CHAPTER I

INTRODUCTION

Background of the Study

Wounds are a major cause of physical disabilities. These

result in an opening or break of the skin that causes a

disruption on the skin’s normal function. Since wounds are

experienced by almost all people, it is only natural that the

proper handling of wounds should be known so that no

infections will occur throughout the course of the healing

process.

Wounds can be classified as acute or chronic depending on

its healing time. Acute wounds heal without complications in

the expected amount of time while chronic wounds take a longer

time to heal which might have the risk of infection.

Furthermore, wounds can also be classified as open or closed.

Open wounds are those with exposed underlying tissue that are

open to the outside environment while closed wounds have

damage that occurs without the underlying tissue exposed.

Wound healing process start from damage of skin. A wound

completely healing depends on degree of injury, human

resistance capacity, infection potential of pathogens and

early effective treatment procedure. The wound healing process

is divided into four parts, mainly Hemostasis, Inflammatory,

Proliferative, and Maturation Phase. Hemostasis, the first

1
phase of healing, begins at the onset of injury, and the

objective is to stop the bleeding. In this phase, the body

activates its emergency repair system, the blood clotting

system, and forms a dam to block the drainage. The

second phase focuses on destroying bacteria and removing

debris—essentially preparing the wound bed for the growth of

new tissue. Once the wound is cleaned out, the wound enters

the Proliferative Phase, where the focus is to fill and cover

the wound. The last phase is the Maturation phase, wherein the

new tissue slowly gains strength and flexibility. Here,

collagen fibers reorganize, the tissue remodels and matures

and there is an overall increase in tensile strength. The

healing process is complex and should not be taken lightly as

infections may occur when immediate and proper action is not

done.

In relation with these, people who are in rural areas do

not have the proper medication for wounds, that’s why the

utilization of medicinal plants is common. Science and

technology advanced the way of life of the people and paved

the way for commercial wound-healing. However, even before

such products were invented, people especially in cultural

groups had their own means of cleaning and treating wounds.

Due to the lack of new and updated resources used in treating

wounds, residents in rural areas rely on traditional medicinal

plants that are found in their surroundings as an alternative

2
to commercial ones. These plants are very important as they

may possess anti-bacterial, anti-microbial or anti-

inflammatory properties which contribute in the proper healing

of wounds. Nevertheless, people should also be wary of certain

effects that may come with these herbal plants when applied

incorrectly.

Kalinga is a province situated in the Cordillera

Administrative Region. Being mountainous, it has a rich source

of herbal plants. Also due to its mountainous terrain,

modernization and development is not as fast as those in plain

terrains allowing for the strong preservation of their

cultures and traditions. In the absence of modern products for

treating wounds, the people make use of resources that are

readily available in their environment, especially in those

areas that are inaccessible by vehicles. Traditional wound

treatment practices were and still are widely utilized in this

province. There is no definite history of the discovery of the

healing potentials of these plants. Only that it was passed on

from generation to generation. The study stems from the

potential of these herbal plants in wound healing and anti-

bacterial.

The researchers have chosen the top 3 traditional

medicinal plants that were suggested by the residents of Tabuk

City. According to the survey, the most common plants in terms

of wound healing are the leaves of Tobacco, Kutsay and Bulak-

3
manok, which are very abundant in the said locality. These

plants are believed to have anti-bacterial, anti- inflammatory

and anti- microbial properties that would greatly contribute

to the wound healing process.

“Tabako” or tobacco is a coarse, erect, viscidly-hairy

annual herb growing to a height of 0.7 to 1.5 meters. The

Leaves are considered antispasmodic, discutient, diuretic,

emetic, expectorant, narcotic, and sedative. In Europe, it is

first reported as a remedy for wound healing and headaches by

Jean Nicot Sieur de Villemain.

“Kutsay” or celery is an upright herb, rarely growing

over 30 centimeters in height. Leaves are pinnate, with large,

deeply-lobed segments, on long petioles. The flowers grow in

umbels, very small, and greenish white, while the fruit is

very small, with slender ridges.  It is considered as anti-

inflammatory, deobstruent, diuretic, resolvent, pectoral,

tonic, carminative, emmenagogue, diuretic, adjunct to

purgatives, lithotriptic, stimulant, antispasmodic and

aphrodisiac. Furthermore, it is considered as an antiseptic

according to the Unani system of medicine.

“Bulak-manok” or billy goat weed is an erect, slender,

perennial branched, hairy and aromatic herb with a height of

15 to 60 centimeters. The leaves are stalked, alternate, oval,

4 to 11 centimeters long, and 1 to 5 centimeters wide, with

the tip and base somewhat pointed, with round toothed margins.

4
It has many small flowering heads of 5 millimeters across, and

borne in dense terminal corymbs. The flowers are pale blue,

purple or white while the fruits are black, with 5 pappus

scales which are awned and often toothed or jagged. A study of

the crude extract showed anti-inflammatory, anti-bacterial and

wound healing effect better than normal saline treated

controls. Also, a study on Wistar rats investigated that the

wound healing properties of the extracts showed significantly

fewer fibroblasts and fewer inflammatory cells. The extracts

showed wound healing activity, with accelerated healing

processes and increased breaking strength. 

The researchers believe that these wound healing

properties should be studied, tested, proven, explored, and

utilized. This is to help the people whether or not to

continue these practices. Moreover, the study was a great

avenue for the contribution to the current knowledge about

wound healing plants. The study is based from the experiences

of the people in using these herbal plants. Hence, to prove

these healing hypotheses, a study using experimental

procedures and research would be conducted.

5
Research Problem

General Problem:

The main objective of this study is to determine if the

traditional medicinal plants found in Tabuk City, Kalinga are

effective wound healing agents.

Specific Problem:

Specifically, it seeks to answer the following questions:

1. What are the most commonly used traditional medicinal

plants in Tabuk City?

2. How effective are these plants in healing wounds?

3. Is there a significant difference between the effectivity

of the traditional medicinal plants and the commercial

drug?

Hypotheses

Null Hypotheses:

There is no significant difference between the

effectivity of the traditional medicinal plants and the

commercial drug.

Alternative Hypotheses:

There is a significant difference between the effectivity

of the traditional medicinal plants and the commercial drug.

6
Significance of the Study

This study will be advantageous to the following

beneficiaries:

Community - This study will raise awareness to the

residents of our community especially those from Tabuk,

Kalinga, about the effectiveness and the validity of the

herbal plants they use for wound healing. This study will

deeply elaborate the aspects and characteristics present

in the herbs.

Future Researchers – This study will serve as their guide

in improving as well as in organizing concepts in their

researchers. This study will also be a ground for ideas

that the researchers can conjugate in their investigatory

projects or researches.

Organizations - This study were a good source of

information and ideas that the Department of Health can

apply in enhancing or even in introducing new medicines

in the health industry. Establishing the study to the

activities and outreach programs of Red Cross

organization were useful too.

School – The study can be applied in schools particularly

in medical services such as the clinic. Since an herbal

7
 medicine is the product of this study, it can be utilized

as a preliminary treatment for wounds and can be used as

in first aids.

Students – Aside from the study being applicable to

different services and organizations, the study can also

be used for educational purposes. Students engaging to

medical courses can coordinate this study to their

subjects and use it as their basis for information.

Scope and Limitations

This study was focused and limited only to the

identification of the wound healing potential of traditional

medicinal plants in Tabuk City. A survey questionnaire on the

most commonly used traditional medicinal plants was

distributed to 20 people in Tabuk City, Kalinga. Heterogeneous

Purposive sampling is used wherein the 20 respondents were

chosen to provide a diverse range of cases relevant to this

study. The purpose of this is to provide as much insight as

possible into the event or phenomenon under examination. This

was done in order to identify the top 3 medicinal plants used

in that locality. The plant samples that were gathered in

Tabuk City are the leaves of Tobacco, Kutsay and Bulak-manok.

The leaf samples were crushed and air-dried. After drying, the

plants will undergo the ethanol method of extraction. To

8
determine the wound healing potential, the extracts were

tested on Sprague Daley rats. The type of wound which was used

is an excision wound.

This study which included sourcing the materials,

preparation of the plant extracts and testing their

effectiveness to the rats were done at the College Lab of St.

Paul University Philippines. The study was conducted from

October 2017 to May 2018.

9
 CHAPTER II

REVIEW OF RELATED LITERATURE

This chapter presents the related literature and the studies

related to the conduct of this research.

A. Bulak - Manok

Bulak-manok is an erect, slender, branched perennial, hairy

and aromatic herb, 15 to 60 centimeters in height. Leaves are

stalked, alternate, ovate, 4 to 11 centimeters long, and 1 to

5 centimeters wide, with the tip and base somewhat pointed,

and with round toothed margins, his idly hairy. Flowering

heads are numerous, small, about 5 millimeters across, and

borne in dense terminal corymbs. Ray flowers are many, pale

blue, purple or white. Disk flowers absent. Fruits (achenes)

are black, with 5 pappus scales which are awned and often

toothed or serrate below. Stuart, G. (2016, May) Bulak-manok,

billy goat weed, ageratum conyzoides: Herbal Medicinal

Plants / Philippine Herbal Therapy / Alternative Medicine in

the Philippines. Retrieved from

http://stuartxchange.com/Bulak.html

Distribution

Native to South and Central America, Bulak - manok is a

common pantropical weed that can extend into subtropical and

temperate zones where it grows during the summer or as an

10
ornamental It has been recognized as an ornamental and in

cultivated crops in Europe since before 1697. The species is

now found in Asia, Africa, North America, Central America, the

Caribbean, South America, Europe and Oceania. Its distribution

is perhaps more extensive than currently published because the

species is also used as an ornamental.

It is a common weed, in open, waste places throughout the

Philippines flowering year-round throughout the Philippines

from sea level to an altitude of 2,000 meters. The seeds are

light, easily dispersed and disseminated by wind. Stuart, G.

(2016, May) Bulak-manok, billy goat weed, ageratum conyzoides:

Herbal Medicinal Plants / Philippine Herbal Therapy /

Alternative Medicine in the Philippines. Retrieved from

http://stuartxchange.com/Bulak.html

Properties

The plant has a characteristic aromatic odour when

crushed. It is considered an antioxidant,

anthelmintic, antibacterial, anti-inflammatory, analgesic,

antispasmodic, febrifuge, tonic, laxative and vulnerary.

Stuart, G. (2016, May) Bulak-manok, billy goat weed, ageratum

conyzoides: Herbal Medicinal Plants / Philippine Herbal

Therapy / Alternative Medicine in the Philippines. Retrieved

from http://stuartxchange.com/Bulak.html

11
Constituents

The plant yields a vegetable proximate principle known as

"coumarin," also found in the allied genus, Eupatorium. The

leaves yield a volatile oil, 0.00054 percent, and yields mono

and sesquiterpenes, chromene, chromone benzofuran and

coumarin, flavonoids, triterpene and sterols, and alkaloids as

well. The essential oil from leaves and flowers yielded

ageratochromene (precocene II, 25.89%), the sesquiterpene

beta-caryophyllene (23.79%); demethoxyageratochromene

(precocene I, 14.76%), and some monoterpene hydrocarbons (2-

5.5%). Chemical profile analyses of leaf, stem, root and

flower was undergone which yielded phytochemicals: alkaloids,

flavonoids, saponins, tannins, glycosides, resins, phenols;

nutrients were proteins, carbohydrates, and reducing forms.,

essential and non-essential amino acids while evaluation of

leaf, stem, root, and flowers for chemical profile yielded

alkaloids, flavonoids and some constituents of flavonoids,

tannins, saponins, glycosides, resins, phenols while proteins,

carbohydrate and its reducing forms were present as nutrients.

To add, a phytochemical screening showed that the composition

of leaves and roots yielded alkaloids, flavonoids, saponins,

tannins, cardiac glycosides, and anthraquinones. Moreover, a

proximate analysis of dried leaves yielded crude carbohydrate

36.84%, protein 14.73%, fiber 23.50%, fat 2.27%, ash 12.64%,

moisture 109.02%. Na, K, Ca, Mg, P, Zn, Mn, and Fe were

12
present in both leaves and roots, with the concentration

slightly higher (p,0.05) in the leaves. The study of the

flowers yielded 0.25% v/w of essential oil. GC-MS analysis

showed the predominance of demothoxyageratochromene. Other

principal constituents were β-caryophyllene (19.5%), β-

cubebene (5.2%), germacrene D (3.9%), α-caryophyllene (2.9%)

and trans-β-farnesene (2.4%). Stuart, G. (2016, May) Bulak-

manok, billy goat weed, ageratum conyzoides: Herbal Medicinal

Plants / Philippine Herbal Therapy / Alternative Medicine in

the Philippines. Retrieved from

http://stuartxchange.com/Bulak.html

Traditional Uses

In the Philippines, juice of fresh leaves is widely used

as a vulnerary, pounded and mixed with salt. The stem, roots,

and flowers of the plant are boiled, the resulting decoction

used for stomach troubles. The whole plant has been used as a

decoction for cough, colds, fever, skin disease, hepatitis,

dysentery, neurasthenia, snake bites, dizziness, high blood

pressure and also for bleeding due to external wounds;

furuncle, eczema, carbuncle. The plant can also be used as

poultices for headaches. Squeezed juice from fresh material

when dropped inside the ears treats otitis media. The leaves

sometimes cooked in coconut oil, and the medicated oil applied

to wounds. Stuart, G. (2016, May) Bulak-manok, billy goat

13
weed, ageratum conyzoides: Herbal Medicinal Plants /

Philippine Herbal Therapy / Alternative Medicine in the

Philippines. Retrieved from

http://stuartxchange.com/Bulak.html

Studies

Antibacterial / Phytochemicals: Phytochemical testing of dried

leaves yielded resins, alkaloids, saponins, tannins,

glycosides and flavonoids while dried stems showed resins,

saponins, tannins, glycosides and flavonoids. In vitro studies

of AC extracts activity against S aureus, Y enterocolitica, S

gallinarum, and E coli, suggesting a potential source for

development of new antibacterials.

Antiulcerogenic / Gastroprotective: Study documents the

beneficial cytoprotective effects of the plant extract against

ethanol-induced gastric ulcers in rats. 

Analgesic / Anti-Inflammatory: Study results suggested that AC

extract exhibited antinociceptive effect and inhibition of

inflammatory reactions induced by neutrophil mobilizing

stimuli. 

Antimicrobial: Crude extract studies demonstrated

antimicrobial properties on S aureus and Methicillin-resistant

S aureus and possible usefulness in skin and wound

infections. 

Wound Healing: Extract study showed wound healing effect

14
better than normal saline treated controls, an effect

attributed to the antimicrobial properties of AC. 

Anti-Inflammatory / Toxicity Study: A hydroalcoholic extract

of leaves was studies for anti-inflammatory effect on sub-

acute (cotton pellet-induced granuloma) and chronic

(formaldehyde-induced arthritis models of inflammation in

rats. Study confirmed the anti-inflammatory properties of A

conyzoides with no apparent hepatotoxicity. 

A 28-day study evaluated the acute and sub-chronic toxicity of

A. conyzoides leaves in Wistar rats. Results showed the

hydroalcoholic extract is relatively safe when administered

orally to rats.

Anti-Cancer / Radical Scavenging Activity: Various extracts of

A conyzoides were screened in some cancer lines including

Human non-small cell lung CA, human colon adenocarcinoma,

human gastric CA, and human breast CA among others. Results

showed A conyzoides possessed anticancer and antiradical

properties. 

Comparative Study / Wound Healing / Polyherbal Formulation /

Roots: Study in rats of root extract showed wound healing

activity, with accelerated healing processes and increased

breaking strength. The wound healing of a polyherbal

formulation, Ageratum conyzoides with Ficus religiosa, C.

longa and T. indica showed better results, attributed to the

synergistic action of the plant constituents. 

15
Chemical Profiles of Leaf, Stem, Root, and Flower: In a study

evaluating the chemical profile of plant parts, the leaf

showed the most concentration of chemicals, followed by the

flower. 

Wound Healing Comparative Study / Vs Honey: Study in Wistar

rats investigated the wound healing properties of methanolic

extracts of Ac compared to honey. Histologically, the day-10

Ageratum sections showed fewer inflammatory cells compared

with honey and controls. Also, healed scar sections of wounds

dressed with herb extract showed more fibrosis. Healed wounds

from Ac group showed significantly fewer fibroblasts.

Inflammatory Bowel Disease / Protective Effects: In a study of

acetic acid-induced colitis and indomethacin-induced

enterocolitis models in rats, pretreatment with an ethanolic

extract produced significant attenuation in biochemical and

histopath parameters. Results suggest a possible benefit for

use in inflammatory bowel disease.  

Toxicity Study: Study evaluated the mean lethal dose of an

ethanolic extract of Ageratum conyzoidesat a daily dose of 500

and 1000 mg kg of extract for 28 days. Results showed no toxic

effects in rates, with an LD50 of 10,000 mg kg. Results

suggest AC s safe for use in ethnomedicine.

Antioxidant / Cytotoxic: Study evaluating a methanolic extract

of A. conyzoides stems for antioxidantactivity using a DPPH

scavenging assay showed dose dependent scavenging of free

radicals. Cytotoxicity evaluation using brine shrimp lethality

16
assay exhibited promising cytotoxicity, comparing with LC50

values of vincristine sulphate. 

Anthelmintic / Schistosomicidal / Essential Oil: Study

evaluated the schistosomicidal effects of the essential oil of

A. conyzoides against adult worms of Schistosoma mansoni. Ac-

EO showed activity, although less effective than the positive

control, praziquantel (PZQ). Ac-EO caused dose-dependent

reduction in the number of eggs. Precocene I and (E)-

caryophyllene were identified as two major constituents. 

Toxicity Study / Leaf Extract: Study evaluated the safety

potential of the leaf extract of Ageratum conyzoides in

Sprague Dawley rats using biochemical, hematological, and

histopatholical indices of toxicity. Histopathological studies

indicated various degrees of hepatocellular necrosis

accompanied by significant increases in liver and spleen

weights. Results suggest the leaf extract significantly alters

biomarkers of cardiac and skeletal muscle disorders, and

higher doses could induce liver injury. 

Antioxidative Potential: Comparative study evaluated the in

vivo effects of two common African herbs: Ageratum

conyzoides stem bark and Zanthoxylum zanthoxyloides leaves

on antioxidant status in the brain, kidney, and liver of

wistar albino rats intoxicated with sublethal concentration of

cadmium chlorides. Results suggest both

possessed antioxidant properties which could provide

protection against oxidative organ damage.

17
Toxicity Studies: Study evaluated acute and subchronic

toxicity potential of Ageratum conyzoides total extract in

rats. An aqueous extract yielded pyrrolizidines alkaloids,

tannins, saponins, flavonoids, and polyphenols. The aqueous

extract showed d dose dependent sedative and analgesic

effects. Subchronic toxicity testing the LD50 of the aqueous

extract was more than 13 g/kbw. Although there was no effect

on body weight, food consumption and water intake,

histological studies showed dose-dependent lesions with

hepatorenal changes associated with high levels of

transaminases and hyperleukocytosis at 800 mg/kg dose. 

Antioxidant / Leaves: Study of an alcoholic extract of

leaves by DPPH assay showed better antioxidant potential

compared to reference standard ascorbic acid. It exhibited

strong antioxidant DPPH radical scavenging activity with C50

of 24.8 mL/ml compared to ascorbic acid of 9.3. The

strong antioxidant activity was attributed to flavonoids and

phenols. 

Stuart, G. (2016, May) Bulak-manok, billy goat weed, ageratum

conyzoides: Herbal Medicinal Plants / Philippine Herbal

Therapy / Alternative Medicine in the Philippines. Retrieved

from http://stuartxchange.com/Bulak.html

18
 B. Tobacco

Tabako is a coarse, erect, viscidly-hairy annual herb

growing to a height of 0.7 to 1.5 meters. Leaves are large,

elliptic-ovate to oblong or obovate, 10 to 30 centimeters long

or longer, narrowed by the base, and sessile or short-stalked.

Inflorescences are terminal. Calyx is green, ovoid or tubular,

1 to 1.5 centimeters long, with five triangular lanceolate

teeth. Corolla is white and pink, linear, funnel-shaped and

about 5 centimeters long. Capsule is ovoid, 1.5 to 2

centimeters long, with very numerous white seeds. Stuart, G.

(2016, September) Tabako, Nicotiana tabacum Linn., TOBACCO -

Herbal Medicine / Philippine Alternative Meeicine /

StuartXchange. Retrieved from

http://www.stuartxchange.org/Tabako.html

Properties

Except for the ripe seeds, all parts are considered

poisonous. The Leaves considered antispasmodic, discutient,

diuretic, emetic, expectorant, narcotic, sedative, maturative,

styptic and sialagogue. Stuart, G. (2016, September) Tabako,

Nicotiana tabacum Linn., TOBACCO - Herbal Medicine /

Philippine Alternative Meeicine / StuartXchange. Retrieved

from http://www.stuartxchange.org/Tabako.html

19
Constituents

Leaves contain active principles, the toxic alkaloid

nicotine (0.6 to 9 %) and three other alkaloids: nicoteine,

nicotelline, and nicotinine. Also yields anabasine, betaine

iamylamine, pyrrolidine, and n-methyl pyrroline, resin,

albumen, gum, extractive matter, and ash containing large

amounts of salt.

Study isolated a pair of sesquiterpene glucosides - 3-

hydroxysolavetivone-beta-D-glucoside A and B - from the

leaves.

Seeds of N. tabacum yielded 30% oil, the rest are

protein, crude fiber, carbohydrate and inorganic material.

Main triglycerides in the seed oil are trilinolein and

palmitodilinolein, accounting for about 90% of the oil

composition. Major individual fatty acids are linoleic acid

(66-76%) oleic acid (17-27%), palmitic acid (7-10%), and

stearic acid (3.1%). Oil is classified as linoleic oil in the

semi-drying oil category.  Stuart, G. (2016, September)

Tabako, Nicotiana tabacum Linn., TOBACCO - Herbal Medicine /

Philippine Alternative Meeicine / StuartXchange. Retrieved

from http://www.stuartxchange.org/Tabako.html

Pharmacokinetic factoids

The nicotine alkaloid is rapidly absorbed from all mucous

membranes, lungs and skin, but more commonly enters through

20
the lung alveoli, quickly reaching the heart and brain

receptors, and largely eliminated by oxidation through the

liver (80 to 90%), a small amount metabolized in the kidneys

and lungs. Elimination half-life is 2 hours.

Nicotine is the primary psychoactive constituent of

tobacco – cigarettes, chewing tobacco, and nicotine patches,

gum, inhalers and sprays.

It acts on the autonomic ganglia. One cigarette causes a

50% rise of noradrenaline and 15% rise of adrenaline. From the

posterior pituitary, vasopressin is released.

A pack per day smoking increases the coronary heart

disease and myocardial infarction three-fold. There is also an

increase in emphysematous lung disease, lung cancer,

peripheral vascular disease and stomach ulcers. In women who

smoke, there is a higher incidence of premature births,

deformities and perinatal infant mortality.

It is used by herbalists as a relaxant, it has long been

abandoned because of its highly addictive nature. Stuart, G.

(2016, September) Tabako, Nicotiana tabacum Linn., TOBACCO -

Herbal Medicine / Philippine Alternative Meeicine /

StuartXchange. Retrieved from

http://www.stuartxchange.org/Tabako.html

21
Studies

Anthelmintic: Study of methanol and aqueous extracts of

Nicotiana tabacum exhibited dose-dependent anthelmintic

activity both in vitro and in vivo, justifying its use in

traditional medicine in Pakistan. (1) Study evaluated the in-

vitro anthelmintic effect of aqueous and alcoholic extract of

N. tabacum against M. marshalli. Results showed considerable

anthelmintic activity with more potent effects observed with

highest concentrations.

Hematologic Effects: Study of the aqueous extract of N tabacum

leaves showed significant decrease in RBC count, PCV, Hb and

platelet count with increase in MCV and MCH. Results suggest

the consumption of the aqueous extract of N tabacum may lead

to some level of anemia despite its "pleasant effects." 

Lipid Studies / Seed Oils: Plant lipids find application not

only in nutrition, but also in the production of lotions,

ointments and various pharmaceutical preparations. ipids Seeds

of N. tabacum and N. rustica were found to contain lipids

41.3% and 36.6% respectively. The lipid class in N. tabacum

were hydrocarbons (1.4%), wax esters (1.7%) sterol esters

(2.4%), triacylglycerols (69.3%), free fatty acids (6.2%),

l,3-diacylglycerols (4.6%), l,2-diacylglycerols (3.5%), free

sterols (2.3%), 2-monoacylglycerols (2.1%), 1-

monoacylglycerols (1.8%), phosphatidylethanolamines (1.7%),

phosphatidylcholines (0.9%), Iysophosphatidylethanolamines

(1.5%) and phosphatidylinositols (0.6%). 

22
Antimicrobial: In vitro antibacterial screening of various

extracts of N. tabacum revealed different ranges of

antibacterial activities. An ethyl acetate extract was more

effective in controlling Bacillus cereus and Erwinia

carotovora followed by a butanol extract against Staph aureus

and Agrobacterium tumefaciens. 

In another study, the antibacterial activity of t the tobacco

leaves extracts on some bacterial strains including

Staphylococcus areus, Bacillu subtillis, Escherichia coli,

Pseudomonas uerauginosa was investigated. Results revealed

that tobacco leaves extracts have effects on bacterial growth.

Ethanolic tobacco extracts are more effective than aqueous

extracts in inhibiting the bacterial growth.

Biotechnological Reduction of Tobacco Toxicity: Genetic

engineering can modify nicotine levels of the tobacco plant.

In the study, through transferring of the blocking gene, the

pathway of nicotine biosynthesis was block to produce

transgenic tobacco with love levels of nicotine. Lower

nicotine levels can reduce toxic effects and facilitate

withdrawl from it.

Review of Properties and Pharmacological Activities: Tobacco

has yielded a spectrum of chemical compounds including

nicotine, solanesol, malic, and citric acid. Pharmacologic

activities are described i.e., antibacterial, antinociceptive,

antimicrobial, antifungal, anthelmintic, CNS activity, and

23
anti-Alzheimer's activity. Review suggest the potential for

use of nicotine in the treatment of Alzheimer disease,

Parkinson disease, depression and anxiety, schizophrenia,

ADHD, pain and obesity. 

Stuart, G. (2016, September) Tabako, Nicotiana tabacum Linn.,

TOBACCO - Herbal Medicine / Philippine Alternative Meeicine /

StuartXchange. Retrieved from

http://www.stuartxchange.org/Tabako.html

C. Kutsay

Kutsay is a kind of leek, ranked-scented, green, growing

20 to 40 centimeters high. Bulbs are small, white and

clustered. Leaves are green, grasslike, narrowly linear,

flattish, 15 to 30 centimeters long, 3 to 6 millimeters wide.

Umbel has a few to many white flowers. The perianth is bell-

shaped. Fruits are on pedicels of 2 to 3 centimeters long,

obovoid, 3-lobed, 5 to 7 millimeters in diameter. Seeds are

black, depressed, globose or reniform, 2.5 to 3 milimeters in

diameter. In the Philippines, the plant seldom flowers.

Stuart, G. (2013, March) Kuchai, allium odorum, chinese chives

/ Philippine Medicinal Herbs / Philippine Alternative

Medicine. Retrieved from

http://www.stuartxchange.org/Kuchai.html

24
 According to wiki, immature leaves and stalks and

buds could be used the same as chives, scallions or garlic.

For stir frying, dumpling, spice and various flavoring

purposes. 

Both leaves and young scapes are used to flavor

vegetables, meat, and seafoods. It can also be eaten raw,

blanched or sauteed. Its medical uses include treatment of

wounds, stomach ache, tumors, intestinal disorders, and

scabies.

Externally, the fresh leaves and bulbs are used as an

antiseptic and vulnerary. The leaves when taken internally,

according to Stuart, act as a cordial. Mensut reports that the

whole plant is used in Indo-China as a diuretic. Kuchai /

Kuchay / Kutsai Uses and Benefits. (2016, June 17) Retrieved

from http://foodrecap.net/health/kuchay-benefits/

Cultivation

Being an easily grown plant, it prefers a sunny position

in a light well-drained soil. The bulbs should be planted

fairly deeply. A very ornamental plant, the flowers are

especially attractive. Very closely related to A. tuberosum .

Grows well with most plants, especially roses, carrots, beet

and chamomile, but it inhibits the growth of legumes. This

plant is a bad companion for alfalfa, each species negatively

affecting the other. Members of this genus are rarely if ever

25
troubled by browsing deer. Stuart, G. (2013, March) Kuchai,

allium odorum, chinese chives / Philippine Medicinal Herbs /

Philippine Alternative Medicine. Retrieved from

http://www.stuartxchange.org/Kuchai.html

Medical Uses

The leaves and bulbs contain sulphur compounds (which give

them their onion flavour), saponins and bitter substances.

They possess antibacterial properties and are used in Vietnam

in the treatment of haemoptysis, epistaxis, cough, sore

throat, asthma, dysentery, dyspepsia etc. When added to the

diet on a regular basis they help reduce blood cholesterol

levels, act as a tonic to the digestive system and also tonify

the circulatory system. The seed contains alkaloids and

saponins. It is used in the treatment of spermatorrhoea,

haematuria, incontinence, lumbago etc. Stuart, G. (2013,

March) Kuchai, allium odorum, chinese chives / Philippine

Medicinal Herbs / Philippine Alternative Medicine. Retrieved

from http://www.stuartxchange.org/Kuchai.html

Studies 

• Antoxidant: Study showed part of the Allium family possess

antioxidant capability. Heat treatments reduced the

antioxidant activity for most foods.

• Antifungal: Study of extracts of 7 Allium plants, including

Chinese chive, were examined for antifungal activity against

three Aspergillus species: A niger, A flavus and A

26
fumigatus. All the plants possessed antifungal activity, the

inhibitory activity decreased with increasing incubation and

heating temperature. Acetic acid plus heat treatment of the

extracts resulted in greater inhibition.

• Hypotensive / Antihypertensive: Study of ethanol extract

of Kutsay showed hypotensive and antihypertensive effect.

Stuart, G. (2013, March) Kuchai, allium odorum, chinese chives

/ Philippine Medicinal Herbs / Philippine Alternative

Medicine. Retrieved from

http://www.stuartxchange.org/Kuchai.html

D. Wound Healing

Wounds are a major cause of physical disabilities. These

result in an opening or break of the skin that causes a

disruption on the skin’s normal function. Wounds can be

classified as acute or chronic depending on its healing time.

Acute wounds heal without complications in the expected amount

of time while chronic wounds take a longer time to heal which

might have the risk of infection. Furthermore, wounds can also

be classified as open or closed. Open wounds are those with

exposed underlying tissue that are open to the outside

environment while closed wounds have damage that occurs

without the underlying tissue exposed. Maynard, J. (2017, July

6) How Wounds Heal: The 4 Main Phases of Wound Healing.

27
Retrieved February 18, 2018, from

http://www.shieldhealthcare.com/community/wound/2015/12/18/how

-wounds-heal-the-4-main-phases-of-wound-healing/

Phase 1: Hemostasis Phase

Hemostasis, the first phase of healing, begins at the

onset of injury, and the objective is to stop the bleeding. In

this phase, the body activates its emergency repair system,

the blood clotting system, and forms a dam to block the

drainage. During this process, platelets come into contact

with collagen, resulting in activation and aggregation. An

enzyme called thrombin is at the center, and it initiates the

formation of a fibrin mesh, which strengthens the platelet

clumps into a stable clot. Maynard, J. (2017, July 6) How

Wounds Heal: The 4 Main Phases of Wound Healing. Retrieved

February 18, 2018, from

http://www.shieldhealthcare.com/community/wound/2015/12/18/how

-wounds-heal-the-4-main-phases-of-wound-healing/

Phase 2: Defensive/Inflammatory Phase

During Phase 2, a type of white blood cells called

neutrophils enter the wound to destroy bacteria and remove

debris. These cells often reach their peak population between

24 and 48 hours after injury, reducing greatly in number after

3 days. As the white blood cells leave, specialized cells

called macrophages arrive to continue clearing debris. These

28
cells also secrete growth factors and proteins that attract

immune system cells to the wound to facilitate tissue repair.

This phase often lasts 4 to 6 days and is often associated

with edema, erythema (reddening of the skin), heat and pain.

Maynard, J. (2017, July 6) How Wounds Heal: The 4 Main Phases

of Wound Healing. Retrieved February 18, 2018, from

http://www.shieldhealthcare.com/community/wound/2015/12/18/how

-wounds-heal-the-4-main-phases-of-wound-healing/

Phase 3: Proliferative Phase

Once the wound is cleaned out, the wound enters Phase 3, the

Proliferative Phase, where the focus is to fill and cover the

wound.

The Proliferative phase features 3 distinct stages: 1) filling

the wound; 2) contraction of the wound margins; and 3)

covering the wound (epithelialization).

During the first stage, shiny, deep red granulation tissue

fills the wound bed with connective tissue, and new blood

vessels are formed. During contraction, the wound margins

contract and pull toward the center of the wound. In the third

stage, epithelial cells arise from the wound bed or margins

and begin to migrate across the wound bed in leapfrog fashion

until the wound is covered with epithelium. The Proliferative

phase often lasts anywhere from 4 to 24 days. Maynard, J.

(2017, July 6) How Wounds Heal: The 4 Main Phases of Wound

29
Healing. Retrieved February 18, 2018, from

http://www.shieldhealthcare.com/community/wound/2015/12/18/how

-wounds-heal-the-4-main-phases-of-wound-healing/

Phase 4: Maturation Phase

During the Maturation phase, the new tissue slowly gains

strength and flexibility. Here, collagen fibers reorganize,

the tissue remodels and matures and there is an overall

increase in tensile strength (though maximum strength is

limited to 80% of the pre-injured strength). The Maturation

phase varies greatly from wound to wound, often lasting

anywhere from 21 days to 2 years.

The healing process is remarkable and complex, and it is also

susceptible to interruption due to local and systemic factors,

including moisture, infection, and maceration (local); and

age, nutritional status, body type (systemic). When the right

healing environment is established, the body works in wondrous

ways to heal and replace devitalized tissue. Maynard, J.

(2017, July 6) How Wounds Heal: The 4 Main Phases of Wound

Healing. Retrieved February 18, 2018, from

http://www.shieldhealthcare.com/community/wound/2015/12/18/how

-wounds-heal-the-4-main-phases-of-wound-healing/

30
 CHAPTER III

Methodology

Materials and methods

Materials

The materials that were used for the study are Dried

tobacco, Kutsay Leaves, Bulak manok Leaves, Distilled water,

Rotary Evaporator , Mortar and Pestle/Blender, Tongs, Round

Bottomed Flask, Erlenmeyer Flask Iris Scissor, Syringe, Curved

Forceps, Straight Forceps, Fixation Forceps, Betadine

Chemical: Ethanol (99.9%),

All the materials that were used are under sterile condition.

Procedure

Floating of questionnaires and tallying of survey results

The questionnaires were given to the residents of Tabuk

City, Kalinga. The plants that were used are Tobacco, Kutsay

and Bulak manok. The part of the plant that was used for the

extraction is the leaves.

Collection of the medicinal plants

The leaf samples were collected in Tabuk City, Kalinga.

Random leaf samples were collected into plastic zip lock bags

31
with appropriate labelling and were stored in an ice cooler

until transportation to the laboratory for extraction.

Preparation of the Plant Samples

70 grams of Tobacco leaves, Kutsay Leaves and Bulak manok

Leaves were washed three times with distilled water to

eliminate dust and foreign particles. The leaves were allowed

for shade drying under room temperature. The leaves were dried

in an oven for 2 hours at 100°C, or until dried. The dried

leaves were ground to fine powder using a blender.

Extraction of oil from the dried and crushed leaves

350 ml, 300 ml and 250 ml of 99.9 % ethanol were used

for the extraction from 60 g ground Kutsay leaves, Tobacco

Leaves and Bulak manok leaves respectively. The ethanol were

poured into the round-bottomed flask and the grounded leaf

samples were put in cheesecloth and soaked in the ethanol for

48 hours. After soaking, the leaf samples were squeezed to

extract the mixture of ethanol and oil. The extracted oil and

ethanol produced from were entered into a rotary evaporator to

remove the impurities in the oil.

Preparation of Testing Animals

15 healthy adult Sprague Dawley Rats (male) weighing 250

– 300 g were obtained from the animal house unit. The rats

were acclimated in in individual cages, in a controlled

32
environment at 22 ± 3 °C and 50% to 60 % humidity under a 12

h–12 h light–dark cycle for at least one week prior to the

experiment. The cages were maintained according to standard

housing conditions, and access to standard diet and water was

provided ad libitum during the experiment. All of the animal

care criteria prepared by the National Academy of Sciences and

outlined in the Guide for the Care and Use of Laboratory

Animals were applied throughout the experiment

Testing of Wound Healing Property of the Plant Extracts

The animals were anesthetized with 0.1 cc of Zolitil.

After anaesthesia, the animal’s right upper back were shaved

and disinfected with Betadine. A lesion was made in the right

upper dorsal region with a scalpel. Subsequently, the animals

were monitored to total recovery and randomly distributed into

5 treatments for the 3 rats. The Treatments were: Treatment 1

(Control Group), Treatment 2 (Bactroban), Treatment 3 (Tobacco

Leaves Extract) Treatment 4 (Bulak manok Leaves Extract) and

Treatment 5 (Kutsay Leaves Extract).

The wounds were topically treated with the 3 different

plant extracts using an applicator for 7 days. The treatments

were administered twice a day for a period of 7 days.

Contraction of the wound area were measured at days 1, 3,

5, and 7 after incision by placing a transparent plastic cover

33
over the wound and tracing it out; area of this impression

were calculated using graph sheet and wound contraction.

Data Analysis

The contraction of the wound area on the transparent

plastic cover was measured and the data gathered were

subjected to ANOVA testing.

These are the following treatments given to the albino

rats:

Treatment 1: Control Group

Replicate 1: No application

Replicate 2: No application

Replicate 3: No application

Treatment 2: Bactroban

Replicate 1: Bactroban

Replicate 2: Bactroban

Replicate 3: Bactroban

Treatment 3: Tobacco Leaves Extract

Replicate 1: Tobacco Leaves Extract

Replicate 2: Tobacco Leaves Extract

Replicate 3: Tobacco Leaves Extract

34
Treatment 4: Bulak manok Leaves Extract

Replicate 1: Bulak manok Leaves Extract

Replicate 2: Bulak manok Leaves Extract

Replicate 3: Bulak manok Leaves Extract

Treatment 5: Kutsay Leaves Extract

Replicate 1: Kutsay Leaves Extract

Replicate 2: Kutsay Leaves Extract

Replicate 3: Kutsay Leaves Extract

35
 CHAPTER IV

RESULTS AND DISCUSSIONS

This chapter presents the data gathered from the experiment

and the analysis of this data.

Table 1: Measurement Of The Contraction Of Wound Area Of The

5 Treatments during Day 0

Day 0 Treatment 1 Treatment 2 Treatment Treatment Treatment

(No (Bactroban) 3 4 5
application) (Billy (Tobacco) (Garlic
goat Chives)
weed)
Replicate 5 18 21.5 25 31.5
1
Replicate 22 20 29.5 26 51
2
Replicate 6 14.5 28.5 24 29
3

Table 1 shows the measurement of the wound area (in

millimeters) of the replicates in each treatment at the start
of the experiment (Day 0). The size of the wounds of the three
replicates in each treatment is measured by placing a
transparent plastic cover over the wound and tracing it out.
The areas of these impressions were calculated using graph
sheet.

36
Table 2: Measurement Of The Contraction Of Wound Area Of The
5 Treatments during Day 2

Day 2 Treatment Treatment Treatment Treatment Treatment

1 2 3 4 5
Replicate 14 12 12 21 29
1
Replicate 22 12 15 24 23
2
Replicate 6 13.5 13 19 26.5
3

Table 2 shows the measurement of the wound area (in

millimeters) of the replicates in each treatment at Day 2. The
size of the wounds of the three replicates in each treatment
is measured by placing a transparent plastic cover over the
wound and tracing it out. The areas of these impressions were
calculated using graph sheet.

Table 3: Measurement Of The Contraction Of Wound Area Of The

5 Treatments during Day 4

Day 4 Treatment Treatment Treatment Treatment Treatment

1 2 3 4 5
Replicate 8 10 9 20 23.5
1
Replicate 13 11 14 21 30
2
Replicate 10 13 10 18 22
3

Table 3 shows the measurement of the wound area (in

millimeters) of the replicates in each treatment at Day 4. The
size of the wounds of the three replicates in each treatment
is measured by placing a transparent plastic cover over the

37
wound and tracing it out. The areas of these impressions were
calculated using graph sheet.

Table 4: Measurement Of The Contraction Of Wound Area Of The

5 Treatments during Day 6

Day 6 Treatment Treatment Treatment Treatment Treatment

1 2 3 4 5
Replicate 8 6 3.5 8 4.5
1
Replicate 7.5 4.5 11 23 6
2
Replicate 6.5 5 5 4 4
3

Table 4 shows the measurement of the wound area (in

millimeters) of the replicates in each treatment at Day 6. The
size of the wounds of the three replicates in each treatment
is measured by placing a transparent plastic cover over the
wound and tracing it out. The areas of these impressions were
calculated using graph sheet.

Table 5: Measurement Of The Contraction Of Wound Area Of The

5 Treatments during Day 8

Day 7 Treatment Treatment Treatment Treatment Treatment

38
 1 2 3 4 5
Replicate 2 4 2 6.5 8.5
1
Replicate 5 4 5 10 9
2
Replicate 5 4 4 3 4
3

Table 5 shows the measurement of the wound area (in

millimeters) of the replicates in each treatment at Day 8. The
size of the wounds of the three replicates in each treatment
is measured by placing a transparent plastic cover over the
wound and tracing it out. The areas of these impressions were
calculated using graph sheet.

Table 6: Sum Of The Three Replicates For Each Treatment

  T1 t2 t3 t4 t5
D0 43 52.5 79.5 75 111.5
D2 42 37.5 40 64 98.5
D4 31 34 33 59 75.6
D6 22 15.5 19.5 20 14.5
D8 12 12 11 19.5 21.5

Table – shows the sum of the three replicates of Treatments 1,

2, 3, 4 and 5 for Days 0, 2, 4,6 and 8.

Table 7: Descriptive Statistics Of The Independent Treatments

Treatment A B C D E Pooled
→ Total
observatio 5 5 5 5 5 25
ns N

39
sum ∑xi∑xi 150.00 151.50 183.00 237.500 321.600 1,043.6
00 00 00 0 0 000
mean x¯x¯ 30.000 30.300 36.600 47.5000 64.3200 41.7440
0 0 0
sum of 5,202. 5,702. 9,510. 13,982. 28,522. 62,919.
squares ∑x 0000 7500 5000 2500 3600 8600
2i∑xi2
sample 175.50 278.07 703.17 675.250 1,959.2 806.492
variance s 00 50 50 0 620 6
2s2
sample 13.247 16.675 26.517 25.9856 44.2636 28.3988
std. 6 6 4
dev. ss
std. dev. 5.9245 7.4575 11.859 11.6211 19.7953 5.6798
of 0
mean SEx¯S
Ex¯

Table 8: One – way ANOVA for the independent treatments

Source Sum of  Degrees Mean F p-

squares SS of  square  statistic value
freedom νν MS
treatment 4,190.7736 4 1,047.6934 1.3817 0.2759
error 15,165.0480 20 758.2524
total 19,355.8216 24

Decision at
Treatments Mean F-ratio p-value
.05 level
30.0000 1.3817 0.2759 Not
1
Significant
2 30.3000 Not
Significant
3 36.6000 Not
Significant
4 47.5000 Not

40
 Significant
5 64.3200 Not
Significant
Table 9: ANOVA Test of the Five Treatments

Table 8 shows that the five treatments labelled as 1, 2, 3, 4

and 5 have no significant difference with the probability
value 0.2759 which is greater than 0.05 significance level.
Therefore, the null hypothesis is accepted.

CHAPTER V

CONCLUSIONS AND RECOMMENDATIONS

Conclusions

According to the data and results gathered, the following

conclusions were derived.

First, the most commonly used traditional medicinal

plants for wound healing utilized by the respondents in Tabuk

41
City, Kalinga are Billy goat weed (Bulak-manok), Tobacco
(Tabako) and Garlic Chives (Kutsay). A survey questionnaire
was given to the respondents in Tabuk City from which the
researchers have gathered the top 3 medicinal plants.

Secondly, after measuring the wound area size of the

albino rats by using a transparent plastic cover and a graph
sheet, the data showed that the contraction of the wounds were
decreasing every other day upon measurement (Day 0, 2,4,6 and
8).

And lastly, basing on the ANOVA results, the five

treatments labelled as 1, 2, 3, 4 and 5 have no significant
difference with the probability value 0.2759 which is greater
than the 0.05 significance level. Therefore, the null
hypothesis is accepted.

Recommendations

The researchers would like to recommend to the future

researchers to further examine the characteristics of the

Traditional medicinal plants to broaden their usage.

42
 They also would like to recommend the uniformity of the

excision wound size so as to clearly see the difference in the

wound contraction of the Sprague Dawley rats.

And lastly, they would like to recommend the analysis of

the rats’ wound tissue.

Bibliography:

https://www.cabi.org/isc/datasheet/3572

http://drfarrahcancercenter.com/portfolio/bulak-manok/

http://www.stuartxchange.org/Tabako.html

43
http://stuartxchange.com/Bulak.html

https://www.herbsia.com/2017/11/ageratum-conyzoides-medicinal-

uses-to.html?m=1

http://foodrecap.net/health/kuchay-benefits/

http://findmeacure.com/2011/01/09/chinese-chives/

http://barefootroamer-

barefootroamer.blogspot.com/2012/02/medicinal-value-of-

chinese-chive-kut.html

http://www.stuartxchange.org/Kutsay.html

http://www.shieldhealthcare.com/community/wound/2015/12/18/how

-wounds-heal-the-4-main-phases-of-wound-healing/

St. Paul University Philippines

TUGUEGARAO CITY 3500
BASIC EDUCATION UNIT

Wound Healing Potential of Traditional Medicinal Plants

Questionnaire

44
Name: __________________________________ Age: ____
Gender: _____

Please answer truthfully.

1. In your locality, give three herbal plants that you

commonly use as an alternative treatment for wounds. (Sa
iyong lugar, magbigay ng tatlong pinaka ginagamit na
halamang panggamot para sa mga sugat)

a. Plant 1 __________________

b. Plant 2 __________________

c. Plant 3 __________________

2. Where do you get these plants? (Saan mo kinukuha itong

mga halaman?)

3. From whom/where have you learned about these plants?

(Saan/kanino mo nalaman itong mga halaman na ito?)

4. Are these plants abundant in your locality? (madami bang

halaman na ganito sa lugar niyo?)

____ YES _____NO

5. How do you use these plants? Indicate the treatment and

application process. (Paano mo ginagamit ang mga halamang
ito? Ibigay ang proseso ng gamutan at aplikasyon.)

6. When you use these plants, how long do you use it for?
(Gaano mo katagal ginagamit tong mga halaman?)
a. Plant 1 _________________

b. Plant 2 _________________

c. Plant 3 _________________

7. Rate the effectiveness of these plants. (5 as the highest

and 1 as the lowest) (Gaano ka-epektibo ang mga halamang
ito? 5 ang pinakamataas at 1 ang pinakamababa)

45
 a. Plant 11 2 3 4 5

2 4
b. Plant 21 3 5

2 3 4 5
c. Plant 31

8. Why do you use these plants as an alternative for healing

wounds? (Bakit mo ginagamit ang mga halamang ito sa
paggamot ng mga sugat?)

46