RESEARCH PROPOSAL ON BIOENGINEERING

SUBMITTED BY

DR.A.GNANAMANI

Central Leather Research Institute (Adyar, Chennai 20)

Tamil Nadu,India

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PROFORMA – I PROFORMA FOR SUBMISSION OF PROJECT PROPOSALS ON RESEARCH

AND DEVELOPMENT, PROGRAMME SUPPORT

(To be filled by the applicant)

PART I: GENERAL INFORMATION 1. Name of the Institute/University/Organisation submitting the Project Proposal:

Central Leather Research Institute

2. State: Tamil Nadu 3. Status of the Institute: Research & Development 4. Name and designation of the Executive Authority of the Institute/University

forwarding the application: Dr. A. B. MANDAL, DIRECTOR, CLRI, ADYAR, CHENNAI 5. Project Title: An early tissue regeneration in burn injured wounds using

biocellulose and collagen (BCC) scaffolds impregnated with natural cooling agent

6. Category of the Project (Please tick):R&D/ Programme Support: R & D 7. Specific Area: Biomaterial and Tissue Engineering 8. Duration: Three. Years 9. Total Cost (Rs.) .Rs. 65, 81,600 lakhs 10. Is the project Single Institutional or Multiple-Institutional (S/M): Single 11. Scope of application indicating anticipated product and processes

The proposal aims to develop bio-cellulose based biomaterial incorporated with cooling agent to have early tissue regeneration in burn wound healing

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13. Project Summary (Not to exceed one page. Please use separate sheet). Wound healing, in general, is a complex process and involves numbers of

biochemical factors. For the past two decades our group involved in the development of

biomaterial to prevent infection and early wound healing along with various herbal

products. Though normal wounds demonstrated early healing in the presence of

external material and herbal extracts, healing of burn wound is always challenging. It

has been found that; invasion of microbes in burn wounds restricts the healing process.

Recently, NIH, USA, promote research on burn wound healing and infection control

using natural materials. In India, although the percentage of burns and death due to

burns is at higher level, the research on burn wound management is very meager.

Followed by the research development on biomaterials, numbers of biomaterial products

are released to the market to satisfy the need. However, in these biomaterials the major

active constituent is collagen, (a bio macro molecule) which was stabilized by

glutaraldehyde, however, recent realization on the adverse effect of this stabilizing

agent, demands agents of natural and non-toxic for stabilization. Further, collagen is

generally extracted from animal sources and fish, the extraction processes consumes,

time, large quantity of chemicals and water. In order to have an alternative, natural

material extracted from plant and microbial sources found suitable. But, extraction of

compounds from plants is also a complex process and the extracellular compounds

secreted by the microbes stood as an alternative and utilization of these compounds

reduces the cost as well as time consumption. Bio-cellulose is one the microbial

extracellular product and recently in limelight. Though bio-cellulose found wide

applications, its use for biomaterial development along with collagen is the first kind of

the study.

In addition, during burns, the irritation and the release of inflammatory signals to

site of wound generate heat to the wound site and make the patient intolerable to the

pain. Because of the continuous release of inflammatory signals, the cross talk between

the other molecules was reduced and necessitates medicating with anti-inflammatory

agents. However, the adverse effect of continuous use of these agents further demands

alternative agents. Application of cooling agents some how offered relief, however, their

continuous supply to large wound area is questionable and impracticable. The present

proposal emphasizes to bring early tissue regeneration in burn wound healing by

biomaterial developed using bio-cellulose of bacterial origin and incorporation of cooling

agent. The science behind the early tissue regeneration will be assessed using suitable

analytical procedures and techniques.

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PART II: PARTICULARS OF INVESTIGATORS Principal Investigator: 14. Name: Dr. A. Gnanamani

Date of Birth: 01/06/1964 Sex (M/F): Female

Designation: Senior Scientist

Department: Microbiology Division

Institute/University: Central Leather Research Institute. Adyar, Chennai 600 020

Telephone: 044-24422024: Fax: 044-24912150: Mobile: 09444829103

E-mail:gnanamani3@gmail.com

Number of research projects being handled at present: One

Co-Investigator:

Name: Dr. A. B.Mandal

Date of Birth: 13/1/ 1952 Sex (M/F): Male

Designation: Director, Scientist H

Institute/University: Central Leather Research Institute. Adyar, Chennai 600 020

Telephone: 044-24437131: Fax: 044-24912150: Mobile: 09940121703

E-mail:abmandal@hotmail.com

Number of research projects being handled at present: Six

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PART III: TECHNICAL DETAILS OF PROJECT 16. Introduction (not to exceed 2 pages or 1000 words) 16.1 Origin of the proposal A damage or wound in human body of any age is unavoidable. One-way or other,

the skin gets damaged and the higher percentage of skin damage occurs during

accidents and fire suicides. Though, nature has beautifully constructed the skin with

self-healing property, nevertheless, it has not been expected from severely damaged

skin. For the skin to self - heal, the minimum requirement of healing agents should be

present. However, in the case of accidents and fire suicides, the damage is deep and

skin could not able to self-heal or restore to its original functions.

Moreover, infection in wound site is also unavoidable because of the presence of

proteinaceous materials. Infection due to bacteria/fungi delay healing of wound and

microbial invasion should be controlled from zero hour of treatment onwards. Because of

the deep damage or deep wound, infection in burn wounds is comparatively higher than

open wound. Most of burn wounds even contain anaerobic organisms, which further

increases the severity of the wound and morbidity.

In order to avoid infection in burn wounds, early tissue regeneration is the need

for the hour. However, because of non-availability of epidermal/dermal components,

the cells responsible for the wound closure could not proliferate at faster rate and

because of the slow proliferation reconstruction of skin correspondingly slows down.

Further, application of biomaterial of any type prevents the invading microbes and also

assists proliferation of cells. However, presence of proteolytic enzymes (MMPS) and its

unregulated expressions, hydrolysis of the applied biomaterials takes place which

releases toxic compounds to the system (from the stabilizing agents used). Hence,

recent research on biomaterial development has been focused on natural stabilizing

agents and the search for natural stabilizing agent is going on.

Furthermore, at the time of burn, immediate release of inflammatory signals to

the wound site adds pain to the patient. Though, anti-inflammatory agents have been

medicated, the irritation and the pain during initial days is intolerable which reduces the

therapeutic processes. Application of cooling agents as suggested by various

researchers, though provided temporary relief, but it is highly challenge to maintaining

the cool environment.

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16.2 (a) Rationale of the study supported by cited literature (b) Hypothesis (c) Key questions. 16.2 (a) Rationale for identified problem

Burn injuries are one of the most expensive catastrophic injuries to treat. For

instance, a burn on 30% of the total body area can cost as much as $200,000 in

initial hospitalization costs and for physician’s fees. For massive burns, there are

additional significant costs that will include costs for repeated hospitalization for

reconstruction and for rehabilitation. Burns are classified either by method or by

degree. Thermal burns are those caused by contact with flames, steam, hot water or

liquids and other sources of intense heat. Light burns are caused by contact with

sunlight or other sources of ultraviolet light. Chemical burns are caused by contact

with an acid or an alkali. Radiation burns are caused by contact with nuclear

radiation or ultraviolet light. According to the degrees, First-degree burn is one

where only the first layer of skin is burned. It is characterized by reddened skin that

may heal in approximately one week and thereafter, peel. Second-degree burn is

one where the first and second layers of the skin are burned. It is characterized by

moist-looking skin and blisters. Third-degree burn is one where all layers of the skin

are burned and the underlying tissue is damaged. It is characterized by a white or

black dry wound. Permanent scarring is inevitable.

Impact of Burn Injuries

Serious burns are complex injuries because it affects a number of other

bodily functions. It can affect muscles, bones, nerves, and blood vessels.

Consequently, they alter the body’s normal fluid level, joint function, body thermal

regulation, body temperature, manual dexterity and physical appearance. In addition

to the physical injuries, patients may also suffer emotional and psychological

problems while recuperating from their injuries.

Management

At the time of injury, the irritation activates the inflammatory signals.

However, in the case of burn injury, the high irritation and fluid loss, makes the

system highly pathetic. However, reducing the burning sensation activate the

inflammatory signal generations. Immediate application of ice, water, oil, honey,

low temperature heating pads and even toothpaste (presence of

peppermint/menthol) are the methods to reduce the burning sensation. However,

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these methods are not advisable by the clinicians, because of the chances of

microbial contaminations at wound site, which unnecessarily delay the healing and in

some times leads to mortality. Till date no biomaterial with cooling as well as healing

potency is in the market. In our laboratory we have developed ICE COOL

BIOMATERIAL using collagen and a novel cooling agent from natural resources.

The animal model studies revealed reduction in IL-6 and IL-1 at significant level

upon the application of this material.

Based on our experience in the Ice Cool Biomaterials, we like to develop a

novel biomaterial which able to regenerate blood vessels and tissues at higher rate

and also to reduce the burning sensation during early hours of burn injury.

(b) Hypothesis

We have chosen Bacterial cellulose (BC) and collagen as base components.

In our laboratory a facultative anaerobe of rumen develops substantial quantity of

bacterial cellulose with in 48 hours. The following figure displayed the production of

cellulose during the growth of the organisms. Extracting the cellulose and prepare a

regenerative and cool biomaterial using type I collagen with the identified ice cool

material (cooler than menthol) is the major objective of the present proposal. The

following schematic hypothesis explain the nature of BCCS for effective acid burn

injuries

+

Schematic representation of preparation of Schematic representation of preparation of Schematic representation of preparation of Schematic representation of preparation of BCCS with impregnated cooling agentBCCS with impregnated cooling agentBCCS with impregnated cooling agentBCCS with impregnated cooling agent++

Schematic representation of preparation of Schematic representation of preparation of Schematic representation of preparation of Schematic representation of preparation of BCCS with impregnated cooling agentBCCS with impregnated cooling agentBCCS with impregnated cooling agentBCCS with impregnated cooling agent

B i o - C e l l u l o s e p r o d u c t i o n B i o - C e l l u l o s e p r o d u c t i o n

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(c) Key Questions 1. Is it possible to have expected volume of bio-cellulose for the manufacture of

biomaterial 2. Is it essential to have blend with collagen 3. What is the percentage level of cooling agents and how long it will offer cooling

property? 4. Are you sure about the toxicity and biodegradability of the material developed? 5. How the in vitro and in vivo assessment will be made for the material? 16.5 Current status of research and development in the subject (both international and national status) International status: The importance of bio-cellulose has been realized a decade and number of

products based on bio-cellulose is available in the market. For example;

Nympheas International Biomaterial Corporation has been a professional

manufacturer & supplier of skincare products, skin care masks since 1999. With

devotion to biomaterial research for years, there was a revolutionary

breakthrough in the research on the bio cellulose (BC) application and

development in 2006. The Outcome of the research launched the marvelous

product called "Hydro-velour" as the biomaterial for high value facial masks.

Nympheas keeps focusing on the BC research, and aims to be a major global

supplier of BC products. Nympheas assists customers in designing unique

products with our professional knowledge, technology and customization

services. Xylos,developed biomaterial using BC, obtained from Acetobacter

xylinum strain. Numbers of companies are involved in the development of facial

masks using bio-cellulose.

National status:

Though developed countries are ahead in the manufacturing of products based

on bio-cellulose for human benefit, we are at initial level to discuss about the

Indian Scenario. Dr.Kalia’s research group in Rajasthan produced nanosized

bacterial cellulose from Brevibacillus parabrevis & Streptomyces albaduncus.

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Research groups as CFTRI identified cellulose producing organisms

Gluconacetobacter hansenii UAC09 and optimized the conditions for the

production. Research groups from Defence food research laboratory working on

biocellulose nanocrystals and its thermal stability. The research group However,

more research concentrations are given to ethanol production and not to

biomaterial development. Sakthi Biotech Company of Indian origin concentrates

on use of bio-cellulose to improve the tastes in Tea.

16.6 The relevance and expected outcome of the proposed study Research on biomaterial development and its wide applications is well

recognized by the clinicians. However, the lacuna on suitable biomaterials, its

nature and the stabilizing agents, needs intensive research. Further, the

reduction of pain and the early tissue regeneration in burn wound healing using

biomaterial is the need of the hour. Since, our research group currently working

on ice cool agents of natural origin, biomaterial development using collagen and

with various stabilizing agents and production of bio-cellulose from bacterial

species, I think it is possible for us to develop biomaterial from bio-cellulose with

cooling agents incorporated for early tissue regeneration of burn wound. Our

recent publications on biomaterial development and burn wound studies

emphasize the knowledge in the subject concerned.

The study will provide a biocompatible, biodegradable, non-toxic, able to

offer coolness to environment for early tissue regeneration of burn wounds,

which in turn reduces the infection rate considerably.

16.7 Preliminary work done so far

• A new bacterial species of rumen origin has been identified for the

production of biocellulose

• Standardized the process of extraction and characterization of type

I collagen from bovine skin

• Identification of suitable stabilizing agents for the required

mechanical and thermal stability

• Identified the natural and synthetic cooling agents to incorporate in

the biomaterial developed.

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17. Specific objectives & brief work plan (i) Separation, extraction and characterization of biocellulose from already

isolated bacterial species

Bio-cellulose produced by the chosen bacterial isolate will be purified and

characterized. Suitable media for the maximum production will be

identified.

(ii) Extraction of collagen from bovine skin

Optimized extraction procedures for collagen from bovine skin were used

for the present study.

(iii) Preparation of biocellulose and collagen scaffold (BCCS) impregnated with

novel cooling material

Biomaterial in the form of sheet or scaffold will be prepared by different

combinations of bio-cellulose and collagen. The one with high mechanical and

tensile strength will be chosen for the impregnation of cooling agent. Further

characterization and optimization studies will be carried out to assess the

presence of cooling agents.

(iv) In vitro cell line models for examination of cytotoxicity cell adhesion and

proliferation

In addition to animal model studies, invitro wound healing assays will be made

using skin fibroblast cells. Cell proliferation and cell adhesion in the presence

of biomaterial (Sheet) developed will be assessed. Further, cell growth in the

scaffold will be assessed to understand how the early tissue regeneration

occurs in the presence of the new biomaterial developed.

(v) In vivo animal model acid burn injury studies to examine the efficacy of the

BCCS based on the analysis on inflammatory signals, wound healing,

regeneration.

Followed by the preparation of scaffolds, assessment on efficacy of the material

will be made using animal model studies. Thermal burn wounds will be created

in rat models and the BCCS scaffolds containing cooling agents were applied on

the top of the wound. Collecting the blood from zero hour of treatment and

subjected to Elisa method to assess the reduction in inflammatory signals.

Wound healing was assessed through reduction in wound size. Cell proliferation

was assessed by H& E staining of granulation tissue. MMP assays will be made

for the granulation tissue to ascertain the earlier remodeling phase.

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18.1 Work Plan:

• The identified bacterial species will be grown in the optimized media

compositions. The extracellular biocellulose will be extracted by

centrifugation and the pellet thus obtained will be treated with sodium

hydroxide and then heated at 80 C in water bath. Followed by cell lysis,

the cellulose solution will be collected by centrifugation.

• Extraction of collagen will be carried out using the standard protocol. In

brief, type I collagen will be extracted from rat tail tendon or bovine skin

using acetic acid and sodium chloride. The sodium chloride precipitated

collagen dialyzed and the pure collagen obtained will be subjected to

lyopholization and stored at 4 C until use.

• Biomaterial in the form of sheet or scaffold will be developed by

incorporating biocellulose and collagen at varied combinations. Here the

stabilizing agent will be of our natural agent (patented).

• Followed by the development, the materials will be subjected to thermal

and mechanical stability tests.

• Followed by the observation on the optimized concentration of biocellulose

and collagen, cooling agent identified will be incorporated during the

preparation.

• The concentration of cooling agent will be decided upon the assessment

through human volunteers.

• Followed by the preparation of biocellulose-collagen- cooling agent

biomaterial in vitro and in vivo assessments will be made.

• For in vitro assessment, skin fibroblast cell lines will be used. Skin

fibroblasts cells will be obtained as a gift from Dr. Mary Babu

• Followed by confluent growth cells will be splitted and at the active

proliferation stage, wound will be created according to the method

followed by Dr. Suvro Chatterjee

• Followed by wound creation, serum samples will be collected and the

biocellulose- collagen –cooling agent solution will be incorporated and

serum samples will be collected at 12, 24 and 36 hours. In addition,

crawling distance of cells will be assessed.

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• The serum samples collected during experimental period will be subjected

to the assessment for IL-1, IL-6, and TNF-alpha.

• For the in vivo studies, albino rats will be employed

• Thermal wounds will be created on the hair free dorsal side of the animal

using brass rod with 1x1 cm circular structure.

• Followed by wound creation, the dead tissue will be removed and the

biomaterial developed in the present study will be applied topically and

allow the wound to heal.

• Dressings will be changed once in two days with new biomaterial

• Serum samples will be collected at 0,2,4,6,8 and till the wound gets

healed

• Wound contraction and wound area measures will also be made during

different healing stages

• Followed by wound closure, the major organs like, liver, kidney and heart

will be collected and subjected to sectioning and H & E staining

• The granulation tissue collected during different days of healing will also

be subjected to H & E staining & Masson Trichrome staining.

• The granulation tissue will also be subjected to MMPs assay to assess the

collagen deposition and remodeling of skin during healing.

• Bioinformatics studies will be carried out to assess the binding energy and

stability of material under dry lab condition.

18.2 Connectivity of the participating institutions and investigators (in case

of multi- institutional projects only) Nil

18.3 Alternate strategies (if the proposed experimental design or method does not work what is the alternate strategy).

The major objective of the present proposal on development of biomaterial

for early tissue regeneration involves bacterial cellulose. If the bacterial

cellulose is not available, then biocellulose of plant origin will be an

alternative, however, it needs higher purification procedures and also it

contains lignin and hemicellulose.

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19. Timelines: (Please provide quantifiable outputs)

Period of study

Achievable targets

6 Months

Extraction of biocellulose and preparation of different types of scaffold or sheet (collagen, Cellulose, collagen + cellulose + cooling compound etc.)

12 Months Characterization of above said scaffolds (Thermal, mechanical, chemical and structural analysis through bio informatics tool software)

18 Months Complete toxicity studies of the scaffolds (MTT assay, cell attachment, cell proliferation, GSH analysis, dead live cells counting assay, wound healing studies.)

24 Months Anti inflammatory signal analysis (IL-1, IL-6.TNF-alpha) using cell lines.

30 Months

Animal model studies (wistar strain albino rats) for burn wound early healing and tissue regeneration

36 Months

Publications and patent work.

20. Name and address of 5 experts in the field

S..No. Name Designation Address

1.

Dr.Mary babu Research

Advisor

Child Trust Hospital,

Chennai

2

Dr.Mukhopadhyay Professor IIT, Delhi

3

P P Kundu Professor University of Calcutta

4

Dr. Jayaraman Surgeon Kilpauk Medical

College, Chennai

5

Dr. D. N.Rao Professor AIIMS, New Delhi

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PART IV: BUDGET PARTICULARS A. Non-Recurring (e.g. equipments, accessories, etc.)

Sub-Total (A) = 17, 00,000 B. Recurring B.1.Manpower (See guidelines at Annexure-III) S.No Position No. Consolidated

Emolument Year 1 Year 2 Year 3 Total

1. Senior Research Fellow

24,000 2,88,000 2,88,000 2,88,000 8,64,000

2 Project Assistant Level-III

14,000 1,68,000 1,68,000 1,68,000 5,04,000

Total 4,56,000 4,56,000 4,56,000 13,68,000 Sub-Total (B.1) = 13,68,000 B.2.Consumables S.No Item Year 1 Year 2 Year 3 Total

1 Glassware 2,00,000 2,00,000 2,00,000 6,00,000 2 Chemicals 4,00,000 4,00,000 2,00,000 10,00,000 3 Other expenses 1,00,000 1,00,000 1,00,000 3,00,000 Total 7,00,000 7,00,000 5,00,000 19,00,000 Sub-Total (B2) = 19, 00,000 Other items Year 1 Year 2 Year 3 Total B.3.Travel 1,00,000 1,00,000 1,00,000 3,00,000 B.4. Contingency 1,00,000 1,00,000 1,00,000 3,00,000 Sub total of B.1+B.2+B.3+B.4

13,56,000 13,56,000 13,56,000 40,68,000

B.5. Overhead (If applicable)

2,71,200 2,71,200 2,71,200 8,13,600

Sub-total of B (B.1+B.2+B.3+B.4+B.5)

16,27,200 16,27,200 16,27,200 48,81,600

Grant total (A+B) 65,81,600

Total = 65, 81,600 (Sixty Five lakhs Eighty one thousand and six hundred)

S.No Item Year 1 Year 2 Year 3 Total

1 Ethylene Oxide Gas Sterilizer

3,00,000 - - 3,00,000

2 Elisa reader 6,00,000 6,00,000 3 ATIR 4,00,000 4,00,000 4 Bioinformatics system +

software 4,00,000 4,00,000

Total 17, 00,000

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Part V: Existing Facilities

A. Infrastructural Facilities

Item Yes

Workshop √

Water & Electricity √

Stand by power √

Laboratory space & furniture √

A/C Room for the Equipment √

Telecommunication √

Transportation √

Administrative & Secretarial

support

√

Library Facilities √

Animal/Glass house √

Computational Facilities √

B. Available Equipment and accessories to be utilized for the project

Availability Name of Equipment of

accessories

1. GC-MS

2. TGA/DTA

3. GPC

4. FT-IR

5. NMR

6. HPLC

7. SEM

8. XRD

Available with in the Institute

9.ESR

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PART VII: PROFORMA FOR BIOGRAPHICAL SKETCH OF INVESTIGATORS

Name: Dr. A. Gnanamani.

Designation: Senior Scientist

Department/Institute/University: Microbiology Division, Central Leather Research Institute, Adyar, Chennai 20.

Date of Birth: 01-06-1964.................. Sex (M/F) Female ... SC/ST: Nil. Education (Post-Graduation onwards & Professional Career)

Sl No. Institution Place

Degree Awarded

Year Field of Study

1 Madurai Kamaraj University

B.Sc 1985 Chemistry

2 Gandhigram University

M.Sc 1987 Applied Chemistry

3 Madurai Kamaraj University

Ph.D 1994 Science

A. Position and Honors Position and Employment (Starting with the most recent employment)

Sl No. Institution Place

Position From (Date)

To (date)

1 Central Leather Research Institute

Senior Scientist

Jan 2009 Till date

2 Central Leather Research Institute

Scientist B Jan 2002 Jan 2005

Honors/Awards

• Received DBT-overseas Associateship fellowship award for the year 2008-

2009 from DBT and spent six months (Feb. 2009 – July 2009) at Albany

Medical College, Albany, New York, USA.

• Received TATA INNOVATIVE fellowship award for the year 2008 for the

research proposal submitted on development of adhesive collagen

biomaterial for clinical applications

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• Received certificate as one of the TOP 10 Scientists of CLRI for the year

2008 based on the impact factor and publications made during the year.

• Received two externally funded project for the amount of Rs. 24 lakhs each

from DBT, New Delhi on, production and application of biosurfactants from

Marine bacterial species and development of new cross – linker for

stabilization of collagen by enzymatic methods

• Received young scientist Reviewer certificate from Biomaterials Journal for

the year 2009 to review the manuscripts submitted for the journal.

• Received second prize in oral presentation in National Seminar at

Ahmedabad during Feb. 2008 for the presentation on wound healing potential

of traditional plant medicine and the scientific validation behind that.

Professional Experience and Training relevant to the Project

• As Honorary Lecturer of Anna University, teaching skin biology, Leather biotechnology, Wastewater Treatment. Etc.

• Supervised and Guided M.Sc and M.Tech Projects on extraction of

collagen and preparation of biomaterials from waste trimmings

• Knowledge on preparation of antimicrobial compounds from traditional medicinal plants to combat microbial infections in animal models

B. Publications (Numbers only) Research Papers: 60 Reports: 4 General articles: 2 Patents: 7 Invited Lectures: 15 National Conferences: 60 International Conferences: 12

Selected peer-reviewed publications (Ten best publications in chronological order) Enclosed as Enclosure - I List maximum of five recent publications relevant to the proposed area of work. Enclosed as Enclosure - II

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Research Support Ongoing Research Projects

Completed Research Projects (State only major projects of last 3 years)

Sl No. Title of Project Funding Agency Amount (Rs)

Date of completion

1 An approach on surfactant production form marine sources for industrial applications BT/PR 8204/AAQ/03 /302/2006; 06.12.2007

DBT, New Delhi 24.55 Lakhs

December

2010

2 Enzymatic conversion of polysaccharide of seaweeds to respective aldehyde for stabilization of collagen BT/PR 10179/AAQ/03/385/2007) 09.07.2008

DBT, New Delhi 23.32 Lakhs

Completed June 2011

Dr. A. Gnanamani Place: Chennai Signature of Investigator Date: 10/8/2011

Sl No.

Title of Project Funding Agency

Amount (Rs)

Date of sanction and Duration

1 Preparation and stabilization of bio adhesives from collagen hydrolysate obtained from enzymatic extraction and hydrolysis of collagen (Bovin/Avian) for wet environment in biomedical application (TATA INNOVATIVE FELLOWSHIP AWARD PROJECT)

DBT, New Delhi

22.20 Lakhs

BT/HRD/35/01/02/2008 1.02.2009 Initially for Three years extendable to additional two years

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Enclosure – I

Selected peer-reviewed publications (Ten best publications in chronological order)

1. Tapas Mitra, G. Sailakshmi, Gnanamani, A., A.B.Mandal. (2011) Cross-linking with acid chlorides improves thermal and mechanical properties of collagen based biopolymer material Thermochimica Acta (in press)

2. Subha TS, Gnanamani A*, Mandal AB. (2011) Pharamcognostic Evaluation of Acorus calamus L. Pharmacognosy Journal, 3 (23)

3. N. Radhakrishnan, V. Kavitha, E. Madhava Charyulu, Arumugam Gnanamani, A.B. Mandal (2011). Isolation, production and characterization of bioemulsifiers of marine bacteria of coastal Tamil Nadu. Indian Journal of Geo-Marine Sciences Vol. 40(1) 76-82.

4. N. Radhakrishnan, V. Kavitha, STK. Raja, A. Gnanamani (2011). Embelin- a natural potential cosmetic agent. Journal of Applied Cosmetology, Vol. 29 (2); 99-107.

5. Tapas Mitra, G. Sailakshmi, A. Gnanamani, A. B. Mandal (2011). Di-carboxylic acid cross-linking interactions improves thermal stability and mechanical strength of reconstituted type I collagen: part I (oxalic acid). Journal of Thermal Analysis and Calorimetry DOI 10.1007/s10973-011-1472-2 (in press).

6. G. Sailakshmi, Tapas Mitra, A. Gnanamani , S. Thirupathy Kumara Raja, T. Thiruselvi, Naga Vignesh Selvaraj, Gopal Ramesh , A. B. Mandal (2011). Bonding interactions and stability assessment of biopolymer material prepared using type III collagen of avian intestine and anionic polysaccharides J Mater Sci: Mater Med. DOI 10.1007/s10856-011-4337-0 (In Press).

7. S. Subathra, Mazher Sultana, Gnanamani A (2011). Collogenolytic activity of serine protease of Perionyx excavatus . Indian Journal of Science and Technology, Vol. 4 (3) 197-200.

8. N. Radhakrishnan, A. Gnanamani, A.B. Mandal (2011). A potential antibacterial agent, Embelin-a natural benzoquinone extracted from Embelia ribes. Biology and Medicine, Vol 3 (2) Special issue 1-7.

9. V. Kavitha, N. Radhakrishnan, Arumugam Gnanamani, A.B. Mandal (2011). Management of chromium induced oxidative stress by marine Bacillus licheniformis. Biology and Medicine Vol 3(2) Special Issue 16-26.

10. T.S. Subha and A. Gnanamani (2011) Topical therapy of 1-2, 4, 5 Trimethoxy phenyl 1' methoxypropionaldehyde in experimental Tinea pedis in Wistar rats. Biology and Medicine Vol 3 (2) Special Issue 81-85.

11. Bikash Sahay, Anju Singh, Arugamam Gnanamani, Rebeca L. Patsey*, J. Edwin Blalock, and Timothy J. Sellati (2011). CD14 Signaling Reciprocally Controls Collagen Deposition and Turnover to Regulate the Development of Lyme Arthritis. American Journal of Pathology. The American Journal of Pathology, Volume 178 (2) 724-734.

12. Tapas Mitra, G. Sailakshmi, A. Gnanamani, S. Thirupathi Kumara Raja, T. Thiruselvi, V. Mangala Gowri, Naga Vignesh Selvaraj, Gopal Ramesh, A.B. Mandal (2011) Preparation and characterization of a thermostable and biodegradable biopolymers using natural cross-linker. International Journal of Biological Macromolecules 48, 276-285

13. Madhava Charyulu, Syamantak Majumder, Suvro Chatterjee, Arumugam Gnanamani (2010). Assessment of Pro-angiogenic Activities of a Free

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Radical Compound Free Radical Biology and Medicine, Volume 49, Supplement 1, 2010, Page S17

14. Arumugam Gnanamani, V. Kavitha, N. Radhakrishnan, A.B. Mandal (2010). Bioremediation of crude oil contamination using microbial surface-active agents: Isolation, production and characterization. Journal of Bioremediation and Biodegradation Vol. 1(2). 107 Pp 1-8

15. E. Madhava Charyulu and Armugam Gnanamani (2010). Condition Stabilization for Pseudomonas aeruginosa MTCC 5210 to Yield High Titers of Extra Cellular Antimicrobial Secondary Metabolite using Response Surface Methodology. Current Research In Bacteriology. Vol. 3 (4): 197-213.

16. V. Kavitha, N. Radhakrishnan, E. Madhavacharyulu, G. Sailakshmi, G. Sekaran, B.S.R. Reddy, G. Suseela Rajkumar, Arumugam Gnanamani (2010). Biopolymer from microbial assisted in situ hydrolysis of triglycerides and dimerization of fatty acids. Bioresource Technology, Volume 101, Issue 1, January 2010, Pages 337-343.

17. Arumugam Gnanamani, V. Kavitha, N. Radhakrishnan, G. Sekaran, G. Suseela Rajakumar, A.B. Mandal (2010). Microbial biosurfactants and hydrolytic enzymes mediates insitu development of stable supra–molecular assemblies in fatty acids released from triglycerides. Colloids and Surfaces B: Biointerfaces, Volume 78, Issue 2, 1 July 2010, Pages 200-207.

18. A. Gnanamani, V. Kavitha, N. Radhakrishnan, G. Suseela Rajakumar, G. Sekaran, A.B. Mandal (2010). Microbial products (biosurfactant and extracellular chromate reductase) of marine microorganism are the potential agents reduce the oxidative stress induced by toxic heavy metals. Colloids and Surfaces B: Biointerfaces, Volume 79, Issue 2, 1 September 2010, Pages 334-339.

19. J. Rajaram , B. Rajnikanth A. Gnanamani (2009). Preparation, Characterization and Application of Leather Particulate-Polymer Composites (LPPCs). J Polym Environ (2009) 17:181–186.

20. Madhava Charyulu E , Sekaran G., G. Suseela Raj Kumar and Arumugam Gnanamani (2009). Antimicrobial activity of secondary metabolite from marine isolate, Pseudomonas sp. against Gram positive and negative bacteria including MRSA. Indian Journal of Experimental Biology (2009) Vol.47 (12), 964-68.

21. T.S. Subha and A. Gnanamani (2009). Candida biofilm perfusion using active fractions of Acorus calamus. Journal of Animal & Plant Sciences, 2009. Vol. 4, Issue 2: 363 – 371.

22. Subha T.S and Gnanamani A (2009). Combating oral candidiasis in albino rats using bioactive fraction of Acorus calamus. Journal of Applied Biosciences Volume 21: 1265 - 1276. Published September 7, 2009.

23. T.S. Subha and A. Gnanamani (2009). In vitro assessment of anti - dermatophytic effect of active fraction of methanolic extracts of Acorus calamus. J. Anim. Plant Sci. [ISSN 2071 – 7024] Volume 5 (1): 450 – 455.

24. Kavitha, V., Radhakrishnan, N., Madhavacharyulu, E., Sailakshmi, G.,Reddy, B.S.R., Sadulla, S. and Arumugam Gnanamani (2009). Production of adhesives from triglycerides for industrial applications. JILTA, 59 (9), 692-702.

25. H. Hemalatha, S. Nagarajan and A. Gnanamani (2008) Synthesis, antibacterial and antifungal activities of some N-nitroso-2,6-diarylpiperidin-4-one semicarbazones and QSAR analysis. Nitric Oxide, Volume 19, Issue 4, December 2008, Pages 303-311.

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ENCLOSURE - II List maximum of five recent publications relevant to the proposed area of work.

1. Tapas Mitra, G. Sailakshmi, Gnanamani, A., A.B.Mandal. (2011) Cross-linking with acid chlorides improves thermal and mechanical properties of collagen based biopolymer material Thermochimica Acta (in press)

2. N. Radhakrishnan, V. Kavitha, STK. Raja, A. Gnanamani (2011). Embelin- a natural potential cosmetic agent. Journal of Applied Cosmetology, Vol. 29 (2); 99-107.

3. Tapas Mitra, G. Sailakshmi, A. Gnanamani, A. B. Mandal (2011). Di-carboxylic acid cross-linking interactions improves thermal stability and mechanical strength of reconstituted type I collagen: part I (oxalic acid). Journal of Thermal Analysis and Calorimetry DOI 10.1007/s10973-011-1472-2 (in press).

4. G. Sailakshmi, Tapas Mitra, A. Gnanamani , S. Thirupathy Kumara Raja, T. Thiruselvi, Naga Vignesh Selvaraj, Gopal Ramesh , A. B. Mandal (2011). Bonding interactions and stability assessment of biopolymer material prepared using type III collagen of avian intestine and anionic polysaccharides J Mater Sci: Mater Med. DOI 10.1007/s10856-011-4337-0 (In Press).

5. S. Subathra, Mazher Sultana, Gnanamani A (2011). Collogenolytic activity of serine protease of Perionyx excavatus . Indian Journal of Science and Technology, Vol. 4 (3) 197-200.

6. N. Radhakrishnan, A. Gnanamani, A.B. Mandal (2011). A potential antibacterial agent, Embelin-a natural benzoquinone extracted from Embelia ribes. Biology and Medicine, Vol 3 (2) Special issue 1-7.

7. T.S. Subha and A. Gnanamani (2011) Topical therapy of 1-2, 4, 5 Trimethoxy phenyl 1' methoxypropionaldehyde in experimental Tinea pedis in Wistar rats. Biology and Medicine Vol 3 (2) Special Issue 81-85.

8. Bikash Sahay, Anju Singh, Arugamam Gnanamani, Rebeca L. Patsey*, J. Edwin Blalock, and Timothy J. Sellati (2011). CD14 Signaling Reciprocally Controls Collagen Deposition and Turnover to Regulate the Development of Lyme Arthritis. American Journal of Pathology. The American Journal of Pathology, Volume 178 (2) 724-734.

9. Tapas Mitra, G. Sailakshmi, A. Gnanamani, S. Thirupathi Kumara Raja, T. Thiruselvi, V. Mangala Gowri, Naga Vignesh Selvaraj, Gopal Ramesh, A.B. Mandal (2011) Preparation and characterization of a thermostable and biodegradable biopolymers using natural cross-linker. International Journal of Biological Macromolecules 48, 276-285

10. Madhava Charyulu, Syamantak Majumder, Suvro Chatterjee, Arumugam Gnanamani (2010). Assessment of Pro-angiogenic Activities of a Free Radical Compound Free Radical Biology and Medicine, Volume 49, Supplement 1, 2010, Page S17.

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