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1.1 This test method provides a protocol for rapid and quantitative measurement of the chemoattractant capacity of a nanoparticulate material (nanoparticles and their aggregates and agglomerates).

1.2 Immune cells recruitment (by chemotaxis) plays a central role in the immune system function especially in the inflammatory process.

1.3 This test method uses an in vitro model. In this model, peripheral blood human acute promyelocytic leukemia cells HL-60 are separated from control chemoattractant or test nanoparticulate material by a 3-µm pore size filter; the cell migration through the filter is monitored and quantified using the fluorescent dye calcein AM (Figs. 1 and 2).

FIG. 1 Chemotaxis Chamber (Boyden Chamber)

FIG. 2 Chemotaxis Assay

a (left)—Parts of the chemotaxis assay assembly.
b (right)—Procedure for testing the chemoattractant capacity of a nanoparticulate material.

1.4 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.

1.5 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.

1.6 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

5.1 This test method will assess whether the test nanoparticulate material has chemoattractant activity.

5.2 This test method will provide a rapid and quantitative measure of the ability of nanoparticulate material to recruit immune cells.

5.3 Recruitment of immune cells by chemotaxis plays an important part in all phases of both humoral and cell-mediated immune responses.

5.4 Testing the capacity of a nanoparticulate material to recruit immune cells in vitro helps in predicting the influence of such material on the immune cell response.