Wound Healing Activity of Latundan (Musa sapientum) Peeling

Ointment on Rats

Submitted as a partial requirement in Practical Research I

Submitted by:

Cedrick Batta

Eirom John Quitang

Tricia Quilang

Ferma Ruiz

Submitted to:

Mrs. Rita Daliwag

Adviser in Practical Research

December, 2017

1
 ABSTRACT

This study entitled "Wound Healing Activity of Latundan

(Musa sapientum) Peeling Ointment on Rats" aimed to determine the

wound healing activity of Latundan (Musa sapientum) peeling

ointment on rats.

The latundan bananas were collected from the public market

of Tuguegarao City, Cagayan. The collection of the extract was

done by first, set up a double boiler, a pot over a bowl on top

of it. Second, pour 300ml of water then boil it in a pot. Third,

put the petroleum jelly or white petrolatum and beeswax in the

bowl to melt it then add the banana peeling oil. Fourth, stir the

mixture in a very low heat. Lastly, pour it into a container. The

mixture was thicken into an ointment as it cools. The experiment

animals was then treated with 3 set ups (latundan banana peeling

extract, terramycin, and no treatment at all) with 3 replicates

each. The phytochemical analysis was showed that the latundan

banana yields Flavanoids and Tannins.

Based on the results, we conclude that banana peel have a

wound healing activity to an inflicted wound in rats used in the

study which is significant on day 5; wounds heal on the 10th day

without infection.

Keywords: Wound healing, Latundan banana peelings and Rat

2
 TABLE OF CONTENTS

I. Approval Sheet . . . . . . . . . . . . . . . . . . . . .i

II. Acknowledgement . . . . . . . . . . . . . . . . . . . .ii

III. Dedication. . . . . . . . . . . . . . . . . . . . . . iii

IV. Abstract . . . . . . . . . . . . . . . . . . . . . . . iv

V. Table of contents . . . . . . . . . . . . . . . . . . . v

VI. Chapter 1

a. Background of the Study . . . . . . . . . . . . . . 1

b. Review of Related Literature . . . . . . . . . . . .4

c. Review of Related Studies . . . . . . . . . . . . .11

d. Statement of the Problem . . . . . . . . . . . . . 26

e. Hypotheses . . . . . . . . . . . . . . . . . . . . 27

f. Conceptual Framework . . . . . . . . . . . . . . . 27

g. Significance of the Study . . . . . . . . . . . . .28

h. Scope and Delimitations . . . . . . . . . . . . . 29

i. Definition of Terms . . . . . . . . . . . . . . . .30

3
 VII. Chapter 2

a. Methodology . . . . . . . . . . . . . . . . . . 31

VIII. Chapter 3

a. Results and Discussion. . . . . . . . . . . . . . 46

IX. Chapter 4

a. Summary . . . . . . . . . . . . . . . . . . . . .50

b. Conclusion . . . . . . . . . . . . . . . . . . . .51

c. Recommendation . . . . . . . . . . . . . . . . . .51

X. Appendix. . . . . . . . . . . . . . . . . . . . . . . .52

XI. Bibliography. . . . . . . . . . . . . . . . . . . . . . .66

4
 CHAPTER 1

Introduction and Review of Related Literature

Background of the Study

A wound is a breakdown in the protective function of the

skin; the loss continuity of epithelium, with or without loss of

underlying connective tissue (i.e muscle, bone, nerves) following

injury to skin or underlying tissues/organs caused by surgery, a

blow, a cut, chemicals, heat/cold, friction/shear force, pressure

or as a result of disease, such as leg ulcers or carcinomas

(Leaper DJ, Harding Kg, Hutchinson J ). A wound may be described

in many ways; by its aetiology, anatomical location, by whether

it is acute or chronical, by the method of closure, by its

presenting symptoms, or indeed by the appearance of the

predominant tissue types in the wound bed. All definitions serve

as a critical purpose in the assessment and appropriate

management of the wound through to symptom resolution or, if

viable healing (Enoch S and Price 2004). Wound occurs when the

skin is broken or damaged because of injury. Causes of injury of

wounds are mechanical, chemical, electrical, thermal, or nuclear

sources. The skin can be damaged in a variety of ways depending

upon the mechanism of injury.

5
 In the United States, chronic wounds affect around 6.5

million patients. It is claimed that an excess of US$25 billion

is spent annually on treatment of chronic wounds and the burden

is growing rapidly due to increasing health care costs, an aging

population and a sharp rise in the incidence of diabetes and

obesity worldwide. The annual wound care products market is

projected to reach $15.3 billion by 2010. Chronic wounds are

rarely seen in individuals who are otherwise healthy. In fact,

chronic wounds patients frequently suffer from “highly branded”

diseases such as diabetes and obesity. This seems to have

overshadowed the significance of wounds per se as a major health

problem (Chandan K. Sendan et. al).

Injuries fatality rates increased by 196% from 14.3 per

100,000 in 1960 to 43.4 per 100,000 in 1995, and in one 11 deaths

in the Philippines are due to injuries. Intentional injuries

account for 48% of all injury deaths and motor vehicles crashes

for 15%. For 15-44 yr. old males, injuries account for 42% of all

deaths, 67% of which are intentional. The proportion of all

deaths attributable to intentional injuries has increased by 925%

and that of motor vehicle crashes by 600% from 1960 to 1995.

Research into risk factors and potential interventions for the

prevention of intentional injuries should be a priority in the

Philippines (Rafael J. Consunji, Adnan A. Hyder).

6
 Terramycin is one of the ointments available in drugstore

nationwide and is used to treat wounds. It contains the active

ingredient oxytetracycline and polymycin B sulfate considered a

tetracycline antibiotic type of medication specifically to cure

wounds. The medication works by interfering with the cell walls

of bacteria to prevent them from producing proteins essential for

their reproductions. The ointment is mainly prescribed to treat

bacterial infections such as acne, scratches, burns, abrasion and

insect bites. It is usually administered topically 1 to 3 times

per day or as directed to doctor. It is usually not tolerated

with known side effects like irritation, nausea, diarrhea, rash,

swelling and etc (John P. Cunha, DO, FACOEP). And because common

commercial drugs are sometimes expensive and out of stock, demand

on the medicinal plant with wound healing increase.

Latundan (Musa sapientum) is widely used for various

ailments including diarrheas, stomach ulcer, diabetes, heart

burn, inflammation as well as to treat wounds. Latundan (Musa

sapientum) peelings as claimed to be able to heal wounds can be

an alternative medicine once the researchers verified its wound

healing potential. This product or test plant may have a lesser

cost and easier preparation either home uses or medical purposes.

The peelings being thrown out at the garbage bin which are

7
considered wastes can already accustomed for a more efficient

intention.

Review of Related Literature

Conceptual Literature

I. Botanical Description of Latundan

The banana plant is the largest herbaceous flowering plant.

The main or upright stem is actually a pseudostem, growing from a

corn, to a height of 6 to 7.6 meters. Leaves are spirally

arranged, as long as 2.7 meters and 60 cm side, fragile and

easily torn by wind, with the familiar frond look. Each

pseudostem produces a single bunch of bananas, the pseudostem

(produces a single bunch of bananas) dies after fruiting, as

offshoots usually develop from the base of the plant. Each

pseudostem produces a single inflorescence, the banana heart,

containing many bracts between rows of flowers. The banana fruits

develop from the heart, in a hanging cluster made up of tiers

(hands), up to 20 fruit to a tier.

8
 Figure 1. Image of Latundan (Musa sapientum)

Taxonomic Classification of Latundan

Ploidy: 3x

Genome group: AAA

Subgroup: Silk

Status: Cultivar

ITC code

www.promusa.org.latundan

II. Albino Mice

Most of the mice used in laboratories are white albino mice

(Mus musculus). The house mouse, a member of the rodent family,

originated in ancient Asia and later spread throughout Europe. In

the 1500's they traveled to North and South America on English,

French and Spanish ships. Today house mice appear both in the

9
wild or commensally (nesting within human dwellings) in nearly

all parts of the United States.

The mouse has been used in biomedical research since the

early 20th century. Today, over 3000 genetically defined strains

of lab mice are used for research purposes. The primary supply

source of laboratory mice is commercial breeders. Relatively few

are captured from the wild or bred in research laboratories,

although the latter is becoming more common with some genetically

engineered mice. Several characteristics have made the mouse an

appealing research subject. These include the mouse's genetic

similarity to humans (at least 80% of DNA in mice is identical to

that of humans), small size, short lifespan and reproductive

cycle, low maintenance in captivity, and mild manner. For these

reasons, house mice constitute the majority of mammals used in

research, testing and education.

The average lifespan of a wild mouse is typically one to two

years, while mice kept as pets usually live about three years and

have survived up to six years. Mice reach sexual maturity at

between 5 and 6 weeks of age and have tremendous reproductive

potential. They breed throughout the year and may produce as many

as eight litters in a single year, with the average litter

consisting of four to seven pups. Female mice care for young pups

that are fully furred at ten days old and weaned at about three

10
 weeks. After this time, young mice venture out of the nest and

leave their mother's territory, although female mice frequently

remain nearby.

III. Wounds

A wound may be described in many ways; by its aetiology,

anatomical location, by whether it is acute or chronica1, by the

method of closure, by its presenting symptoms, or indeed by the

appearance of the predominant tissue types in the wound bed. All

definitions serve a critical purpose in the assessment and

appropriate management of the wound through to symptom resolution

or, if viable healing.

A wound is a breakdown in the protective function of the

skin; the loss continuity of epithelium, with or without loss of

underlying connective tissue (i.e. muscle, bone, nerves)

following injury to skin or underlying tissues/organs caused by

surgery, a blow, a cut, chemicals, heat/cold, friction/shear

force, pressure or as a result of disease, such as leg ulcers or

carcinomas.

Wounds heal by its primary intention or secondary intention

depending upon whether the wound may be closed with sutures or

left to repair, whereby damaged tissue is restored by the

formation of connective tissue and re-growth of epithelium.

11
 An open wound is an injury involving an external or internal

break in body tissue, usually involving the skin. Nearly everyone

will experience an open wound at some point in their lives. Most

open wounds are minor and can be treated at home. Fall, accidents

with sharp objects or tools, and car accidents are the most

common causes of open wounds. In the case of a serious accident,

call 911 or seek immediate medical attention, particularly if

there’s a lot of bleeding of if bleeding last for 20 minutes.

Types of open wounds, which are classified depending on their

cause:

Abrasion

An abrasion occurs when the skin rubs or scrapes against a

rough or hard surface. Road rash is an example of an abrasion.

There’s usually not a lot of bleeding, but the wound to be

scrubbed and cleaned to avoid infection.

Incision

A sharp object, such as knife, shard of glass or razor

blade, causes an incision. Incision bleed a lot and quickly. A

deep incision can damage tendons, ligaments and muscles.

Laceration

12
 A laceration is a deep cut or tearing of the skin. Accidents

with knives, tools and machinery are frequent causes of

lacerations. The bleeding is rapid and extensive.

Puncture

A puncture is a small hole caused by along pointy object,

such as a nail, needle or ice pick. Sometimes bullet can cause a

puncture wound. Punctures may not bleed much, but these wounds

can be deep enough to damage internal organs. If you have a

puncture wound (even just a small one), visit your doctor to get

a tetanus booster shot and prevent infection.

Avulsion

An avulsion is a partial or complete tearing away of skin

and tissue. Avulsions usually occur during violent accidents,

such as body-crushing accidents, explosions and gunshots. They

bleed heavily and rapidly.

IV. Developing Concepts of Wound Healing

Wound repair is a fundamental process that is required for

tissue homeostasis and regeneration following damage. Most

studies of wound healing have focused on changes in the leading

edge of wounded cells, but here William Razzell, Will Wood and

Paul Martin show that morphogenetic cell shape changes that occur

in multiple cell rows back from the wound are important for

13
efficient wound re-epithelialisation. Using laser-induced

wounding of the Drosophila embryo epidermis as a model, the

researchers first show that multiple row of cells around the

wound stretch towards the closing tissue. They further reveal

dramatic shrinking of the cell-cell junctions that are

perpendicular to the pulling force of the wound. This shrinking

which is driven by pulses of myosin that are directed to the cell

junctions, leads to cell intercalations. Importantly, these

morphogenetic changes, which resemble those observed during the

developmental event of germband extension, are essential for

wound closure; blocking myosin activity in cells behind the

leading edge results in delayed wound contraction. This work

highlights an important role for cells surrounding the wound in

its closure, and suggests that the cellular morphogenetic

mechanisms used during development are recapitulated during wound

healing.

Wounds heal by its primary intention or secondary intention

depending upon whether the wound may be closed with sutures or

left to repair, whereby damaged tissue is restored by the

formation of connective tissue and re-growth of epithelium.

Whether wounds are closed by primary intention, subject to

delayed primary closure or left to heal by secondary intention,

the wound healing process is a dynamic one which can be divided

14
into three phases. It is critical to remember that wound healing

is not linear and often wounds can progress both forwards and

back through the phases depending upon intrinsic and extrinsic

forces at work within the patient.

Related Studies

This chapter presents a resume of literature and studies in

the Philippines and foreign countries that relevant to the study.

Foreign Literatures

Based on the study entitled “From a Traditional Medicinal

Plant to a Rational Drug: Understanding the Clinically Proven

Wound Healing Efficacy of Birch Bark Extract” by Ebeling, Sandra;

et. al (2014), birch bark has a long lasting history as a

traditional medicinal remedy to accelerate wound healing.

Recently, the efficacy of birch bark preparations have also been

proven clinically. As active principle pentacyclic triterpenes

are generally accepted. They, conducted a comprehensive study on

the underlying molecular mechanisms of the wound healing

properties of a well-defined birch bark preparation named as TE

(triterpene extract) as well as the isolated single triterpenes

in human primary keratinocyles and porcine ex-vivo wound healing

15
models. Results show positive wound healing effects of TE and

betulin in scratch assay experiments with a primary human

keratinocyles and in a porcine ex-vivo wound healing model (WHM).

Mechanistically studies elucidate that TE and betulin transiently

up-regulate pro-inflammatory cytokines, chemokines and

cyclooxygenase-2 on gene and protein level. For COX-2 and IL-6,

this increase of mRNA is due to an mRNA stabilizing effect of TE

and betulin, a process in which p38 MAPK and HuR are involved. TE

promotes keratinocyte migration, putatively by increasing the

formation of actin filopodia, lamellipoda and stress fibers.

Detailed analyses show that the TE components betulin, lupeol and

erythrodiol exert this effect even in nanomolar concentrations.

Targeting the actin cytoskeleton is dependent on the activation

of Rho GT Pases. Our result provide insights to understand the

molecular mechanism of the clinically proven wound healing effect

of birch bark. TE and betulin address the inflammatory phase of

wound healing by transient up-regulation of several pro-

inflammatory mediators. Further, they enhance migration of

keratinocytes, which is essential in the second phase of wound

healing. Our results, together with the clinically proven

efficacy, identify birch bark as the first medical plant with a

high potential to improve wound healing, a field which urgently

needs effective remedies.

16
 Another study of the “Wound Healing and Antibacterial

Properties of Methanolic Extract of Pupalia Lappacea Juss in

Rats” by Udegbunan, Sunday Ositadinna et. Al (2014), investigated

the wound healing activities of 80% methanolic leave extract of

Pupalia lappacea using excision, incision and dead space wound

models. Also it effects on three common wound contaminants were

investigated. Excision wounds were created, contaminated with

microbes and treated with ointments (10% and 20%) prepared from

Pupalia lappacea. Incision and dead space wounds were also

created in rats which were subsequently dosed orally with the

extract. The wound healing activities of Pupalia lappacea

ointment on excision wound was assessed by rates of wound

contraction and epithelialization as well as its antibacterial

effects. The effects of Pupalia lappacea on incision and dead-

space wounds were determined by the wound breaking strengths and

weights of the granuloma tissues formed, respectively. Pupalia

lappacea ointments significantly (p<0.05) accelerated wound

healing with 20% ointment having the highest percentage wound

contraction and rate of epithelialization. At 4, 7 and 14 days

post treatment, mean total viable bacterial count of excision

wounds of the extract treated groups were significantly (p<0.05)

lower compared against the control. Wound breaking strengths and

weights of granuloma tissues formed in the extract treated groups

were significantly (p<0.05) higher than those of the control

17
group. The minimum inhibitory concentration values obtained for

the Pupalia lappacea extract against Pseudomonas aeruginosa,

Staphylococus aureus and Bacillus subtiles were 9 mg/ml, 4 mg/ml

and 3 mg/ml, respectively, while the corresponding minimum

bactericidal concentrations were 10 mg/ml, 8 mg/ml and 7 mg/ml.

the results obtained showed that Pupalia lappacea has good wound

healing and antibacterial activities. These findings validate the

use of this plant in traditional medicine for treatment of

wounds.

A study on the “Wound Healing Activity of Argyreia nervosa

Leaves Extract” by AK Singhal, H Gupta, and VS Bhati in 2011,

aimed to evaluate the wound healing property in normal and

diabetic animals by oral and tropical administration of ethanolic

extract of leaves. Phytochemical investigations showed the

presence of various biochemicals (alkaloids, flovonoids,

carbohydrates, triterpenoids, proteins, saponins, steroids,

tannins). A single injection of alloxan monohydrate (20 mg/kg,

i.p.) prepared in citrate buffer (0.1 M, pH 4.5) was to produce

diabetes in rats and mice, after overnight fasting. Excision

wounds (sized 300 mm2 and of 2 mm depth) were used for the study

rate of contraction of wound and epithelization. The means of

wound area measurement between groups at different intervals were

compared using one-way analysis of variance (ANOVA), followed by

18
Dunnet’s test. Extracts of A. nervosa showed significant wound

healing effect in normal (topically treated) and diabetic rats,

the topically treated group showed more significant effect than

the orally treated groups. The present study demonstrates that A.

nervosa leaves extract applied topically promotes healing of

wounds more significantly as compared to oral application, in

both normal rats and alloxan induced diabetic rats, where healing

is otherwise delayed.

Another study on the “Evaluation of the Wound Healing

Activity of an Ethanolic Extract of Ceylon cinnamon Mice” by M.R

Farahpour and M. Habibi in 2012 was conducted to verify the

effect of Ceylon cinnamon on experimentally induced excision

wounds in rats. Thirty-two rats were divided into four groups of

eight rats each. Group A received a placebo containing 1.5% of

cinnamon and Group B a placebo containing 3%. Group C, as the

control group, did not receive any treatment and finally Group D

received a blank placebo as the reference standard group. Wound

healing was monitored on Days 3, 6, 9 and 14 and histological

evaluation was carried out on the samples. The results show that

cinnamon extract served to accelerate the wound healing process

and specifically increased epithelialization in treatment groups

compared to other groups. Thus, this study demonstrates that

19
Ceylon cinnamon may be effective in stimulating the enclosure of

wounds.

In the research conducted by Vipin Kumar Garg and Sarvesh

Kumar Paliwal in 2006 entitled: “Wound-Healing Activity of

Ethanolic and Aqueous Extracts of Ficus benghalensis” was

designed to explore the same. The wound healing activity was

evaluated in excision and incision wound models. The parameters

studied include rate of wound contraction, period of complete

epithelialization, and tensile strength of incision wound.

Student’s test was used to analyze the results obtained from the

present study and P<0.05 was considered significant. Both the

ethanolic and aqueous extracts of F. benghalensis were found to

process significant wound healing activity, which was evidenced

by decrease in the period of epithelialization, increase in the

rate of wound contraction and skin-breaking strength. The present

study has demonstrated that the ethanolic and aqueous extracts of

F. benghalensis have properties that render them capable of

promoting accelerated wound-healing activity compared with

placebo control.

Another study on the “Evaluation of Wound Healing Activity

of Allamanda cathartica L. and Laurus nobilis L. Extracts on

Rats” by Shivananda Nayak, et. al investigated the wound healing

20
activity of wound healing activity of aqueous extracts of

Allamanda and Laurus nobilis in rats.

Excision and incision wound models were used to evaluate the

wound healing activity of both the extracts on Sprague Dawley

rats. In each model, animals were divided into four groups of 10

animals each. In both the model, group 1 served as control and

group 2 as reference standard. In an excision wound model, group

3 animals were treated with Allamanda (150 mg/kg day) and group 4

animals were treated with Laurus nobilis (200 mg/kg b.w day) for

14 days respectively. In the case of incision wound, group 3 and

group 4 animals were treated with Allamanda and Laurus

respectively for 10 days. The effect of vehicles on the rated of

wound healing were assessed by the rate of wound closure, period

of epithelialisation, tensile strength, weights of the

granulation tissue, hydroxyproline content and histopathology of

the granulation tissue.

The aqueous extract of Allamanda promoted wound healing

activity significantly in both the wound models studied. High

rate of wound contraction (p<.001), decrease in the period of

epithelialisation (10.2 + 0.13), high skin breaking strength

(440.0 + 4.53), significant increase in the weight of the

granulation tissue (P<.001) and hydroxyproline (P<.001) content

were observed in animals treated with the aqueous extract of

21
Allamanda. Histological studies of the granulation tissue from

the Allamanda treated group showed the presence of a lesser

number of inflammatory cells, and increased collagen formation

than the control. In Laurus nobilis treated animals, the rate of

wound contraction, weight of the granulation tissue and the

hydroxyproline content were moderately high (P<.05). The

histological study of the granulation tissue of the Laurus

nobilis treated animals showed larger number of inflammatory

cells, and lesser collagen when compared with the Allamanda

treated group of animals. However, it was better than the control

group of animals.

The data of this study indicated that the leaf extract of

Allamanda possesses better wound healing activity than the Laurus

nobilis and it can be used to treat different types of wounds in

human beings too.

In the research conducted by S Mageswari, S Karpagam and G

Aadenaath Reddy in 2015 entitled: “Evaluation of Wound Activity

of the Plant Carmona retusa (Vahl) Masam, in Mice” the effect of

various extracts of Carmona retusa (Vahl) Masam was studied. The

ointment was prepared from the alcohol extract of root, stem and

leaves with petroleum jelly as a base in 5% and 10%

concentrations. Swiss albino mice were used for the evaluation of

wound healing activity. The ointment prepared from various parts

22
exhibited considerable response when compared with standard drug

Nitrofurazone (0.2%) ointment in terms of wound contracting

ability and wound closure time. Remarkable wound healing activity

was observed with the root extract ointment at 5% and 10%

concentration. This result showed that the ointment of plant

extracts accelerate the wound healing process and specifically

increased the epithelization in treatment groups when compared

with control groups. Thus, this wound healing evaluation study

demonstrates that C. retusa is effective in stimulating the

closure of wounds.

Another study on the “Investigations on the Phytochemical

Activities and Wound Healing Properties of Adhatoda vasica Leaf

in Swiss Albino Mice” by Subshani s and Kantha D. Arunachalam in

2010 evaluate the wound healing activity of five different

extracts of the leaves of the plant. Methanol, ethanol, ethyl

acetate, chloroform and hexane were used for the extraction of

the active ingredients. Excision wound model on Swiss albino mice

was used to assess the wound healing activity of the leaves.

Remarkable wound healing activity was observed with the ointment

formulation of the methanol extract at 1% concentration. Wound

contraction was calculated as percentage of the reduction in

wound area. A specimen sample of tissue was isolated from the

healed skin of each group of mice for the histopathological

23
examination. The results of histopathological examination also

supported the outcome of excision wound models. Hematoxylin and

Eosin stained sections and Van Gleson’s stained metachromatic

staining of mast cells. The present study demonstrated that the

aerial parts of A. vasica promote wound healing activity in mice

as a preclinical study.

Sumantha, Meera; Bharvagi and Y in 2014 evaluated the “Wound

Healing Property of Ethanol Extract of Ziziphus mauritania L.

Leaf (EEZM) on Rats Using Excision, Incision and Dead Space Wound

Models”. Wistar albino rats (150-200 gm) were divided into four

groups (n=6). Group I (control) animals received no treatment,

Group II animals were given EEZM 300 mg/kg orally. Group III,

animals were applied EEZM Gel, 100 mg/ cm2 and group IV animals

were applied Framycetin skin cream, 100 mg/ cm 2 topically. In

incision and dead space wound models, the treatment was given for

10 days only from the day of wound, skin-breaking strength and

dry granulation tissue weight were estimated in incision wound

model. Hydroxyproline content was estimated in dead space wound

model. In excision wound model, treatment continued till the

complete healing of the wound. Rate of wound contraction and

period of episthelisation was evaluated thereafter. The data was

analyzed by one-way ANOVA, followed by Dunnet’s test. P<0.05 is

considered significant. Results: EEZM increased the rate of wound

24
contraction, decreased period of episthelisation, increased skin-

breaking strength and dry granulation tissue weight, evaluated

hydroxyproline content. Conclusion: Ziziphus mauritania possesses

wound healing activity.

Local Literature

According to Marzha Ancheta and Liwayway Acero in 2016

entitled “Wound Healing Property of Carica papaya Stem in Albino

Rats” focused on which remains in this field after the harvest is

often neglected study. Carica papaya stems were dried cut and

turned into ointment form. The final product was applied to

inflicted wounds of five albino rats which are assigned in the

treatment group (treatment 2). The Albino rats served as

experimental animals. They are randomly assigned in two groups.

The first group or treatment 1 has 5 animals. The second group,

treatment 2 served as the experimental animals where the ointment

from papaya stem was applied. Initial wound size in millimeter

after fifteen days showed significant result. The result revealed

that the stem of Carica papaya has the wound healing property in

Albino rats. The result of this study attested that the stem of

Carica Papaya can be utilized as source of herbal plat,

specifically to heal wounds.

25
 Another study on the “Wound Healing Potential of Tinospora

crispa (Willd) [Menispermaece] Stem on Diabetic Mice” by Ryan O

Arcueno, et. al(2015) aimed to compare the effect of Tinospora

crispa on healing of diabetic wound among albino mice when given

intraperitoneally and when administered along with an ointment

topically applied to the wound. Results showed significantly

reduction in blood glucose level among mice given the plant

extract. However, it may be suggestive that the additional

ointment had not contributed significantly to wound healing as

the wound healing time and percent wound contraction between the

two groups are not significantly different.

Another study on “Xanthine oxidase Inhibition of Selected

Philippine Medicinal Plants” by Karmella L. Apaya and Christine

L. Chichioco-Hernandez in 2010 examined selected species of

Philippine plants traditionally used for their anti-inflammatory

properties for their ability to inhibit the action of Xanthine

oxidase. The degree of inhibition using Adenanthena payonina,

Antegonon leptopus, Blumea balsamifera, Calophyllum olercea,

Pogostemon cablin, Solanum tomum, Tinospora rumphill and vitex

negundo extracts were determined by measuring the increase in

absorbance at 295 nm which is associated with uric acid formation

which is linked to gout. B. balsamifera has the highest percent

inhibition at 79.67% followed by M. pudica with 62.36%

26
inhibition. The Xanthine oxidase inhibitory potential and IC 50

values of the extracts are reported.

Still another study entitled: “The Effect of Mangifera

indica (Mango) Leaf Crude Extract in the Wound Healing Properties

of Rattus norvegicus (Sprague Dawley)” by Jose Jurel M. Nuevo and

Emerson Ronald K. Bernanindo 2012-2013 evaluated the effects of

Mangifera indica (Mango) leaf crude extract on the wound healing

properties of Rattus norvegicus (Sprague Dawley).

The male Rattus norvegicus was divided into three groups and was

inflicted full thickness wound created on the predetermined

dorsal nape area via surgical procedure. The three groups were

treated topically with Bactroban, sterilized distilled water and

the leaf crude extract of Mangifera indica once a day from day 0

to 14. Macroscopically, the wounds were evaluated and the

measurement were compared by using Mann-Whitney Test to show the

significance between the wound size treated with Bactroban and

Mangifera indica leaf crude extract and also by comparing the

wound size treated with Mangifera indica leaf crude extract and

the negative control (sterile distilled water). The Mangifera

indica leaf crude extract had significantly (p<0.05) promoted

wound healing compared to the positive and negative controls.

27
 Another study on “The Burn Wound Healing Potential of Bixa

Orellana Linn [Bixaceae] Leaf Extracts on Albino Mice” by Albert

A Espiritu, Shaira Nicole L Lao and Jonathan Jaime G Guerrero in

2015 investigated the healing potential of the plant on burn

wounds using five replicates of mice per treatment as the animal

model. Different preparation procedures were employed: crude,

ethanolic and ointment. These were compared to the commercially

available silver sulfadiazine (SSD). Results suggest that Bixa

orellana, regardless of preparation procedure, were effective in

treating burn wounds and is comparable to SSD. Percent wound

contraction, re-epithelialization time, and histological features

of the wound treated with Bixa orellanaextracts were comparable

to the control (SSD).

Another study on “The Wound Healing Action of Kakawati Gel

from Gliricidia sepium (Jacques) Steudel (Family Fabaceae)” by

Rogie Royce Carandang, Kristal Cielo Beumio and Adelaine Lopez in

2015 states the Objective: Gliricidia sepium, locally known as

kakawati is recognized for its wound healing property. The study

aimed to determine the wound healing action of a safe, stable,

low cost formulation from the lyophilized sap of G sepium.

Methods: The organoleptic, physical, and chemical properties were

determined. Short term stability testing, as well as

compatibility study with common excipients was performed. Wound

28
healing property was determined by three methods: (1) measurement

of the rate of wound contraction, (2) tensile strength of the

healed tissue using a tensiometer and (3) histological

examination of collagen deposition. The 7.5% gel formulation was

also tested for its efficacy with wound healing-impaired rats.

Results: The pilot-scale manufacture of a gel-based formulation

using water extract of G sepium was feasible. The extract was

compatible with glycerin, sodium CMC, sodium benzoate and sodium

citrate. Stability testing with common excipients showed that the

gel formulation was stable at a temperature of 30ºC. It was

stable at pH values of 5 to10 and in natural and artificial

light. The formulated product is a green gel of moderate

consistency possessing a natural fragrance with faint fetid odor.

The gel formulation was found to be effective at 7.5%

concentration as a wound healing agent. Conclusion: The gel

formulation has been proven to have wound healing properties and

was found to be safe, effective, stable and low.

Statement of the Problem

29
 The study aimed to determine the wound healing activity of

Latundan (Musa sapientum) peeling ointment on rats.

Specifically, it sought to answer the following questions:

1) . What are the secondary metabolites present in the peeling

of the test plants?

2) . What is the mean length of the wounds in the test animal

before and after administration of the test drug?

3) . Is there a difference in the mean length of the wounds in

the test animals before and after the administration of the

test drug?
4) . What is the mean length of the wounds in the test animals

before and after administration of the commercial drug?

5) . Is there a difference in the mean length of the wounds in

the test animals before and after administration of

commercial drug?
6) . Is there a difference between the mean length of the

wounds after administration of test drug and commercial

drug?

Hypotheses

30
  There is no difference in the mean length of the wounds in

the test animals before and after the administration of the

test drug.
 There is no difference in the mean length of the wounds in

the test animals before and after administration of

commercial drug.
 There is no difference between the length of the wounds

after the administration of test drug and commercial drug.

Conceptual Framework

The study was based on the following framework showing the

relationship of the variables to assess the wound healing

activity of Latundan (Musa sapientum) peeling ointment on rats.

INDEPENDENT VARIABLE DEPENDENT VARIABLE

Latundan (Musa sapientum) Mean length of wound

Peeling extract

The paradigm shows the relationship between the variables.

The independent variable is Latundan (Musa sapientum) peeling

extract and the dependent variable is mean length of wound.

31
Significance of the Study

If the study is proven to be effective, it will benefit the

following:

 Patients – It helps the wound suffering patients in

decreasing expenses used on buying commercial ointments.

 Pharmaceutical companies – The study will give an advantage

on providing an alternative and inexpensive means of

treating wounds.
 Community – This study helps the community in providing

knowledge about the use of latundan as wound healing.

Moreover, instead of throwing the peelings of banana, it

will be used for something beneficial.

 Banana Farmers – It helps the farmers who plant banana to

have a source of income.

 The Researchers – The research provides a better grasp of

understanding and experiences in different laboratory

methods thus enhancing their laboratory skills. In addition,

new ideas and information related to this study which can be

useful for pursuing their Science-related careers.

 Future Researchers – The study will serve as a future

reference for other researchers who would be interested to

conduct similar study.

32
Scope and Delimitation

The study was limited to the usage of ripened Latundan

banana (Musa sapientum) peeling ointment on rats. It was also

limited to the evaluation of its efficacy by observing: rate of

healing, wound length decrease, and presence of wound scores

(tissues) in comparison to the commercial one (Terramycin) via

extra dermal application in albino rats incisional wounds. During

the course of the study, the administration of products was

further limited to twice a day only. The plant was obtained from

the public market of Don Domingo. Plant authentication was done

at the National Quarantine Services Division, Station 02, Region

Government Center, Carig Sur, Tuguegarao City, Cagayan and the

pythochemical analysis of the plant was done at the Department of

Science and Technology in Bagay, Tuguegarao City, Cagayan. The

infusion and preparation of ointment was done at research

laboratory of SPUP. The bioassay was conducted at the Cagayan

Valley Herbal Processing Plant, Carig Sur, Tuguegarao City,

Cagayan.

The study was conducted on December-March 2016.

Definition of Terms

33
 The study entitled: “Wound Healing Activity of Latundan

(Musa sapientum) Peeling Ointment on Rats” uses this following

terminologies:

 Infusion – a process whereas the vegetable oil infuses with

peelings of latundan.
 Incision wound - a cut on the rats administered by blade.
 Keratinization – a thick layer of skin usually seen after

the healing of wound of the rats.

 Musa sapientum – a kind of banana used in the study.
 Terramycin – the positive control of the study.

CHAPTER 2
METHODOLOGY
This chapter presents the methods and procedures used in

assessing the wound healing activity of Latundan banana (M.

Sapientum) peeling ointment on rats.

A. Chemicals

The standard drug that was used for the comparison of

the commercial drug from the Latundan banana peeling

ointment is Terramycin. It was bought from the

pharmaceuticals.

34
B. Plant Material

Peelings of Latundan banana (M. Sapientum) were

collected from the public market of Don Domingo. The

specimen was authenticated at the Cagayan Valley Herbal

Processing Plant, Carig Sur, Tuguegarao City, Cagayan.

C. Phytochemical Procedures

The extract was evaporated to incipient dryness and

tested for the presence of the stated phytochemicals following

Guevarra et al (2005).

2.5.1 Test tube screening methods

 An equivalent of 10g plant material was taken from the stock

plant extract prepared in Section 1.1 and evaporated to

incipient dryness over a steam bath;

 It was cooled to room temperature; defatted by taking up the

residue with 9mL hexane and water (2:1) or with petroleum

ether. The hexane or petroleum ether extract was discarded.

35
  The defatted aqueous layer with 10Ml of 80% ethyl alcohol

was diluted;

 The filtrate was filtered and divided into four test tubes;

 One portion was taken as control.

2.5.1.1 Test for leucoanthocyanins: Bate-Smith and Metcalf method

 One portion of the above alcohol filtrate was treated with

0.5 Ml conc, hydrochloric acid (12M);

 It was observed for any color change;

 It was warmed for 15 minutes in a water bath. Observed for

further color change within an hour and compared with the

control;

 The results were recorded.

2.5.1.2 Test for y-benzopyrone nucleus: Wilstatter “cyadin” test

 Another portion of the alcohol filtrate above, refer to

section 2.5.1, were treated with 0.5 ml conc, hydrochloric

acid (12M);

 3-4 pieces of magnesium turnings were added;

36
  Observed within 10 minutes for any change in color and

compared with the control tube;

 If definite coloration occurs, dilute with an equal volume

of water and add 1mL of ethyl alcohol;

 Shake well and allow to stand;

 Note the color in each layer. Record your results.

2.5.2 Two-dimensional thin-layer chromatographic (TLC) screening

method

Two Dimensional Chromatographic

 Glass plates 12x12 cm coated with silica gel G were used;

 Spot the extract about 3-4 cm from the left corner of the

silica coated plate;

 Developed in equilibrated chamber using the first solvent up

to the solvent front;

 Air-dried; turned the plate 90° counter clockwise and

developed in the second solvent allowing the solvent to

travel in a perpendicular direction as that of the first

solvent;

 Air-dried and sprayed.

37
Preparation of rutin standard

A 0.05% solution of rutin in distilled water was prepared.

Acetic acid, 5%

17 mL of glacial acetic acid were dissolved in enough water

to make 100 mL.

 The last portion of the alcoholic filtrate above (refer to

Section 2.5.1) was applied, as a spot on a silica gel G-

plate for thin-layer chromatography.

 Applied the rutin standard as a spot adjacent to the

alcoholic filtrate;
 Developed the chromatogram with 1-butanol-acetic acid-water

(4:1:5), upper phase;

 Dried the chromatogram and turned the plate 90º counter-

clockwise and developed with 5% acetic acid;

 The chromatogram was air dried, then observed under visible

and UV light for colored spots;

 Sprayed the chromatogram with ammonia, and observed for

color changes in the spots comparing with that obtained with

the rutin standard;

 Document the results with tracing paper.

2.6 Saponins

2.6.1 The screening methods for saponins

38
 2.6.1.1 The froth test

Saponins in plant materials can cause a persistent

foam when the aqueous solution is agitated.

 Take a volume of the plant extract prepared in Section

1.1equivalent to 2 g plant material and transfer this to a

test tube;

 In a separate test tube have 1 mL of the plant extract to

stand as the standard;

 Add 10 mL of distilled water to each of the test tubes,

stopper and shake both test tubes vigorously for 30 seconds;

 Allow to stand for 10 min. and observe for a ‘honeycomb’

froth;

 Compare the result of the plant extract with that of the

standard.

2.6.1.2 The capillary tube test

Saponins tend to lower the surface tension in water.

Capillary tubes specifications

39
 Use a pair of 15 x 1 mm O.D. capillary tubes.

 Load a capillary tube with the plant extract from Section

1.1 by immersing the tube to a height of 10 mm in the plant

extract;

 Likewise load another capillary tube with distilled water;

 Lift the capillary tubes and keep both in a vertical

position to allow the liquid inside to flow out freely;

 After some time, compare the height of the liquids in the

two tubes.

2.6.1.3 The hemolytic test: the agar cup semi-quantitative method

The most characteristic property of saponins is their

ability to cause hemolysis. When added to a suspension of blood,

saponins produce changes in erythrocyte membranes causing

hemoglobin to diffuse into the surrounding medium.

2.6.1.4 Determination of saponins in the presence of tannins

Tannins and certain plant acids have been shown to interfere

with the hemolytic test for the detection of saponins. If

saponins and tannins were both present in a plant extract, no

hemolysis would occur. A false-negative saponin test results.

40
 If tannins and other plant polyphenolic compounds are found

to be present in the plant extract then these are removed by

preliminary treatment of the plant extract with magnesium oxide.

Refer to Section 2.7.1. The latter has been found to be effective

in removing interfering tannings and other plant polyphenolic

compounds.

Sodium chloride, 10%

Dissolve 10 g NaCl in enough distilled water to make

100 mL solution.

 A volume of the plant extract prepared equivalent to 10 g

plant material was taken and evaporated into incipient

dryness over a steam bath;

 The residue was extracted with 20 mL of hot distilled water.

Five drops of 10% sodium chloride solution were added and

filtered;

 The saline filtrate with 2 g of magnesium oxide, to form a

slurry and heat, was treated over a steam bath for about 10

min;

 The slurry was extracted with hot 80% ethanol, then

filtered;

41
  The detannated aqueous-ethanol filtrate was subjected to the

hemolytic test, given in Section 2.6.1.3.

2.6.2 Semi-quantitative assays for saponins (WHO, 1998)

The hemolytic activity of plant material is determined by

comparison with a reference material, Saponin R, a commercially

available reference material. Saponin R has a hemolytic activity

of 1000 units per gram.

A suspension of erythrocytes is mixed with equal volumes of

serial dilution of the extract of the plant material. The lowest

concentration to effect complete hemolysis is determined after

allowing the mixtures to stand for a given period of time.

2.6.2.1 Preliminary test

 A serial dilution of the plant extract with phosphate buffer

pH 7.4 solution and 2% blood suspension in four test-tubes

were prepared. Refer to Table P1.

Table P1. Serial dilution of the plant extract with phosphate

buffer

Tube No.
1 2 3 4
Plant extract (mL) 0.10 0.20 0.50 1.00
Phosphate buffer pH 7.4 0.90 0.80 0.50

solution (mL)
2% Blood suspension (mL) 1.00 1.00 1.00 1.00

42
  As soon as the tubes have been prepared, they were gently

inverted, avoiding the formation of foam.

 They were gently inverted again after a 30-min interval and

were allowed to stand for 6 hours at room temperature.

 The tubes were examined and the dilution was recorded at

which total hemolysis had occurred, indicated by a clear,

red solution without any deposit of erythrocytes.

2.6.2.2 Main test

 A serial dilution of the plant extract was prepared,

undiluted or diluted as determined by the preliminary test,

with phosphate buffer pH 7.4 solution and 2% blood

suspension using 13 test tubes as shown in Table P2.

Table P2. Serial dilution of the plant extract, undiluted or

diluted with phosphate buffer

Tube 1 2 3 4 5 6 7 8 9 10 11 12 13

No

Particular

s
Plant extract: 0.40 0.45 0.50 0.55 0.60 0.65 0.70 0.75 0.80 0.85 0.90 0.95 1.00
diluted if

necessary (mL)
Phosphate buffer 0.60 0.55 0.50 0.45 0.40 0.35 0.30 0.25 0.20 0.15 0.10 0.05 ---
pH 7.4 solution

(mL)
2% Blood 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00

suspension (mL)

43
  The dilutions and evaluations as in the preliminary test

were carried out but the results were observed after 24

hours;

 The amount of plant material in grams (g) that produces

total hemolysis were calculated;

 To eliminate the effect of individual variations in

resistance of the erythrocyte suspension to saponin

solutions, a series of dilutions of saponin R in the same

manner as derived above for the plant extract was prepared.

 The quantity of saponin R in grams that produces total

hemolysis were calculated.

2.6.3 The Liebrmann-Burchard test for unsaturated sterols and

triterpenes

The Liebermann-Burchard test for unsaturated sterols and

triterpenes gives a range of colored from blue to green, red,

pink, purple or violet.

 The equivalent of 10 g plant material from the stock plant

extract prepared was taken and was evaporated to dryness

over a steam bath. Cool to room temperature;

 The material as in Section 2.2.1 was defat.

44
  The aqueous layer with 10 mL chloroform was treated

cautiously, then, the mixture was gently shaken.

 Allow to stand and pipette off the chloroform extract;

 The chloroform extract was dried by filtering the mixture

through about 100 mg anhydrous sodium sulfate held over dry

filter paper.

 The filtrate was divided into two portions. One portion was

used for control;

 The other portion was treated with 3 drops of acetic

anhydride, then one drop of conc. sulfuric acid;

 Any immediate color change was observed and let stand for an

hour;

 Further color changes were observed;

 It was compared with the control and the results were

recorded.

2.6.4 Thin-layer chromatographic (TLC) test for sapogenins

45
 The equivalent of 10 g plant material from the stock extract

prepared was taken and places in an Erlenmeyer flask;

 2 mL of 1M HCL were added and was stirred thoroughly;

 The flask was covered with a funnel and was refluxed for 2-6

hours over a water bath to hydrolyze the saponins. It was

cooled to room temperature;

 The hyrolyzate with dilute ammonia (6M) was extracted and

evaporated to incipient dryness over a steam bath;

 The residue was extracted twice with 3 mL portions of

hexane, discarding the hexane-insoluble residue;

 The hexane extract was washed by partitioning with 2 mL

water. The hexane layer was pipette off.

 The hexane layer was evaporated to incipient dryness over a

steam bath. The residue contained the sapogenins;

 The sapogenin residue were dissolve in 5 drops of chloroform

cautiously;

 It was subjected to thin-layer chromatography in silica gel

G plates;

46
  The developed chromatogram was sprayed with antimony (III)

chloride in acetic acid and the sprayed chromatogram was

heated at 100ºC for 10 min.;

 The results were recorded by tracing the spots observed in

the chromatogram tracing paper.

2.7 Tannins and Polyphenols

2.7.1 Screening methods for tannins

2.7.1.1 The test tube method

 An equivalent of 10 g plant material from the stock plant

extract prepared was taken and evaporated to incipient

dryness over a steam bath;

 The residue with 20 mL of hot distilled water was extracted.

5 drops of 10% sodium chloride solution were added;

 The filtrate was filtered and divided into three test tubes.

One portion was taken as the control;

 An aqueous solution of tannic acid was taken as reference

standard.

Gelatin test

47
  One portion of the filtrate of the plant extract with three

drops of gelatin salt reagent was treated. The same was done

to the tannic acid solution, the reference standard;

 The control and the reference standard were compared.

Ferric Chloride test (Confirmation)

 Another portion of the plant filtrate with three drops of

ferric chloride reagent was treated. The same was done to

the tannic acid solution;

 The control and the reference standard were compared.

D. Infusion

Bananas were washed with running water. The fruits were

peeled; only the peels were used. One hundred grams (100g)

of peels were diced into cubes. It was fry on a vegetable

oil (100g) for 5-6 minutes. Then get only the oil.

E. Peel Ointment Preparation

First, set up a double boiler, a pot over a bowl on top of

it. Second, pour 300ml of water then boil it in a pot.

Third, put the petroleum jelly or white petrolatum and

48
 beeswax in the bowl to melt it then add the banana peeling

oil. Fourth, stir the mixture in a very low heat. Lastly,

pour it into a container. The mixture was thicken into an

ointment as it cools.

F. Preparation of Test Animals

1. Restrain the animals.

2. Wipe injection site (back, dorsal region) with cotton

soaked with 70% alcohol.

3. Make one (1) cm wound by cutting its skin.

G. Administration of the Test drug and Control

There were three set-ups with three replicates. Set-up A was

treated with banana peeling ointment. Set-up B was treated with

Terramycin while no treatment was given to the test animals in

Set-up C. A small amount (an amount equal to the surface area of

the tip of a finger) of experimental and control drugs were

applied twice a day and observation of changes in every five (5)

days.

H. Statistical Analysis

To determine the Wound Healing Activity of Latundan (Musa

sapientum) Peeling Ointment on Rats, the Analysis of Variance

(ANOVA) was used with 0.05% level of significance.

49
 CHAPTER 3

Results and Discussion

This chapter deals with the presentation, interpretation and

analysis of data.

Results

Table 1. Result of the Phytochemical Analysis of Latundan (Musa

sapientum) Peeling Extract

Sample Parameter Result Method Used

Distribution
Ethanolic of Phytochemical

Latundan Screening: Guevara et. al

Banana Peel Flavanoids + (2005)

Tannins +

Saponins -

50
The table shows the results of the phytochemical analysis of

Latundan (Musa sapientum) peeling extract following the Guevara

et. al (2005) method where flavanoids and tannins are present.

A total of 9 rats inflicted incision wound on the thigh part

were used in the study. Each rat were measured their individual

wound size upon incision and thereafter measured 5 and 10 days

later after application of corresponding test drug in ointment

form.

CHAPTER 4

Summary OF Findings, Conclusion and Recommendation

This chapter presents the summary of findings, conclusions,

and recommendations of the study.

Summary of Findings

Nine rats were used in the evaluation of latundan ointment wound

healing activity. The objective was to compare the wound size of

rats medicated with the ointment preparation and compare it with

terramycin ointment and none application of any drug for the

third group following completely randomized design. The rats

wounds were inflicted with the same size initially were treated

with the ointment daily for 5 and 10 days where the evaluation of

51
the wound size were obtained thereon. Results showed the initial

wound size were not significantly different suggesting that the

wounds inflicted were homogenous from the beginning of the study.

The 5th and 10th day however did show significantly different mean

size effect between treatments after the application of

treatments (appendix table 3c and 4c).

Conclusion

Based on the results, we conclude that banana peel have a wound

healing activity to an inflicted wound in rats used in the study

which is significant on day 5; wounds heal on the 10 th day

without infection.

Recommendation

Based on the results of the study, the researchers recommend the

following:

1. A parallel study should be conducted using other parts of

Latundan (Musa sapientum) plant (e.g. fruits, roots, stem,

etc).

2. Toxicity studies of Latundan peeling extract should be

conducted to determine the safety of the test drug.

3. Use other types of wounds such as puncture, abrasion and

etc.

52
 BIBLIOGRAPHY

References

1 Enoch S and Price P (2004) Cellular, Molecular and biochemical

differences in the pathophysiology of healing between acute

wounds, chronic wounds and wounds in the aged. Worldwidewounds.

2 Leaper DJ and Harding KG. (1998) Wounds: Biology and

Management. Oxford University Press.

3 Hutchinson J (1992). The Wound Programme. Centre for Medical

Educational: Dundee

4 Cooper P. (2005) A review of different wound types and their

principles of management in Wound Healing: A systematic approach

to advanced wound healing and management. Cromwell Press, UK

(Chandan K. Sendan et. al)

Diversity of banana cultivars portal >> AAB genome group >> Silk

subgroup >> Latundan

53
Evaluation of the Wound Healing Activity of an Ethanolic Extract

of Ceylon Cinnamon Mice by M.R Farahpour and M. Habibi in 2012

Evaluation of Wound Healing Activity of Allamanda cathartica L.

and Laurus nobilis L. Extracts on Rats by Shivananda Nayak, et.al

From a Traditional Medicinal Plant to a Rational Drug:

Understanding the Clinically Proven Wound Healing Efficacy of

Birch Bark Extractby Ebeling, Sandra; et.al (2014)

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

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

Investigations on the Phytochemical Activities and Wound Healing

Properties of Adhatoda vasica Leaf in Swiss Albino Mice

(John P. Cunha, DO, FACOEP)

Maynard J (2015) How wounds heal. Four main phases of wound

healing.Swiss albino Mice by Subshanis and Kantha D. Arunachalam

in 2010

Marzha Ancheta and Liwayway Acero in 2016 entitled “Wound Healing

Property of Carica papaya Stem in Albino rats”

(Rafael J. Consunji, Adnan A. Hyder)

S Mageswari, S Karpagam and G Aadenaath Reddy in 2015 Evaluation

of Wound Activity of the Plant Carmona retusa (Vahl) Masam, in

Mice
54
Sumantha, Meera; Bharvagi and Y in 2014 evaluated the wound

healing property of ethanol extract of Ziziphus Mauritania L.

leaf (EEZM) on rats using excision, incision and dead space wound

models

The Burn Wound Healing Potential of Bixa Orellana Linn [Bixaceae]

Leaf Extracts on Albino Mice by Albert A Espiritu, Shaira Nicole

L Lao and Jonathan Jaime G Guerrero in 2015

The Effect of Mangifera indica (Mango) Leaf Crude Extract in the

The Wound Healing Action of Kakawti Gel from Gliricidia sepium

(Jacques) Steudel (Family Fabaceae) by Rogie Royce Carandang,

Kristal Cielo Beumio and Adelaine Lopez in 2015

Vipin Kumar Garg and Sarvesh Kumar Paliwal in 2006 Wound-Healing

Activity of Ethanolic and Aqueous Extracts of Ficus benghalensis

Wound Healing Activity of Argyreia nervosa Leaves Extract by AK

Singhal, H Gupta, and VS Bhati in 2011

Wound Healing Potential of Tinospora crispa (willd)

[menispermaece] Stem on Diabetic Mice by Ryan O Arcueno, et.al

(2015)

Wound Healing Properties of Rattus norvegicus (Sprague Dawley) by

Jose Jurel M. Nuevo and Emerson Ronald K. Bernanindo 2012-2013

55
Xanthine oxidase Inhibition of Selected Philippine Medicinal

Plants by Karmella L. Apaya and Christine L. Chichioco-Hernandez

in 2010

www.wikihow.comUse-Bananapeel

56