Solid materials are extracted in cell culture medium and the extracts are then placed in triplicate containers of L-929 mouse fibroblast cells. Cells are examined at 24, 48 and 72 hours for signs of toxicity.

Solid materials are extracted in cell culture medium and the extracts are then placed in duplicate containers of L-929 mouse fibroblast cells. Cells are examined at 24 and 48 hours for signs of toxicity.

Solid materials are extracted in cell culture medium and the extracts are then placed in a single container of L-929 mouse fibroblast cells. Cells are examined at 24 hours for signs of toxicity.

When a test material is shown to be toxic by the MEM elution test, four 2-fold dilutions are made to determine the toxic endpoint. Performance of the test will result in an estimation of the relative "strength" of the cytotoxic substance in the material.

Solid materials are extracted in cell culture medium and the extracts are then placed in triplicate containers of MRC-5 human embryonic lung cells. Cells are examined at 24, 48 and 72 hours for signs of toxicity.

Solid materials are extracted in cell culture medium and the extracts are then placed in duplicate containers of MRC-5 human embryonic lung cells. Cells are examined at 24 and 48 hours for signs of toxicity.

Solid materials are extracted in cell culture medium and the extracts are then placed in one container of MRC-5 human embryonic lung cells. Cells are examined at 24 hours for signs of toxicity.

When a test material is shown to be toxic by the MEM elution test, four 2-fold dilutions are made to determine the toxic endpoint. Performance of the test will result in an estimation of the relative "strength" of the cytotoxic substance in the material.

A sample is placed in direct contact with L-929 mouse fibroblast cells. Cells are examined at 24 hours for signs of toxicity.

SAMPLE REQUIREMENTS 
3 samples of 1 cm2 solid (sterile)

TURNAROUND TIME 4 days

A sample is placed in direct contact with MRC-5 human embryonic lung cells. Cells are examined at 24 hours for signs of toxicity.

SAMPLE REQUIREMENTS 
3 samples of 1 cm2 solid (sterile)

TURNAROUND TIME 4 days

This test evaluates the cytotoxic response of L-929 mouse fibroblast cell line when exposed to the extract of the materials under test. It is designed to utilize the sensitivity typical of low cell density cultures to determine material or device cell toxicity.

SAMPLE REQUIREMENTS 
1.0 gram or 50 cm²

TURNAROUND TIME 3 weeks

The Ames Mutagenicity test is performed on extracts of solid materials. To perform this test, five tester strains of bacteria (five Salmonella typhimurium or four Salmonella typhimurium and one Escherichia coli), in which specific mutations can be induced, are exposed to the test material and to a number of positive controls. This test can only be used for screening purposes.

The ISO Bacterial Reverse Mutation test is performed according to ISO 10993-3 using OECD test method 471. Tester strains of bacteria (five Salmonella typhimurium or four Salmonella typhimurium and one Escherichia coli or five Salmonella typhimurium and one Escherichia coli) are exposed to extracts of the test material in the presence and absence of an exogenous metabolic activation system. One dose level of the test article per extract and both positive and negative controls are used.

This Bacterial Reverse Mutation test is performed according to ISO 10993-3 using OECD test method 471. Tester strains of bacteria (five Salmonella typhimurium or four Salmonella typhimurium and one Escherichia coli or five Salmonella typhimurium and one Escherichia coli) are exposed to an extract of the test material in the presence and absence of an exogenous metabolic activation system. One dose level of the test article extract and both positive and negative controls are used.

The purpose of this study is to evaluate the mutagenic potential of the test article (or its metabolites) by measuring its ability to induce back mutations at selected loci of four strains of Salmonella typhimurium and one strain of Escherichia coli in the presence and absence of microsomal enzymes. For the assay, the test article is extracted according to specialized procedures involving the use of solvents and evaporation in order to obtain a residue for testing. When residues are obtained, they are tested at five dose levels along with appropriate vehicle and positive controls. Alternate methodologies are outlined in the study protocol for test articles that do not yield a residue.

Test materials yielding positive results on the Ames Mutagenicity plate incorporation test should be repeated with the strains that showed positive results. The confirmation test is run with appropriate positive and negative controls and in the presence and absence of metabolic activation.

Mammalian cells are exposed to the test material or extract in the presence and absence of metabolic activation and blocked in metaphase using a spindle poison. Visualization of chromosomes is performed microscopically after hypotonic swelling, fixation and staining. Test code 190828 (two extracts) satisfies the ISO requirement for assessing in vitro chromosomal aberrations.

Mammalian cells are exposed to the test material or extract and blocked in metaphase using a spindle poison. Visualization of chromosomes is performed microscopically after hypotonic swelling, fixation and staining the cells. For the assay, the test article is extracted according to specialized procedures involving the use of solvents and evaporation in order to obtain a residue for testing. When residues are obtained, they are tested at five dose levels along with appropriate vehicle and positive controls. Alternate methodologies are outlined in the study protocol for test articles that do not yield a residue.

TURNAROUND TIME 70 days (GLP)

Mouse Lymphoma cells are used to determine whether a test material has the capacity to induce either point mutations or clastogenic (chromosomal breakage) events in a cultured mammalian cell line. Mutants can be selected and mutant frequencies derived by including a thymidine analogue (trifluorothymidine, TFT) in the culture medium of cells after exposure to the test material. Test code 190850 (two extracts) satisfies the ISO requirement for assessing in vitro DNA genotoxic effects.

Sister chromatid exchanges (SCE) are visualized by allowing Chinese Hamster Ovary cells (CHO) to replicate twice in the presence of the nucleotide analog, 5-bromo-2-deoxyuridine (BrdUrd) and the test article extract. Through the use of special staining techniques, the two chromatid arms (sister chromatids) display differential staining, permitting enumeration of SCE.

The assay is designed to determine whether a test material or extract damages DNA in a fashion that provokes "long patch" DNA repair. Primary rat hepatocytes are exposed to test materials in the presence of tritiated thymidine. Labeled thymidine is incorporated in "patches" where damaged DNA has been excised and new DNA resynthesized. Cell are fixed on slides and evaluated autoradiographically for the presence and quantity of repair sites.

The mammalian in vivo micronucleus test is used for the detection of damage induced to the chromosomes or the mitotic apparatus of erythroblasts by analysis of erythrocytes sampled from bone marrow of mice. For this assay, male mice are injected with 1 dose level of the test article extract or positive or negative control. Cells are collected at 24 and 48 hours after dosing.

The mammalian in vivo micronucleus test is used for the detection of damage induced to the chromosomes or mitotic apparatus of erythroblasts by analysis of erythrocytes from bone marrow of treated mice. In this assay, male and female mice are injected with 1 dose level of the test article extract or positive or negative control. Cells are collected for analysis at 24 and 48 hours after dosing.

The activation of complement resulting from the use of a medical device has been associated with many adverse clinical findings. An enzyme immunoassay is used to screen for complement component(s) in human serum that has been incubated with the test article. Elevated levels of complement components C3a and SC5b-9 indicate activation of the complement system. Both the C3a and SC5b-9 assays are available. 

Sample Requirements 
6 cm² per assay

The PTT assay is a general screening test for the detection of coagulation abnormalities in the intrinsic coagulation pathway. The test determines the time it takes citrated human plasma to form a clot when it is exposed first to the test material, then to calcium chloride and finally to partial thromboplastin. (Partial thromboplastin is a phospholipid suspension extracted from rabbit brain cephalin.) Test results report the "partial thromboplastin time," which is the time it takes the recalcified citrated plasma to clot once the partial thromboplastin has been added. Shortening of the PTT following contact with a material under standardized conditions indicates activation of the contact phase of blood coagulation. 

Sample Requirements 
12 cm²

TURNAROUND TIME 21 days (GLP)

Platelet and leukocyte counts are evaluated before and after exposure to the test material in human blood. Counts are evaluated for changes that may indicate activation, adhesion, aggregation, or lysis. 

Sample Requirements 
36 cm²

TURNAROUND TIME 21 days (GLP)

Test article (e.g., tubing or catheter) is implanted in the jugular veins of two (2) dogs. Usually the test article is implanted in the jugular vein on one side and a sponsor-supplied comparative control article on the contralateral side for up to 3 days. The test article and implant sites are removed and examined for the presence of thrombi, and the vein is examined for patency (occlusion). These observations are augmented with photographs.

Sample Requirements 
2 test products and 2 control products

TURNAROUND TIME 25 days (GLP)

This test is intended for materials that directly contact the bloodstream or compromised tissues. It is performed in triplicate and uses rabbit blood in direct contact with the test material. The degree of hemolysis is measured spectrophotometrically.

TURNAROUND TIME 2 days
Tests set up on Tuesday.

This test is intended for materials through which fluids pass before entry into the body. It is performed in triplicate and uses saline to extract leachable substances. The material is removed and rabbit blood is added to the extract. The degree of hemolysis is measured spectrophotometrically.

Specify one set of extraction conditions: 
121°C/1hour,   70°C/24 hours,   37°C/72 hours,    50°C/72 hours

TURNAROUND TIME 2-5 days
Tests set up on Tuesday.

Biomaterial is implanted intramuscularly into rabbits to assess the reaction of the surrounding tissue. The implants remain in the muscle for the sponsor-designated time period. If the animals exposed to the test article do not show significant signs of irritation above that observed in the concurrent test control, the test article passes the test. Scoring of the sample and control reactions will be by gross observation. Histopathologic evaluation and photomicrographs (gross and histologic) will also be provided at the request of the sponsor. Additional fees apply. 

SAMPLE REQUIREMENTS
The sample used in the test is 1 mm by 10 mm dimension. The sponsor can submit the sample in this form or the lab will sub-divide a larger sample. Somewhat larger samples can be surgically implanted. (Additional fees apply.) A minimum of 15 implant samples must be submitted. The sponsor can submit the sample pre-sterilized or the lab will sterilize according to sponsor directions.
[Note: For ISO, sample preparation complies with ISO10993-12.] 

TURNAROUND TIME (GLP) 
Implant        Gross        With Histopathology
1 wk              27 days           62 days
2 wk              34 days           69 days
4 wk              48 days           83 days
8 wk              76 days           111 days
12 wk            104 days         139 days

A portion of the material or device is implanted subcutaneously into rabbits to assess the reaction of the surrounding tissue. The implants remain for the sponsor-designated time period. If the animals exposed to the test article do not show significant signs of irritation above that observed in the concurrent test control sites, the test article passes the test. The sample and control tissue sites will be evaluated grossly. Histopathologic evaluation and photomicrographs (gross and histologic) will also be provided at the request of the sponsor. Additional fees apply.

SAMPLE REQUIREMENTS 
Representative portions of the device. (Minimum 12)
[Note: For ISO, sample preparation complies with ISO10993-12.] 

TURNAROUND TIME (GLP) 
Implant        Gross        With Histopathology
1 wk              27 days           62 days
2 wk              34 days           69 days
4 wk              48 days           83 days
8 wk              76 days           111 days
12 wk            104 days         139 days

This implantation test is most applicable for biomaterials that are exposed to the body for an extended duration. This test is used to access not only the local tissue effects but also any long-term systemic effects of the implanted material. A sample device or biomaterials are implanted into the muscle of rabbits for the sponsor-designated time period. Blood and body weights will be collected at specified intervals from each rabbit throughout the study. Target organs will be collected at termination and evaluated microscopically. All implant sites will be observed for gross reaction and microscopic evaluation. The results will be based on the overall assessment, including statistical analysis. Gross and histologic photomicrographs will also be provided at the request of the sponsor. Additional fees apply. 

Sample Requirements
The sample size best suited for this test is 10 mm by 1 mm (length : diameter). The sponsor can submit the sample in this form or the lab will subdivide a larger sample. Somewhat larger samples may be surgically implanted. (Additional fees apply.) The client should discuss the number of samples during protocol development. 

TURNAROUND TIME Inquire

AppTec’s professional staff can assist in the design and implementation of studies to investigate medical device biocompatibility issues. These studies help determine whether device surface characteristics, polymeric composition and physical geometry may effect local tissue responses such as inflammation, tissue ingrowth, vascularization and fibroplasia. Gross and histologic photomicrographs can also be provided at the request of the sponsor.

For other custom implant studies, such as bone, intracranial and intraperitoneal implants, contact your Account Manager.

Histopathology assessment is conducted for muscle or subcutaneous tissues if part of the protocol. (Additional fees apply.)

Note: ISO guidelines assume histopathology will be conducted.