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The choice of lesion types has resulted in a broad scope of wound dressings with new merchandises frequently begun to aim different facets of the lesion mending procedure. The perfect dressing has to carry through fast mending at tolerably monetary value with minimum irritation to the patient. Besides of import is the usage of biological polymers as tissue engineered scaffolds and skin transplants. Direct bringing of these agents to the lesion site is preferred, chiefly when systemic bringing could do organ hurt because of toxicological concerns combined with the preferable agents ( Boateng, Matthews, Stevens, & A ; Eccleston, 2008 ) .

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Figure 1. Structure of the Human tegument

They used rough drug infusions ( largely of works beginning ) , carnal fat and honey to mend lesions. For illustration in Senegal, the people used the foliages of Guiera senegalensis to set on lesion. In Ghana the people used infusions of Commelina diffusa herb and Spathodea campanulata bark to set on lesion and mend it. The interesting point is that the researches have shown that some of these infusions and herbs have so antibacterial and antioxidant consequence ( Boateng et al. , 2008 ) .

In order to stipulate optimum characteristics of ideal lesion dressing, dressings must incorporate a all right balance of features ; they should ease the quick and successful healing of lesions, be safe and biocompatible, and, if possible, incorporate a remedy to heighten the healing procedure.

An ideal stuff to be applied to injure should be atoxic and biocompatible, heighten cellular interaction and tissue development ( Huang and Fu, 2010 ) . done

Some of wound dressings can do allergic reactions when applied to the defected

country. There are three sort of allergic reactions that can look via wound dressings:

Irritant reactions that originally have mechanical grounds. These reactions can

go on because of occlusion or strong adhesion of dressing to the lesion

Immediate allergic reaction ( contact urtication )

Delayed allergic reactions ( reach eczema ) ( Goossens and Cleenewerck,

2010 ) . done

1.1.1Classification of dressings

Dressings can be classified in a figure of ways. They can be classified based on

their map in the lesion ( antibacterial, absorbent ) , type of stuff employed to

bring forth the dressing ( collagen, hydrocolloid ) , physical signifier of the dressing

( unction, movie and gel ) , traditional and modern dressings. Some dressings can be

placed in several categorizations because they fit standards in several groups. The

simplest categorization is as traditional and modern dressings and peculiar focal point will

be given to hydrogels, one of the most common modern dressings ( Boateng et al,

2008 ) . done

Hypothesis? ? ? ?

To run into this demand, we have created a novel wound dressing from a composite of collagen and Polyhema which are biocompatible, bioabsorbable, biodegradable, and has curative efficaciousness. Here, we present the methods by which this dressing was fabricated, and we describe the consequences of an probe of the dressing ‘s physiological belongingss.

1.2 Materials used for wound dressings ( belongingss / chief types of stuffs:

1.2.1Natural Polymers:

Many natural polymers and their man-made parallels are applied as biomaterials, nevertheless the features of collagen as a biomaterial are clear from those of man-made polymers largely in its manner of interaction in the organic structure ( Lee, et al. , 2001 ) . Done

Naturally happening stuffs such as purified collagen and hyaluronic acid have been considered as options to man-made scaffolds ( Bao et al, 2008 ) .

Alginate dressings

This type of dressing is produced from Ca and Na salts of alginic acid, a polyose consisting mannuronic and guluronic acid units. When alginate dressings are applied to the lesion, ions present in the alginate fibres are exchanged with those present in exudations and blood. This makes it possible to keep an optimum moist environment and an optimum temperature for the lesion during mending procedure. They can be used for moderate to to a great extent exudating lesions ( Boateng et al. , 2008 ) . Alginates can keep H2O up to 20 times of their weight. Alginates contain

Ca and Na salts of alginic acid. An ion exchange occurs between Ca from alginate and Na from lesion fluid, organizing sodium-calcium alginate which gives a gelatinlike mass and can maintain the moist environment ( Fonder et al. , 2008 ) . done

Figure 7: A classical hydrocolloid dressing

Hydrocolloid dressings: unwichtig

This group of dressings is a combination of hydrocolloid stuffs ( gel organizing agents ) and other stuffs such as elastomers and adhesives. They are widely clinically used because they can adhere to both dry and damp surface. They are used largely in visible radiation to reasonably exudating lesions ( Boateng et al. , 2008 ) .

In contrast to hydrogels, hydrocolloids have the absorptive ability. They absorb wound exudation and organize a hydrophilic gel that helps to keep a moist environment. These types of dressings are adhesive, occlusive and comfy dressings. They have less moisture vapor transmittal compared to movies and manage absorb exudation well ( Fonder et al. , 2008 ) . done

Chitosan

cellulose

Chitosan has been helpful for many biomedical applications, including lesion dressing, tissue technology applications, and drug bringing, because it has good biocompatibility, low toxicity, and debasement by enzymes such as chitosanase. Chitosan is readily soluble in the presence of acid and is indissoluble in impersonal conditions and most organic dissolvers. Many chitosan derived functions have been heightening solubility and processing. Furthermore, chitosan was found to injure healing and it is besides an ideal lesion dressing. ( Huang & A ; Fu, 2010 ) done

Chitin/Chitosan from triumph

Chitin is the 2nd abudant natural polymer in the universe after cellulose

Chitin occurs as a constituent of crustaceous shells, insect exoskeletons, fungous cell walls, micro zoology and plankton

Chemically chitin is a polymer formed chiefly of reiterating units of N-acetylglucosamine

Most commercial applications use of deacetylated derivative, chitosan

Chitosan is a polyose formed chiefly of reiterating units of D-glucosamine

Man-made polymers

Man-made stuffs such as poly ( Image ) – ( lactic acid ) and poly ( glycolic acid ) have received considerable attending for tissue technology applications. Man-made stuffs have predictable and consistent mechanical and physical belongingss, for illustration tensile strength and pore size and can be manufactured with great truth. On the other manus, man-made stuffs tend to pull out a foreign stuff type of response in the host, in peculiar, a hempen connective tissue deposition taking to formation of dense cicatrixs and fibrosis. Naturally happening stuffs such as purified collagen and hyaluronic acid have been considered as options to man-made scaffolds ( Bao, et al. , 2008 ) .

Biodegradable man-made polymers provide a figure of benefits over other stuffs for developing scaffolds in tissue technology. The chief advantages include the ability to orient mechanical features, debasement dynamicss to fit diverse applications and in add-on they are attractive since they can be fabricated into diverse forms with favoured pore morphologic public presentations conducive to weave in-growth. Furthermore, polymers can be designed with chemical functional groups that can get down tissue in-growth.

Biodegradable man-made polymers such as poly ( lactic acid ) , poly ( glycolic acid ) and their copolymers, poly ( p-dioxanone ) , and copolymers of trimethylene carbonate have been applied in a figure of clinical applications. The main applications include resorbable suturas, drug bringing systems and orthopedic arrested development devices such as pins, rods and prison guards. And besides the polyesters have been attractive for these applications because of their easiness of debasement by hydrolysis of ester linkage, debasement merchandises being resorbed through the metabolic tracts in some instances and the possible to orient the construction to change debasement rates. Polyesters have been considered for development of tissue technology applications ( Pathiraja A.Gunatillake and Raju Adhikari ) done

Properties of Collagen

Collagen is the major hempen protein of extracellular connective tissues, and it is besides the most omnipresent and plentiful protein in the carnal land. Collagen is synthesized by fibroblasts and degenerated by metalloproteinases ( e.g. collagenases ) . They are the most abundant type of protein in the human organic structure consisting more than 30 % of the entire organic structure protein. The word collagen is derived from Grecian word goora nut ( gum ) plus cistron. The map of about all systems and variety meats of the organic structure is dependent on collagenic constructions. About 70 per centum of the dry weight of the tegument is collagen. Use of collagen for lesion healing has drawn enormous involvement from the scientist in recent old ages. ( endarbeit ) .done

In add-on collagen has been joined with other stuffs for application, for case collagen microsponges can be merely impregnated into before prepared man-made polymer scaffolds to better mechanical public presentation. Collagen has good biocompatibility, weak antigenicity and debasement and H2O consumption belongingss. In malice of its biocompatibility belongings, collagen is automatically weak and goes through fast integrating upon nidation ( Huang & A ; Fu, 2010 ) .

The efficaciousness of collagen in biomedical application is that collagen can organize fibres with more strength and stableness through its self-aggregation and cross-linking. Largely drug bringing systems prepared of collagen, in vivo soaking up of collagen is checked by the usage of crosslinking agents, such as glutaraldehyde. ( Lee, et al. , 2001 ) . Done

Applications of collagen

The application of collagen as a drug bringing system is really comprehensive.

The most of import applications of collagen as drug bringing systems are collagen shields in ophthalmology, sponges for lesions, and tablets for protein bringing, as commanding stuff for transdermic bringing, and tissue technology including basic matrices for cell civilization systems ( Lee, et al. , 2001 ) . Done

The Structure of Collagen

The collagen molecule is a ternary spiral assembled from three single protein ironss

The collagen molecules contribute a structural model to other issues, such as blood vass and most variety meats. Six types of collagen have been isolated. ( I-VI )

Collagen is the chief protein constituent of skin sinews, bone and blood vass.

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Figure 1: Ternary Helical Structure of Collagen

Types of Collagen

The collagens represent a big household of proteins and at least 19 different collagen types have been described so far ( Prockop and Kivirikko 1995 ; van der Rest and Garrone 1991 ) . These are divided approximately into three groups, based on their abilities to organize filaments.

The most easy recognized signifiers of collagens are those that signifier banded filaments, and these are called fibril-forming collagens. Type I, II, III, V, and XI collagens belong to this group. In these molecules, the ternary coiling sphere contains an uninterrupted stretch of 338 to 343 GIy-X-Y threes in each alpha concatenation, and the molecule measures 15 ten 3000 A ( Ramachandran and Ramakrishna ; Peiz 1976 ) .

The 2nd group of collagens consists of proteins in which collagenic spheres are interrupted by noncollagenous sequences ( Fibril associated collagens with the interrupted three-base hit spirals – FACIT ) , and includes collagen types IX, XII, XIV, and possibly XVI. The type IX, XII, and XIV collagens are alone as they contain glycosaminoglyscan constituents covalently linked to the protein. All other nonfibrillar collagens form the 3rd group, including web forming collagens ( types IV, VII and X ) those organizing beaded ( type VI ) and grounding filaments ( type VII ) and spineless cuticle collagens. These collagens form

sheets of proteins integrating short ternary coiling collagen spheres. This group of collagen includes the Clq constituent of Cl complement, lung wetting agent protein, acetycholinesterase, and mannose adhering protein.

1.4 PolyHEMA

( Paulo et al. , 2010 ) hypertext transfer protocol: //www.chemicalregister.com/upload/cr/1991.png

Poly ( 2-hydroxyethyl methacrylate ) ( pHEMA ) hydrogels were foremost studied and prepared for biological usage by Wichterle and Lim. They have so been normally explored and applied in biomedical applications. PolyHEMA hydrogel is flexible, biocompatible, non-toxic and has no antigenic belongingss ( Hsiue, Guu, & A ; Cheng, 2001 ) .

It is used to fabricate skin coatings, immune isolation membranes, polyHEMA hydrogel nowadayss light weight. Although polymers are considered comfortable as biomaterials, it may convey jeopardy along due to its debasement and interaction with the tissue. ( Paulo et al. , 2010 ) . done

Poly ( 2-hydroxyethylmethacrylate ) [ poly ( HEMA ) ] is a widely used biomaterial which does non let cell adhesion and growing on its surface, restricting its usage in biomedical applications in which cell coherence is damaging ( Santin et al. , 1996 ) .

Applications of polyHEMA hydrogel are various and it has been used in the post-surgical Reconstruction of rhinal gristles, artifcial corneas and wound dressings in the control of wound infection. The chief disadvantage of the polyHEMA hydrogels in application is its bad

mechanical belongings after swelling. Methods for promotion have been good documented, for illustration majority copolymerization, grafting onto cotton cellulose or polymers, for case, styreneA± butadieneA±styrene ; and organizing permeating composite web with natural biopolymers like collagen. Done

These modified complexs showed better strength than the pure 1s, their hapless mechanical belongingss still remained. We have antecedently observed that the extent of H2O in the gels from different batches of the freshly synthesized pure polyHEMA fail to exhibit proper mechanical belongingss. This observation prompted us to speculate that the sum of H2O ab initio added to the monomer mixture may subsequently find the tensile strength of the membranes of the freshly synthesized polymers. In this survey, the consequence of initial H2O content in the monomer mixture and the equilibrium H2O content in the polymer on the physical belongingss of the polyHEMA merchandises such as polymerisation grade, wettability by H2O, dimensional alteration during swelling and the tensile strength are discussed. ( Young, Wu, & A ; Tsou, 1998 ) .

An Overview of PolyHEMA

Poly ( 2-hydroxyethylmethacrylate )

2-Hydroxyethylmethacrylate ( HEMA ) is an of import functional monomer, which was foremost used in biomedical applications and now widely used in the industry of soft contact lenses ( Montheard et al, 2002 ) ( Dumitriu et, al 1994 ) .

Applications of PolyHEMA

Applications are due to high mechanical stableness, high refractile index and O permeableness. It forms non-ionic hydrogels and exhibits no pH dependent puffiness. The monomer is water-soluble while the polymer has limited solubility. This stage behavior allows the formation of a macroporous sponge construction when reacted in dilute monomer solutions. The H2O content can be regulated by copolymerization with hydrophobic or hydrophilic comonomers. Mizuno et Al. have investigated the construction of H2O sorbed into HEMA copolymers by attenuated entire contemplation infrared spectrometry ( ATR-IR ) . Their consequences indicate that the interaction between the primary hydration H2O around the polymer ironss and the H2O environing the primary hydration H2O is really weak. This confirms the formation of cold crystallisable H2O generated by caging H2O molecules in a little infinite by the polymer ironss. Although high molecular weight HEMA homopolymer is hydrophilic and has comparatively high grade of hydration, it is normally known as H2O swellable, instead than water-soluble [ 99 ] . This unsolvability may be linked to the presence of low degree of ethylene ethanediol dimethacrylate found in HEMA monomer, which could take to some crosslinking reaction during polymerisation. HEMA can non be polymerized by anionic polymerisation due to the presence of labile proton on the hydroxyl group. Furthermore in extremist polymerisation, the cardinal difference between propenoate and methacrylate polymerisation, which can be pronounced in ATRP procedures, is that, like methyl methacrylate ( MMA ) , the propagating groups of HEMA are more stable and propagate more easy. Unlike MMA, dissolvers required to fade out HEMA polymer are really polar, which may dramatically impact the polymerisation procedure. Beers et al. [ 45 ] and Weaver et Al. [ 100 ] have reported efficient and controlled polymerisation of

HEMA utilizing atom transportation extremist polymerisation in assorted dissolvers at lower temperatures.

1.5 Bioadhesive belongingss of lesion dressings ( generalizations on adhesion / theories of adhesion / importance for wound dressings )

The term bioadhesion is specified as adhesion between two stuffs where at least one of the stuffs is of biological beginning. In the instance of bioadhesive drug bringing systems, bioadhesion on a regular basis relates to the adhesion between the excipients of the preparation and the biological tissue. The facet of using bioadhesive stuffs in the advancement of pharmaceutical preparations emerged in scientific articles in the early 1980s ( Edsman & A ; Hagerstrom, 2005 ) .

Bioadhesives are used for tissue adhesion and haemostasis in surgery. The most normally used surgical adhesive is fibrin gum. However, it is readily separated from the adhered tissue since its hapless bonding strength ( Edsmann ) . Done

Bioadhesives are used for tissue adhesion and haemostasis in surgery. The most normally used surgical adhesive is fibrin gum. However, it is readily separated from the adhered tissue since its hapless bonding strength ( Sung, Huang, Chang, Huang, & A ; Hsu, 1999 ) .

L. Maggi New 19.12. done

Bioadhesion occurs under assorted grade of hydration and about in every application has to be reversible. The stuffs used should non alter tissue activity and functionality. Bioadhesion is reasonably a complex happening since a batch of physiochemical variables are involved in the process. Consequently, a new standardised method is required to test the bioadhesive belongingss of assorted stuffs. It must be various adequate to be modified to assorted experimental conditions, particularly to imitate the biological environment and at the same clip, able to bring forth similar consequences.

The method most widely used for this intent is the fracture trial.

Edsmann done

The five theories that are most normally presented in concurrence with bioadhesion are the soaking up, diffusion, electronic, break and wetting theories.

New 12.1.2011

Bioadhesive Strength

Another of import physical belongings of dressings meant for application to moist lesion surfaces, is adhesive strength both in vivo ( bioadhesivity, mucoadhesion ) and in vitro ( adhesivity ) . Adhesivity has been defined as the force required to detach a sample from the surface of excised porcine skin292 ( utilizing a Texture Analyser, a common type of mechanical proving equipment ) . The trial is adopted from word picture of bioadhesive polymeric preparations meant for application to other damp surfaces such as vagina, buccal and rhinal cavities.271, 293-296 Sakchai et al.220 have determined the bioadhesive belongingss of Eudragit-chitosan movie dressings by mensurating the force required to detach the movie from hog big bowel washed in physiological solution. Peppas and Buri297 have discussed the surface and interfacial phenomena that occur during bio-adhesion of polymeric molecules to soft tissue including lesion surfaces. Adhesivity can besides be determined by measuring assorted tensile responses of different gels.296 Adhesivity is of import in lesion healing where dressings should be self adhesive with the lesion, easy removed and painless ( i.e. it must hold reduced adhesiveness with clip ) .296 The force of adhesion depends on factors such as hydrophobicity which is reported to better bioadhesion,105 degree of hydration and rate of polymer eroding in contact with the hydrating surface.298 A fresh drug-loaded lesion dressing with optimised adhesive drug let go ofing belongingss was developed by adhering self-adhesive Eudragit E ( cationic copolymer based on dimethylaminoethyl methacrylate and impersonal methacrylic esters ) movie with antibacterial loaded poly ( N-isopropyl-acrylamide ) microgel beads to accomplish adhesive, absorbent and easy to skin functions.299. ( Boateng et al. , 2008 )

2. AIMS AND OBJECTIVES

The purpose of the present survey is to measure the bioadhesive belongingss of fresh polymeric stuffs prepared from collagen poly ( HEMA ) which were designed for tegument regeneration and drug bringing.

The chief aims of the present survey:

were peculiarly to mensurate the adhesive belongingss such as entire work of adhesion and force of withdrawal of a assortment of stuffs in contact with unreal membranes by utilizing an machine-controlled Texture Analyser.

Raman spectrometry will perchance be applied in a attempt to derive an penetration into the intimate structural facets responsible for the adhesive behavior of these stuffs.

3.2. Equipment

3. Experimental

3.1. Materials

Was obtained from Fisher Scientific ( UK )

The collagen movies used in this survey were supplied by the Faculty of Biomaterials and Bioengineering, University of Iasi, Romania.

Fixing of Phosphate buffer ( PB ) , pH 7.0

A liter of BP solution was prepared utilizing the undermentioned solutes and their corresponding measures: xxxxxxxxxxx

Distilled H2O ( milliliter ) was added and the solutes were allowed to fade out utilizing a magnetic scaremonger for 15 proceedingss. The solution was made up to a liter by adding distilled H2O.

Water bath

The temperature of the H2O bath of the phosphate buffer was kept at 37 EsC.

3.2.2. Preparation of collagen samples

Texture Analysis

A package controlled penetrometer, TA-XT2 Texture Analyzer ( Stable Micro Systems, UK ) , with a 5 kilogram burden cell, a force measuring truth of 0.0025 % and a declaration of 0.0025 millimeter was applied in texture analysis ( Tamburic & A ; Craig, 1997 ) .

This setup is able to find the opposition that a investigation comes across to perforate into a semisolid stuff as a map of distance, nevertheless can besides transport out tensile experiments. The chief difference is that the Texture Analyzer can set and maintain the preload applied to the viscoelastic colony of the sample. The control unit, by uninterrupted monitoring of the force used, drives the motor to travel the sliding base upwards or downwards to keep the selected preload value ( 500g in this instance ) stalls for all the preload clip. The force required to detach the polymer from the substrate is recorded as a map of elongation. With the package equipped both maximal strength and work of adhesion can be obtained ( Maggi et al. , 1994 ) .

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Apparatus: A-XT2 Texture Analyzer ( Stable Micro Systems, UK )

The pre-test velocity was set up at 1 mm/s, the trial velocity at 2 mm/s a and the incursion deepness at 5 millimeters, with an acquisition rate of 100 points/s. The investigation used was a fictile cylinder with a diameter of 13 millimeter. The survey was carried out at room temperature ( 37 C ) with at least at three repetitions obtained for each sample ( Tamburic and Craig, 1997 )

Text process: The sample to be examined is fixed to the sliding base utilizing a dual side tape. The sliding base is conveyed into contact with the collagen samples and a preload is used. A given clip is left for the constitution of adhesion bonding called preload clip. The sample holder is so lowered at changeless velocity ( 0.1 mm/s ) , the measuring ends when the two substrates are wholly detached. The force calculated by the transducer, during the procedure, is plotted versus distance. A negative extremum is obtained whose maximal value, normalized by the 1 cm2 is considered as bioadhesion strength. This process is validated and standardized for the in vitro findings in a old survey in which the chief variables, able to act upon the phenomenon, were besides discussed. Half done. Maggi

Fig. 1. Typical secret plan obtained from the Texture Analyser ( with an applied

force of 12.5N and a contact clip of 10 s ) .

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