CHAPTER ONE

INTRODUCTION

1.1 Background of Study

The skin is the largest organ in human body and plays a crucial role in different processes
such as hydration, protection from chemicals and pathogens, vitamin D synthesis initialization,
excretion and thermal regulation. Severe skin damage can therefore be life-threatening (Erika et
al, 2020).

From a microbiological perspective, the primary function of normal, intact skin is to control
microbial populations that live on the skin surface and to prevent underlying tissue from
becoming colonized and invaded by potential pathogens. Exposure of subcutaneous tissue
following a loss of skin integrity (i.e., a wound) provides a moist, warm, and nutritious
environment that is conducive to microbial colonization and proliferation (P. G. BOWLER et al
2001). Infection is the leading cause of death after thermal injury (Leopoldo C. Cancio 2021).
Globally, burns are the fourth most frequent type of injury, after traffic accidents, falls and
physical violence (Agniezka et al 2022 [1]). They are complicated wounds that are difficult to
heal and are associated with high mortality rates and hence the need for serious consideration.

Since prehistoric times, humans have used natural products from plants, animals,
microorganisms, and marine organisms, in medicines to alleviate and treat diseases (Haidan Y,
Qianqian M 2016). Plants are just like the ‘treasure box’ hidden with a lot of active components
that are essential for the process of developing new medicine. These active components are
known as secondary metabolites, such as phenols, tannins, saponins, carbohydrates, flavonoids,
gylcosides, and alkaloids (Carlina K, Nabila P 2021.). Plants are one of the greatest inventions of
God to serve as the ‘lungs’ of our lovely planet. And God said, Let the earth bring forth grass,
the herb yielding seed, and the fruit tree yielding fruit after his kind, whose seed is in itself, upon
the earth: and it was so(Genesis1:11).

Carica papaya is considered as one of the most common fruit related to human
consumption and provides a favourable cost benefit in consideration of its nutritional
value.Papaya (Carica papaya Linn.) is well known for its exceptional nutritional and medicinal
properties throughout the world.(Dr. Neethu S K &Sreeja D P 2016). Folk tales have it that the
whole plant including its fruit, leaves, seed, root, bark, juice and latex are used nutritionally,
medicinally and for various other purposes. All these reported medicinal uses either from fruit,
seed or leaves ofCarica papaya, prompted the investigation for its antimicrobial and wound-
healing activity of the seed extract.The taxonomical classification includes Kingdom (Plantae),
Order (Brassicales), Family (Caricaceae), Genus (Carica) and Species (papaya). (Dr. Neethu S
K &Sreeja D P 2016).Carica papaya is a tropical fruit which is one of the major fruit crops
cultivated in tropical and sub-tropical zones due to its tasty and juicy flesh. Other than that, the
increment in demand is because of the high medicinal and nutritive value possessed by papaya
fruit. (Subenthiran S, Choon TC, Cheong KC, Thayan R, Teck MB, et al. 2013). Carica Papaya,
known as pawpaw (English), is the most cultivated species of the Caricaceae family, although
considered to be an economic tree because of its fruits, which is common in tropical Africa.

The seeds are found in the fruits which are very offensive, peppery, and as such
unpalatable when eating as food (Reuben et al 2020). However, the seeds seem to have more
potent medicinal values than the flesh(Jyotsna et al. 2014).Medicinal uses of papaya seed are
carminative,anti-fertility agent in males, counter irritant, as a paste in the treatment of ringworm,
psoriasis,emmenagogue, vermifuge, liver cirrhosis and abortifacient. Seed juice is used for
bleedingpiles, enlarged liver and pectoral properties. Seed paste is used as anthelmintic,
stimulation ofmenstruation or abortion (Dr. NeethuS Kumar &Sreeja Devi PS 2016). Studies
have shown thatC. papaya has antibacterial effects that could be useful in treating chronic skin
ulcers to promote healing. It is widely used in developing countries as an effective and readily
available treatment of various wounds, particularly burns (Bijoor et al., 2012(6)Dawkins et al,.
2003)).

1.2 Medicinal Plants

Plants are an essential source of chemical compounds with different biological properties that
man can use to his advantage. These substances are mainly produced as a result of chemical
conversions of secondary metabolism(Irina Francesca González Mera et al., 2019).

A medicinal plant is any plant which in one or more of its organs, contains substances that
can be used for therapeuticpurposes, or which are precursors for chemo-pharmaceutical semi-
synthesis. When a plant is designated as medicinal, it isimplied that the said plant is useful as a
drug or therapeutic agent or an active ingredient of a medicinal preparation (P. Yudharaji et al.,
2016).Medicinal plants represent the most ancient form of medication, used for thousands of
years in traditional medicine in many countries around the world. The empirical knowledge
about their beneficial effects was transmitted over the centuries within human
communities(MariangelaMarrelli 2021 [1]).The term of medicinal plants include a varioustypes
of plants used in herbalism and some of these plantshave medicinal activities. Besides that,
theseplants play a critical role in the development of humancultures around the whole world (P.
Yudharaji et al., 2016).

Many phytochemicals with established or potential biological activity have been identified in
plants.The compounds found in plants (phytochemicals) are of several kinds, but most are in four
mainbiochemical classes: terpenes, alkaloids, glycosides, and polyphenols. Medicinal plants are
usedwidely in non-industrialized societies and developing countries in Africa, Asia, and
SouthernAmerica, mainly because they are thought to be very effective, cheaper than modern
medicines,and readily available (AwuchiChinazaGodswill, 2019). Moreover, some plants are
considered as important source of nutrition and because ofthat, these plants are recommended for
their therapeutic values.These plants include ginger, green tea, walnuts and someothers plants.
For other plants, their derivatives are considered asimportant source for active ingredients, which
are used inaspirin and toothpastes.These medicinal plants areconsidered as rich resources of
ingredients, which can be used in drug development and synthesis (Bassam Abdul Rasool
Hassan, 2012).

1.2.1 Brief History of Medicinal Plants

The earliest historical records of medicinal plants (herbs) are found from the
Sumeriancivilization, where hundreds of medicinal plants, opium inclusive, are listed on the clay
tablets. Ebers Papyrus from the ancient Egypt, c. 1550 BC, defines over 850 plant
medicines.Dioscorides, a Greek physician, who worked in the Roman army, acknowledged and
documented over a thousand recipes for medicines with over 600 medicinal plants in the De
materiamedica, c. 60 AD; this preformed and laid the foundation for the basis of
pharmacopoeias for 1500 years (AwuchiChinazaGodswill, 2019). From ancient times to
present,Ayurvedic medicines as documented in the SushrutaSamhita, the Atharva Veda, and the
RigVeda, have used hundreds of pharmacological active spices and herbs such as turmeric,
whichcontains curcumin(Aggarwal et al., 2007; Girish and Shridhari, 2007). The pharmacopoeia
ofthe Chinese, the Shennong Ben Cao Jing record plant medicines such as chaulmoogra
forephedra, leprosy, and hemp (Sumner, 2000). This was extended in the Tang Dynasty
YaoxingLun (Wu, 2005). In the 4th century BC, an Aristotle's pupil Theophrastus composed the
firstsystematic botany text, known as Historiaplantarum (Grene, 2004).

The place and roles of plants in medicine were radically altered in the nineteenth century
by theapplication and use of chemical analyses. Alkaloids were extracted and isolated from
successionof medicinal plants, beginning with morphinefrom the poppy, and soon followed by
strychnosand ipecacuanha, quinine from the cinchona tree, and several others. As chemistry
advanced,more classes of pharmacologically active compounds and substances were discovered
in themedicinal plants (Atanasov et al., 2015).Drug related research makes use of theethnobotany
to searchfor the pharmacologically active natural substances, and has in this manner discovered
hundredsof beneficial compounds. These include the popular drugs aspirin, opium, digoxin, and
quinine (AwuchiChinazaGodswill, 2019).

Medicinal plants have a promising future becausethere are about half million plants around
the world, andmost of them their medical activities have not investigateyet, and their medical
activities could be decisive in thetreatment of present or future studies (P. Yudharaji et al.,
2016[3]).

1.2.2 Characteristics of Medicinal Plants

 Synergic medicine: The ingredients of plants allinteract simultaneously, so their uses can
complement ordamage others or neutralize their possible negative effects.
 Support of official medicine: In the treatment of complexcases like cancer diseases the
components of the plantsproved to be very effective.
 Preventive medicine: It has been proven that thecomponent of the plants also
characterize by their ability toprevent the appearance of some diseases. This will help
toreduce the use of the chemical remedies which will be usedwhen the disease is already
present i.e., reduce the sideeffect of synthetic treatment studies (P. Yudharaji et al.,
2016).
1.3 Phytochemicals
 Plants are autotrophic organisms. In addition to the primary metabolism present in all living
beings, they have secondary metabolism that allows them to produce and accumulate compounds
of a very diverse chemical nature. The compounds derived from secondary metabolism in plants
are called secondary metabolites (Irina Francesca González Mera et al., 2019).

Plant secondary metabolites are produced in the form of phytochemicals in various plant
parts as a natural defense system against attack of various microorganisms and environmental
stresses (Satish et al. 2022).{All plants produce chemical substances and compounds which give
them evolutionaryadvantages, such as defense against herbivores or, in the instance of salicylic
acid, as hormone inplant defenses (United States Department of Agriculture, 2017; Hayat and
Ahmad, 2007) (AwuchiChinazaGodswill, 2019)}.

The secondary metabolites of the plants constitute a large and varied group of organic
compounds that are synthesized in small quantities; they have no direct function in essential
processes such asphotosynthesis, respiration, solute transport, protein synthesis, nutrient
assimilation, and the differentiation or formation of carbohydrates, proteins, and lipids. They
appear in plants asa result of chemical conversions and even when many of their functions
areunknown, it is believed that secondary metabolites are related to the defense of the plant
against predators and pathogens. They also act as allelopathic agents that influence growth,
survival, and reproduction of other plants, attract seed pollinators and serve to face adaptation to
sudden changes in temperature, humidity, light intensity and drought(Irina Francesca González
Mera et al., 2019).

The role of these compounds is beyond providing protection, as they are linked to many
biochemical pathways inside and outside the plants and possess various well known therapeutic
applications (Satish et al. 2022).{Thesephytochemicals have the potential for use as drugs, and
also the contents and knownpharmacological activities of these biochemical substances in
medicinal plants formed thescientific basis for their uses and applications in modern medicine, if
scientifically proved (Ahn,2017)(AwuchiChinazaGodswill, 2019)}.

1.3.1 Classification of Phytochemicals

Phytochemicalsare usually classified according to their biosynthetic pathways in three

principal groups: phenolics, terpenes, and alkaloids (IlariaChiocchio et al 2021[ 24]). Several
criteria have been considered forthe classification of secondary metabolites: chemical structure
(presence of ring or sugar), composition (containing nitrogen or not), their solubility in organic
solvents or water, and the biosynthetic pathway. Of them, the most common criterion used for
grouping the secondary metabolites in plants has been the biosynthetic pathway (Irina Francesca
González Mera et al., 2019).The main classes of pharmacologically active phytochemicals are
explained below.

1.3.1.1 Phenolics

They are chemical compounds containing a hydroxyl group directly attached to an

aromatic hydrocarbon (Irina Francesca González Mera et al., 2019). Phenolic compounds are a
group of small molecules characterized by their structures having at least one phenol unit.
Phenols show their presence in almost every plant, hence a widespread compound among the
plant genera which significantly contribute to their taste, color, and flavor
(AwuchiChinazaGodswill, 2019).

{The classification of phenolics can be done according to their biosynthetic origin or

structure.Phenolic compounds are synthesized in plant cells by the shikimic acid pathway or the
malonate/acetate pathway (or both, for example, flavonoids)36.The polyacetate pathway
provides quinones and xanthones.alonate/acetate pathway (or both, for example, flavonoids)36.
The shikimic acid pathway provides the synthesis of phenylalanine and cinnamic acids and their
derivatives (simple phenols, phenolic acids, coumarins, lignans, and phenyl propane
derivatives)37,38.The mixed pathways combine precursors of both the shikimic acid pathway
and the polyacetate pathway. This is the case of flavonoids39,40.pathway(Irina Francesca
González Mera et al., 2019).}

{Phenols also accumulate on surfaces of leaves, capturing up to 90% of UV radiation46.

Phenols confer aromas and colors to the fruits making them appetizing for herbivores, which
favors the dispersion of seeds through feces47. Plants compete with each other to preserve their
territories, and in this process (allelopathy) the phenols participate48. Plants also defend
themselves against the attack of pathogens by synthesizing phytoalexins that are toxic to
microorganisms and their presence prevents infections49. Phenols also protect plants by
generating bitter flavors or textures that are unpleasant for herbivores50.pathway(Irina Francesca
González Mera et al., 2019).}. {The pharmacological activities of phenols are noteworthy such
as anti-inflammatory activity of quercetin is well reported, while silybin is greatly recognized for
its antihepatotoxic property. Other examples include phytoestrogenic activity of daidzein and
genistein, while naringenin shows insecticidal effects (Goławska et al. 2014). Phenolic
compounds are also responsible for the antioxidant property of many plants, especially
flavonoids.(AwuchiChinazaGodswill, 2019)}.

1.3.1.2 Terpenes

Terpenes, also termed terpenoids constitute a large family of natural

productsextremelydiversifiedintheirstructure,functions,andproperties. Terpenes are derived from
mevalonic acid biosynthetic pathway. However, since their decomposition generates isoprene
units (C5), this compound has been defined as terpene’s basic constituent. The broader term of
terpenoids is applied to terpene-related molecules that, in addiction to isoprene units, contain
other substituents, for instance, oxygenated functional groups [102]. (IlariaChiocchio et al.,
2021).

Terpenes are also knownisoprenoids, since the basic structural unit that forms them is the
isoprene molecule13(Irina Francesca González Mera et al., 2019).}. For this reason, these
secondary metabolites are classified based on the number of isoprene units present in their
structure, and generally condensed head-to-tail. Following this rule they are divided into
hemiterpenes (C5H8), monoterpenes (C5H8)2, sesquiterpenes (C5H8)3, diterpenes (C5H8)4,
sesterterpenes (C5H8)5, triterpenes (C5H8)6, etc. . (IlariaChiocchio et al., 2021).

Terpenes may serve a wide spectrum of functions in plants, including attracting

pollinators or protecting injured tissues from herbivore, insects and parasites attack. Due to their
numerous bioactivity, terpenes are applied in several industrial sectors, such as pharmaceutical,
food, cosmetics, perfumery, and agriculture, and are used as drugs, food supplements, flavors,
fragrances, biopesticides, etc..(IlariaChiocchio et al., 2021).
{Terpenesaremostlyknownfortheiressentialoil components which can be obtained from steam
distillation, hydro-distillation, or dry distillable fractions having characteristic smell or odor
(Ludwiczuk et al. 2017). Terpenoids have positive effects on health like anxiolytic, analgesia,
antiinflammatory, cancer hemopreventive, antimicrobial,anti-hyperglycemic, and antidepressant
effects and provide protection to the skin. Most of these effects were reported by preclinical
studies with animal or in vitro methods (Baron 2018). .(AwuchiChinazaGodswill, 2019)}.
1.3.1.3 Alkaloids

Alkaloids are bitter-tasting chemicals which are often toxic and very widespread in
nature, found in many medicinal plants(AwuchiChinazaGodswill, 2019 (Aniszewski, 2007).
{Historically, alkaloids have been defined as metabolites containing one or more nitrogen
atom(s) within heterocyclic ring(s) [159]. However, N-containing compounds where
theNatomisnotheterocyclic,suchashordenine,ephedrine,colchicineandcapsaicin,were further
included into this secondary metabolite group and classified as proto-alkaloids or amino-
alkaloids. ForthisreasonheterocyclicN-containingcompoundsareoftenregardedas “truealkaloids.”
Since “true alkaloids” biosynthetically derive from amino acids, they are classified on the basis
of the biogenetic origin. (IlariaChiocchio et al., 2021).}.

Alkaloids have remarkable physiological properties and toxicologythat are exerted

primarily on thecentral nervous system, with predominance in some of its levels. For these
reasons, they can be used as drugs. Prolonged use of any of these compounds produced in man
accustoming, which constitute true drug addictions, with physical and psychic dependence and
an increase in the tolerance(Irina Francesca González Mera et al., 2019). { Medicines of different
classes include scopolamine, hyoscyamine, and atropine (all from nightshade), caffeine (Coffea),
cocaine (Coca), the traditional medicine berberine (from the plants such as Mahonia and
Berberis), ephedrine (Ephedra),morphine (opium poppy), quinidine and quinine (Cinchona),
nicotine (tobacco), reserpine (Rauwolfiaserpentina), vincristine (Catharanthusroseus), and
vincamine (Vincaminor) (Elumalai and Eswariah, 2012; Gremigni et al., 2003). The opium
poppy Papaversomniferum is a source of the alkaloids codeine and morphine. Deadly
nightshade, Atropa belladonna, gives tropane alkaloids including scopolamine, hyoscyamine,
and atropine. The alkaloid nicotine from tobacco directly binds to the Nicotinic acetylcholine
receptors of the body, accounting for its pharmacological and biochemical effects.
(AwuchiChinazaGodswill, 2019}.

1.4 Carica papaya as a Medicinal Plant

Papaya is one of the affordable, nutritionally, and medicinally important fruit which belongs
to the Caricaceae family (Shama Kakkar et al., 2021). The fruit is a part of the human diet
and it is also use in cosmetics and medical field (Marzha Ancheta1 & Liwayway Acero 2016).
Carica papaya is a potential natural medicinal source. Several studies and methods have been
used in extracting Carica papaya materials from different parts of the plant. Extracts from
different parts of Carica papaya plant have shown protective effects against many diseases such
as intestinal worms infection and different types of wounds (MOHAMED ABD ELGADIR et
al., 2014). Papaya is well known for its exceptional nutritional and medicinal properties
throughout the world. The whole plant including its fruit, leaves, seed, root, bark, juice and latex
obtained from papaya plant are nutritionally and medicinally used for various other purposes.
Papaya is a source of vitamins A, C, and E; minerals (viz., potassium, magnesium, etc.),
carotenoids; phenolic compounds, fiber, and folate. C. papaya is used traditionally by many
countries due to its nutritional and health benefits(Ashutosh Sharma et al., 2020).

The papaya seed contain fatty acids, crude protein, crude fibre, papaya oil, carpaine, caricin,
glucotropaeolin, benzyl glucosinolates, benzyl Isothiocyanate, benzyl thiourea, hentriacontane,
ß-sitostrol, caressing and an enzyme myrosin. The seeds and the pulp of Carica papaya
contain benzyl glucosinolate which can be hydrolyzed by myrosinase to produce benzyl
isothiocyanate. Seed extracts have profound bactericidal activity. The seeds of unripe fruits are
rich in benzyl isothiocyanate, a sulphur containing chemical that has been reported to be an
effective germicide and insecticide (Dr. Neethu S Kumar & Sreeja Devi PS, 2017).

Seven flavonoids were obtained from the papaya leaves, named as quercetin,
kaempferolkaempferol 3‐rutinoside, quercetin3‐(2G‐rhamnosylrutinoside), quercetin 3‐
rutinoside, kaempferol 3‐(2Grhamnosylrutinoside), myricetin 3‐rhamnoside {(Nugroho et al.,
2017) (Ashutosh Sharma et al., 2020)}. Another study by (Musa, 2015(Ashutosh Sharma et al.,
2020)) reported that n‐hexane extract and methanolic extract of leaves showed the presence of
different components such as anthraquinone, glycoside, tannin, saponin, flavonoid, steroid and
resin. Chromatography (column chromatography) and spectroscopy methods (NMR, IR and
mass spectrometry) were used for the isolation and characterization of carpaine from the leaves.
One research investigation concluded that leaves contains various phytochemicals such as
carpaine, kaempferol 3-(2G-glucosylrutinoside), kaempferol 3-(2”-rhamnosylgalactoside), 7-
rhamnoside, kaempferol 3-rhamosyl-(1-2>)-galactoside-7-rhamnoside, luteolin 7-galactosyl-(1-
>6)-galactoside, orientin 7-Orhamnoside, 11-hydroperoxy-12,13-epoxy-9-octadecenoic
acid,palmitic amide, and 2‐hexaprenyl‐6‐methoxyphenol (Soib et al., 2020 (Ashutosh Sharma et
al., 2020)).
 Gas chromatography‐mass spectrometry (GC–MS) technique was used to recognize the
phyto‐components present in unripe fruit aqueous extract and showed the presence of fifteen
phytochemical compounds. The major phytochemical compounds were hexadecenoic
acid, Z‐11, methyl ester, and octadecanoic acid (Ezekwe and Chikezie, 2017). The GC–MS
technique was used to identify phytocomponents present in unripe fruit aqueous extract which
showed that fifteen phytochemical components were present. The major phytochemical
compounds were hexadecenoic acid, Z‐11, methyl ester, and octadecanoic acid. Minor
phytochemical compounds present in the unripe fruit aqueous extract of this plant was within the
range of 0.78–5.38% (Ezekwe and Chikezie, 2017 (Ashutosh Sharma et al., 2020)).

1.4.1 HEALTH SUPRISING BENEFITS OF PAPYA SEED

C. papaya is tremendously efficient in different types of ailment cure due to availability

of wide varieties of phytoconstituents in almost all parts of plant. Some of the significant benefits
of C. papaya seeds are further discussed below.

1.4.1.1 WOUND HEALING ACTIVITY

Wound is the most common injury. Wound is an injury involves the breaking of the skin
caused by cuts, animal bites, burns, splinter, thorns or puncture. A break in the skin
could cause infections including pus, swelling, redness and even fever. Recent studies have
stated that fresh papaya has a wound healing property. One of the most important ingredients in
papaya is the enzyme papain, and vitamin C which are important for wound healing (Marzha
Ancheta & Liwayway Acero 2016). Various skin disorders as well as wounds can be cured by
papaya. The ethanolic papaya seed extract was tested in Sprague‐Dawley rats, for its wound‐
healing activity. Results showed that the seed extract assists wound healing in rats (Nayak et al.,
2012 (Ashutosh Sharma et al., 2020).

The use of Carica papaya L. (Caricaceae) in traditional medicine relies on papain, a

proteolytic enzyme, is the active principle which exerts an ulcer protective effect . The C. papaya
possesses antimicrobial antioxidant and anti-inflammatory activities. C. papaya has antibacterial
effects that could be useful in treating chronic skin ulcers to promote healing. C. papaya is
traditionally used to treat various skin disorders, including wounds. It is widely used in
developing countries as an effective and readily available treatment of various wounds,
particularly burns. A decoction made from the seeds of C. papaya has been used
to treat skin ulcers and inflammation (Bijoor Shivananda Nayak et al., 2012).

1.4.1.2 ANTI-OXIDANT ACTIVITY

Free radical causes many chronic health problems. Antioxidants can help us by
preventing the formation of free radicals.

Carica papaya help counteract oxidative stress via various mechanisms of action closely
related to its antioxidant properties and eventually improving the management of various
oxidative stress-related health conditions. Carica papaya is a topical plant species discovered to
contain high amounts of natural antioxidants that can usually be found in their leaves, fruits and
seeds. It contains various chemical compounds demonstrate significant antioxidant properties
including caffeic acid, myricetin, rutin, quercetin, α-tocopherol, papain, benzyl isothiocyanate
(BiTC), and kaempferol. Therefore, it can counteract pro-oxidants via a number of signaling
pathways that either promote the expression of antioxidant enzymes or reduce ROS production.
These signaling pathways activate the antioxidant defense mechanisms that protect the body
against both intrinsic and extrinsic oxidative stress. To conclude, Carica papaya can be
incorporated into medications or supplements to help manage the
health conditions driven by oxidative stress and further studies are needed to investigate the
potential of its chemical components to manage various chronic diseases. (Yew Rong Kong et
al., 2021).

{One recent research investigation reported that the antioxidant activity of methanolic
extract of seeds was determined by DPPH free radical scavenging activity (Singh et al., 2020a,
2020b, 2020c). In another study, papaya seed extracts were used to examine the antioxidant
activity and results revealed that the highest DPPH free radical scavenging ability was found in
hexane extract and the least activity in the aqueous extract (Agada et al., 2020). (Ashutosh
Sharma et al., 2020}.

1.4.1.3 ANTI-MICROBIAL AND ANTI-BACTERIAL ACTIVITY

{Infectious desease are the biggest threat for human health and almost 50,000 death cases
occure daily due to the infectious diseases[1]. The resistance of various infection agents over
synthetic drugs can be the main reason of the antibiotic alternatives development [2]. Plants have
main advantage to be effective and cheaper drug altenative[3].( Masfufatun ET AL., 2019}

{Published reports are available which showed that the various parts of papaya have
significant antimicrobial activity. Oleic acid isolated from the seed and was tested for its chemo‐
preventive action on live fish which was infected with pathogenic bacteria named Klebsiella
PKBSG14 and it was found that drug bioavailability increases with an increase in oleic acid
(Ghosh et al., 2017). Recent research investigation reported that methanolic extract of papaya
seed was tested for its antibacterial activity against E. coli, Klebsiella pneumonia,
and Pseudomonas vulgaris by using the Agar disc diffusion method. The extract showed
antimicrobial activity in the following order E. coli > P. vulgaris > K. pneumonia (Singh et al.,
2020a, 2020b, 2020c). (Ashutosh Sharma et al., 2020}. Papaya seed extract possessed high
antimicrobial activities against V. cholerae and low antimicrobial activities against C. albicans.
The inhibitory abilities of papaya seed ethanol extract against the growth of C.albicans and V.
cholerae could be due to its active compounds with antimicrobial activities such as alkaloids,
flavonoids, stereoids, polyphenols and tannins (Masfufatun ET AL., 2019)

1.4.1.4 ANTI-CANCER ACTIVITY

{In vitro investigation of C. papaya suggested that it has anti‐cancer properties. The plant
contains an enzyme, namely papain which is a constituent of papaya and very helpful in cancer
treatment. Fibrin breaks down by papain which coats the tumor cells into amino acid. The
pigment lycopene is found inside the papain which is highly reactive towards free radical and
oxygen. Papaya also contains isothyocynate which protects the breast, prostate, pancreas, lung,
leukemia, and colon cancer (Fauziya and Krishnamurthy, 2013). In a research study, it was
revealed that the leaf extract of C. papaya can prevent the progression of cancerous cells.
Various markers such as CA15‐3 and LDH are the important biochemical parameter for the
detection of cancerous cells; and it was found that the leaf extract in a dose of 200 mg/kg body
weight has a significant effect to decrease both markers in the treatment of cancer (Gurudatta et
al., 2015). Petroleum ether, ethyl acetate, chloroform, and methanol (80%) extracts of aerial parts
C. papaya were analyzed for their anticancer effect against three types of cancer cells such as
UACC62 (melanoma) TK10 (renal) and MCF7 (breast) cancer cells. It was concluded that the
petroleum extract of papaya aerial parts had a significant effect on MCF7 (breast) cancer cells
(Khaled and Gerda, 2013). One research report concluded that black seed from yellow ripe
papaya has a direct effect to reduce the growth of prostate cancer cells. Methanolic extract of
black seed (ripe papaya) and white seed (unripe papaya) was tested against the prostate cancer
cell line. It has been reported that the black seed extract is effective against prostate cancer
cells whereas the white seed shows a stimulating effect on pre‐existing prostate cancer cells
(Alotaibi et al., 2017). Another research study reported that papaya leaf juice has also an anti‐
proliferative effect on prostate cancer cells (Pandey et al., 2018).( Ashutosh Sharma et al.,
2020)}. Flavonoids are rich in water extract of papaya seeds, act as an anticancer agent ( Shama
Kakkar. Et al., 2021). An aqueous extract of Carica papaya was also examined for its effect on
growth of various tumor cell lines and on human lymphocytes and have shown positive
significant results [9]. The results also showed significant growth inhibitory activity of Carica
papaya extract on tumor cell lines [9] (MOHAMED ABD ELGADIR et al., 2014).

1.4.1.5 ACTIVITY AGAINST HUMAN INTESTINAL PARASITOSIS

{Human intestinal parasitosis constitutes a significant global health problem with

enormous financial implications. unfortunately, for reasons mainly attributable to poor hygiene,2
the burden is more in the tropics and subtropics where, particularly among children,3,4 parasites
cause noteworthy morbidity such as anemia, diarrhea and dysentery, malnutrition, apathy, and
underdevelopment as well as severe acute abdominal and surgical conditions.2 As most patients
particularly from the tropics and particularly from Africa are from low socioeconomic groups
who can ill-afford imported and expensive medicines,5 the need for affordable and readily
available local alternatives cannot be overemphasized. Carica papaya, a fruit plant also called
papaya, papaw, pawpaw, mamao, or tree melon, is found in virtually every tropical and
subtropical country.6 C. papaya contains proven antihelminthic7 chemical agents such as benzyl
isothiocyanate and papain8,9 and has other antiparasitic properties.10,11 In folk medicine, C.
papaya seeds have been used to treat helminthiasis. (John A.O. Okeniyi et al., 2014)}

1.4.1.6 ANTI-INFLAMMATORY ACTIVITY

Inflammation is a localized physical condition in which part of the body becomes redden,
swollen, hot and often painful, especially as a reaction to injury or infection. Inflammation is a
complicated pathway of the body’s own protective mechanism against pathogens. In short, the
pathogenesis of inflammation starts with tissue injury, which causes infiltration and activation of
macrophages and relevant antigen-presenting cells (APCs). This causes the release of
proinflammatory cytokines such as tumour necrosis factor-α (TNF-α) and interleukins (ILs).
Cytokines stimulate the release of chemokines, which further recruit and activate lymphocytes
and leukocytes. ROS are produced to eliminate invaders whereby activates Nuclear factor kappa-
B (NF-κB). NF-κB is a transcription factor and plays a role in inducing inducible nitric oxide
synthase (iNOS) activity and, thus, nitric oxide (NO) production. Excessive ROS upregulated
prostaglandin E2 (PGE2) synthesis and, hence, cyclooxygenase-2 (COX-2) expression, which
eventually leads to oxidative stress, that causes tissue damage and worsen inflammation (Yew
Rong Kong et al., 2021).

Papaya contains an extensive range of secondary metabolites such as alkaloids, tannins,

flavonoids, saponins, which have been shown to a marked effect to reduce chronic inflammatory
reaction (Ashutosh Sharma et al., 2020). High phenolic and flavonoid content in papaya seed
extracts were proposed to act as free radical scavengers and metal ion chelators [15].
Phytochemicals including tocopherols and quercetin are showed to enhance AMP-activated
protein kinase (AMPK) activation, as well as the inhibition of COX-2 expression [17]. In
addition, a range of phytochemicals with great strength of anti-inflammatory effect, such as
benzyl isothiocyanate (BiTC), β-carotene, lycopene, and vitamin C could be found in various
parts of papaya fruits, in either pulp or seeds. These phytochemicals were proven to inhibit pro-
inflammatory cytokines including TNF-α, IL-6 and monocyte chemoattractant protein-1 (MCP-
1) [18,19]. In addition, polyphenols within Carica papaya could act as free radical scavenger and
at the same time exerting its effects in upregulating the antioxidant enzymes activities [18].
(Yew Rong Kong et al., 2021).

1.4.1.7 ANTI-MALARIA ACTIVITY

{There were several reports available that support the anti‐malarial activity of C. papaya.
According to Suleman et al. (2018) papaya along with other plants used by peoples in the
treatment of malaria and related symptoms. Another study also revealed about antimalarial
effect of methanolic extract of C. papaya in mice on Plasmodium berghei NK65 strain (Longdet
and Adoga, 2017). Also, leaf extract was tested against Plasmodium falciparum 3D7 and Dd2
strains. Carpaine was the most active alkaloid extract in dichloromethane leaf extract and
displayed good activity against both strain of Plasmodium falciparum IC50 of 2.01 ± 0.18 μg/mL
(4.21 μM) and 2.19 ± 0.60 μg/Ml (4.57 μM). This alkaloid is highly selective against the parasite
and non‐toxic to healthy uninfected R.B.C (Teng et al., 2019). Further, methanol, chloroform,
petroleum ether extract of fruit rind, and roots of papaya were tested against Plasmodium
berghein mice for their antiplasmodial activity. The result showed that petroleum ether and
chloroform extract of C. papaya fruit rind has considerable antiplasmodial activity in a dose‐
dependent manner but petroleum ether extract had the highest antimalarial activity (Zeleke et al.,
2017).( Ashutosh Sharma et al., 2020)}.

1.5 PETROLUEM JELLY AS A TOPICAL VEHICLE

Topical vehicles are used as carrier systems, which aid in the delivery of an active drug
(e.g., corticosteroid, calcineurins, vitamin D derivatives, retinoids) across the SC and into deeper
regions of the skin, namely the epidermis and dermis, whilst minimizing absorption
into the systemic circulation (Tanya M. Barnes et al., 2021). Many dermatological conditions,
such as eczema, burn and psoriasis, are treated with topical drug delivery systems. Instead of
applying the active drug directly onto the skin, it is combined with a vehicle to aid in its delivery
across the stratum corneum (SC) and into deeper regions of the epidermis and dermis. In so
doing, systemic absorption and circulation are minimized.

Petroleum jelly (also called petrolatum) is a mixture of mineral oils and waxes, which form a
semisolid jelly-like substance. This product hasn’t changed much since Robert Augustus
Chesebrough discovered it in 1859. Chesebrough noticed that oil workers would use a gooey
jelly to heal their wounds and burns. He eventually packaged this jelly as Vaseline. Petroleum
jelly’s benefits come from its main ingredient petroleum, which helps seal your skin with a
water-protective barrier. This helps the skin to heal and retain moisture (Deborah Weatherspoon,
2019).

Petroleum jelly is now used for various purposes to:

 Heal minor skin scrapes and burns.

 Moisturize the face and hands.
  Prevent diaper rash.
 Remove eye makeup.
 Preserve perfume scents.
 Prevent skin stains from hair dyes or nail polish.
1.6 WOUND (BURN) HEALING

Skin wound healing is a complex process involving interrelated and overlapping mechanisms
of cell migration and proliferation, synthesis of extracellular matrix, growth factors and
cytokines that coordinate the healing process (Lucas Fernando Sérgio Gushiken et al., 2021).
Classically, there are four major phases in wound healing; hemostasis, inflammation, tissue
proliferation, and tissue maturation or remodeling [23,24]. The underlying connective tissue is
exposed upon injury to the skin, and the collagens attract platelets to the injury
site. This triggers platelet aggregation to deliver clotting factors such as prothrombin
and fibrinogen, which initiates platelet clotting, coagulation, and a complement cascade
through extrinsic and intrinsic pathways (Thambirajoo Maheswary et al., 2021).

The human body is susceptible to numerous local and systemic conditions which can
negatively affect skin repair through various mechanisms, leading to a delay in the process. The
major conditions that interfere with wound healing are stress, infection, nutrition, hypoxia, sex
hormones, aging, smoking, genetics, medications and alcohol (Lucas Fernando Sérgio
Gushiken et al., 2021).

A burn is defined as damage to the skin and underlying tissues caused by heat, chemicals, or
electricity (America Burn Association, 2018). According to the World Health Organization
(WHO), it is estimated that each year approximately 11 million people suffer from burn wounds,
180,000 of whom die because of such injuries. Regardless of the factors causing burns, these are
complicated wounds that are difficult to heal and are associated with high mortality rates
(Agnieszka Markiewicz-Gospodarek et al., 2022) . Globally, burns are the fourth most frequent
type of injury, after traffic accidents, falls and physical violence [1] (Agnieszka Markiewicz-
Gospodarek et al., 2022). The process of burn wound healing is a complex long-lasting process
that involves a few repair processes dependent on the immune system [26], and involves the
reconstruction of broken tissue continuity resulting from a random event, e.g., a fire. For a burn
wound to heal, three stages must occur successively: inflammation, granulation tissue
formation (proliferation), and remodeling (which may result in scarring) (Agnieszka
Markiewicz-Gospodarek ET AL., 2022).

1.6.1 PHASES OF THE WOUND HEALING PROCESS

Wound healing, sometimes called the healing cascade, is generally described in three
distinct phases: inflammation, tissue proliferation, and tissue maturation or remodeling.

1.6.1.1 INFLAMMATION PHASE (THE FIRE FIGHTER)

The inflammation phase starts immediately after the injury is caused and lasts 0-3 days.
It is similar to the fire-fighting phase in that it is the body’s emergency response to the injury
(Annemarie Brown, 2015). The main purpose of the inflammatory response is to prevent further
blood loss by vasoconstriction, which results in a blood clot (Ng, 2010(Annemarie Brown,
2015).). Without the inflammation phase, no healing can take place. The inflammatory phase of
wound healing begins with the activation of platelets, which synthesize the compounds
responsible for fibrin clot formation, restoring the local hemostasis and acting as a provisional
extracellular matrix for the migration of blood cells [5]. Simultaneously, the platelets and injured
cells release cytokines and growth factors such as IL-1β (interleukin-1β), TNF-α (tumor necrosis
factor-α), FGF (fibroblast growth factor), and PDGF (platelet derived growth factor) which
attract leukocytes to the region [4,6]. Initially, neutrophils migrate to the injured area, starting the
debridement of necrotic tissue and the phagocytosis of pathogenic antigens. Furthermore, the
neutrophils release pro-inflammatory cytokines as IL-1β, TNF-α, IL-6 (interleukin-6) and IL-8
(interleukin-8), which attract other inflammatory cells to the wounded area (Table 1) [7]. The
neutrophils also release VEGF (vascular endothelial growth factor) and IGF-1 (insulin growth
factor- 1) which activate the local proliferation of fibroblasts, keratinocytes, and endothelial cells
[7,8]. After a few days, macrophages start to migrate to the wound region to continue the
debridement of necrotic tissue and phagocytosis of deleterious antigens, and secrete
growth factors and cytokines which coordinate the subsequent mechanisms of the wound healing
(Lucas Fernando Sérgio Gushiken et al.,2021).

1.6.1.2 TISSUE PROLIFERATION PHASE (THE BUILDER)

After the debris from a house fire has been cleared, builders are called in to restore the
house to its former structure; in wound healing, this is known as the proliferative
phase and usually lasts 3-24 days (Annemarie Brown, 2015).
The re-epithelization process, characterized by the proliferation and migration of keratinocytes
towards the core part of the lesion, originates in this phase as the area between the bottom and
the edges of the wound is filled with granulation tissue. This represents the matrix in which
keratinocytes, residing on lesion edges, migrate and proliferate [14]. Skin re-epithelization
structural organization can be explained by two models: sliding and rolling models. According to
the sliding model, keratinocytes of the basal layer suffer a modification of their anchoring joints
(desmosomes and hemidesmosomes), allowing their detachment and lateral migration into the
core part of the lesion. According to the rolling model, keratinocytes go through a morphological
and functional modification, together with desmosomes, resulting in them rolling towards basal
keratinocytes, which instead remain anchored to the basal membrane [16]. Basal layer
regeneration leads keratinocytes to proliferate and differentiate vertically, restoring the
physiological features of the multilayered epithelial tissue (Erika Maria Tottoli et al., 2020).

1.6.1.3 REMODELING OR MATURATION PHASE (THE DECORATION)

The maturation phase can be likened to redecorating the house after a fire; it lasts
between 21 days and two years. Epithelial cells, located in intact hair follicles, sweat
glands and around the edges of the wound, move over the newly formed granulation
tissue and reduce the size of the wound by contracting, thereby pulling the edges
together (Annemarie Brown, 2015). The remodeling phase starts about three weeks after an
injurious advent and lasts for over a year. During this phase, all processes activated in previous
phases are silenced and macrophages, isolated endothelial cells and myofibroblasts run into
apoptosis or they are relocated from the wound, leaving a region rich in collagen and other
extracellular matrix deposition (ECM) proteins. Interactions between the epidermis and dermis,
together with additional feedback, allows the continuous regulation of skin integrity and
homeostasis. Type III collagen, located in ECM, is gradually replaced in 6–12 months [17]
(Erika Maria Tottoli et al., 2020).

1.6.2 WOUND HEALING AND INFECTION

Infection is the leading cause of death after thermal injury. Optimal prevention and
treatment of burn wound infection is enabled by an in-depth understanding of burn wound
treatment modalities not only from a technical standpoint, but also from the standpoint of the
clinical context in which these modalities were originally developed (Leopoldo C. Cancio,
2021). Wound infections are associated with morbidity and mortality in patients, especially in
developing countries, regardless by the type of wound [5,6,8]. Failure in the treatment implies an
increase in the healthcare costs, since they involve a prolonged hospitalization due to diagnostic
tests, a huge administration of antibiotics and, sometimes, invasive surgery [9].( Valentina Puca
et al., 2021). A lot of pathogens, like bacteria, viruses and fungal parasites, can be responsible
for skin infections, since they find a suitable environment for their colonization and proliferation
in the deeper tissues of the skin [4,10,11]. The most common bacterial species that cause wound
infections are Pseudomonas aeruginosa, Staphylococcus aureus, Klebsiella pneumoniae,
Enterococcus faecalis and Acinetobacter baumannii. In particular, in the initial phase of
infections, within the first week, Gram-positive bacteria, especially S. aureus, appear to be the
most frequent colonizers [16,17]. From the beginning of the second week, Gram-negative
bacteria, such as P. aeruginosa and A. baumannii, start to colonize the wound, provoking sepsis
if they enter the lymphatic system and blood vessels (Valentina Puca et al., 2021).

1.7 LITERATURE REVIEW

Many research works have been carried out on Carica papaya. These researches have been
targeted into discovering and exploring the antioxidant properties of the plant, the phytochemical
composition, anti-inflammatory and immunomodulatory activity, anti-bacterial activity, wound
healing activity, the effect of extraction solvent on papaya plant or plant part, the health benefits,
ant-microbial activity, activity against human intestinal parasitosis and lots more.

Masfufatun et al (2019) carried out a research to determine the Minimum Inhibitory

Concentration (MIC) of the ethanol extract of papaya seed that inhibit the growth of Candida
albicans and Vibrio cholera. They also analyzed the active compounds present in ethanol seed
extract. This research was performed using experimental design laboratory. The ethanol extract
of papaya seed was obtained by maceration. The antimicrobial assay of papaya seed ethanol
extract was performed using agar plate diffusion. The MIC determination of papaya seed ethanol
extract was performed by 20, 15, 10, 5 and 1% concentration. The MIC of papaya seed ethanol
extract against Candida albicans and Vibrio cholerae were in 5% concentration (diameter of
inhibition zone=0.75 mm) and 15% (diameter of inhibition zone =14.75 mm), respectively. The
phytochemical assay of papaya seed ethanol extract showed there were alkaloids, flavonoids,
steroids, polyphenols, tannins and saponins. It means that papaya seed ethanol extract has a good
potential as antimicrobial therapy agent.

Anibijuwon and Udeze (2009) worked on the Antimicrobial Activity of Carica Papaya
(Pawpaw Leaf) on Some Pathogenic Organisms of Clinical Origin from South-Western Nigeria.
The bioactive compound of leaf and root extracts of Carica papaya was extracted, using water
and organic solvents, and were investigated for antibacterial activity against some human
pathogenic bacteria using the agar diffusion method. The aqueous extracts of the root extracts
did not show significant activity, but the organic extracts had significant activity with the
methanol extracts demonstrating the highest activity against the test bacteria. The root extracts
demonstrated higher activities against all the gram-positive bacteria than the gram-negative
bacteria tested, with the highest activity (14 mm zone of inhibition) demonstrated against
Pseudomonas aeruginosa while the aqueous leaf extract showed pronounced inhibition
demonstrating higher activities against the test bacteria than the organic solvents. The extracts
demonstrated higher activities against all the gram-positive bacteria than the gram-negative
bacteria tested, with the highest activity (4.2 mm zone of inhibition) demonstrated against
Pseudomonas aeruginosa. Increase in temperature enhanced the activity of
the extracts, while alkaline pH decreased the activity. The Minimum Inhibitory Concentration
(MIC) and Minimum Bactericidal Concentration (MBC) of the root extracts ranged between 50-
200 mg/ml. Preliminary phytochemical analyses showed that the extracts contain alkaloids,
tannins, saponins, glycosides and phenols. Carica papaya may be used for the treatment of
gastroenteritis, uretritis, otitis media and wound infections.
 Soib et al (2020) researched on the wound healing properties and effect of extraction
solvents on antioxidant of Carica papaya leaves extract. They noted that selection of proper
extraction solvent plays a pivotal role in extracting the bioactive compounds from the plant.
Therefore, the study was aimed to examine the effect of three types of extraction
solvents (methanol, ethanol and aqueous) on the antioxidant activity and wound healing
potential of C. papaya leaves. In the study, the effect of different solvents of C. papaya
leaves extracts were determined through 1,1-Diphenyl-2-picrylhydrazyl (DPPH) radical
scavenging activity, cytotoxicity assay and scratch migration assay on Human Skin
Fibroblast cells (HSF1184). The result showed that the methanolic extract of C. papaya
possessed a significant antioxidant activity as compared to ethanol and aqueous extract.
The result also demonstrated that methanolic extract significantly stimulates the highest
migration rate of HSF1184 cell at a concentration of 3.9 µg/mL, 7.8 µg/mL and 15.6 µg/
mL (p  0.05) after 48 hrs with no cytotoxicity observed at the concentration lower than
1000 mg/mL as compared to other solvents. HPLC analysis of methanol extract identified
the presence of two flavonoids (catechin, quercetin) and two phenolic acids (caffeic acid,
cinnamic acid). The findings suggest that the methanolic extract of C. papaya is effective
in fighting free radicals and it has good wound healing activities. It also has the potential
to be further explored for its medicinal values owing to the abundance of bioactive
compounds from the extract.

Rachmi Fanani Hakim et al (2019) worked on the effect of Carica papaya extract toward
incised wound healing process in mice (Mus musculus) clinically and histologically. Papaya
extract was obtained by a maceration method. It contains papain enzyme, flavonoid, saponin, and
alkaloid, which act as an anti-inflammatory, astringent substance, vasodilator, antioxidant,
analgesic, with antifungal, and antibacterial properties, and increase the collagen synthesis. The
experimental laboratory study was performed using 24 mice weighing between 30 and 40 grams
and 12 and 14 weeks of age as experimental animals. Each group was incised along 5 mm at the
labial gingiva under both of mandible anterior teeth with a depth reaching alveolar bone. Papaya
extract was applied topically in the morning and evening for 14 days. The clinical result was
obtained by assessing the length of wound closure measured every day for 14 days. Based on the
statistic test result, it showed that the papaya extract has a significant effect (p ≤ 0.05) toward the
healing process of an incised oral wound in mice. Histologically, at day 14, 75%
concentration papaya extract showed perfect epithelial layer and fibrillation.

Marzha Ancheta and Liwayway Acero (2016) studied the Wound Healing Property of
Carica papaya stem in albino rats. Carica papaya stem were dried cut and turned to ointment
form. The final product was applied to inflicted wounds of five Albino rats which are assigned in
the treatment group (treatment 2).Ten Albino rats served as experimental animals. They are
randomly assigned in two groups. The first group, or treatment 1, (5 animals) as the control.
Inflicted wounds of Albino rats in treatment1 was applied by the usual antiseptic (Betadine). The
second group, treatment 2 served as the experimental animals where the ointment from
papaya stem was applied. Initial wound size and mean size of the wound every two days interval
showed no significant result. The final 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 even the stem of Carica papaya can
be utilized as a source of herbal plant, specifically to heal wounds.

Kong et al (2021) researched on the Beneficial Role of Carica papaya Extracts and
Phytochemicals on Oxidative Stress and Related Diseases. They noted that it contains various
chemical compounds which demonstrate significant antioxidant properties including caffeic acid,
myricetin, rutin, quercetin, α-tocopherol, papain, benzyl isothiocyanate (BiTC), and kaempferol.
Therefore, it can counteract pro-oxidants via a number of signaling pathways that either promote
the expression of antioxidant enzymes or reduce ROS production. These signaling pathways
activate the antioxidant defense mechanisms that protect the body against both intrinsic and
extrinsic oxidative stress. To conclude, Carica papaya can be incorporated into medications or
supplements to help manage the health conditions driven by oxidative stress and further studies
are needed to investigate the potential of its chemical components to manage various chronic
diseases.

Okeniyi et al (2007) reported the effectiveness of dried Carica papaya seeds against human
intestinal parasitosis. To determine the effectiveness of air-dried C. papaya seeds on human
intestinal parasitosis, 60 asymptomatic Nigerian children with stool microscopic evidence of
intestinal parasites received immediate doses (20 mL) of either an elixir composed with air-dried
C. papaya seeds and honey (CPH) or honey alone (placebo) in two randomized treatment groups.
Repeat stool microscopic examinations were conducted 7 days postintervention for intestinal
parasites. Significantly more subjects given CPH elixir than those given honey had their stools
cleared of parasites [23 of 30 (76.7%) vs. five of 30 (16.7%); z 4.40, P .0000109]. There were
no harmful effects. The stool clearance rate for the various types of parasites encountered was
between 71.4% and 100% following CPH elixir treatment compared with 0–15.4% with honey.
Thus, air-dried C. papaya seeds are efficacious in treating human intestinal parasites and without
significant side effects. Their consumption offers a cheap, natural, harmless, readily available
monotherapy and preventive strategy against intestinal parasitosis, especially in tropical
communities. Further and large-scale intervention studies to compare C. papaya with standard
antiparasitic preparation are desirous.

Carlina Kong Jia Ying et al (2021) In their study, the Carica Papaya Seeds (CPS) were
extracted by using 95% ethanol with Soxhlet extraction method. After the plant extract was
subjected for the sterility test and results showed no growth of bacteria, the plant extract was
examined for the phytochemical screening. The antibacterial test was carried out with agar well
diffusion via examination of nutrient and tryptic soy agar. MIC test is used to determine
whether the bacteria can be inhibited by plant extract concentrations that are achievable
and safe in the blood stream, it was conducted by culturing microorganism in liquid
media. The antioxidant activity and total phenolic content were carried out to identify
the IC50 value and the total amount of phenolic content of CPS. For the phytochemical
screening test, the results showed the presence of alkaloids, anthraquinone, flavonoids,
and carbohydrate. The DPPH radical scavenging assay and total phenolic test were shown
positive for the antioxidant activity of the CPS. As a basis, Folin-Ciocalteu reagent was
used to measure the phenolic content of different concentrations of plant extract. In this
test, the Gallic acid was used as a standard and the total phenolic content was calculated
by using the formula C = (A/B) x dilution factor. In the present study, the total phenolic
content found was 6.420 mg GAE/g and 6.097 mg GAE/g for the concentrations of 50 and
200µg/ml respectively. According to the results obtained, the IC50 was calculated for the
BHT as well as CPS extract, the value was 84.04 µg/ml and 182.82 µg/ml, respectively.
Besides that, the percentage scavenging value obtained for the ethanolic C. papaya seed
was inconsistent and not precise, therefore the results cannot be considered or equivalent
to the antioxidant activity exerted by the CPS as the results are not reliable. However, it
can be concluded that the CPS are exhibiting antioxidant effect as it shows positive results
for the Total Phenolic Content (TPC) as well as DPPH scavenging method.

1.8 STATEMENT OF PROBLEM

All it takes is one quick accident or injury and a serious, chronic wound can form. The
immense social and economic impact of wounds worldwide is a consequence of their high rate of
occurrence in general and their increasing frequency in the ageing population. In addition to a
high number of acute wounds, there are also a large number of chronic, hard-to-heal wounds
associated with diseases and abnormalities that directly or indirectly culminate in damage of the
cutaneous coverage, including arterial, venous, diabetic and pressure ulcers. The prevalence of
these chronic wounds increases with age. For example, it has been estimated that chronic wounds
affect 120 per 100 000 people aged between 45 and 65 years and rises to 800 per 100 000 people
> 75 years of age (T VELNAR et al 2009).

Numerous factors can make wound healing a difficult process. One out of the many factor
are infections, which increases complications and cost. Wound infections are associated with
morbidity and mortality in patients, especially in developing countries, regardless by the type of
wound [5].( Valentina Puca et al., 2021). Previous studies have noted the increase of microbial
resistance to many contemporary drugs. With the surge of multi-resistant bacteria, health care
providers have become increasingly challenged in effective wound management (3) The
multiple antibiotic resistance of isolates calls for an immediate action on the control of
antimicrobial usage and the need to monitor resistance. ( O. M. Ogba et al 2014). There are also
risks in the use of contemporary antibiotics in pregnancy especially in the first trimester. This
can lead to drug side/adverse and teratogenic effects.

1.9 JUSTIFICATION OF THE STUDY

The resistance of various infection agents over synthetic drugs can be the main reason of the
antibiotic alternatives development [2]. Plants have main advantage to be effective and cheaper
drug altenative[3].( Masfufatun ET AL., 2019}. Studies have shown the wound healing and
antimicrobial properties of C. papaya seed. One of the most important ingredients in papaya is
the enzyme papain, and vitamin C which are important for wound healing (Marzha Ancheta &
Liwayway Acero 2016). Hence the need for natural sources of drugs as they present with low
risk of adverse effects and drug resistance.

1.10 AIMS AND OBJECTIVES OF THE STUDY

The specific aims objectives of this study are as follows;

1. To synthesize Carica papaya methanol seed extract, as an ointment using petroleum jelly
as a vehicle.
2. To determine the wound healing and anti-microbial activity of Carica papaya methanol
seed extract.
3. Compare the wound healing effect of Carica papaya ointment using petroleum jelly as a

vehicle to a known standard say Neurogesic cream.

4. To determine the quantitative and qualitative phytochemical composition of Carica

papaya seed.

5. To the synergistic effect of Petroleum jelly and Carica papaya in wound healing and

antimicrobial acticity.