Animal Test for Dental Materials

Prepared by: Faten Ahmed Samer Abdelalhalem

Researcher in Pedodontics department

Submitted by: biomaterial department

Under supervision: Dr. /Mohamed Bahaa-Eldin

Lecturer of dental materials faculty of dentistry Minia University

Dr. /Hanaa Farouk

Lecturer of dental materials faculty of dentistry Minia University

Dr. /Asmaa Abdelhakim

Lecturer of dental materials faculty of dentistry Minia University

2020
Contents
Introduction ........................................................................................................... 1
Animal tests or secondary tests .............................................................................. 2
Advantages ......................................................................................................... 2
Disadvantages ..................................................................................................... 2
Types for these tests ............................................................................................ 2
mucous membrane irritation test ............................................................................ 3
skin sensitization test ............................................................................................. 3
Implantation test .................................................................................................... 5
Systemic toxicity test ............................................................................................. 6
inhalation test ......................................................................................................... 7
hemolysis test ........................................................................................................ 7
References ............................................................................................................. 9
Introduction
Biocompatibility is the ability of a material to perform with an appropriate

host response in a specific situation (1).

Which means that the material harmless to human body.

Every dental material has to undergo certain tests in order to be certified as
biocompatible material.
These tests have different levels and these levels are,
1-In vitro tests (primary tests): apply in test tube, cell culture dish or outside a living
organism. These tests are done to evaluate cell response (figure 1).
2-Animal tests in vivo (secondary test): apply in mammals such as mice, hamsters
or rats. These tests to evaluate tissue response (figure 2).
3-Usage tests: apply in large animals then in human volunteer body (figure 3) (2).

Figure 1 (primary test) Figure 2 (secondary test ) Figure 3(usage test)

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Animal tests or secondary tests:
These tests are usually done in mammals such as mice, rats, hamsters, or
guinea pigs (2).
Advantages:
1-more relevant than in vitro test.

2-allow complex systemic interactions.

Disadvantages:
1-difficult to control.
2-time consuming.
3- expensive.
4-involve ethical concern.
Types for these tests:
1-Mucous membrane irritation test.
2-skin sensitization test.
3-implantation test.
4-systemic toxicity test.
5- inhalation test
6- hemolysis tests.

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1-mucous membrane irritation test
This test determines if a material causes inflammation to the mucous
membranes.
During this test, test materials are placed into contact with hamster cheek- tissue or
rabbit oral tissue.

After several weeks of contact, test sites are examined, and the gross tissue
reactions in the living animals are recorded and photographed in color (figure 4).
The animals are then sacrificed, and biopsy specimens are prepared for
histological evaluation of inflammation (5).

Figure 4 (mucous membrane irritation test)

2-skin sensitization test

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 In guinea pigs, the test materials are injected intradermally to test the
development of skin hypersensitivity reactions.
This injection is followed by secondary treatment with adhesive patches containing
the test substance. If the hypersensitivity is developed from initial injection, the
patch will elicit an inflammatory response.
The skin-patch test can result in a spectrum from no reaction to intense redness and
swelling (figure 5) (2).

Figure 5 (steps of sensitization test)

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3-Implantation test
These tests are used to evaluate materials that will contact subcutaneous
tissue or bone. The location of the implant site is determined by the use of the
material and may include connective tissue, bone or muscle (figure 6)
Materials tested in this way are the ones that directly contact soft tissue like implant
materials, and materials used for endodontic and periodontal therapy.
Amalgams and alloys are also tested because the margins of the restorative materials
contact the gingiva.
Short term implantation test:
Studied by placing the compounds in small, open ended polyethylene tubes into the
tissue. The test samples are placed and allowed to remain for 1 to 11 weeks (figure
7).
At the appropriate time the areas are excised and prepared for the microscopic
examination.
The tissue response can be evaluated by normal histological methods.
For implantation tests of longer duration, identification of either chronic
inflammation or tumor formation is the same as that of short-term tests except the
materials remain in place for 1 to 2 years before the examination (3).

figure6(implantation test).

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Figure 7(formation of an abscess between a material and connective tissue).

4-Systemic toxicity test

Most substances or materials are administered orally (feeding of an extract
of the test material, mostly finely ground).
Tests such as LD50 (median lethal dose), test sample administrated orally or by
dietary inclusion to rats for 14 days, if 50% of animals survive, the material has been
passed the test.
It may be acute, subacute or chronic.
Acute toxicity: will be determined by administering the material within 24 hours
(figure 8).
Subacute toxicity: will be determined by administering the material during a period
of 14 to 28 days.
Chronic toxicity: will be determined by administering the material or extract over
several months.
Tests are sometimes extended over the lifetimes of the experimental animals. At the
end of these studies, survival rates of the animals and histopathological alterations
are studied (1).

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Figure 8(rat used in acute toxicity test)
5-inhalation test
An aerosol of the test material is sprayed around the head of experimental
animal in a chamber periodically (figure 9).
If the animal dies in a few minutes, or hours, the materials are considered toxic (4).

figure 9(tubes used in inhalation test)

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6-hemolysis test:
Is used to evaluate the acute hemolytic activity of a material in prolonged
contact with bone and soft tissue.
Material or their extracts are incubated in saline for a 90 -minute period, with rabbit
blood present for the last 60 minutes (figure 10).
Both the physical surface of the test material (affecting adherence and activation of
the plasma protein systems and cellular components) and its soluble, leachable
components contribute to the hemolytic activity (6).

Figure10 (hemolysis test)

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References:
1-schmalz G, dorthe arenholt-Bindslev.Biocompatibility of Dental Material.2009;
13-22.
2-Ronald L.Sakaguch, John M. Craige’s Restorative Material. 13th edition.2012;
133-115.
3-Thomas, Grohens Y, Ninan N.Nanotechnology Applications for Tissue
Engineering.1st edition.2015; 21-44.
4-Goldberg NB,Goldberg AF,Gerhans GA,Logas,Taschinip,Molnar ZV.A rabbit
lung model for testing reaction to inhaled dental restorative particles.Chest.1992
Mar;101 (3) :829-32.doi:10.1378/chest.101.3.829.PMID:1541152.
5-B.B. Harsanyi, W.C.Foong,RE.Howell,Aidi and P.W.Jones.Hamster Cheek
Pouch Testing of Dental Soft Polymers. Journal of Dental Research.1991; 991-996.
6-David Charles Thom. The Hemolysis and Cytotoxicity of a Zeolite-Containing
Root Filling Material in Vitro.2001; 49-52.