US 5,294,541 AGranted

Real-time monitoring of oxidative products from in vitro cell-biomaterial interaction using chemiluminescence

AssigneeThe United States of America, as represented by the Secretary, Department of Health and Human Services

InventorsDavid S. Kaplan, Grace L. Picciolo, Edward P. Mueller

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Abstract

A chemiluminescence method for continuously monitoring in real time the generation of oxidative products such as hydrogen peroxide and superoxide from in vitro cell-biomaterial interactions using cell lines such as Human Leukemic cells (HL-60); tumor cell line hybridomas; cells lacking the respiratory burst such as Chronic Granulomatous Disease cells as controls; Monocytic cell lines; Primary Human cells such as monocytes, pmns, fibroblasts, endothelial cells; and whole and isolated blood cells. The oxidative products have the potential for the degradation of the biomaterial and thus this information can be used to aid in predicting the functional lifetime of the biomaterial when it is used to fabricate an implanted medical device. Further, this method may be used to determine the amount of activation of biological cells which is inferred from the amount of oxidative products. This activation level aids in the determination of the degree of the inflammatory response and thus influences the degree of acceptance of the biomaterial. Thus, toxicity and biocompatability tests could be supported by these measurements.

Claims21

Dependent claims

A method for analyzing in real time in vitro interactions between biological cells and biomaterial proposed for implantation which comprises:

introducing biological cells and biomaterial proposed for implantation into an environment for interaction, wherein said environment includes chemiluminescent probes which emit chemiluminescent light in the presence of oxidative products produced by said cells;
monitoring over real time the amount of chemiluminescent light produced by oxidative products of said cells in the presence of said biomaterial as a first measurement;
introducing a biological activator into said environment, wherein said biological activator causes the production of oxidative products;
monitoring over real time the amount of chemiluminescent light produced by oxidative products of said cells as a second measurement;
repeating the above steps under the same conditions except without the introduction of said biomaterial;
and
measuring the amount of said oxidative products produced by the interaction between said biological cells and said biomaterial in said environment over real time based on a comparison of each of the first chemiluminescence measurements, respectively, and each of the second chemiluminescence measurements, respectively.

4 more elements

Dependencies are inferred from the claim text.

Facts

Application: 07/912,590Filed 13 Jul 1992 · continuation
Priority: 21 Sep 1989
Granted: 15 Mar 1994
Term ends: Not estimatedThe source records don’t support an estimate.

Classification3 codes

G01N 33/5008
Involving human or animal cells: for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
G01N 33/5014
For testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics: for testing toxicity
G01N
Investigating or analysing materials by determining their chemical or physical properties