Preparation of Novel Biocompatible Honey Hydrogel with
Turmaric and Aloe vera.
Rohan R. Vakhariya*, Rutuja R. Shaha, Archana R.
Dhole, Dr. C.S Magdum.
Rajarambapu College of Pharmacy, Kasegaon. Tal-Walwa,
Dist Sangli, Maharashtra
*Corresponding Author E-mail: rohanwakhariya@gmail.com
ABSTRACT:
There is an ever-present need for non-allergenic
antibacterial and antifungal wound dressing with a superior healing property
for chronic ulcers. Among the entire modern wound healing dressings, hydrogel
has a good capacity to donate moisture or absorb exudate and thereby providing
a moist environment to facilitate wound healing process and at the same time
protect the wound too. In the present study, povidne iodine loaded acrylamide
based biocompatible biodegradable hydrogel dressings incorporating alginate,
chitosan and gelatin showed good fluid absorbance capacity. The addition of honey
showed improved tensile strength and moisture absorbance capacity of the
hydrogel sponge. Addition of turmeric and aloe vera give additional wound
healing properties. Chitosan and Alginate
films with different Aloe vera contents (5, 15, 25 and 35%) and turmeric
contents (5, 15, 25 and 35%) were prepared and its properties evaluated in
terms of thickness, transparency, swelling behavior and in vitro
degradation.
KEYWORDS: Chitosan, Hydrogel,
Aloe vera, Turmeric.
1.
INTRODUCTION:
A hydrogel dressing prevents the
wound from microbial contamination, inhibits the loss of body fluids, provides
free flow of oxygen to the wound, and accelerates the healing process. Wound healing is a complex dynamic process that involves
many cascades of events like hemostasis, inflammation, proliferation and
remodeling of tissues in order to fill the damage area and reestablish the skin
barrier Honey gives tensile strength and
moisture content to the dressing. Both these parameters are very essential for
the dressings which are used for a prolonged period. Also, honey has been well
proven anti microbial and anti-bacterial since time immemorial 1,2.
Aloe Vera promoted complete healing
of burn wounds. Several studies and clinical trial have assessed the
effectiveness of aloe vera in the treatment of skin burns. It was found that
aloe vera was found to exhibit anti-inflammatory effects. A huge investigation
exposed that turmeric and curcumin has an extensive variety of curative
property such as anti-inflammatory, antibacterial, antifungal, anticancer,
antispasmodic, antioxidant, antiamoebic. Chitosan is nontoxic, biocompatible and
biodegradable3. It is used in drug delivery, cell delivery systems,
and orthopedics,
wound healing, ophthalmology, pharmaceuticals and bone healing. 4
2. MATERIALS AND METHODS:
2.1. MATERIALS:
Turmeric and Pure honey were obtained as gift sample
from Phyto Pharma, Kolhapur .All the other excipients were of analytical grade.
2.2.
METHODS:
2.2.1. PREPARATION OF BIOCOMPATIBLE HYDROGEL:
A: EXTRACTION METHOD:
1 Hot Continuous Extraction (Soxhlet):
In this method, the finely ground crude drug is placed
in a porous bag or “thimble” made of strong filter paper, which is placed in
chamber of the Soxhlet apparatus. The extracting solvent in flask is heated,
and its vapors condense in condenser. The condensed extract drips into the
thimble containing the crude drug, and extracts it by contact. When the level
of liquid in chamber rises to the top of siphon tube, the liquid contents of
chamber E siphon into fl ask. This process is continuous and is carried out
until a drop of solvent from the siphon tube does not leave residue when
evaporated. The advantage of this method, compared to previously described
methods, is that large amounts of drug can be extracted with a much smaller
quantity of solvent. This affects tremendous economy in terms of time, energy
and consequently financial inputs. At small scale, it is employed as a batch
process only, but it becomes much more economical and viable when converted
into a continuous extraction procedure on medium or large scale. The apparatus
used to carry out extraction process is as shown in fig. 1.
Fig. 1 Soxhlet Apparatus Extraction of turmeric and Aleo
vera was carried out by this method.
B: Preparation of hydrogel by using extract:
The acrylamide hydrogel base was used .The proportion
of monomer, cross linker and other ingredients were 4% of acrylamide, 0.08% of
ammonium persulphate (APS) and 0.08% N, NÊ-methylene bis acrylamide (MBA). The
preliminary screening for optimization of concentration of honey for good
tensile strength sponge was carried out. Concentrations of honey screened were
5%, 7%, 10% and 15% v/v. From the results of tensile strength and stickiness of
the sponge, it was concluded that 10%v/v honey shows best optimized hydrogel
sponge characteristics. Different amounts of natural polymer like chitosan /
alginate/ gelatin were dispersed separately into 25 ml of double distilled
water for 24 hours. Into this soaked polymer the above mentioned acrylamide
based was added accordingly. To this mixture 10% vv honey was added at the end
and homogenized using a high speed homogenizer. The resultant mixture was
transferred into a petridish .The petridish were kept at 600C for 24
hrs. Various batches were prepared by using polymers at different concentration,
alone and in combination. The detailed methodologies for each of these
evaluation parameters are described below.5
FORMULATION TABLE
Table No. 1
|
Ingredients
|
Batches
|
|
|
F1
|
F2
|
F3
|
F4
|
|
Turmeric %
|
35
|
25
|
15
|
5
|
|
Aleo vera
%
|
5
|
15
|
25
|
35
|
|
Chitosan %
|
3
|
3
|
3
|
3
|
|
Honey %
|
9
|
9
|
9
|
9
|
|
Alginate %
|
3
|
3
|
3
|
3
|
|
Gelatin %
|
3
|
3
|
3
|
3
|
|
AM solution %
|
2
|
2
|
2
|
2
|
3. EVALUATION OF PREPARED HYDROGEL
3.1. Determination of viscosity:
The viscosity microemulsion was measured at 25°C with
a Brookfield viscometer. (Brookfield DV–E) Viscosity of the samples was
determined using a Brookfield digital viscometer with spindle number 63.
3.2. Physical appearance and pH:
The pH of hydrogel formulations was determined by
using digital pH meter. The measurement of pH of each formulation was done in
triplicate and average values were calculated, using calibrated digital pH
meter at 250c.
3.3. Spreading Coefficient:
The apparatus was made of wooden block with scale and
glass slide having a pan mounted on a pulley. Excess formulation was placed
between glass slide and the smooth polish board. A 100g weight was placed on
the upper glass slide for 5 min to compress the formulation to uniform
thickness. Weight (100 g) was added to the pan. The time in seconds required to
separate the slides was taken as a measure of spread ability (Margaret et
al., 1956). The spread ability was calculated by using the following
formula:
S = (m × l)/t
Where S is spread ability; m is weight
tied to the upper slides; l is Length of glass slide and; t is
time taken in seconds 3.
3.4. Extrudability Study of Topical Gel:
The method adopted for evaluating gel formulation for
extrudability was based upon the quantity in percentage of gel and gel extruded
from lacquered aluminum collapsible tube on application of weight in grams
required to extrude at least 0.5 cm ribbon of gel in 10 seconds. More quantity
extruded better was extrudability. The extrudability was than calculated by
using the following formula:
Extrudability = Applied weight to extrude gel from
tube (in gm) / Area (in cm2) 6
3.5. Drug Content Determinations:
Drug content of gel was determined by dissolving
accurately weighed 1gm of gels in 0.1N NaOH. After suitable dilution absorbance
was recorded by using UV- visible spectrophotometer at 392 nm. Drug content was
determined using slope of standard curve. 2,11The drug content was determined
by using following equation:
Drug Content = (Concentration × Dilution Factor ×
Volume taken) × Conversion Factor 7
3.6. Swelling Index Study of Topical Gel:
Swelling of the polymer depends on the concentration
of the polymer, ionic strength and the presence of water. To determine the
swelling index of prepared topical gel, 1 gm of gel was taken on porous
aluminum foil and then placed separately in a 50 ml beaker containing 10 ml 0.1
N NaOH. Then samples were removed from beakers at different time intervals and
put it on dry place for some time after it reweighed. Swelling index was
calculated as follows:
Swelling Index (SW) % = [(Wt – Wo) / Wo] × 100.
Where, (SW) % = Equilibrium percent swelling, Wt =
Weight of swollen gel after time t, Wo = Original weight of gel at zero time. 8,9
3.7. In-Vitro Drug Release Study:
The in vitro drug release studies were
performed by using Franz diffusion cell with cellophane paper. The water jacketed
recipient compartment had total capacity of 30 ml and it had one arms for
sampling and other side for water inlet and outlet. The donor compartment had
internal diameter of 2.8 cm2. The donor compartment was placed in such a way
that it just touches the diffusion medium in receptor compartment. The receptor
compartment contained phosphate buffer solution pH 7.4. That was maintained at
37°C ± 1°C. The membrane was equilibrated before application of the micro
emulsion based Hydrogel equivalent to 8 mg of drug onto the donor side. 1ml of
Samples were periodically withdrawn from the receptor compartment, replacing
with the same amount of fresh PBS solution, and assayed by using a
spectrophotometer at 378 nm.10 The apparatus used to carry out In-Vitro
Drug Release Study is as shown in fig. 2.
Fig. 2. Franz diffusion cell with cellophane paper.
3.8. Wound Healing Activity:
For the wound healing study, healthy wistar rats were
selected and divided into six groups. The wound healing protocol was sanctioned
by the Institutional Animal Ethical Committee. Excision wounds were used for
study of rate of contraction of a wound on a wistar rat. These wistar rats were
anaesthetized by giving ketamine injection (intra peritoneal) dose of 50 mg /
kg. Back side area of each wistar rat was shaved to create an excision.
Excision wound sized 2 and 2 mm depth was made by cutting out a layer of skin
from the shaved area by using a surgical scissor. The 2 cm area of PVI hydrogel
sponge was cut and fixed it on the excision area with the help of blunt
forceps. Wound areas were measured on days 2, 4, 8 and 12 for calculation of
wound contraction.
4. RESULT
AND DISCUSSION:
4.1. Results
of evaluation parameters from no.1 to no.6 are as in table no.2
4.3. Wound Healing:
The optimized batch F2: was subjected to the wound
healing study on male wistar rats. The rate of wound contraction was taken as a
measure of the wound healing process.11
4.2. In Vitro Drug Release Study
Table No. 3: In vitro Drug Release of Hydrogel.
|
Time in Hour
|
F1
|
F2
|
F3
|
F4
|
|
1
|
30.22
|
20.18
|
22.33
|
19.11
|
|
2
|
41.69
|
35.23
|
28.21
|
25.23
|
|
3
|
47.32
|
46.24
|
45.27
|
41.76
|
|
4
|
52.57
|
54.11
|
52.91
|
40.33
|
|
5
|
65.73
|
62.23
|
65.75
|
55.62
|
|
6
|
69.72
|
71.44
|
68.32
|
63.54
|
|
7
|
74.80
|
82.23
|
70.47
|
75.33
|
|
8
|
79.41
|
95.54
|
82.33
|
84.22
|
DAY 0
DAY 4
DAY 12
Fig. 3: Wound healing activity of PVI honey hydrogel
sponge on Wister rat
Table No. 4: % of wound contraction of PVI honey
hydrogel chitosan alginate sponge (F2)
|
Days
|
% of wound contraction
|
|
|
Control
|
Test
|
|
1
|
25.28±1.09
|
22.22±1.3
|
|
2
|
40.30±0.22
|
39.22±0.42
|
|
4
|
62.90±1.22
|
60.50±1.40
|
|
6
|
87.02±1.33
|
89±0.22
|
|
8
|
91.11±2.11
|
92±1.30
|
|
10
|
94.33±0.22
|
95±0.30
|
|
12
|
97.55±0.31
|
99.28±1.22
|
5. CONCLUSION:
·
Summary, in this research paper
hydrogels were developed and evaluated for their potential as topical delivery
systems for turmeric and aloe vera.
·
The results showed that the
content of based hydrogels components (Aloe Vera and Turmeric 15%) had
significant effect on their physical, rheological and in vitro drug
release characteristics. It was considered as most desirable formulations of
the hydrogels F2.
·
Animals treated with formulations
(F2) showed a smaller wound area than the control groups, however, the
reduction of wound area are statistically insignificant till the 12thday,
the period of study.
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