compared in order to know the difference in terms of the bacterial count
before and after the experiment.
The outcomes of the Stillman's and Modified Bass methods were compared following the
experiment. This was accomplished by obtaining the average bacterial count after the experiment
36
and comparing it to the average of the other brushing method to see whether there was a significant
difference between the two, hence rejecting or failing to reject the null hypothesis.
Employing the t-test, the value of using Carabao Mangifera indica twigs with two different
brushing techniques to suppress bacterial development was evaluated. If the numerical data are
significantly different, the null hypothesis may be rejected. However, suppose the value calculated
falls under the non-reject zone. In that case, it may indicate that there was little to no significant
difference between the Stillman’s and Modified Bass brushing method when using Carabao
Mangifera indica twigs in terms of the inhibition ability of bacterial growth, leading to the
possibility that the null hypothesis is true.
37
Ethical Considerations
The participants' comfort, security, and safety were given priority during the research.
When volunteering for the research study, participants were required to sign a consent form before
the experiment is conducted. If at any time the participant decides they would like to stop taking
part in the experiment, they may do so without further delay. Additionally, no physical or
psychological harm or danger of any kind may be inflicted onto anyone who participated in the
activity. To avoid harming the participants psychologically, the questions provided by the
researchers must avoid any personal questions that may trigger emotional traumas that cause
distress. The research must be regarded as invalid if any are harmed because of the
experimentation. Lastly, all the information gathered by the researchers from the respondents shall
be kept strictly confidential.
For the duration of the research, the following ethical considerations were implemented:
1. The researchers will seek the participants' informed consent that they are willing to participate
in the experiment.
2.Research participants will be provided with pertinent information regarding the research.
3. Every participant has the option to participate or not when the researchers explain the process of
the experiment.
4. The researchers will acquire the participants' consent to use either their true identities or
nicknames in the research report.
5. The researchers will not collect personal data without the consent of the participants.
38
6. The well-being and dignity of the participants were always upheld.
7. Any harm—physical, social, psychological, or otherwise—w will be avoided or limited as much
as practicable and,
8. Assurance that the work is original and devoid of research misconduct or plagiarism.
39
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