TABLE OF CONTENTS

INTRODUCTION 2
LITERATURE RIVIEW. 3
JUSTIFICATION 5
OVERALL GOAL. 6
OBJECTIVES. 6
PLAN OF STUDY 7
METHODOLGY. 8
PLACE OF WORK. 10
REFERENCES. 11

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 INTRODUCTION

The topical administration of ash along with local defence mechanism appears to be superior to
other types of therapies as it reduces super infection and enhance epithelization in rabbit skin
wound healing.

Wood ash were prepared and collected by complete burning of the parent material i.e. wood
chips and dead branches of wattle (Acacia) from local supplier respectively. Ashes were
analysed and applied besides the Polymyxin B-Bacitracin Zinc ointment as control.

Typically between 0.43 and 1.82 percent of the mass of burned wood (dry basis) results in
ash. Also the conditions of the combustion affect the composition and amount of the residue ash,
thus higher temperature will reduce ash yield.

Much wood ash contains calcium carbonate as its major component, representing 25 or even 45
percent.Less than 10 percent is potash, and less than 1 percent phosphate; there are trace
elements of iron, manganese, zinc, copper and some heavy metals. However, these numbers
vary, as combustion temperature is an important variable in determining wood ash composition.
All of these are, primarily, in the form of oxides

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 LITERATURE REVIEW
In Sardinia (Italy),Piluzza et al. (2015)have documented the use of ash in water, wrapped
with a cloth around the wounds from red fox – Vulpes vulpes (Linnaeus, 1758)– or dog bites. In
fact, several tests conducted with animal models have confirmed that the topical administration
of ashes (corresponding to complete burning of woody materials) accelerates the wound healing
process, enhances epithelialisation and reduces infection (Hamid and Shaikh, 2009;Shaikh and
Hamid, 2009).
The analysis of different types of wood ashes revealed that they contain the appropriate
combination of metals required to heal a wounded area faster (Hamid and Shaikh, 2009; Shaikh
and Hamid, 2009).
In fact, several tests conducted with animal models have confirmed that the topical
administration of ashes (corresponding to complete burning of woody materials) accelerates the
wound healing process, enhances epithelialisation and reduces infection (Hamid and Shaikh,
2009;Shaikh and Hamid, 2009). The analysis of different types of wood ashes revealed that they
contain the appropriate combination of metals required to heal a wounded area faster (Hamid and
Shaikh, 2009; Shaikh and Hamid, 2009).
Some industrial by-products have been studied for use as supplementary cementitious
materials (SCM) viz. fly ash, silica fume , pulverized fuel ash , volcanic ash , rice husk ash and
corn cob ash (CCA).
However, literature is scarce on the utilization of saw dust ash or wood waste ash. Elinwa
and Ejeh (2004) worked on to know the performance of cement based products like paste,
mortar and concrete by using ash as a partial replacement additive produced by incinerating solid
waste substance collected from the municipal garbage’s. Cheah and Ramli (2011) also reviewed
the properties of mortar and concrete used in construction of structural members produced by
using WWA as a mineral admixture in partial replacement levels.
Elinwa et al. (2008) investigated on the performance of self compacting concrete
prepared by incorporating ash produced from incinerating sawdust during fresh stage as a
mineral additive and from the studies carried out by Elinwa and Mahmood (2002) on utilisation
of ash obtained from incineration of timber waste products as cement replacement material.
Based on the measured physical, chemical, morphological properties.

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 Naik et al. reported that wood waste ash can used as a substantial mineral admixture
material as a pozzolanic and an activator in materials manufactured with cement as a major
product. Further, it is indicated that ash produced from incinerating waste wood substance gives
more potential to use in production of cement based materials possessing low to medium strength
products used in works, where the compressive strength is paid less attention. These material is
suggested to be used in construction works like masonry, pavement works both in surface and
sub surface works and it can be used as a cement blended with rich siliceous product i.e., ash to
increase durability.

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 JUSTIFICATIONS LIKELY BENEFITS
Wound healing is a complex biological process that consists of hemostasis, inflammation,
proliferation, and remodeling. Large numbers of cell types—including neutrophils, macrophages,
lymphocytes, keratinocytes, fibroblasts, and endothelial cells—are involved in this process.

Multiple factors can cause impaired wound healing by affecting one or more phases of the
process and are categorized into local and systemic factors.

The influences of these factors are not mutually exclusive. Single or multiple factors may play a
role in any one or more individual phases, contributing to the overall outcome of the healing
process.
That’s why start study on this topic and want to know the effect of wood ash paste on healing of
wound.

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 Overall Goal
To study the effect of wood ash on healing of chemically injured skin in rat .

Objectives
1. To prepare the optimize dose of wood ash .
2. To establish rat model of chemically injured skin.
3. To apply ash paste topically on chemically injured skin of rat.
4. To evaluate the effect of wood ash in healing of wound.

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 PLAN OF STUDY
MONTH 1:
 Preparation of wood ash .
 Formation of ash paste by mixing it with water.

MONTH 2:
 Development of rat model of chemically injured skin of rat.

MONTH 3:
 Apply wood ash paste topically on chemically injured skin of rat.

MONTH 4;
 Assessment of skin regeneration.
 Statically analysis of data obtained.
 Thesis write up and submission.

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 Methodology
1. Wood ash preparation:
Firstly, the wood ash will be prepared by burning the wood and then mixing it with water to
make the paste of wood ash.

2. Optimization of chemical injury of Rat:

Normal skin of rat will be injured with three different concentrations HCL to optimize the skin
chemically injury model.

The skin chemically injured model will be evaluated with Microscope, PCR and Trypan Blue
Assay.

3. Application of Wood Ash paste :

Apply wood ash paste topically on chemically injured skin of rat.

: 4. Assessment of regeneration
I. Real Time PCR;
The Cytoprotective effect of wood ash paste in response to chemical injury model will be
evaluated by real time PCR. RNA will be extracted from both the normal and chemically injured
skin of rat. The cDNA will be prepared from 1µ gms of RNA and amplification target gene will
carried out using relevant primers through real time PCR, which will be under optimized
conditions for each gene normal and chemically injured groups.

II. Wound Size Reduction:

Wound size reduction will be evaluated by tracking wound area at different days, first at slides
and then at graph paper. The digital photography of wound skin will be carried out at different
days.

III. The Complete Rehabilitation

The period of complete re habilitation will be determined by observing the complete
regeneration of each wound.

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IV. HISTOLOGICAL EVALUATION:
Wood ash treated tissue of skin will be immunohistologically analyzed for differentiation
through various molecular markers using appropriate anti bodies the morphology of skin will be
evaluated by hematoxlin and Eosin staining.

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 PLACE OF WORK AND FACULTIES AVAILABLE

Zoological lab of Ghazi University D.G.Khan is well equipped with microscope ,graph
papers , glass slides , chemicals, animal house facility for the experimental animals. These
facilities will prove to be a backbone for achieving the proposed out-comes of this research
project.

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 REFFERENCES

1. Piluzza et al. (2015) This article retrieved from

http://shodhganga.inflibnet.ac.in/bitstream/10603/5140/6/06_chapter%202.pdf dated: 09-
01-2019
2. Linnaeus, 1758 This article retrieved from
http://shodhganga.inflibnet.ac.in/bitstream/10603/5140/6/06_chapter%202.pdf dated: 09-
01-2019
3. Hamid, 2009 This article retrieved from
http://shodhganga.inflibnet.ac.in/bitstream/10603/5140/6/06_chapter%202.pdf dated: 09-
01-2019
4. Shaikh, 2009 This article retrieved from
http://shodhganga.inflibnet.ac.in/bitstream/10603/5140/6/06_chapter%202.pdf dated: 09-
01-2019
5. Elinwa and Ejeh (2004) This article retrieved from
http://shodhganga.inflibnet.ac.in/bitstream/10603/135326/9/09_chapter%202.pdf dated:
09-01-2019
6. Cheah and Ramli (2011) This article retrieved from
http://shodhganga.inflibnet.ac.in/bitstream/10603/135326/9/09_chapter%202.pdf dated:
09-01-2019
7. Elinwa et al. (2008) This article retrieved from
http://shodhganga.inflibnet.ac.in/bitstream/10603/135326/9/09_chapter%202.pdf dated:
09-01-2019
8. Elinwa and Mahmood (2002) This article retrieved from
http://shodhganga.inflibnet.ac.in/bitstream/10603/135326/9/09_chapter%202.pdf dated:
09-01-2019
9. Naik et al. This article retrieved from
http://shodhganga.inflibnet.ac.in/bitstream/10603/135326/9/09_chapter%202.pdf dated:
09-01-2019
10. Hume E (11 April 2006). "Wood Ashes: How to use them in the Garden". Ed Hume
Seeds.
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11. ^ Jump up to:a b Tarun R. Naik; Rudolph N. Kraus & Rakesh Kumar (2001), Wood Ash:
A New Source of Pozzolanic Material, Department of Civil Engineering and Mechanics,
College of Engineering and Applied Science, The University of Wisconsin – Milwaukee
12. ^ Jump up to:a b Etiegni L, Campbell AG (1991). "Physical and chemical characteristics
of wood ash". Bioresource Technology. 37 (2): 173–178. doi:10.1016/0960-
8524(91)90207-Z.
13. ^ Wolff, Emil (1871). Aschen-Analysen. Berlin: Wiegandt und Hempel.
14. ^ Jump up to:a b c d Misra MK, Ragland KW, Baker AJ (1993). "Wood Ash Composition
as a Function of Furnace Temperature" (PDF). Biomass and Bioenergy. 4 (2): 103–
116. doi:10.1016/0961-9534(93)90032-Y.
15. ^ Jump up to:a b c Lerner BR (16 November 2000). "Wood Ash in the Garden". Purdue
University, Department of Horticulture and Landscape Architecture. Retrieved 2008-10-
01.
16. ^ Sholto Douglas, James (1985). Advanced guide to hydroponics: (soiless cultivation).
London: Pelham Books. pp. 345–351. ISBN 9780720715712.
17. ^ Demeyer A, Voundi Nkana JC, Verloo MG (2001). "Characteristics of wood ash and
influence on soil properties and nutrient uptake: an overview". Bioresource
Technology. 77 (3): 287–95. doi:10.1016/S0960-8524(00)00043-2. PMID 11272014.
18. ^ Rosenfeld, P. & Henry, C. (2001). "Activated Carbon and Wood Ash Sorption of
Wastewater, Compost and Biosolids Odorants". Water Environment Research. 7 (4):
388–393.
19. ^ Rogers, Phil (2003). Ash Glazes (2nd ed.). London: A&C Black. ISBN 978-0-7136-
57821.
20. ^ "Making lye from wood ash". Journey to Forever. 14 May 2009. Retrieved 2008-10-01.
21. ^ Geoffrey Michael Gadd (March 2010). "Metals, minerals and microbes:
geomicrobiology and bioremediation". Microbiology. 156 (Pt 3): 609–
643. doi:10.1099/mic.0.037143-0. PMID 20019082.

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