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TEQIP SUMMER INTERNSHIP

RESEARCH WORK REPORT

ON

Honey Aloevera loaded electrospun PVA nanofibers as a

potential wound dressing material

SUBMITTED TO:

THE TEQIP OFFICE, IITH

Under the guidance of

Dr. Chandra Shekhar Sharma Associate Professor

Creative & Advanced Research Based On Nanomaterials (CARBON) Laboratory

Department of Chemical Engineering,

Indian Institute of Technology, Hyderabad

Submitted by:

Apeksha Sharma

B.E Chemical Engineering

Madhav Institute of Technology and Science,Gwalior

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CERTIFICATE

This is to certify that the candidate Miss Apeksha Sharma, a 3rd year Bachelor of

engineering student from the Department of Chemical Engineering, of Madhav

Institute of Technology and Science, Gwalior has satisfactorily completed the

Internship from 1st June 2018 to 30th June 2018 under my guidance. During her

training period she has submitted a research work report entitled “Honey Aloevera

loaded electrospun PVA nanofibers as a potential wound dressing material”

which has been properly examined and found to be satisfactory.

The candidate has fulfilled all the prescribed requirements.

Dr. Chandra Shekhar Sharma Associate Professor

Creative & Advanced Research Based On Nanomaterials (CARBON) Laboratory

Department of Chemical Engineering,

Indian Institute of Technology, Hyderabad

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ACKNOWLEDGEMENT

With great pleasure, I wish to put my deep gratitude and indebtedness to

my respected summer internship guide, Dr. Chandra Shekhar Sharma, Associate Professor ,

Dept. of Chemical Engineering , IIT Hyderabad for all the advice and valuable guidance that

I received from him throughout this work.

I am also grateful to Ms. Mrunalini Gaydhane, Research scholar, Dept. of Chemical

Engineering , IIT (Hyderabad) who showed her great efforts and guidance at required times

without which it would have been very difficult to carry out my research work. I am thankful

to NPIU-MHRD TEQIP IIT Hyderabad which provided me such a great opportunity of

internship.

I would like to thank my friends and family members for providing me with the moral support in course of my program.

- Apeksha Sharma

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INDEX

1. INTRODUCTION TO ELECTROSPINNING

2. WOUND HEALING AND WOUND DRESSING

3. MATERIAL SELECTION

4. EXPERIMENTAL PROCEDURE

a) Samples Preparation

b) Characterization

Optical Imaging

FTIR (fourier transform infrared spectroscopy)

5. RESULTS AND DISCUSSION

Optical images

FTIR plots

Anti-oxidant Assay

6.CONCLUSION

7.REFERENCES

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Electrospinning :

Many techniques can be used to obtain nanofibers, such as drawing, template synthesis, phase

separation and self-assembly, but electrospinning is the only process that can control fiber size,

can be scaled for larger production, and is easily replicable. Electrospinning is a process that

creates nanofibers through an electrically charged jet of polymer solution or polymer melt.

Process description: The electrospinning process, in its simplest form consisted of a pipette to

hold the polymer solution, two electrodes and a DC voltage supply in the kV range.The polymer

drop from the tip of the pipette was drawn into a fiber due to the high voltage. The jet was

electrically charged and the charge caused the fibers to bend in such a way that every time the

polymer fiber looped, its diameter was reduced. The fiber was collected as a web of fibers on the

surface of a grounded target [1].

Schematic diagram of the electrospinning process

Wound Healing and Wound Dressing

Wound healing is a special biological process which is related to physiological parameters.

Selection of a suitable wound dressing material for a specific type of wound requires

comprehensive knowledge of the wound healing process [2].

Phases of wound healing are:

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The first stage includes hemostasis and inflammation which occurs soon after there is

damage to the skin.

Another stage, In the migratory phase, the new and live cells called epithelial move towards

skin injury to replace dead cells.

The proliferation stage consists of the complete coverage of wound by epithelium.

The final stage in the healing process of a wound is tissue remodeling.

Wound Dressings

Ordinary dressings, such as gauze, act as a common cover on a wound. The second types of

wound dressings are interactive materials containing polymeric films or foams which are

transparent and permeable to water vapor and atmospheric oxygen. These materials are good

barriers against permeation of bacteria to the wound environment. The last types of wound

dressings are bioactive materials or in other words active wound dressing materials (AWD)[3].

Material selection:

Polyvinyl alcohol (PVA)

Poly(vinyl alcohol) is a water-soluble synthetic polymer. It has the idealized formula

[CH2CH(OH)]n. It is used in papermaking, textiles, and a variety of coatings. It is white

(colourless) and odorless. It is sometimes supplied as beads or as solutions in water.

Biodegradable and non- toxic, high tensile strength [4].

Aloevera: sometimes described as a "wonder plant," is a short-stemmed shrub. Aloe is a genus

that contains more than 500 species of flowering succulent plants. Some uses: Soothes Rashes

and Skin Irritations, Treats Burns and provides Antioxidants and Reduces Inflammation,Fast

healing [5].

Honey: Honey is a sweet liquid made by bees using the nectar from flowers. Honey also has

antiseptic and antibacterial properties. Modern medical science has managed to find uses for

honey in chronic wound management and combating infection [6].

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EXPERIMENTAL PROCEDURE:

a) Samples Preparation:

1. PVA (polyvinyl alcohol) solution: [PVA].6 wt % of PVA salt in 10 ml water (solvent)

2. PVA and aloevera solution:[PA].3ml PVA solution with 2ml aloevera solution

3. PVA and honey solution :[PH].4ml PVA solution with 1ml honey

4. PVA aloevera and honey solution:[PAH].3.5ml PVA with 1ml aloevera solution and 0.5 ml

honey.Mats of fibers of above samples by electrospinning and also with some other parameters

for optimization.

b) Characterization

1. Optical imaging: Optical microscopy is a technique employed to closely view a sample

through the magnification of a lens with visible light. We get the optical images of nanofibers

through optical microscope for optimization. Optical microscopy was done using Zeiss optical

microscope.

2. FTIR (fourier transform infrared spectroscopy)

Fourier Transform-Infrared Spectroscopy (FTIR) is an analytical technique used to identify

organic (and in some cases inorganic) materials. This technique measures the absorption of

infrared radiation by the sample material versus wavelength

FTIR was done by using Bruker Tensor-27 model.

RESULTS AND DISCUSSION

Optical images are as follows :- F=flowrate ,V=voltage

Samples: 1. PA nanofibers (PVA+aloe)

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[F=10ul,V=10KV,50x] [F=14ul,V=10KV,50x] (fused and beaded)

2. PH nanofibers (PVA+honey)

[F=10ul,V=12KV,50x] [F=14ul,V=10KV,50x] (wet fused)

3.PAH nanofibers (PVA + Aloe +honey]

[F=8ul, V=14KV,50x] [F=14ul,V= 14KV,50x]

4.Plain PVA: [F=10ul,V=10KV]

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FTIR PLOTS(wavenumber vs transmittance):

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0

0.2

0.4

0.6

0.8

1

1.2

5001000150020002500300035004000

PVA FTIR

PVA Aloe vera Honey PH PA PAH

Wave Number

Analysis Wave number

Analysis Wave Number

Analysis Wave number

Analysis Wave number

Analysis Wave number

Analysis

3454 -OH (alcoholic)

3341 Alcohol

OH Stretch

3292 Alcohol

OH stretch

3411 Alcohol

OH stretch

3439 Alcohol

OH Stretch

3428 Alcohol

OH Stretch

2926 -CH Alkyl

stretch

1638 C=C 2933 Carboxylic

acid OH

stretch

2925 -CH Alkyl

stretch

2923 -C-H

aldehydic 2934 -C-H

aldehydic

2851 -CH Alkyl

stretch

552 C-Br 1644 C=C 1639 C=C 2853 -C-H

aldehydic 1733 C=O

aldehyde

2362 -CH 484 C-I 1418 N02

stretch 1258 C-O-C

stretch 1642 C=C 1640 C=C

1640 C=C 467 C-I 1028 C-F 1077 C-F 1384 C-F 1425 NO2

stretch

1410 C=C 449 C-I 554 C-Br

1260 C-F 1257 C-O-C

stretch

1253 C-OH 427 C-I 488 C-I

1077 C-F

1123 C-O 412 C-I 466 C-I

1020 C-OH

447 C-I

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The FTIR analysis showed the presence of characteristics peaks from natural ingredients into the

encapsulated nanofibers.

ANTIOXIDANT ASSAY:

Importance of Antioxidants

Antioxidants are nature's way of fighting off potentially dangerous molecules in the body.

Every day tens of thousands of free radicals are generated within the body, causing cell damage

that can lead to chronic and degenerative diseases if left unchecked.

The body sometimes creates its own free radicals in order to destroy viruses or bacteria. To

balance out these unruly molecules, the body also creates antioxidants, which have the sole

purpose of neutralizing free radicals [7].

Anti-oxidant activity:

1. 10 mg of fiber is dissolved in 10 ml of 80% ethanol aqueous solution.

2. 1000 micro litre of above sample solution is reacted with 3 ml of DPPH (0.1 mM solution in

methanol) separately for 30 min in dark.

3. The absorbance is recorded spectrophotometrically at 516 nm.

4. Anti-oxidant activity is calculated from the absorbance data;

%Anti-oxidant Activity = [(Acontrol - Asample) / (Acontrol)]*100 %

where Acontrol and Asample are the absorbance values of the DPPH solution without and with

the presence of the sample solutions.

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CONCLUSION:

A nanofibrous dressing loaded with aloe vera and honey was successfully fabricated.

The optical images confirms the smooth and uniform encapsulation of honey and aloe vera into

the PVA nanofibers. The FTIR analysis confirms the presence of active groups from aloe and

honey into their corresponding nanofibers. The anti-oxidant testing helped to determine the anti-

oxidant activity of aloe – honey loaded nanofibers. It is found that the composite mat shows 56.6

% of activity against DPPH radical.

The ultrathin nanofibrous dressing protects the wound from mechanical trauma, absorbs the pus

and provides therapeutic cure to the wound.

21.19

39.69 40.11

56.63

28.35

0.000.00

10.00

20.00

30.00

40.00

50.00

60.00

Aloe H PH PAH PA dpph

% Anti-oxidant activity

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REFERENCES:

[1] Ramakrishna, Seeram. An introduction to electrospinning and nanofibers. World Scientific,

2005.

[2] Velnar, Tomaž, Tracey Bailey, and Vladimir Smrkolj. "The wound healing process: an

overview of the cellular and molecular mechanisms." Journal of International Medical

Research 37, no. 5 (2009): 1528-1542

[3] Cai, Zeng-xiao, Xiu-mei Mo, Kui-hua Zhang, Lin-peng Fan, An-lin Yin, Chuang-long He,

and Hong-sheng Wang. "Fabrication of chitosan/silk fibroin composite nanofibers for wound-

dressing applications." International journal of molecular sciences 11, no. 9 (2010): 3529-3539.

[4] Peresin, Maria S., Youssef Habibi, Justin O. Zoppe, Joel J. Pawlak, and Orlando J. Rojas.

"Nanofiber composites of polyvinyl alcohol and cellulose nanocrystals: manufacture and

characterization." Biomacromolecules 11, no. 3 (2010): 674-681

[5] https://en.wikipedia.org/wiki/Aloe_vera

[6] https://en.wikipedia.org/wiki/Honey

[7] Sunil Kumar. / Asian Journal of Research in Chemistry and Pharmaceutical Sciences. 1(1),

2014, 27 - 44. www.uptodateresearchpublication.com January - March 27 Review Article ISSN:

2349 – 7106 THE IMPORTANCE OF ANTIOXIDANT AND THEIR ROLE IN

PHARMACEUTICAL SCIENCE - A REVIEW