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biocompatibility of dental materials

biocompatibility of dental materials

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Published by Preethi Iyengar

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Published by: Preethi Iyengar on Jan 21, 2011
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  • Biocompatibility HISTORICAL BACKGROUND
  • Dentin and pulp:
  • y Dentin rein interface
  • The bio compatibility of a restoration is altered by the leakage process which
  • may cause a number of undesirable events:-
  • Oral immuno system:
  • Mutagenic Reactions
  • Osseo integration and bio integration
  • Osseointegration-
  • Biointegration :
  • Immunotoxicity:
  • Types of Tests
  • Disadvantages
  • Advantages of cytotoxicity assays are
  • Disadvantage include
  • Mucous membrane Irritation test
  • Skin sensitization Test ( Guinea pig maximization test)
  • Advantage
  • Disadvantage
  • Dental pulp irritation tests
  • Slight
  • Moderate
  • Severe
  • Difficulties include
  • Bacterial plaque
  • Using in vitro Animal and Usage Tests Together
  • ISO STANDARD 10993
  • Chronic form


Introduction History Requirement of dental materials Biologic response in the dental environment Enamel Dentin Pulp Pediodntium Gingival and mucosa The oral immune system Adverse Effects from dental materials
y y y y

Toxicity Inflammation Allergy Mutagenic radians.

Local and systemic effects of materials Measuring the biocompatibility of materials Types of Tests In Vitro tests
y y y y y

Cytotoloxicity assays Cell number and growth Membrane permeability tests Tests for cell metabolism or cell function Tests that USC barriers.

y y y y y y y y

Other Assays for cell functions Mutagenesis assays Animal tests Usage tests Dental implants in bone Using in vitro, animal and usage tests together. Standards that regulate the measurement of biocompatibility. Biocompatibility of dental materials

Reaction of pulp
y y y y y y y y y y

Microleakage Amalgam Direct filling gold Dental casting alloys Non metallic restorative materials Bonding agents Composite resins Liners and varnishes Ceramics Conclusion

INTRODUCTION When a biomaterial is placed in contact with the tessiues and fluids of the human body, there is inivariable some form of interaction between the material and biological environment this interaction is referred to as biocompatibility. Biocompatibility is defined as the ability of a material to function in a specific application in the presence of an appropriate host response. Dentistry shares concerns about biocompatibility as with other fields of medicine, such as orthopedics, cardiology and vascular biology among others. In the development of any biomaterial, one must consider the strength, esthetics and functional aspects of the material, as well as its biocompatibility furthermore, demands for appropriate biological responses are increasing as we requires that materials perform more sophisticated functions in the body for longer time periods Thus, considerations of biocompatibility. sophisticated functions in the body for longer time periods. Thus considerations of biocompatibility are important to manufacturers practitioners, scientists and patients. The field of biocompatibility, is interdisciplinary and draws on knowledge from materials science, bioengineering biochemistry, molecular biology tissue engineering and other fields. Biocompatibility HISTORICAL BACKGROUND Although the concept of ethical treatment of patients extends back to the line of Hippocrates ( 460 ± 377 B.C) the idea that new materials must be tested for safety and efficacy before clinical use is more recent. As late as the mid 1800¶s dentists tried new materials for the first time by putting patients mouths. Even G.V.Black used patients to test many of his new ideas for restorative materials such as early amalgams. The concept of protecting the patient as a research

subject is only 30-40 years old and many of the regulations and ethics in this area are still being challenged and defined today. In most cases a committee of clinicians, basic scientists and laypersons regulate and oversee the testing of new materials in humans. Using humans as research subjects today without some previous testing or knowledge of the biological proportions is unethical and illegal still over new material must be inserted into a human for the first time at same point. Therefore many alternative tests have been developed to try to minimize the risks to humans. The current philosophy about testing the biological properties of dental materials in a systematic way evolved in the 1960¶s as the need to protect patients became politically acute and the number of new materials increased. The oversight for this testing in U.S now rests largerly with the Food and Drug Administration ( FDA), but these activities are regulated by the American National Standards Institute ( ANSI). American Dental Association ( ADA) Biological testing of materials has significantly evolved over the past 40 years. Since initially being used on animal models. Many studies between the 1950¶s and the 1970¶s involved the use at premolar teeth that were scheduled for orthodontic extraction. As well as cell solutions techniques and biological responses to material today the field of biocompatibility has reached a point where some prediction of biological properties is possible and the future will likely provide the ability to design materials that elicit customized biological responses. Some of the pioneering works are

Use of in vitro techniques to study the toxicity of various synthetic materials began some 30 yrs. After tissue culture was first established as a technique in 1926.

REQUIREMENT OF DENTAL MATERIALS y y y y Be nontoxic Be non irritant to the oral or other tessiues It should not produce allergic reactions. d) Those which affect the hard tessuies of the mouth. y Leirskar and Helgeland (1972) did the first study of cell culture tests using human epithelial cells on and around standard size disks of dental materials. y Kawahara. .y Various investigators began to apply organ and tissue culture techniques to toxicological problems in 1950¶s and 1960¶s. It should not be mutagenic or Coriogenicity Classification of materials Matrials may be classified into following types from the perspective of biological compatibility a) those which contact the soft tissue within the mouth. y One of the earliest quantitative in vitro tests was a membrane permeability assay for release using L. y Schmalz in 1982 was one of the first dental investigators to apply the agar overlay techniques to test the cyloloxicity of dental materials.929 cells by spangbeng in 1973. including silver amalgam and copper amalgam resins silicate and gold alloy.s lab began to use cell culture methods to investigate dental materials in the 1950¶s these investigators reported the cytotoxicity of pure metals. dental cements and medicaments. b) Those which could affect the health or vitality of the dental pulp/ c) Those which are used as used as root eanal filling materials.

are handled and may be accidentally engisted or inhaled. silicious particles in investment materials and fleeces containing fluorides. b) Polymer based filling materials may contain irritating chemicals such as unreacted monomers. Examples of hazards a) some dental cement components are acidic and may cause irritation. f) Some dental porcelain powder contain uranium.such as cyanaids solution for electroplating. physical condition ±like stresses. c) Mercury is used in dental amalgam. y Evaluation of the biocompatibilities-before adopting any methods for biocompatibility fests the following aspects must be careful considred: 1. 6.e) Those used in the dental laboratory.salvia blood 4. d) Dust from alginate impression materials may be inhaled. location of the material 2.vapoures from low fucing metal dies. ANATOMICAL AND PATHOLOGICAL ASPECTS OF ORAL TESSUIES Oral anatomy influencing the biologic response . type of contacts. and mercury vapour is toxic. g) Laboratory materials have their hazards. exposure of the materials to the oral situations. which though not used I the mouth. nature of the tessiues-soft or meniralized hard tessuie 3. direct or indirect through a barriears like a epithelvies 5.fatigue resistancs.degradations. chemical nature ±compositions .some products contain lead compounds/ e) Some people show allergic reactions to alloy containing nickel.

the periodontal attachment and to the peripical environment that have influences on the biologic response to materials and all are sites of interface between materials and tissue in dentistry. Dentin and pulp: The dentin and pulp to be a single tissue the dentinial matrix (both ciassified and unclassified forms the greatest bulk of the tooth collagen constitutes approximately 85% of the organic portion of dentin and hydroxy apatite is the main unorganic compound the dentenail matrix contains protein type 1 type 5 collagens. . The differential etching that occurs is a consequence of the different orientation of enamel rods. and it seals the tooth to outside agents. The full effects of the oral anatomy on the biocompatibility of materials are not known but these effect will be a major focus of anatomy of the tooth.and the orientation provides maximal strength because of its high mineral (hydroxylapatile) content. phosphoryjns dentinsealoprotien and dentin matrix protein. enamel is much more bittele than dentin and is soluibied to a greater extent by acid solution this property is used to advantage with bonding agents. However recent evidence indicates that enamel is not totally emperineable. permeability of enamel to most oral molecules is quite low. osteoclasin and osteopontien. Peroxides in bleaching agents has been shown to penetrate intact enamel in just a few seconds.Several aspects of oral anatomy influence the biocompatibility of dental restorative materials. when acids are used to etch the enamel to provide micromechanical retention of resin composites. ENAMEL Mature human enamel is highly meneialyid enamel rods have a specific orientation with each other.

A serum like fluid fills the dentenial tubules this fluid has conteneity with the extracellular of the pulp tissue. .The dentin matrix surrounds dentenial tubles that are filled with odontoblastic processes. which replace any odontoblasts desriyed during cavity preparation or material placement and allowes the tooth to form secondary or separation dentin. Smear layer is formed by the action of the beer or hand instruments on the classified dentin matrix. pulpal circulation maintains an intercellular hydraulic pressure. These effects are the basis of hydrodynamic theory of hyperlagesisa (pulpal hypersensitivity). The dentenial tubules establish continuity with the pulpal fluid to facilitate the diffusion of molecules both natural or from materials into and out of the pulp. The positive or negative displacement of this fluid through exposed denture tubules is capable of affecting either odontoblasts or pulpal nerve endings. The pulp of the tooth is a connective tessuie containing normal elements such as fibroblasts. which causes fluid flow into tubules to be directed from the pulp outward towards the DEJ when enamel is removed.external hydrostatic and osmotic pressure can also cause fluid movement toward or from the pulp.dentenial tubules and dentenial matrix are all important in the application of dentenial bonding agents and the ability of components of the bonding agents to reach and affects pulpal tessuies. Which also deminiralyses the openings of the tubules. The smear layer . The smear layer can be removed by acid etching. The tubules traverse the region between the dentino enameljunction (DEJ) and the pulp.collages. this occludes the dentinal tubules to some extent and it is quite effective in reducing hydrostatin pressures but less effective in reducing hydrosstative pressures but less effective in reducing diffusion.capillarises and nerve pulp supplies the cells.

The periodontial pocket is the site of the development of periodontal pocket is the site of the development of periodontal desease. .the periodontal desease process or the body.s ability to defend against bacteria that cause periodontal desease periodontal desease periodontal pocket is a unique microenvironment that allow concentration of components from materials to reach higher levels than are seen in the rest of the oral cavity the influence of dental materials on the periodontal disease and the biting forces that strain the periodontal ligament and supporting bone.the gingivial epithelium is also attached to the cementum of the tooth by a specialized junctional epithelium.PERIODONICAL ATTACHMENT: Junction between the outside of the body (oral cavity) and the inside of the body. It is likely that this approach will expand as better drug delivery materials are developed. Because many dental restorations are near or in the periodontal attachment area the biocompatibility of these materials may influence the normal periodontal architecture. the periodontal pocket has also been used as a site to place materials that release therapeutic agents to combat periodontal disease. It is often difficult to determine with certainty whether inflammation in this area is caused by periodontal disease. The dentin of the root of the tooth is covered by a thin layer of the root of the tooth is covered by a thin layer of cementum that may seal the dentenial tubules cementum attaches the collages fubers of the periodontal ligament the genguia normally extends above the level of the cementum and forms a potential space against the enamel called periodontal pochet. the material or some combination of these factors.periodontal ligament and supporting aluolar bone.which can destroy the junctional ipetheluim. occlusal trauma.

endodontic materials are placed in the pulpal space and these materials interface with the body through the apex of the tooth. The use of the retrograde approach to seal the apical foramen. The delineation between the bacterial and material threats are not clear . and the ability of materials to influence the body¶s immunological response in the perapical area has only recently been studied. SPECIAL BIOLOGICAL INTERFACE WITH DENTAL MATERIALS: y y Dentin rein interface Implant bone interface Dentin resin interface:-when resin based restorations material bonds to dentin. the filling materials will be in direct contact with the perapical tessuies. The apex of the tooth is the junction of the pulp of the tooth and the alueolar bone nerves and blood vessels entire through the apical foramen. If the endodontic procedure is not performed correctly the filling materials may extruded from the apex into the periapical area and cause additional physical damage. The ability of the root canal material toward the apex is also an important consideration.PERIAPICAL AREA: Another interface between materials and the inside of the body. When the pulp of the tooth is destroyed by infection or during restoration of a tooth. the release of substances from root canal filling materials may cause adverse responses around the apex or may alter the body¶s reaction to bacterial products that have contaminated the area. The composite nature of the denture allows the mineralized matrix to be dissolved away . As with the periodontal attachment area. Thus biological responces to these materials must be critically examined.

by acids which preserving the collagen network. If the resin penetrate the collagen network of the dentin but does not penetrate it completely a much smaller gap will exist between the mineralized matrix of the dentin and collagen resin hybrid layer. Although nanoleakage probably doesnot allow bacteria or bacterial products to penetrate the marginal gaps of the restoration and the pulp. which may happen if the dentin is dessicated it may be embedded by a resin ± containing material thereby mechanically bonding the resin material to dentin. this also occurs in enamel this few missions wide gap allows bacteria and oral fluids to pucolate from the pulp outward or from the oral cavity onward. Osseointegration: . That has been uncompletely embedded with resin. If the network doesnot collapse. This leakage is termed as microleakage. The bio compatibility of a restoration is altered by the leakage process which may cause a number of undesirable events:1. a gap will form between the resin and the dentin. Nanoleakage. thereby reducing the longevity of the dentin resin bond this degradation process may also gradually increase the gap size until microleakage begins to occur. If the resin material doesnot penetrate the collgenous network or debonds from it as the resin shrinks during polymerization. It encourages the breakdown then exposes the margins of the restorations making the tooth restoration complex aesthetically unacceptable.fluid exchange most likely occurs that can degrade the resin or the collagen network. 2. It may allow bacteria or bacterial products to reach the pulp and cause infection.

the success of these implants relieves on the ability of the material to promote osseointegration and allow a close approximation of bone with the material.the material is said to biointegrate with the bone.fatigue of the materials or function of the implant.biointegration appears to require a degradation of the ceramic to promote bone formation .The use of endosseous dental implants has uncriased tremendulously over the past decade.no known desirabily of osseointegration over biointegration has been established.it is also important to understand that neither osseointegration nor bio integration munic the normal ligamentous connection between a tooth root and alerolar bone. 3.they are commercially purelitanum.tantalum 4. relatively few materials allow osseointegration 1. several types of ceramics. Oral immuno system: It plays an important role in the biological response to any material. Materials that allow osseointegration have very low osseointegration have very low degradation rates.when this integration occurs.and they tend to form surface oxides that promote bony approximation some materials such as bioglass-ceramics. promote an integration between the bone and material with no intervening space at all.it appears to behave some what differently in oral epitheluium and connection tessuie than in the rest of the body and the biological responses to materials in the mouth may not always parallel those seen in other locations the oral environment is not always equivalent in ..like all biocompatibility phenomena osseointegration and biointegration are dynamic processes that may be altered by changes in the host. 2. titanium-aluiminium-vanaduim alloy.the ability of a material to allow osseointegration is closely related to its biocompatibility in dentistry.

. it involves the activation of the host immune system inflammation may result toward off some threat. berylluim released into the body.the inflammation may be caused by toxic or allergic materials. which can cause overt bixicity above a certain amount. The pulpal and periodontal disease are largely chronic inflammatory responses to long term infections. Example-some substances like mercury . And sometimes may preceed toxicity. HYPERSENSITIVELY AND ALLERGY Hypersensitively is the abnormal reaction that occurs when the body is exposed to a foreign material this materials sensitive the immune system. so that when the person is repiately exposed to the same material the body responds with a hypersensitivity reaction which is not dose dependent. This is a close related potential of a material resulting in cell or tissue death. toxicity or from allergy and often the inflammatory response precedes toxicity. INFLAMMATION It is a second fundamental type of biological response to a material. The allergic or hypersensitive reaction developes only in persons whose immune system recognize the material as foreign the allergic reactions can manifest as localized reaction in the tessuie which is directly in contact with the material allergy .nickel. It in the first screening test used for all dental materials.structure or function to other areas of the body and that differences may alter the biological response to materials ADVERSE EFFECT FROM DENTAL MATERIALS TOXICITY Material release certain substances which in convert concentration can lead to the toxicity.

bisphenol A.as the immune system supplies lot of cellular energies and mechanics to repair the mutated DNA.fortunately these may not be (arcinogenic.composite restoratibe materials and few other series are found to be esteogenic.breathing difficulties. According to gell and coomb¶s classifications the allergic reaction or immune responses can be-: Type I-immedaite TypeII-cyto-toxic hypersensitivity TypeIII-immune complex hypersensitivity Type 4 delayed.can be a systemic manifestations in the form of itching skin eruptions.Be.to act in the body as estrogens.altering the base-pair sequence of DNA in cell (mutation) some resin-based restoration.the resin. In most cases the surface characterstics such as composition.Cu) are found to be mutagens.it is the key phenomena in the implantology.cons of metals (Ni.sneezing.roughness and corrosion degradation products affect the biocompatibility. cell-medicated hypersensitivity The allergic reaction are due to there individuals emmuine systems. Osseo integration and bio integration It is the dynamic interaction between the body and materials. .recognizing a substance as foreign and are initially dose dependent Mutagenic Reactions These are caused by the materials or its components.erythema. OestroginicityIt is the ability to produce materials like xeno-istrogens.realants.the starting substance of BISGMA.

cellular organelles. The biological system may consist of mammalian cells. These tests maybe conducted in a test tube. flash. this practice has not been acceptable for many years and current materials must be extensively screened for biocompatibility before they are used in humans. cell culture dish.HEMA EVALUATION TEST OF BIOCOMPATIBILITY: Historically new materials were simply tested in human to assess their biocompatibility.palladium. These are classified as a) in vitro test b) animal test usage test a) in vitro test: these tests are performed outside of an organism. Biointegration : Certain materials like bioglasses which undergo biointegration with the bone directly any intervening space. Immunotoxicity: it is due to the alteration in the cells of immune system by the matrials. or other contains but they are performed separately from an intact organisms. tessuies .OsseointegrationFormation of living body tissues with in 10 mm space from the implant materials surface without any fibrous connection tessuis Eg: Tantalum ceramics. Several varieties of tests are currently used to unsure that new m aterials are biologically acceptable.this cause either increase or decrease of cellular of functions. Eg-mercury.bacteria or some text of .

With some biologic system the biologic system may consist of mammalian cells. the animal test. which is placed in contact. a membrane filter. The contact involves the exposure of a material and biological system are a)direct b) indirect direct contact involves the exposure of a material or an extract from a material directly with the biological system where as indirect contact occurs through a barrier of some sort such as agar. and the usage test performed either in animals or in humans. Types of Tests There are three basic types of tests : the in vitro test. or dentin. However this practice has not been acceptable for many years and current materials must be extensively screened for biocompatibility before they are used in humans several varieties of tests are currently used to ensure that new materials are biologically accepted. These tests may be conducted in a test tube. but they are performedseperately from an intact organism. flask or other containers. In vitro tests are performed outside f an organism historically. Historically new materials were simply tested in humans to assess their biocompatibility.enzyme. in vitro tests have been used as the first screening test to evaluate a new material. cellular . In vitro test can be subdi MEASURING THE BIOCOMPATABILITY OF MATERIALS Measuring the biocompatibility of a material is not simple and the methods of measurement are evolving rapidly as more is known about the interactions between dental materials and oral tissues as technologies for testing improve. cell culture dish.

Such as agar a membrane filter or death. those that determine cellular function of some type. Disadvantages y y Relevance to in vivo is questionable Lack of complex co-ordination of systems that is present in an organism such as immune.organells tissue bacteria or some sort of enzyme. The contact between the biological system and the material may be direct or indirect. ADVANTAGES y y y y y y Relatively fast Less expensive Can be standardized Large scale screening Good experiment control Tightly controlled conditions for quality. Direct contact involves the exposure of a material or an extract from a material directly with the biological system where as indirect contact occurs through a barrier of some sort. IN VITRO TESTS SYTOTOXICITY ASSAYS By these tests the effect of the material or its leachable products are studied on the metabolic functions of the test cell system. In vitro tests can be subdivided into tests that measure cell growth or death. In general cytotoxicity tests measure the effect of a material on y y Cell number or growth Integrity of cell membranes . and those that evaluate the integrity of the genetic materials of the cell. inflammatory and circulatory system.

y y y Screening large number of samples quickly and inexpensively Quantifying results Having potential for standardization of test methods Disadvantages include y y y Limitation of testing to only one cell type at a time Dissimilarity of test cells to host cells Lack of inflammatory and other tissue protective mechanism in tissue culture. Two types of cells can be used for in vitro assays. Continuous cells or cell lines are primary cells that have been transformed previously to allow them to grow more or less indefinitely in culture because of this . These cells will grow for only a limited time in culture but retain many of the characteristic of cells in vivo. Advantages of cytotoxicity assays are y Testing for a specific function of cell metabolism in isolation from other events. The biological systems used in vitro cytotoxicity tests may be y y y Organ cultures Cells in culture or Cell organelles. The most widely used biological systems for in vitro toxicity testing of dental materials are cells in culture.y y Biosynthesis or enzyme activity or The genetic material of the cell. Primary cells are those cells taken directly from an animal and cultured.

This test used on the basis that a loss in membrane permeability is equivalent to or very nearly equivalent to cell death. sterile and reproducible so as not to interfere with the analysis of the material.) There are two basic types of dyes used. In all cytotoxicity tests the test system itself must be nontoxic. Neutral red Na2 Cr o4 Non vital dyes are not actively transports and are only taken up if membrane permeability has been compromised by cytotoxic. These cells do not retain all in vivo characteristic but they do consistently exhibit any features that they do retain. Eg.transformation. The material is then placed in the test system if the material is not . Eg. This feature is important because it is possible for cells to be physically present but dead (when materials fix the cell. The main advantage of membrane permeability is It identifies cells that are dead or alive under the microscope. It is important to establish that the dye itself does not exhibit cytotoxicity during this frame of the test. vital dyes are actively transported into viable cells where they are retained unless cytotoxic efforts increase the permeability of the membrane. The main disadvantages of this test is the managing of radioactive isotopes (a gamma radiation emitter. CELL NUMBER AND GROWTH TESTS These assess the cytotoxicity of a material by measuring cell number or growth after exposure to a material cells are plated in a well of a cell culture dish where they attach. Trypan blue and propidium iodine. MEMBRANE PERMEABILITY TESTS Membrane permeability is the ease with which a dye can pass through a cell membrane.

use the biosynthetic or enzymatic activity of cells to assess cytotoxic response. If the material is a solid. TESTS FOR CELL METABOLISM OR CELL FUNCTION Some in vitro tests for biocompatibility. If the dehydrogenases are not active because of the cytotoxic effects. The cell density can be assessed either quantitatively. the cells may stop growing exhibit cytopathic features or detach from the well.cytotoxic. they can also be localized around the test sample by light or electron microscopy. (eg. via several cellular reducing agents. Production of Formazan is quantified by dissolving if and measuring the optical density of the resulting solution. which converts a chemical MTT. the cells will remain attached to the well and will proliferate with time. . If the material is cytotoxic. then the density of cells may be assessed at different distances from the material and a zone of inhibited cell growth may be described. the formazan will not form. insoluble formazan compound. This test measures the activity of cellular dehydrogenases. Tests that measure DNA or protein by cells is usually analyzed by adding radio isotope labeled precursors to the medium and quantifying the radioscope incorporated into DNA or protein. to a blue. 3H ± thymidine) A commonly used enzymatic test for cytotoxicity is the MTT test. Recently developed indicator dyes such as alam blue have been formulated to quantitatively measure cell proliferation using and oxidation reduction (redox) indicator that yields a calorimelic change and fluorescent signal in response to metabolic activity and permit continous monitoring of cells overtime. semi quantitatively or quantitatively substances such as Teflon can be used as negative ( non cytotoxic) controls where as materials such as plasticized polyvinyl chloride can be used as positive cytotoxic controls.

it is difficult to correlate the intensity of color or width of the zone around or material with the concentration of leachable toxic products. The culture medium is then replaced with medium containing 1% agar and this mixture is allowed to get over the cells. This technique establishes a mondayer of cells on filters made of cellulose esters. Thus several in vitro barrier tests have been developed to mimic in vivo conditions. After exposure to the test samples the filter is removed and an assay is used to determine the effect of the sample on acellular metabolic activity toxicity in the Millipore filter test is assessed by the width of the cytotoxic zone around each test sample. furthermore because of variability of the agar¶s diffusion properties. One such test is the agar overlay method in which a monolayer of cultured cells is established before adding 1% agar plus a vital stain such as neutral red to a fresh culture media. finally the filter monolay get is detached and turned over so that the filter is on top for placement of solid or soluble test samples for 2 or more hours. However the agar may not adequately represent barriers that occur in vivo. A second barriers assay is the Millipore filter assay. . It has the drawback of arbitrarily influencing the diffusion of leachable products from the test materials. The agar forms a barriers between the cells and the material which is placed on top of the agar sold test sample or liquid samples adsorbed onto filter paper can be tested with this assay upto 24 hrs.TESTS THAT USE BARRIERS Indirect tests researches have long recognized that in vivo direct contact does not exist between cells and the materials separation of cells and materials may occur from keratinized epithelium dentin or extracellular matrix.

The use of dentin disks offers the added advantage of directional diffusion between the restorative material and the culture medium. chemotaxis or T. Other tests measure the ability of a material to alter cell cycle or activate complement. These assays measure cytokine production by lymphocytes macrophages. Each chemical may be associated with a specific type of DNA mutation Genotoxic chemicals may be mutagens in their native states or may require activation or biotransformation to be mutagens. in which case they are called promutagens . The thickness of the dentin correlates directly with the protection offered to the pulp thus assays have been developed that incorporate dentin disks between the test sample and the cell assay system.Dentin barriers tests have shown improved correlation with the cytotoxicity of dental materials in usage tests in teeth and are gradually being developed for screening purpose. it is possible that activation of complement by resins or metals or their corrosion products may prolong inflammation in the gingival or pulp. Material that activate complement may generate inflammation or thrombi and may propagate a chronic inflammatory response whereas concerns about complement activation by dental materials are fewer. There is a wide range of mechanisms by which materials can affect the genetic material of the cell. A number of studies have shown that dentin forms a barrier through which toxic materials must diffuse to reach pulpal tissue. lymphocyte.cells. MUTAGENESIS ASSAYS They assess the effect of materials on a cells genetic material. Genotoxic mutagens directly alter the DNA of the cell through various types of mutations. proliferation. OTHER ASSAYS FOR CELL FUNCTIONS In vitro assays to measure immune functional or other tissue reaction have also been used.

This test on mammaion cells was developed to offer an alternative to bacterial tests ( Ames test) which may not be relevant to mammation systems. Thus the qualification and relevance of tests that attempt to measure mutagenesis and carcinogensis are extremely complex. The use of an animal allows many complex . It uses mutant stocks of salmonella typhimurium that require exogenous histidine native stocks of bacteria do not require expgenous histidine. The Ames test is the most widely used short term mutagenesis test and the only short term test that is considered thoroughly validated. A second test for mutagenesis is the styles cell transformation test. Chemicals that significantly increase the frequency of reversion back to the native state have a reportedly high probability of being carcinogenic in mammals because they significantly alter genetic material performance of this test requires experience in the field and special strains of salmonella to produce meaningful results.mutagens may or may not be carcinogens and carcinogens may or may not be mutagens. ANIMAL TESTS After screening by initial tests of the materials developed for a specific use animal tests for biocompatibility are used in mammals such as mice rats hamsters or guinea pigs. This characteristic that correlate with the ability of cells to produce tumors in vivo. This assay quantifies the ability of potential carcinogens to transform standard cell lines so that they will grow in soft agar untransformed fibroblasts normally will not grow within an agar get whereas genetically transformed cells will grow below the gel surface. However there has been some difficulty in reproducing these results. Animal tests are distinct from usage tests in that the material is not placed in the animal with regard toxic use. Exclusion of histidine from the culture medium allows a chemical to be tested for its ability to convert the mutant strain to a native strain.

the materials are injected intradermally to test for the development of skin hypersensitivity reactions. Skin sensitization Test ( Guinea pig maximization test) Used in guinea pigs. . Mucous membrane Irritation test This determines a material causes inflammation to mucous membrane or abraded skin. After several weeks of contact the control and test sites are examined and the gross tissue reaction in the living animals are recorded and photographed in color. especially in estimating the appropriateness of an animal species to represent a human. This injection is followed by secondary treatment with adhesive patches containing the test substance. changes. specimens are prepared for histological evaluation of inflammatory. The animals are then sacrificed and biopsy. The biological responses in animal tests are more comprehensive and may be more relevant than in vital tests. Disadvantage include y y y y y They can be difficult to interpret and control Expensive Time consuming Often involve significant ethical concerns and paperwork Relevance of the test to the in vivo use of a material can be quite unclear.interactions between the material and a functioning complete biological system to occur. This test is conducted by placing the test materials and positive and negative controls into contact with hamster¶s cheek pouch tissue or rabbit oral tissue.

the areas are excised and prepared for microscopic examination and interpretation. Animal tests that measure the mutagenic and carcinogenic properties of materials have been developed by toxicologists. the patch will elicit an inflammatory response. The degree of reaction in the patch test and the percentage of animals that show a reaction are the basis for estimating the allergenicity of the material. Although amalgams and alloys are tested because the margins of the restorative materials contact the gingival most subcutaneous tests are used for materials that will directly contact soft tissue during implantation endodontic or periodontal treatment. The location of the implant site is determined by the use of the material and may include connective tissue. for identification of either chronic inflammation or tumor formation are performed in a manner similar to that of short term tests except the materials remains in place for 1 to 2 years before examination. The implant site is then reopened and the test material is placed into this healed site or is packed into the tube and the test material is placed into this healed site or is packed into the tube that was placed previously. Short term implantation is studied by aseptically placing controls are placed at separate sites and allowed to remain for 1 ± 11 weeks. IMPLANTATION TESTS These tests are used to evaluate materials that will contact subcutaneous tissue or bone. The tissue response can be evaluated by normal histological. At the appropriate time. Those tests are employed w a ith . histochemical. The skin patch test can result in a spectrum from no reaction to intense redness and swelling. bone or muscle.If hypersensitivity develops from the initial injector. or immunohistochemical method. Alternatively an empty tube is embedded first and the inflammatory reaction from surgery is allowed to subside. Implantation tests of larger duration.

The usefulness of a usage test for predicting biocompatibility is directly proportional to the fidelity with which the test mimics the clinical use of the material in every regard include time location en vironment and placement techniques. They are distinct from other animal tests because they require the material be placed in a situation identical to its intended clinical use. y Long term invivo tests are performed by keeping the chemical in contact with the animals over the majority of its lifetime. with which the test mimics USAGE TESTS These tests may be done in animals or human volunteers. Fidelity. These tests are gold standard in that they give the ultimate to whether or not a material will be biocompatibility. If humans are used. the usage test is dentical to the clinical trial.strategy tests are applied in a specific order and testing is stopped when any indicates mutagenic potential of the material or chemical. In dentistry dental pulp periodontium and gingival or mucosal tissue are generally the largest of usage tests. For this reason usage tests in animals employ larger animals that have similar oral environment to humans such as dogs or monkeys. Advantage Relevance to use of material is assured. The validity of any of these tests may be affected by tissue of species tissue gender and other factors. . Tests are generally divided into y Limited in vivo tests that measure altered lives function or increased tumor induction when animals are exposed to the chemicals for a fraction of their lifetimes.

the teeth are removed and sectioned for microscopic examination. At the conclusion of the study. Zinc oxide eugenol and silicon cements have been used as negative and positive control materials respectively. . According to the intensity of the response the thickness of the remaining dentin and reparative dentin for each histological specimen is measured with a phalomicrometer and recorded. The materials are left in place from 1 to 8 weeks. The response of the pulp is evaluated based on its appearance after treatment. The severity of the lesion is based on disruption of the structure and the number of inflammatory cells present. Dental pulp irritation tests Materials to be tested on the dental pulp are placed in class V cavity preparations in intra noncarious teeth of monkeys or other suitable animals. After careful uniformly sized cavity preparations under sterile conditions the compounds are placed in equal number of anterior and posterior teeth of the maxilla and mandible to ensure uniform distribution in all types of teeth. The tissue sections are evaluated by the investigation without knowledge of the identity of the materials and necrotic and inflammatory reaction are classified.Disadvantage y y y y y Very expensive Very time consuming Major legal / ethical issues Can be difficult to control Difficult to interpret and quantify.

Responses categorized as Slight characterized by a few mononuclear inflammatory cells (mainly lymphocytes) in the epithelium and adjacent connective tissue. Moderate Definite increase in number of inflammatory cells hyperemia slight disruption of odontoblastic zone. MUCOSA AND GINGIVAL USAGE TESTS Since various dental materials contact gingival and mucosal tissues.Pulpal response is classified as either Slight Mild hyperemia. Usage tests that study teeth with induced pulpils allow evaluation of types and amount of reparative dentin formed. Moderate characterized by numerous mononuclear cells in the connective tissue and a few neutrophils in the epithelium. The materials effects on gingival tissues are observed at 7 days and again after 30 days. Severe evokes a significant mononuclear and neutrophilic infiltrate and thinned or absent epithelium . Severe Decided inflammatory infiltrate hyperemia total disruption of odontoblastic lay and even localized abscesses Efforts have been made to develop techniques that identify bacterial insults to pulp. the tissue response to these materials must be measured. Materials are placed in cavity preparation with subgingival extensions. few inflammatory cells slight hemorrhage in the odontoblastic zone.

However. Radiographs indicating either osseous integration or radiolucency around the implant. Bacterial plaque Secondary factors are the surface roughness of the restorative material open or overhanging margins and over contouring or under contouring of the restoration. Currently for implants in bone. Using in vitro Animal and Usage Tests Together Generally no single test is used to evaluate the biocompatibility of a new material. They argued that this was actually an attachment similar to the periodontal ligament and should be considered a sign of an acceptable material. Previously investigators argued that formation of a fibrous connective tissue capsule around a subperiosteal implant or root cylinder was the natural reaction of the body to a material. 1. in most cases it resembled the wall of a cyst. Mobility of the implant 3.Difficulties include Presence of some degree of preexisting inflammatory in gingival tissue. The ways by which these tests are used together however are controversial . 2. implants should be completely encased in bone the most differentiated slab of that tissue fibrous capsule formation is a sign of irritation and chronic inflammation. Penetration of periodontal probe along the side of the implant. which is the body¶s attempt to isolate the implanted material as the material slowly degrades and leaches its components into tissue. DENTAL IMPLANTS IN BONE The best estimations of the success and failure of implants are gained from three tests.

The philosophy was similar to the first scheme except the types of tests were broadened to encompass biological reactions other than toxicity such as immunogenicity and mutagenicity. The final lie was a clinical trial of the material.and have evolved over the years as knowledge has increased and new techniques developed. . The next layer shows specific toxicity that presumably death with conditions more relevant to the use of the material. Early combination schemes proposed a pyramid testing protocol in which all materials were tested at the bottom of the pyramid and materials were weekend out as the testing continued towards the top of the pyramid. Number of tests Number of tests Later another pyramid schemes was proposed that divided tests into initial secondary and usage tests. The concept of a usage test in an animal was also added first only materials that passed the first lier of tests were graduates to the second lier and only those that passed the second lier were graduated to the clinical trials. Tests at the bottom of the pyramid were unspecific toxicity tests of any type with conditions that did not necessarily reflect those of the material use.

Since then standardization has been difficult and a length. The committee that developed this documents recognized the need for standardized methods of testing and for sequential testing of materials to reduce the number of compounds that would need to be tested clinically. Firstly all tests continue to be of value in assessing biocompatibility of a material during its development and even in its clinical service. STANDARDS THAT REGULATE THE MEASUREMENT OF BIOCOMPATABILITY The first efforts of the ADA to establish guidelines for dental materials came in 1926 when scientists at the national Bureau of standards development specifications for dental amalgam.Two newer schemes have evolved in the recent past with regards to using combinations of biocompatibility tests to evaluate materials. Secondly these new schemes recognize the inability of current test methods to accurately and absolutely screen in or out a material. the council on dental materials. In 1972. Tests in animals for inflammatory response may be useful not only during development of a material but also if a problem is noted with the material after it has been on the marked for a time. instruments and equipments of ANSI / ADA approved document no 41 for recommended standard practices for biological evaluation of dental materials. . Eg. They incorporate the philosophy that assessing the biocompatibility is an ongoing process. process made more difficult by disagreement on the appropriateness and significance of particular tests.

it is not restricted to dental materials. sensitization and systemic toxicity and supplementary tests for chronic toxicity carcinogenicity and biodegradation. rather it is up to the manufacturer to select the tests and defined the selection to the ANSI /ADA and later FDA when applying for approval of the materials. This document is periodically documents. ISO STANDARD 10993 The ISO 10993 document is the international standard for testing the biocompatibility of materials unlike ANSI / ADA documents 41. BIOCOMPATABILITY OF DENTAL MATERIALS Reaction of pulp . Two types of tests for cytotoxicity. The initial tests include in vitro assays for cytotoxicity red blood cell membrane lysis mutagenesis and carcinogenesis as well as animal tests for inflammatory or immune responses secondary tests include animal tests for inflammatory or immune responses. As the ANSI/ADA standard the selection of test is left to the manufacturer who must defined the selection upon application for approval.ANSI / ADA DOCUMENT 41 The original ANSI/ ADA documents 41 for biological testing was updated in 1982 to include tests for mutagencity. This specification uses the linear paradigm for materials screening and divides testing into initial secondary and usage tests. The required tests for a given material are not listed specifically. In addition some specialized tests for devices are addressed such as the dentin barrier test for restorative dental materials. In this standard usage tests are part of supplementary tests. In 2002 consisted of 16 parts each addressing a different area of biological testing. Usage tests include tests for pulpal and bone response.

This intimate relationship has far reaching clinical application. The nature of pulp reaction that follows peripheral injury of the dentin which depends on y y The nature of the causative agent It¶s proximity to the pulp. Clinical components of the material Injurious products generated during setting of the material Pulpal reactions can also be caused by Bacterio y Either residual bacteria left behind in the cavity. Iatrogenic injuries y May result in inflammation of pulp As pulp inflammation can also be iatrogenic in origin Do not Harm is the basic principal should be followed by all members of the health profession. 1 Attrition abrasion Responses   Mild irritation Reparative dentin 2. . Progressive dental caries  Inflammation of pulp 3. Iatrogenic pulp injury can develop y y y y y During the preparation of a tooth for a restoration During the insertion of the restorative material It can be due to inherent irritational properties of the material either.y Dentin protects the pulp and owes its validity and its sensitivity to stimulation of the dental pulp.

several recent studies hypothesized that it was often the products of microleakage. Subsequently numerous studies should that bacteria were present under restoration and in dentinal labours which might be responsible for pulpal irritation. Microleakage can lead to y y y y Secondary caries Stains or discoloration Pulpal pathology Post operative sensitivity Restorative materials depending on their chemical nature can be grouped as Metallic restorative materials Amalgam Non. food debris or saliva may be drawn into the gap between the restoration and the tooth by capillary action. The importance of microleakage in pulpal irritation has been extensively studied early studies reported that various dental restoration materials irritated pulp tissue in animal tests. not the restorative materials that caused pulpal irritation. y Pulpal response to cast metal restoration. However. proximation of the pulp and techniques of cementation.metal restorative materials Different restorative materials . MICROLEAKAGE There is evidence that restorative materials may not bond to enamel or dentin with sufficient strength to resist the forces of contraction during polymerization wear or thermal cycling when debonding occurs. bacteria. porcelains etc mainly depends on the comenting agents. This effect has been termed microleakage.y Or by the bacteria that gain access to the cavity after restoration as a result of microleakage.

Direct filling Gold Dental casting Alloys Techniques Alloys Gold Base metal alloys Acrylic Composite Porcelain AMALGAM Dental amalgam has been used extensively for dental restoration Biocompatability of amalgam is thought to be determined largely by the correction products released while in service. Lichenoid reaction representing a long term effect in the oral mucous membrane adjacent to amalgam restoration is quite often Buccal mucosa and lateral border of the longer being the areas affected. does not inhabit cell growth. the response of the pulp to amalgam in shallow or in deeper but lined cavities is minimal and amalgam rarely causes invisible damage to the pulp however. pain results from using amalgam is deep unlined cavity preparations( 0. Corrosion in turn depends on the type of amalgam whether it contains the y2 phase and its composition In cell culture screening tests. Implantation tests show that low copper amalgams are well tolerated but the high copper amalgams can cause severe radiations when in direct contact with tissue. .5 mm or less) Firstly because of its thermal conductivity Secondly margins of newly placed amalgam restorations show significant microleakage marginal leakage of corrosion and microbial products is probably enhanced by the natural daily thermal cycle in the oral cavity. In usage tests. free or non leaded mercury from amalgam is toxic with the addition of copper amalgams become toxic to cells in culture but low copper amalgam that has set for 24 hrs.

The symptoms of chronic Hg.For many years a controversy has raged over the biocompatibility at amalgam restoration because of the presence of element hg. but appeared to be more cytotoxic than high copper amalgams chemically these materials show much higher corrosion rates than standard amalgam leading to roughness and discolouration. Poisoning ( elemental) are y y y y y y y y Weakness Fatigue Anorxia Weightloss Insomnia Irrilability Tremois in extremities Shyness The signs and symptoms of meth Hg poisoning ( Sea food) y y y Alaxia (gait disturbances) Paresthesia of extremities lips and tongue Constriction of visual fields Amalgams based on gallium rather than mercury have been developed to provide direct restorative materials that are mercury free.7 mg/ml . y y y Accepted Hg levels Patient with amalgam ± over Hg level Normal 0.

It may result in Acute form To date there have been no documented cases of Be toxicity at . y y y y Under extremely rare conditions patients have been reported with Burning sensation Lichenoid lesions of oral mucosa Generalized systemic reactions. whether with hand instrument / with mechanical pneumatic instrument. Co. The responses develop when the condensation occurs over freshly cut dentinal lubules. And the biocompatibility of each metal varies to different degree of tissue tolerance. Cr. However. the presence of be constitutes recognized health hazard.( minimum of 2 mm sapiapulpal dentin thickness) but not when the dentinal tubules are lined with pre-operatively formed reparative dentin induced from previous episodes of disease / restorative procedures. DENTAL CASTING ALLOYS We have to select alloys based on individual patients specific functional and economics requirement ± There is no one alloy suitable for all applications e. Apparently it was found that when compact properly into sound tooth structure produce only minimal pulp response showing least marginal leakage. Ni. when inhalation of dust and fumes can be anticipated. 1) Beryllium : dental origin.g certain base metal alloys contain Be.DIRECT FILLING GOLD The pulp responses from the insertion of cohesive and compacted gold are associated with condensation. under controlled conditions.

y y y Cobalt alloys have a potential for Dermatologic Systemic effect that may result from patient and personal exposure to cobalt alloys. And Ni. Compound as carcinogenic. The use of nickel has been controversial for many years because of the biological properties of nickel ions and compound. Compounds are carcinogenic when admixture by oral cutaneous route. The major hazardous route is aspiration.y Response vary from contact dermatitis to severe chemical Chronic form y y y y Symptoms range from coughing.Berylliosis Which result in lung cancer. It causes dermatitis ( contact) because it is a potential sensitizing agent and in sensitized patients intra orally. 2) NICKEL : Epidermiologic studies on workers in non dental industries have identified Ni. However there is no correlation found between the incidence of Ni hypersensitivity and the presence of intra oral Ni alloy restoration. chest pain and general weakness To pulmonary dysfunction Chronic inflammatory condition . . There is no experimental evidence that Ni. y y Burning and tingling sensation during the first 24 hours and. Later exhibited a slight erythematous reaction in the mucosa.

because resin components dissolve the film of vanish. the Ca(OH2 ) become less irritating and are able to stimulate reparative dentin bridge formation mere quickly than Ca(OH)2 suspension alone with no zone of necrosis and reparative dentin is laid down adjacent to the liner. The alkaline pH also helps to coagulate any hemorrhagic exudates of the superficial pulp shortly after necrosis occur neutrophils infiltrate into the subnecrotic zone. The liners such as copal vanishes and polystyrenes are not generally used under resin based materials. The initial response after exposing pulp tissue to these highly alkaline pulp capping agents is necrosis to a depth of 1 mm or more.Inhibition of cell metabolism is reversible in tissue culture by high levels of serum proteins suggesting that protein binding or buffering in inflamed pulp tissue may play an important role in detoxifying these materials in vivo. y The vanish also prevent penetration of corrosion products of amalgams into the dentinal tubules. After 5-8 weeks only a slight inflammatory response remains within weeks to months however the necrotic zone undergoes dystrophic calcification which appears to be a stimulus for dentin bridge formation. This indicates that replacement odontoblasts form the dentin bridge in contact with the liner. . y Because liners are used in such thin layers. they do not provide thermal insulation but do initially isolate the dentinal tubule contents from the cavity preparation. When resins are incorporated. y They may also reduce penetration of bacteria or chemical substances for a time.

5 the P-11 rapidly increases thereafter approaches neutrality in 24 hours. GROUP II ZINC PHOSPHATE CEMENT y y y It has an irritational potential intermediate to Z no E and silicate cement As a base it is not highly toxic As a lulling agent on pressure causes a widespread three dimensional lesion involving all the coronal pulp tissue as the phosphoric acid within the mix of 2n phosphate cement is forced in the dentinal tubules. y The P-11 of the cement 3 minute after mixing is 3.CERAMICS Among the most biocompatible materials used for dental restorations are ceramics and the use of dental constructions manufactured using ceramics has increased during the past few decades. new ceramics materials have been used both as high strength care materials and as veneers. This biocompatibility of dental ceramics has been largely assumed on the basis of studies of traditional feldspathic porcelains and the low corrosion rates of feldspathic materials studies showed clearly that all ceramics materials are not equivalent in their initial state of fabrication with aging after polishing procedure. this . y A young tooth with wide open dentinal tubules is more susceptible to such an intense inflammatory response compared to an older tooth which has sclerotic / Reparative dentin that blocks dentinal tubules and prevent the acid from reaching the pulp. Thus damage to the pulp occurs during the first few hours after insertion of the cement.

y In screening tests freshly prepared ionomer is mildly cytotoxic but this effect is reduced over time. y y The fluoride release from these materials is of therapeutic value The overall pulpal biocompatibility of glass ionomer cement has been attributed to the weak nature of the polyacrylic acid. y Histological studies in usage tests show that any inflammatory intitrak from ionomer is minimal or absent after a month. y Tendency to form complexes with proteins would limit its diffusion through the tissues y y In this regard polycarboxylate cements are equivalent to ZOE cements. Post operative sensitivity effects are negligible for cements. GLASS IONOMER CEMENT Glass ionomers have been used as both cement ( luting agent) and as a restorative material light cured ionomer systems have been introduced these systems use BISGMA or other oligomers as pendant chains on the polyacrylate main chain.can be prevented by using appropriate varnishes and dentin bonding agents. It is unable to diffuse through dentin due to its high molecular weight. . GROUP III SILICATE CEMENT y y It has high irritational potential It is an ideal material for the control in studies that evaluate pulp reactions to restorative materials.

y Repeated prolonged contact with monomer may result in contact dermatitis. y Clinical experience indicates that true allergic reactions to acrylic resins seldom occurs in the oral cavity. Theoretically. y The amount of residual monomer in processed polymethyl is externally low.y The ptt is below 3 at the time of insertion and the ptt remains below neutrality even after 1 month calcium hydroxide base provides adequate pulp protection from the cement. y Residual monomer that reaches the blood stream is rapidly hydrolyzed to methacrylic acid and excerted. RESINS y Those is no indication that commonly used acrylic resins produce systemic effects in humans. y Clinically most patients reported denture induced SORE MOUTH which on evaluation indicates tissue irritation which is generally related to unhygiene conditions / trauma caused by ill fitting prosthesis. COMPOSITE RESINS . y The oral mucosa and underlying tissues function as barriers that significantly diminish the volume of monomer reaching the blood stream. the use of monomers should be restricted to well ventilated areas. such reaction( toxic and allergic) could occur after contact with polymer residual monomer benzyl peroxide hydroquinone. y Finally inhalation of monomer vapour may be detrimental therefore.

of exposure although several newer systems seem to have minimal toxicity.5 mm of dentin and cause significant suppression of cellular metabolism for upto 4 week after application suggesting that residual unbound reagents may cause adverse effects. however when placed on dentin and rinsed with water between applications of subsequent reagents as prescribed by the manufacturer the cytotoxicity is reduced. Many of these reagents are cytotoxic to cells if tested alone. The cytotoxicity is significantly reduced 24-48 hrs. freshly set chemically cured and light cured resins often cause moderate cytotoxic reaction in cultured cells over 24 to 72 hrs. Long term in vitro studies suggest that compounds of the bonding agent may penetrate upto 0. however that adverse effects of resins occur at much lower concentrations when exposure times are increased to 4-6 .The material whether conventional / microfilled auto polymerizing photo activated (UV/VL) are found irritating to the pulp. In vitro. after setting and by the presence of a dentin barrier. Cytotoxicity is thought to be medicated by resin compounds released from the materials. Hydroxyethyl methacrylate (HEMA) a hydrophilic resin contained in several bonding systems is at least 100 times less toxic in tissue culture than BIS GMA studies using long term in vitro systems have shown. BONDING AGENTS Numerous bonding agents have been developed and are applied to cut dentin during restoration of the tooth. Evidence indicates that the light cured resins are less cytotoxic than chemically cured systems. It appears likely that reactive radicals generated during polymerization of the resin are responsible for pulp injury. but this effect is highly dependent on curing efficiency of the light and the type of resin system used.

In cytotoxicity assays by siogren et al in 2000. In the end. Many cytotoxic effects of resin compounds are reduced significantly by the presence of a dentin barrier studies have shown that combinations of HEMA and other resins found in dentin bonding agents may ad synergistically to cause cytotoxic effects in vitro. The high PH of calcium hydroxide in suspensions leads to extreme cytotoxicity in screening tests calcium hydroxide cements containing resins cause mild to moderate cytotoxic effects in both cut and long term set conditions. ranging from saline suspensions with a very alkaline PH( above 12) to modified forms containing Zinc oxide. there was no evidence of cytotoxicity in dental ceramics indicating good biocompatibility in vitro break down in products of dental ceramics . The location of a material in the oral cavity. further more the clinician must rely on clinical judgement. Diverse biological response to these materials depends on whether they release their components and whether those components are toxic immunogenic or mutagenic at the released concentrations.weeks. titanium dioxide and resins. CONCLUSION Biocompatibility of a dental material depends o its composition location and interaction within the oral cavity. Elements such as potassium and sodium appear to be the most liable from the ceramics but these elements are not particularly hazardous. no material can be shown to be 100% safe or risk free. Partly determinesits biocompatibility finally the interactions between the material and the body influence the biocompatibility of the material. data available and analyze the risk benefit ration before choosing a dental material. LINERS AND VARNISHES Calcium hydroxide cavity liner come in many forms.

.have not been reported to have known toxic effects and several of the ions in dental ceramics are considered non toxic.

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