TABLE OF CONTENTS INTRODUCTION

INTRODUCTION 2

LITERATURE REVIEW 4

JUSTIFICATION 6

OVERALL GOAL 7

OBJECTIVES 7

PLAN OF STUDY 8

METHODOLGY 9

PLACE OF WORK 11

REFERENCES 12

1
 INTRODUCTION
Azadirachta indica, commonly known as neem, nimtree or Indian lilac, is a tree in
the mahogany family Meliaceae. It is one of two species in the genus Azadirachta, and is native
to the Indian subcontinent, i.e. India, Nepal, Pakistan, Bangladesh, Sri Lanka, and Maldives. It is
typically grown in tropical and semi-tropical regions. Neem trees also grow in islands located in
the southern part of Iran. Its fruits and seeds are the source of neem oil.

In the world of Ayurveda, neem is a popular medicinal herb that’s been part of traditional
remedies that date back almost 5000 years. Also known as Azadirachta Indica in English or
‘Neemba’ in Sanskrit, the neem tree is a really good example of how nature holds both the
problem and the cure. It’s home to more than 130 different biologically active compounds! No
wonder it’s such an effective anti-viral and anti-bacterial, along with being a powerful immuno-
stimulant. Murli Manohar, author of the book 'Ayurveda For All: Effective Ayurvedic Self Cure
for Common and Chronic Ailments' suggests that the primary purpose of neem leaves is the
treatment of vaata disorders or neuromuscular pains. Then come the other benefits: purify the
blood, prevent damage caused by free radicals in the body, remove toxins, treat insect bites and
ulcers. Neem leaves have anti-bacterial properties which is why it works wonders on infections,
burns and any kind of skin problems. It destroys the bacteria that causes infections, stimulates
the immune system and encourages rapid healing. We tell you some benefits of neem.

Wound healer: Make a paste out of the neem leaves and dab it on your wounds or insect
bites a few times a day till it heals.

Goodbye dandruff: Boil a bunch of neem leaves till the water turns green, allow it to cool.
After washing your hair with shampoo, c leanse it with this water.

Eye Trouble: Boil some neem leaves, let the water cool completely and then use it to wash
your eyes. This will help any kind of irritation, tiredness or redness.

Treat that zit: Grind a few neem leaves, make a paste and apply it daily till the acne dries
out. The paste also helps any kind of eruptions, dark spots and chronic ulcers.

2
  Ear ailments: Blend some neem leaves and add some honey to it. Use a few drops of this
mix to treat any ear boils.

 Other skin disorders: Turmeric combined with a paste of neem leaves can also be used
for itching, eczema, ring worms and some mild skin diseases.

 Boost immunity: Crush some neem leaves and take them with a glass of water to
increase your immunity

3
 LITERATURE REVIEW
The active principles from bark, leaves, seeds, and other parts of neem are exploited for
their proven immunomodulatory, antiseptic, diuretic, antipyretic, antiparasitic, antimicrobial,
analgesic, antifeedant, contraceptive, pediculicide, antiulcer, antimutagenic, and anticancer
effects represents the synthesis of all the beneficial effects of neem extracts.

The active principles from bark, leaves, seeds, and other parts of neem are exploited for
their proven immunomodulatory, antiseptic, diuretic, antipyretic, antiparasitic, antimicrobial,
analgesic, antifeedant, contraceptive, pediculicide, antiulcer, antimutagenic, and anticancer
effects represents the synthesis of all the beneficial effects of neem extracts. The beneficial
activity of neem in general health, disease prevention, and treatment (adapted from Tiwari.

Acharya and Raj reported that the botanical research on medicinal plants is getting much
importance and it is included among the advanced research in India during the recent years.
Plants have much relevance on Socio-economic, Socio-religious aspects of human life inIndia.
They described various medicinal plants with their uses. They described that the use of karanj
Seed oil is good for curing skin infections.

Pankaj Oudhia reported that the reference literatures related to different systems of medicine in
India specially related to Ayurveda are full of miraculous therapeutic properties of Karanj. He is
astonished then why we are ignoring this useful tree? He reported that the karanj is better than
Jatropha.

Mondal and Islam investigated on the toxicity study of Derrisindica, which belongs to family
Fabaceae, and is inhabitants of India, Srilanka, Malaysia, North Australia and Polynesia, occurs
in the tidal forests, river and canal banks, along the water edge in all districts in Bangladesh. It is
a medicinal plant. The dried flowers are used in decoction to quench thirst in diabetes. Extracts
of the leaves were active against Micrococcus pyogenes Var. aureus. The juice of the leaves is
prescribed in flatulence dyspepsia, diarrhoea and cough.

4
 They have mentioned karanjin or pongapin, kanugin and dimethoxy kanugin, pongamol,
etc. to be present in the seeds of the test plant D. indica. Review of LiteratureShailja Vohra (4)
isolated chemical compound using methanolic extract of Pongamia glabra and analysed for its
karanjin and other flavonoid and hydrocarbon content by electrospray ionisation (ESI) mass
spectroscopy. It showed the presence of six flavonoids namely karanjin, desmethoxykanugin,
pongachalcone, pongapin, glabrachromene I, glabrachromene II and six hydrocarbons namely
octadecane, undecane,decane, octane, undecanol and tetradecanol.

The karanj based products exhibited outstanding antifungal activity against the soil-borne
phytophagous fungus Sclerotium rolfsii (Sacc.). Karanj oil in the karanjin was more active than
karanj oil without karanjin. Karanjin, however, exhibited moderate antifungal activity. Karanjic
acid and their three esters exhibited significant antifungal activity. Karanjic acid showed the
highest antifungal activity.

JUSTIFICATION AND LIKELY BENEFITS

5
 The injuries has always been a feature of human society. Before the present modern
medical remediation of such exposures, the classical, natural homeopathic remedies have been
used for these tragedies. Neem is a verified antibacterial and cleaning agent. This plant can be
used for the treatment of such injuries. This can easily be obtained and used for the medications.
This plant can be found world widely and is easily available. Neem can be used for the treatment
of the skin injuries due its healing ability.

OVERALL GOAL
To study the effect of Neem Plant leaves on chemically injured skin of rat.

6
 Objectives
1. To prepare the optimize dose of Neem Plant Leaves Paste.

2. To establish the rat model of chemically injured skin.

3. To Neem plant Leaves Paste topically on chemically injured skin.

4. To evaluate the effect of Neem Plant Leaves Paste on chemically injured skin of rat.

PLAN OF STUDY

MONTH 1 :

7
 Making paste of the leaves of Neem plant.

MONTH 2 :

Development of rat model of chemically injured skin .

MONTH 3 :

Appllication of Neem plant leaves paste topically on chemically injured skin of rat.

MONTH 4 :

Assessment of skin regeneration.

Statistical analysis of data obtained.

Thesis write up and submission.

Methodology
Collection of Neem Plant Leaves and paste formation: Firstly, the neem plant leaves will be
collected and then paste of the leaves will be make.

8
 OPTIMIZATION OF CHEMICAL INJURY OF RAT:

Normal skin of rat will be injured with three different concentration of HCL to optimize
skin chemically injury model . The skin chemical injury model will be evaluated by microscope.
Real Time PCR:

The cytoprotective effect of neem plant leaves paste in response to chemical injury will be
evaluated by Real Time PCR. RNA will be extracted from normal as well as skin injured with
fire. Then cDNA will be prepared 1µ gm of RNA and amplification of the target genes will be
carried out under optimized conditions for each gene from normal and pretreatment groups.

Wound Healing Assay:

Scratch wound for the normal and pretreated skin cells six well plates will be made by sterile
white tips. A sharp streak will be made in the middle of plate with a sterile tip. After different
days, the cells will be stained with Crystal Violet. The healing of empty spaces in the plates will
be observed and images will be taken under phase contrast microscope to evaluate the wound
healing potential both the normal and the burnt skin.

APPLICATION OF NEEM PLANT LEAVES PASTE:

Apply neem plant leaves paste topically on chemically injured skin .

ASSESSMENT OF SKIN REGENERATION:

Wound size reduction will be evaluated by tracking wound area at different days . First at
glass slide and then at graph paper . The digital photography of wounded skin will be carried out
at different days . The period of complete re -epithelialization will be determined by observing
the complete regenetation of each wound.

9
 PLACE OF WORK AND FACULTIES AVAILABLE
Zoloogical lab of Ghazi University D.G.Khan is well eqipped with microscope ,graph
papers , glass slides , chemicals, animal house facility for the experimental animals. These
facilities will prove to be a backbone for achieveing the proposed out-comes of this research
project.

10
 References
1. Mondal and Islam This article retrieved from
http://shodhganga.inflibnet.ac.in/bitstream/10603/5140/6/06_chapter%202.pdf dated:
09-01-2019
2. Pankaj Oudhia This article retrieved from
http://shodhganga.inflibnet.ac.in/bitstream/10603/5140/6/06_chapter%202.pdf dated:
09-01-2019

11
3. Acharya and Raj This article retrieved from
http://shodhganga.inflibnet.ac.in/bitstream/10603/5140/6/06_chapter%202.pdf dated:
09-01-2019
4. Tiwari This article retrieved from
http://shodhganga.inflibnet.ac.in/bitstream/10603/5140/6/06_chapter%202.pdf dated:
09-01-2019

5. Jump up to:a b "Azadirachta indica". World Checklist of Selected Plant

Families (WCSP). Royal Botanic Gardens, Kew. Retrieved 14 December 2016 –
via The Plant List.

6. Jump up to:a b "Azadirachta indica". Germplasm Resources Information

Network (GRIN). Agricultural Research Service (ARS), United States Department of
Agriculture (USDA). Retrieved 9 June 2017.

7. Compact Oxford English Dictionary (2013), Neem, page 679, Third Edition 2008
reprinted with corrections 2013, Oxford University Press.

8. Henry Yule and A. C. Burnell (1996), Hobson-Jobson, Neem, page 622, The Anglo-
Indian Dictionary, Wordsworth Reference. (This work was first published in
1886) Encarta World English Dictionary (1999), Neem, page 1210, St. Martin's Press,
New York.

9. Plant Risk Assessment, Neem Tree, Azadirachta indica (PDF). Biosecurity

Queensland. 2008. Retrieved 24 January 2014.

10. Neem has been declared: what you need to know (PDF), Department of Land
Resource Management, 2015, archived from the original (PDF) on 24 March 2015,
retrieved 17 March 2015

11. Neem Azadirachta indica (PDF), Department of Land Resource Management, 2015,
archived from the original (PDF) on 24 March 2015, retrieved 17 March 2015

12. Jump up to:a b c Anna Horsbrugh Porter (17 April 2006). "Neem: India's tree of life".
BBC News.

13. "Neem Baigan". Jiva Ayruveda. Archived from the original on 9 July 2014. Phyllyppo
Tum.

14. D.P. Agrawal (n.d.). "Medicinal properties of Neem: New Findings".

12
15. ^ S. Zillur Rahman and M. Shamim Jairajpuri. Neem in Unani Medicine. Neem
Research and Development Society of Pesticide Science, India, New Delhi, February
1993, p. 208-219. Edited by N.S. Randhawa and B.S. Parmar. 2nd revised edition
(chapter 21), 1996

16. "Neem". Tamilnadu.com. 6 December 2012. Archived from the original on 11 April

17. Jump up to:a b c Neem, WebMD.

18. M.V. Bhaskara; S.J. Pramoda; M.U. Jeevikaa; P.K. Chandana; G. Shetteppa (May 6,
2010). "Letters: MR Imaging Findings of Neem Oil Poisoning". American Journal of
Neuroradiology. 31 (7): E60–E61. doi:10.3174/ajnr.A2146. PMID 20448012.

19. Material Fact Sheets — Neem Archived 12 February 2013 at the Wayback Machine.

20. YashRoy, R.C.; Gupta, P.K. (2000). "Neem-seed oil inhibits growth of termite
surface-tunnels". Indian Journal of Toxicology. 7 (1): 49–50.

21. Development of PU Coatings from Neem Oil Based Alkyds Prepared by

Monoglyceride Route, Journal of American Oil Chemist’s
Society, doi:10.1007/s11746-015-2642-3

22. Nardi, Isabella (2007). The Theory of Citrasutras in Indian Painting. Routledge.
p. 121. ISBN 978-1134165230.

23. "Make A Neem Toothbrush (Neem Tree Home Remedies)". Discover Neem. Birgit
Bradtke. Retrieved 16 July 2013.

24. Bhambal, Ajay; Sonal Kothari; Sudhanshu Saxena; Manish Jain (September
2011). "Comparative effect of neemstick and toothbrush on plaque removal and
gingival health – A clinical trial" (PDF). Journal of Advanced Oral Research. 2 (3):
51–56. ISSN 2229-4120. Archived from the original (PDF) on 5 November 2013.

13
25. Callahan, Christy (Oct 11, 2010). "Uses Of Neem Datun For
Teeth". Livestrong.com. Demand Media. Retrieved 16 July 2013.

26. Schroeder, Paul. "Carbon storage potential of short rotation tropical tree
plantations." Forest Ecology and Management 50.1-2 (1992): 31-
41. doi:10.1016/0378-1127(92)90312-W

14