Bone Fracture Healing with Ulasimang-Bato Extract

St.
Paul University Philippines 1

Tuguegarao City, Cagayan 3500
Chapter 1
THE PROBLEM AND REVIEW OF RELATED LITERATURES AND STUDIES
Introduction
The human skeleton is the basic structure of the body.
It composes of 270 bones at birth- this total decrease to 206
bones by adulthood after some bones have fused together (Kids
Video Show, 2015). Therefore, we need to take good care of our
body because if one of our bones get injured or even fractured,
the whole body is affected.
A bone fracture is a break in the continuity of the bone
(Odokuma E, Akpoghene O, Osemeke I, Eronimeh O, 2015). Bone
fractures can either be of traumatic origin (motor accidents
or falls) or of pathological origin (mainly osteoporosis) in
most cases, as a result of bone fragility (Doblare M, Garcia
JM, Gomez MJ, 2004).
A bone as an organ has the ability to regenerate naturally
when a fracture occurs. Generally, in modern medicine,
treatment of bone fractures involves the use of a cast or
plaster to immobilize the fractured limb, followed by the
administration of analgesics (pain killers) and anti-
inflammatory drugs to patients. The broken bone is then
allowed to regenerate naturally on its own (Dimitriou R, Jones
SPUP School of Nursing and Allied Health Sciences

St. Paul University Philippines 2
E, McGonagle D, Giannoudis P., 2011), since there is no orally
active pharmacological agent available for rapid fracture
repair (Ngueguim FT, Khan MP, Donfack JP, Siddiqui JA, Tewari
D, et al, 2012). Even though fractures heal naturally, it
would be of great interest to discover alternatives to
accelerate a bone healing process, thereby reducing the
healing period, reducing the cost of treatment and helping
patients to quickly regain their good health.
Ulasimang-bato (Peperomia pellucida) from the family
Piperaceae is an annual herb, shallow rooted, may reach 40 cm
high, with succulent stems. Leaves are alternate, heart-
shaped and turgid, as transparent and smooth as candle
wax. Tiny dot-like flowers are scattered along solitary
and leaf-opposed stalk (spike); naked; this grass grows in
moist areas in Southeast Asia. Its heart-shaped leaves, about
1.5cm diameter, are shiny and watery and are very soft, easily
destroyed. Maturing gradually from the base to the tip and is
turning brown when ripe (A guide to the use of Philippine
medicinal Plants as an alternative Medicine, 2009).
Several studies were conducted to study the fracture-
healing property of Ulasimang-bato (Peperomia pellucida).
Positive results were collected which means that this plant
has fracture-healing property.

One study is entitled Aqueous extract of Peperomia
pellucida (L.) HBK accelerates fracture healing in Wistar rats
by Odokuma E, Akpoghene O, Osemeke I, Eronimeh O, 2015.
Phytochemical studies were also done by several researchers
to find out the different components of this plant specially
the responsible constituent regarding its fracture-healing
property. In an article entitled Philippine Herbal Plants and
Their Uses, phosphorus, potassium, calcium, sodium, magnesium,
alkaloids, flavonoids, tannins, steroids and carbohydrates are
the chemical constituents of the plant. With the help of these
studies, the researchers opt to study the fracture healing
effect of Ulasimang-bato (Peperomia pellucida) aqueous extract
and by testing it to hamsters.
Review of Related Literature
According to Res J Pharm Biol Sci. 2011, Ulasimang-bato
(Peperomia pellucida) is an herbaceous plant found mainly in
America, Africa and Asia, which grows during rainy periods
and thrives in damp, humid soils and under the shade of
trees.
In addition to that, Obenaus, Hayes, 2011 described that
the said plant is consumed as vegetable and as a traditional

medicine for the management of diseases and for fracture
healing. This is because of the mineral contents of the extract
that is used in the treatment of bone fracture.
Florence, et al., 2017 revealed the presence of minerals
such as potassium, phosphorus, magnesium, calcium and sodium
after conducting analysis on the mineral content of the dry
extract of the said plant. Furthermore, potassium was found
to be the most abundant mineral element with a mass of 16.640
μg/g, followed by phosphorus with 1.405 μg/g. Calcium was
found to weigh 0.454 μg/g while sodium was the least with a
weight of 0.181 μg/g.
Aside from the dry extract of the plant, Ethnopharmacol,
2013 states that the ethanolic extract of Ulasimang-bato
(Peperomia pellucida) by an anabolic effect on osteoblasts
promotes fracture healing. Osteoblast differentiation and
mineralization as said by Stuart, 2016, has a stimulatory
effect on the increase of fracture repair. In a study conducted
by Iqbal S., 2012, the plant contains calcium and some
metabolites as flavonoids- which are known respectively to
contribute in matrix deposition during osteogenesis and
promote bone formation.
In 2010, Emily L. Miedel, VMDa, F. Claire Hankenson, DVM,
MS, DACLAMb, 2015, Laboratory Animal Medicine reported that

approximately 146,000 hamsters were used in research in United
States. Hamsters are classified as members of the family
Cricetidae. Animals in this family are characterized by large
cheek pouches, thick bodies, short tails, and an excess of
loose skin. They also have incisors that erupt continuously
and cuspidate molars that do not continue to grow ((I 1/1, C
0/0, PM 0/0, M 3/3) × 2 = 16).
Ulasimang-bato (Peperomia pellucida)
Ulasimang-bato (Peperomia pellucida) is a fleshy shallow
rooted herb that has been used as food item as well as a
medicinal herb. Moreover, the whole plant is edible either
cooked or raw. In the folkloric herbal medicine providing
health benefits for gout, arthritis and other inflammatory
conditions, the plant has taken also niche. The plant grows
about 15 to 45 cm in height in damp and lightly shaded areas.
Also, it has been used as a medicinal herb for its analgesic,
anti-arthritic, diuretic activity.
Ulasimang-bato (Peperomia pellucida) can grow wild but
can also be grown as ornamental foliage. It is characterized
by its shiny heart shaped leaves about 4 cm in length, growing
from an erect translucent green stalk.

In addition to that, the said plant has tiny dot-like
flowers that grow from erect and slender green spikes that
turn brown when matured. The fruits are small, round to oblong
in shape, ridged, first green later black and tiny seeds drop
off that grows easily in groups.
The plant is widely used as folkloric herbal medicine.
It is known for its benefit in some health conditions such as
eye and skin inflammations, sore throat, diarrhea, prostate
problems, high blood pressure, arthritis, gout, skin boils,
wounds, burns, abscesses, pimples, headache, fever, abdominal
pains, renal and mental excitement disorder.
Ulasimang-bato (Peperomia pellucida) – role in bone
mineralization (fracture healing)
Calcium
99% of the body’s calcium resides in the skeleton. The
said mineral element is one of the main bone-forming minerals
in the body. Adequate dietary calcium maximizes peak bone mass
early in life and prevents bone loss later.
According to Bauer, 1954; Herman and Richelle, 1961;
Lemaire, 1966 in American Journal, calcium and vitamin D play
pivotal roles in bone remodeling and mineralization.

Mineralization is an important part of fracture callus
formation. However, both calcium and vitamin D influences
fracture-healing process. Calcium is essential for callus
mineralization.
Phosphorus
The second most abundant mineral in the body is
phosphorus. It makes up a full 25% of all the mineral material
in the body.
Also, of special interest to everyone is the fact that
phosphorus combines with calcium to form a mineral crystal
that gives strength and structure to our bones and teeth. Of
all the phosphorus in the body, 80% of it is found in the
teeth and bones in the form of crystalline
bone, hydroxyapatite.
Magnesium
Approximately 60% of the magnesium in the body is in
bone. Magnesium influences mineral metabolism indirectly
through its role in ATP metabolism and as a cofactor for over
300 enzymes, and by direct effects on bone quality by
decreasing hydroxyapatite crystal size, thereby preventing
larger, more perfect mineral crystals that result in brittle
bone.

Phosphate
Approximately 85%, the majority of body phosphate is
present in the mineral phase of bone as with calcium. The
balance of body phosphate is present in a variety of inorganic
and organic compounds distributed within both intracellular
and extracellular compartments. Normal blood concentrations
of phosphate are very similar to calcium.
Sodium
In the form of sodium chloride, elevates urinary calcium
excretion and, evokes compensatory responses that may lead to
increased bone remodeling and bone loss.
Potassium
A high potassium intake promotes an alkaline environment,
thus reducing the demand for skeletal salts to balance the
endogenous acid generated from acid producing foods, such as
meats. By preserving calcium in bones, which is mobilized to
maintain normal pH, potassium rich foods may help prevent
osteoporosis.

Hamster (Cricetinae)
Hamsters (Cricetinae) are approximately four to seven
inches long, with short tails, small eyes and fur in a variety
of coat colors, like black, brown, grey, white, yellow or red.
They also have large cheek pouches where they use to collect
food to transport from one place to another. Wide and hairless
are used to describe their feet with four toes on their front
and five toes on their back feet. In addition to that, hamsters
have large, chisel-shaped incisors that grow throughout their
lives.
Hamster breeds are arranged into giant, medium and dwarf
hamsters. The most popular type of pet hamster is called the
Syrian hamster- a median hamster which has a golden color and
is usually 6 or 7 inches in length. Another popular breed is
the dwarf hamster. These hamsters are unusual in that they'll
peacefully live with other hamsters of the same sex. To add
information, some hamsters are long-haired while others are
short-haired. Long-haired hamsters have thick, fluffy fur,
while the short-haired ones are short and velvety.
The said animal are small rodents that are commonly kept
as house pets. They are distinguishable from other rodents due
to their short tails, stubby legs and small ears.

Hamsters have many different colors. This includes black,
grey, brown, white, yellow, red or a mixture of several colors.
With 24 species of hamsters, these animals come in a wide
range of sizes. The European breed can grow as large as 13.4
inches (34 centimeters) long and are some of the biggest
varieties of hamsters. The dwarf hamster lives up to its name.
These tiny hamsters grow to around 2 to 4 inches (5.5 to 10.5
cm) long. The most common pet hamster, the Syrian hamster,
also known as the teddy bear hamster or golden hamster, usually
grows to about 6 inches (15.24 cm) long.
Hamsters are nocturnal, according to the ASPCA, which
means they like sleeping during the day. In the wild, they dig
burrows, which are a series of tunnels, to live and breed in.
Hamsters will also store food in their burrows. Living
underground keeps wild hamsters cool in hot climates.
Some hamsters are very social, while others are loners.
For example, the Syrian hamster doesn't like living near other
hamsters. They are very territorial and should never be put
in a cage with other hamsters. It will bite the other hamster,
and may even kill it. Dwarf hamsters, on the other hand, are
social and like to have a friend nearby.

Wild hamsters will hibernate if the weather gets cold
enough. Hamsters will wake up from their hibernation
periodically to eat. If there isn't enough food stored,
hamsters will wait to hibernate until their surplus is to
their liking.
Male and female hamsters waste no time when mating with
one another. If you put two opposite gender hamsters into a
cage together, the female will become pregnant very quickly.
Their gestation period lasts 15 to 20 days.
After the birth, the litter is blind until they are two
weeks old and at 3 to 4 weeks the young are weaned. Females
tend to have two to three litters per year. Hamsters typically
live one to two years, but can live up to three years in
captivity.
Hamsters have very poor eyesight. Their scent glands on
their backs secrete an easily identifiable smell. To find
their way around, they leave a trail of scent by rubbing their
backs against objects. When a hamster needs to find their way
back home, they will use this scent trail as a guide.

Hamster's teeth grow continually. Chewing on wood or
twigs keeps their teeth short. Without something to chew on,
their teeth would grow so long they would injure the roof of
the mouth and lips.
Typically, a fracture takes at least 4 weeks to heal, but
some hamsters could take longer, up to 12 weeks. Once you've
given him some time, watch for signs of healing, like walking
on the leg, or no swelling over the fracture site. When you
gently handle the leg, he won't react with pain and there
shouldn't be any grating if the fracture has healed. If he
reacts with pain, stop handling him immediately.
Fracture
A fracture is a break, usually in a bone. If the broken
bone punctures the skin, it is called an open or compound
fracture. Fractures commonly happen because of car
accidents, falls, or sports injuries. Other causes are low
bone density and osteoporosis, which cause weakening of the
bones. Overuse can cause stress fractures, which are very
small cracks in the bone.

Symptoms of a fracture are intense pain, deformity (the
limb looks out of place), swelling, bruising, or tenderness
around the injury, numbness and tingling and problems moving
a limb.
Fractures (Broken Bones)
A fracture is a broken bone. A bone may be completely
fractured or partially fractured in any number of ways
(crosswise, lengthwise, in multiple pieces).
Types of Fractures
Bones are rigid, but they do bend or "give" somewhat when
an outside force is applied. However, if the force is too
great, the bones will break, just as a plastic ruler breaks
when it is bent too far.
The severity of a fracture usually depends on the force
that caused the break. If the bone's breaking point has been
exceeded only slightly, then the bone may crack rather than
break all the way through. If the force is extreme, such as
in an automobile crash or a gunshot, the bone may shatter.
If the bone breaks in such a way that bone fragments
stick out through the skin, or a wound penetrates down to the
broken bone, the fracture is called an "open" fracture.

This type of fracture is particularly serious because
once the skin is broken, infection in both the wound and the
bone can occur.
Common types of fractures include:
• Stable fracture. The broken ends of the bone line up
and are barely out of place.
• Open, compound fracture. The skin may be pierced by
the bone or by a blow that breaks the skin at the
time of the fracture. The bone may or may not be
visible in the wound.
• Transverse fracture. This type of fracture has a
horizontal fracture line.
• Oblique fracture. This type of fracture has an angled
pattern.
• Comminuted fracture. In this type of fracture, the
bone shatters into three or more pieces.
FIGURE 1. Common Types of Fracture
A bone can fracture in different ways.

Traction
Traction is usually used to align a bone or bones by a
gentle, steady pulling action.
External Fixation
In this type of operation, metal pins or screws are placed
into the broken bone above and below the fracture site. The
pins or screws are connected to a metal bar outside the skin.
This device is a stabilizing frame that holds the bones in the
proper position while they heal.
In cases where the skin and other soft tissues around the
fracture are badly damaged, an external fixator may be applied
until surgery can be tolerated.
FIGURE 2. An external fixator applied to a broken femur
(thighbone).

Open Reduction and Internal Fixation
During this operation, the bone fragments are first
repositioned (reduced) in their normal alignment, and then
held together with special screws or by attaching metal plates
to the outer surface of the bone. The fragments may also be
held together by inserting rods down through the marrow space
in the center of the bone.
FIGURE 3. A specially designed metal rod, called an
intramedullary nail, provides strong fixation for this
thighbone fracture.
FIGURE 4. In this x-ray, the broken bones of the forearm
are held in position by plates and screws while they heal.

Recovery
Fractures take several weeks to several months to heal,
depending on the extent of the injury and how well you follow
your doctor's advice. Pain usually stops long before the
fracture is solid enough to handle the stresses of normal
activity.
BONE HEALING PROCESS
FIGURE 5. Bone healing Process
(Pountos I, Giannoudis P, 2018, Fracture Healing: Back to
Basics and Latest Advances)

TYPES OF BONE FRACTURES
FIGURE 6. Types of Bone Fracture
(Orthopedic-institute.org)
“Aqueous extract of Ulasimang-bato (Peperomia pellucida) HBK
accelerates fracture healing in Wistar rats”
The same study was conducted which were entitled “Aqueous
extract of Ulasimang-bato (Peperomia pellucida) HBK
accelerates fracture healing in Wistar rats”. This study aimed
to evaluate the effects of the aqueous whole plant extract of
Ulasimang-bato (Peperomia pellucida) on fracture healing in
female Wistar rats. The method used on this study was a drill
hole method. A drill hole injury was created by inserting a
drill bit in the diaphysis of the femur.

The aqueous extract of the whole plant of Ulasimang-bato
(Peperomia pellucida) was administered orally at the doses of
100, 200 and 400 mg/kg to adult female Wistar rats. The vehicle
(distilled water) was given to the control. Besides these
rats, one group of rats without fracture received the extract
(400 mg/kg). After 14 days of treatment, the rats were
sacrificed under anesthesia and the effects of the extract
were evaluated on body weight, the relative weights of organs
(femurs, uteri and ovaries) and on hematology. Bone (calcium,
phosphorus, alkaline phosphatase) and serum biochemical
parameters (calcium, phosphorus, alkaline phosphatase) were
also evaluated. Radiological and histological tests were
carried out on the femurs. The mineral content of the plant
extract was also investigated. Their results showed that the
extract induced an increase in body weight at high dose and
in WBCs count at low doses. Aqueous extract from Ulasimang-
bato (Peperomia pellucida) increased bone calcium at lowest
dose but maintained this parameter at normal range at high
dose in fractured rat. Alkaline phosphatase and phosphorus
concentrations reduced significantly (p < 0.01) at the dose
of 400 mg/kg as compared to fractured rats.

The radiological tests revealed a dose dependent
formation of callus at the level of the fracture gap, confirmed
by the formation of a highly dense and compact
fibrocartilaginous callus. The mineral content of the plant
extract revealed the presence of calcium, phosphorus,
magnesium, sodium and potassium. In conclusion, the aqueous
extract of Ulasimang-bato (Peperomia pellucida) accelerates
bone healing due partly to the mineral content of the extract.
These results confirm its traditional use in the treatment of
bone fractures. (Florence NT, Huguette ST, Hubert DJ, et al.,
2017)

CONCEPTUAL FRAMEWORK
Figure 7. Conceptual Framework of the Study
The Ulasimang-bato (Peperomia pellucida) extract in 5
treatments: Treatment 1 includes the positive control while
Treatments 2, 3, 4, and 5 consists of Ulasimang-bato
(Peperomia pellucida) aqueous extract in different
concentrations namely 100mg/kg, 200mg/kg, 400mg/kg, and
500mg/kg sample respectively.
Statement of The Problem
Generally, the study aimed to determine the fracture-
healing effect of Ulasimang-bato (Peperomia pellucida) aqueous
extract on hamsters (cricetinae) with Oblique fracture.

Specifically, the study aimed to answer the following
questions:
1. What are the mineral constituents present in Ulasimang-
bato (Peperomia pellucida) aqueous extract?
2. What is the extent of the fracture-healing effect of the
different concentrations of the Ulasimang-bato
(Peperomia pellucida) aqueous extract and the positive
control after 14 days?
3. Is there a significant difference on the fracture-healing
effect of the different concentrations of the aqueous
extract of Ulasimang-bato (Peperomia pellucida) and the
positive control?
Hypotheses
The study tested the following hypotheses at 0.01 level
of significance:
There is no significant difference on the fracture-
healing effect of the different concentrations of the aqueous
extract of Ulasimang-bato (Peperomia pellucida) and the
positive control.

Significance of The Study
The results of this study will serve as basis for future
pharmacological screening and will lead to accessibility to
the community.
Students. This research study will encourage students to
conduct more researches for the development of previous
studies.
Community. This research study will provide a better
understanding on Ulasimang-bato’s therapeutic value and to see
the value of the said plant and that they will discover freedom
from costly commercial drugs.
Future researchers. The ideas presented maybe used as
reference data in conducting new researches or in testing the
validity of other related findings.
Pharmacists. They may formulate dosage forms that would
require less expensive natural raw materials rather than
synthetic ones such as Ulasimang-bato (Peperomia pellucida),
thus producing cheaper medicines.

Scope and Limitations
This study focused on the determination of the fracture-
healing effect of Ulasimang-bato (Peperomia pellucida).
It started with the collection and preparation of the
parts of Ulasimang-bato (Peperomia pellucida) including the
different qualitative analysis on the mineral constituents
which is responsible for the fracture-healing effect of the
plant sample. Biological testing will be performed by oblique
manual fracture method using hamsters. Pre-test and post-test
x-ray were done only once. Post-test x-ray was done after 14
days of treatment.
Definition of terms
The following terms are defined to facilitate an easy
understanding of the study.
Aqueous Extract. It is a liquid preparation obtained by
decoction. (Nafiu MO, Adeyemi SB, 2017)
Bone. It forms the skeleton of the body. They support and
protect internal organs (for example, the skull protects the
brain and the ribs protect the lungs). (Sheil W, 2018)

Calcium. A mineral found mainly in the hard part of bones,
where it is stored. It is essential for healthy bones and is
also important for muscle contraction, heart action, and
normal blood clotting. (Sheil W, 2018)
Decoction. It is the action or process of extracting the
essence of something. (Collins Dictionary)
Fracture. It is a break in bone or cartilage. Although
usually a result of trauma, a fracture can be the result of
an acquired disease of bone, such as osteoporosis, or of
abnormal formation of bone in a congenital disease of bone.
(Sheil W, 2018)
Fracture healing. It is the bridging of fracture site by
callus, obliteration of the fracture line, and continuity. (Li
J, Stocum D, 2014)
Oblique Fracture. It is a complete fracture that occur
at a plane oblique to the long axis of the bone. (Murphy A,
Jones J, et al.)
Osteoblast. It is cell that develops bone. (Setiawati R,
Rahardjo P, 2018)
Osteogenesis. It is the formation of bones (Setiawati R,
Rahardjo P, 2018)

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St.

Paul University Philippines 1

THE PROBLEM AND REVIEW OF RELATED LITERATURES AND STUDIES

The human skeleton is the basic structure of the body.

It composes of 270 bones at birth- this total decrease to 206

bones by adulthood after some bones have fused together (Kids

Video Show, 2015). Therefore, we need to take good care of our

body because if one of our bones get injured or even fractured,

the whole body is affected.

A bone fracture is a break in the continuity of the bone

(Odokuma E, Akpoghene O, Osemeke I, Eronimeh O, 2015). Bone

fractures can either be of traumatic origin (motor accidents

or falls) or of pathological origin (mainly osteoporosis) in

most cases, as a result of bone fragility (Doblare M, Garcia

JM, Gomez MJ, 2004).

A bone as an organ has the ability to regenerate naturally

when a fracture occurs. Generally, in modern medicine,

treatment of bone fractures involves the use of a cast or

plaster to immobilize the fractured limb, followed by the

administration of analgesics (pain killers) and anti-

inflammatory drugs to patients. The broken bone is then

allowed to regenerate naturally on its own (Dimitriou R, Jones

SPUP School of Nursing and Allied Health Sciences

E, McGonagle D, Giannoudis P., 2011), since there is no orally

active pharmacological agent available for rapid fracture

D, et al, 2012). Even though fractures heal naturally, it

would be of great interest to discover alternatives to

accelerate a bone healing process, thereby reducing the

healing period, reducing the cost of treatment and helping

patients to quickly regain their good health.

Ulasimang-bato (Peperomia pellucida) from the family

Piperaceae is an annual herb, shallow rooted, may reach 40 cm

high, with succulent stems. Leaves are alternate, heart-

shaped and turgid, as transparent and smooth as candle

wax. Tiny dot-like flowers are scattered along solitary

and leaf-opposed stalk (spike); naked; this grass grows in

moist areas in Southeast Asia. Its heart-shaped leaves, about

destroyed. Maturing gradually from the base to the tip and is

turning brown when ripe (A guide to the use of Philippine

medicinal Plants as an alternative Medicine, 2009).

Several studies were conducted to study the fracture-

healing property of Ulasimang-bato (Peperomia pellucida).

Positive results were collected which means that this plant

has fracture-healing property.

SPUP School of Nursing and Allied Health Sciences

One study is entitled Aqueous extract of Peperomia

pellucida (L.) HBK accelerates fracture healing in Wistar rats

by Odokuma E, Akpoghene O, Osemeke I, Eronimeh O, 2015.

Phytochemical studies were also done by several researchers

to find out the different components of this plant specially

the responsible constituent regarding its fracture-healing

property. In an article entitled Philippine Herbal Plants and

Their Uses, phosphorus, potassium, calcium, sodium, magnesium,

alkaloids, flavonoids, tannins, steroids and carbohydrates are

the chemical constituents of the plant. With the help of these

studies, the researchers opt to study the fracture healing

effect of Ulasimang-bato (Peperomia pellucida) aqueous extract

and by testing it to hamsters.

Review of Related Literature

According to Res J Pharm Biol Sci. 2011, Ulasimang-bato

(Peperomia pellucida) is an herbaceous plant found mainly in

America, Africa and Asia, which grows during rainy periods

and thrives in damp, humid soils and under the shade of