Pomegranate (Punica granatum)

Flower used as Wound

Healing Cream

TABLE OF CONTENTS

TITLE

TABLE OF CONTENT

CHAPTER I: INTRODUCTION

Background of the Study

Objective

Statement of the Problem and Research Questions

Significance of the Study

Literature Review

Hypotheses

Research Paradigm

CHAPTER II: METHODOLOGY

Specific method and Research Design

Instruments and Procedures

Data Analysis

CHAPTER III: RESULT

CHAPTER IV: DISCUSSION

CHAPTER V: CONCLUSION

CHAPTER VI: RECOMMENDATION

CHAPTER VII: REFERENCE

CHAPTER VIII: APPENDICES

Literature Matrix

Variable matrix

Synthesis Tally

Data Collection Forms

Data Matrix

General Tables

Other Supporting Materials

CHAPTER I

INTRODUCTION

Pomegranate has been in traditional medicinal use for more than 3000 years.

Ancient and mystical, it has been written about in the Old Testament of the Bible, the

Jewish Torah, and the Babylonian Talmud as a sacred fruit with powers that bestow

fertility, abundance, and good luck. It is also part of the iconography of rituals, art, and

mythology - the personal emblem of the Roman Emperor, Maximilian, the symbol and

heraldic device of the ancient city of Granada in Spain.

More than 80% of the world is population depends upon traditional medicines for

various skin diseases. Recently, the traditional use of plants for wound healing has

received attention by the scientific community. Approximately one-third of all traditional

medicines in use are for the treatment of wounds and skin disorders, compared to only

1-3 % of modern drugs. Wound healing is a complex process characterized by

homeostasis, reepithelization, and granulation tissue formation and remodeling of the

extracellular matrix. Reports about medicinal plants affecting various phases of the

wound healing process, such as coagulation, inflammation, fibroplasia, collagenation,

epithelization and wound contraction are abundant in the scientific literature. A survey of

the ethnobotanical studies, carried out in other contries, indicated the use of several of

plant species by the inhabitants of the area, especially by those habiting the rural areas,

for wound healing purpose.

Punica granatum Linn., known locally as Granada. Granada is a shrub growing 2

to 3 meters high. Branchlets are slender and 4-angled. Leaves are oblong-lanceolate to

oblong-elliptic; 4 to 6 centimeters long, short-stalked, and pointed at both ends. Flowers

are red and showy, usually with six segments in the calyx which are 2.5 to 3 centimeters

long. Petals are obovate, about 2 centimeters long. Stamens are numerous. Fruit is

rounded, reddish-yellow or purplish, 7 to 10 centimeters in diameter. Rind is thin, tough,

and brittle. Fruit contains numerous seeds, each seed surrounded by a watery,

translucent, flavorful pinkish-red pulp. Granada is an important medicinal plant in Iran

whose flowers are used as astringent, hemostatic, antibacterial, antifungal, antiviral and

as a remedy for cut wound, bronchitis, diarrhea, digestive problems, man sex power

reconstituent, dermal infected wounds and diabetes in Unani medicinal (Iranian

Traditional Medicine) literature. This flower was also used for the treatment of injuries

from falls and grey hair of young man in the traditional Chinese medicine.

Punica granatum contains polyphenol compound named pomegranatate, ellagic

acid, 3,3,4-tri-Omethylellagic acid, ethyl brevifolincarboxylate, urolic and maslinic acids,

and daucosterol.

BACKGROUND OF THE STUDY

Herbal drugs are considered alternative agents and have been used for several

years around the world. Recurrent aphthous stomatitis (RAS) is one of the most

common problems recognized by dentists and skin specialists. This problem is

characterized by recurring, painful, small oral mucosal ulcers with a round or oval aspect

that mostly appear in keratinized mucosa, cheeks, and on the surface of the mouth

under the tongue.

Chronic wounds represent a significant burden to patients, health care

professionals, and the US health care system, affecting 5.7 million patients and costing

an estimated 20 billion dollars annually. To effectively manage these problems, one

must understand the normal healing process and engineer a salubrious physical and

biochemical environment. This article outlines normal healing biology, describes the

factors that facilitate or impair wound healing, surveys common types of problem

wounds, and discusses emerging concepts in chronic wound management.

Rural areas in Cagayan don’t know that this plant is also a remedy for wound

healing. According to Perez et al., pericarp extract of Punica granatum possess strong

antibacterial activity against the multiple resistance of Salmonella typhi. Pomegranate is

a garden plant here in the Philippines and what we are trying to search was what are the

other part can have a wound healing property of the plant and is it more effective than

the leaves which are researched before.

OBJECTIVE

The study aims to achieve the following:

a. Cite the factors that causes chronic wounds.

b. Provide a cream remedy for wound healing.

c. To determine the effectiveness and the benefit of Pomegranate as

would healing cream.

STATEMENT OF THE PROBLEM

The study aims to determine the efficacy of wound healing cream using

pomegranate flower.

Specifically it aims to answer the following questions:

a. Can pomegranate effectively kills bacteria in wound?

b. What bacteria can pomegranate kill?

c. What are the diseases that pomegranate can cure?

SIGNIFICANCE OF THE STUDY

This research in one way or another would greatly help the following in the future.

a. Community

The flowers used in this cream are readily available everywhere so many people

especially in rural areas in Cagayan.

b. Businessman

This could be a source of income to small businessman.

c. Educators

A good source of information to discuss with the students and to promote

experiment of undiscovered property of plants.

d. Future Researchers

This researcher could be a Literature Review for the future researchers who will

improve or make a related study to these.

REVIEW OF RELATED LITERATURE

Pomegranates have been known for hundreds of years for their multiple health benefits,

including antimicrobial activity. The recent surge in multidrug-resistant bacteria and the

possibility of widespread global virus pandemics necessitate the need for additional preventative

and therapeutic options to conventional drugs. Research indicates that pomegranates and their

extracts may serve as natural alternatives due to their potency against a wide range of bacterial

and viral pathogens. Nearly every part of the pomegranate plant has been tested for

antimicrobial activities, including the fruit juice, peel, arils, flowers, and bark. Many studies have

utilized pomegranate peel with success. There are various phytochemical compounds in

pomegranate that have demonstrated antimicrobial activity, but most of the studies have found

that ellagic acid and larger hydrolyzable tannins, such as punicalagin, have the highest

activities. In some cases the combination of the pomegranate constituents offers the most

benefit. The positive clinical results on pomegranate and suppression of oral bacteria are

intriguing and worthy of further study. Much of the evidence for pomegranates’ antibacterial and

antiviral activities against foodborne pathogens and other infectious disease organisms comes

from in vitro cell-based assays, necessitating further confirmation of in vivo efficacy through

human clinical trials.

Creams are used for patients prone to skin breakdown from pressure, shear, or

incontinence. This study intend for prevention and for resolving new-onset problems. Apply

creams several times a day on the skin over bony prominences or on areas prone to

breakdown. Gallic acid and catechin are the major components of Punica granatum which are

responsible for the healing activity. This properties were found in leaves and we must prove that

the flower part must also have these properties.

A skin wound that doesn't heal, heals slowly or heals but tends to recur is known as a

chronic wound. Some of the many causes of chronic (ongoing) skin wounds can include trauma,

burns, skin cancers, infection or underlying medical conditions such as diabetes. Wounds that

take a long time to heal need special care.

HYPOTHESIS

The researchers assume the following:

a. There is a significant effectiveness of Pomegranate flowers as wound healing cream.

b. There is a significant relationship between the number of hours that the wound heals

and the type of cream used.

c. The cream may not be effective because the flower part use is not yet tested and not

prepared yet as cream.

SCOPE AND DELIMITAION

A. Subject Matter Discussed

This study is going to focus on the determination of the effectiveness of

pomegranate flowers in wound healing.

B. Locale of the study

The study is going to conduct in the Pharmacy Laboratory M-Building University of

Saint Louis Tuguegarao, Lecaros Ext. Ugac Sur, Tuguegarao City, Cagayan.

RESEARCH PARADIGM

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CHAPTER II

METHODOLOGY

I. MATERIALS AND METHODS

a. Plant materials

The male abortive flowers of Punica granatum L. (Punicaceae), is a shrub or small tree and

considered to be a native of Cagayan . Punica granatum will be collected on the slopes of the

Solana , District of Cagayan.

b. Preparation of the extract

Ethanol successive extract of Punica granatum, yield: 10% for flower, were prepared. Ethanolic

extract of flower samples tested positive for polyphenols in flower.

c. Animals

Male Wistar rats (180-200 g) of 2-3 months of age were used. The animals were housed in

standard environmental conditions of temperature (22 ± 3OC), humidity (60 ± 5%) and a 12-h

light/dark cycle.

d. Wound healing activity

Wound induction and evaluation extracts for properties wound healing before the beginning of

the wound healing experiments, the dorsal skin of the Wistar rats were shaved. Animals were

anesthetized with 1.5 mg/kg, i.p. of Ketamin and Xylazine. A full thickness of the excision wound

(circular area about 150 mm2 and 2 mm depth) was created along the markings using toothed

forceps, a surgical blade and pointed scissors. The animals were divided randomly into four

groups of nine rats each. Group 1 was treated with nitrofurazone ointment and served as a

reference standard; groups 2 and 3 were treated topically with the simple ointment prepared

from extract of Punica granatum (200 mg/kg/day), respectively, and group 4 was treated

topically with the simple ointment (Control). The percentage of wound closure was calculated as

follows using the initial and final area drawn on glass slides during the experiments (15): % of

wound closure = (wound area on day + wound area on day n)/wound area on day 0 × 100

where n is a number of days (6th, 4th, and 16th day).

During the wound healing period and at the presented time intervals, the wound area will trace

manually and photograph. The wound area was calculated using Auto CAD RL 14 software. At

days 6th, 9th and 16th the experiment was terminated and the wound area was removed from

the surviving animals for histological examination. The excision skin biopsies were fixed in 4%

formaldehyde solution 48 h during the experimentation period.

e. Statistical analysis

The relative wound area was statistically analyzed using one-way ANOVA by the program SAS

ver. 6.12 fully and comparison of the means of the wound areas at different days evaluated by

Duncan’s test at p < 0.05 level.

DEFINITION OF TERMS

For clearer understanding of the terms used in the study, below are their meaning:

Pomegranate is a neat, rounded shrub or small tree that can grow to 20 or 30 ft., but more

typically to 12 to 16 ft. in height. Dwarf varieties are also known. It is usually

deciduous, but in certain areas the leaves will persist on the tree. The trunk is covered

by a red-brown bark which later becomes gray. The branches are stiff, angular and

often spiny. There is a strong tendency to sucker from the base. Pomegranates are

also long-lived. There are specimens in Europe that are known to be over 200 years

of age. The vigor of a pomegranate declines after about 15 years, however.

Wound is a breakdown in the protective function of the skin; the loss of continuity of epithelium,

with or without loss of underlying connective tissue (i.e. muscle, bone, nerves)2 following

injury to the 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

Creams are semisolid dosage forms that contain one or more drug substances dissolved or

dispersed in a suitable base. This term traditionally has been applied to semisolids that

possess a relatively soft, spreadable consistency formulated as either water-in-oil or oil-

in-water emulsions. However, more recently the term has been restricted to products

consisting of oil-in-water emulsions or aqueous microcrystalline dispersions of long-chain

fatty acids or alcohols that are water washable and more cosmetically and aesthetically

acceptable.

Wound healing is an intricate process where the skin or other body tissue repairs itself after

injury. In normal skin, the epidermis (surface layer) and dermis (deeper layer) form a

protective barrier against the external environment. When the barrier is broken, an

orchestrated cascade of biochemical events is quickly set into motion to repair the

damage.

CHAPTER VII

REFERENCES

E. A. Hayouni, K. Miled, S. Boubaker, et al., “Hydroalcoholic extract based-ointment from Punica granatum L. peels with enhanced in vivo healing potential on dermal wounds,” Phytomedicine, vol. 18, no. 11, pp. 976–984, 2011.

Butterfield, Harry M. A History of Subtropical Fruits and Nuts in California. University of California, Agricultural Experiment Station. 1963.

USP NF 2009

Nguyen, D.T., Orgill D.P., Murphy G.F. (2009). Chapter 4: The Pathophysiologic Basis for Wound Healing and Cutaneous Regeneration. Biomaterials For Treating Skin Loss. Woodhead Publishing (UK/Europe) & CRC Press (US), Cambridge/Boca Raton, p. 25-57. (ISBN 978-1-4200-9989-8/ISBN 978-1-84569-363-3)

http://www.crfg.org/pubs/ff/pomegranate.html

http://www.clinimed.co.uk/Wound-Care/Education/Wound-Essentials/What-is-a-Wound-.aspx