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FORMULATION OF OI NTMENT
10.1. Introduction
179
contains an extract of the herb to be incorporated into the mixture.
agent, the two phases quickl y form a stable emulsion. The mixing
congeal. Upon mixing the two phases together, the mixture initiall y
to mix the extract with a small quantit y of lanolin (wool fat) using a
porcelain mortar and then incorporating the resulting mixt ure into
mixing.
to the base separately and named as extract ointment (EO) and Nano
ointment (NO).
The ointment of the Urena lobata was prepared by using the simple
180
Table: 10.1 Composition for the formulation of U rena lobata
White soft
90 White soft paraffin 90
paraffin
Procedure
A A - Original Extract
B C B - Extract ointment
C - Nano ointment
181
10.2. Quality control of the formulated Ointment
Loss on drying
noted.
182
viscosit y values are expressed as Mean ± Stan dard deviation
modified in the laboratory and used for the study. The experiment
S = MxL/T.
Acute skin irritation study: This test was performed on albino rats
animal feed and had free access to water ad libitum. The total mass
was separated into four groups, each batch containing five animals.
Dorsal hair at the back of the rats were removed one day prior to the
contact with the other rats. Two groups of each were used for
control and standard irritant. Other two groups were used as test.
The 50mg of the each formulation were applied over one s quare
The animals were observed for seven days for any signs of oedema
183
Stability studies: The stabilit y studies were carried out in all
different time intervals i.e., 15, 30, 60 and 90th days ( Shinde et al.,
Table: 10.2 Stability studies of the ointment of the Urena lobata extracts
Physicochemical
Formulation
parameters
Odour Characteristic
buffer pH 6.0, Whatman filter paper no.41 was mounted on cell. The
184
temperature was maintained at 37 ± 0.5°C. The formulation was
Absorbance: 304 nm
NANO OINTMENT
Time Absorbance
15 mts 0.114
30 mts 0.183
45 mts 0.214
1 hour 0.302
2 hours 0.41
3 hours 0.522
4 hours 0.602
185
5 hours 0.64
6 hours 0.712
EXTRACT OINTMENT
Time Absorbance
15 mts 0.232
30 mts 0.262
45 mts 0.284
1 hr 0.362
2 hours 0.488
3 hours 0.52
4 hours 0.612
5 hours 0.632
6 hours 0.702
nano ointment
186
10.3. RESULTS AND DISCUSSION
¾ All the formulations did not produce any skin irritation, i.e.,
erythema and edema for about a week when applied over the
skin.
Discussion
187
an idea about measureme nt of strength and the result of
erythema and edema for about a week when the ointments are
applied over the skin for skin irritation test indicates patient
188
References:
1, Appendix 4: A. 88 -A.11.
¾ Kim, J.Y., J.Y. Song, E.J. Lee and S.K. Park, 2003.
pp: 193-210.
189
¾ Mutimer, M.N., C. Riffikin, J.A. Hill, E. Marry and C.N.G.
212-212.
¾ Shinde, A.J., S.B. Bhise, R .J. Jarag and N.R. Jadhav, 2005.
190
¾ United States Pharmacop oeial Convention. Pharmacop oeial
191
11. WOUND HEALING STUDY OF THE FORMULATIONS OF
Urena lobata
11.1. Introduction
up the gaps between the wound edges and are associated with
third intention are usuall y those wounds left for three to five days
192
wound healing have also been identified -inflammatory,
processes.
ointment (NO) from the plant Urena lobata were evaluated using the
Chemicals
Experimental animals
All animals were kept in pol yacrylic cages and main tained
humidit y 60Ǧ65% with 12:12 light: dark cycles. Food was provided
193
All experiments involving animals complies with the ethical
Model design
Male Wister rats were used for excision and incision wound
models and the ointments were applied topicall y and animal were
(6 animals)
GroupǦ III: 5%w/w Extract ointment (EO) was applied once dail y.
i) Excision-4 animals
i) Excision- 4 animals
i) Excision-4animals
194
1) Excision wound model
excised.
model was studied under light ether anesthesia the anim al was
alcohol soaked with cotton swabs. They were kept in separat e cages.
195
All the sutures were removed on the 9 t h post wounding day. On 10 t h
suppl y technique.
All the above mentioned treatments were started from the day
cervical dislocation on day 30. Liver, spleen, stomach and skin were
¾ Body weight
¾ Tensile strength
Marklund, 1974)
1979)
196
11.2.1 Physical evaluation
healed wound at a distance 0.5 cm. the left clam p was fastened
position of the board was adjusted so that, the pol ythene bag was
constant rate from the reservoir until the wound opened. Amount of
water in pol ythene bag was measured (in ml) and was considered as
197
11.2.2 Antioxidant Studies
tris HCl buffer (0.4 M), 200µl K.EDTA (0.4 mM) along with 100 µl
(DTNB) were added and the colour intensit y formed was absorb ed at
198
412 nm using Thermo Scientific Multiskan Spectrophotometer,
8.0) were added and mixed well.The absorbance was read at 412 nm
et al.,1979).
4. Activity of Catalase:
mixture was stirred well and incubated at 37° C for 5 min and then
glacial acetic acid mixed in 1:3 ratio) was added and absorbance
5. Myeloperoxidase activity
199
of dinizidine. The absorbance was read at 450nm at 0 sec and 60 sec
tubes were removed and cooled for 5 minutes in cold water. 0.6 m L
The test tubes were once again placed in a hot water bath at 75 ° C
for 15 minutes and then cooled for 5 minutes under running stream
200
Results
All values are mean SEM ± n =6, **P < 0.001 indicates
201
Fig: 11.1 Tensile strength of the tissue of wound treated rats
600
500
400
300
200
100
0
Control STD NO EO
³.UXVNDO-:DOOLV7HVW´
Square
202
Day 7 Group 2 60.7777 46.6074 1.580* 0.664*
203
1. Effect of Extract ointment and Nano ointment on Protein and
EO 305.6±11.3* 203.6±9.9**
EO 66.20±2.66* 12.9±0.29**
204
Fig 11.2 Wound healing progression in treated rats
al 2013).
205
The wounds treated with ointments showed significant
Values are mean SEM of 6 rats in each group. * -P value is <0. 05,
**-P value is <0. 01 and *** -P value is <0. 001 compared to control by
.UXVKNDOZDOOL¶VWHVW
206
Fig: 11.3 LPO on wound treated rats
207
Fig: 11.5 Catalase on wound trea ted rats
208
11.3. Histopathology
Procedure:
method. On day 30, all rats from each group were euthanized using
anesthetic ether. The skin from the incised and excised region were
micron thickness were obtained and stained with Hematox ylin and
ointment
Fig: 11.7
Hypodermis
Muscle
Hair follicles
Dermis
209
Fig: 11.7.1
Hair follicles
Skin erosion
H& E 40 X
Fig: 11.7.2
Fig: 11.7.3
210
Fig: 11.7.4
Fig: 11.7.5
dermis H & E 40 x
Fig: 11.7.6
211
Fig: 11.7.7
Hair follicles
Gland
Fig: 11.7.8
Fig: 11.7.9
212
Fig: 11.7.10
Fig: 11.7.11
Epidermis
Dermis
Secretory glands
method. On day 30, all rats from each group were euthanized using
anesthetic ether. The skin from the incised and excised region were
micron thickness were obtained and stained with Hematox ylin and
213
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proliferation.
214
inflammatory cells was reduced than those observed in the standard
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215
216
SCORING OF WOUND FROM 0TH DAY TO 21ST DAY
217
Each area represents values in sq.cm
Each row represents groups
218
Each area represents values in sq.cm
Each row represents groups
219
Each area represents values in sq.cm
Each row represents groups
220
Each area represents values in sq.cm
Each row represents groups
221
Each area represents values in sq.cm
Each row represents groups
222
Each area represents values in sq.cm
Each row represents groups
223
Each area represents values in sq.cm
Each row represents groups
224
Each area represents values in sq.cm
Each row represents groups
225
Each area represents values in sq.cm
Each row represents groups
226
Each area represents values in sq.cm
Each row represents groups
227
Each area represents values in sq.cm
Each row represents groups
228
Each area represents values in sq.cm
Each row represents groups
229
Each area represents values in sq.cm
Each row represents groups
230
Each area represents values in sq.cm
Each row represents groups
231
Each area represents values in sq.cm
Each row represents groups
232
References:
279-282.
Analytical biochemistry,47,389-394.
Livingstone
10.1128/CMR.14.2.244 -269.2001
233
neutrophil content with an enz yme marker. J. Invest.
1963,268:1367,
1-4.
436.
234
¾ Harkness, R. D , ³Biological Functions of Collagen ´. Biol.
1929, 92:42.
01Ǧ07.
235
¾ Mustafa MR, Mahmood AA, Sidik K, Noor SM. ³Evaluation o
306.
¾ 5 ( 1HZPDQ DQG 0 $ /RJDQ ³7KH GHWHUPLQDWLRQ RI
1, 299±306, 1950.)
796
236
tensile strength evaluation of burn wound tissue healing ´.
237