METHODS FOR PRODUCTION AND USE OF SUBSTANCE-LOADED ERYTHROCYTES (S-IEs) FOR OBSERVATION AND TREATMENT OF MICROVASCULAR HEMODYNAMICS

Abstract

Disclosed herein are methods, kits, and compositions for medical imaging relating to fluorescent dyes entrapped in erythrocytes. Also disclosed therein are methods, and compositions further comprising erythrocytes entrapping at least one therapeutically active agent, as well as methods for releasing the entrapped therapeutically active agent(s). Disclosed herein also are methods for preparation of the cells entrapping dye and therapeutically active agent(s) in a freeze-dried form that makes them readily available and easy to use in a clinical environment.

Description

RELATED APPLICATION[0001]This application is a continuation-in-part of PCT App. No. PCT / US2009 / 042606, filed May 1, 2009, which claims the benefit of priority under 35 U.S.C. §119(e) to U.S. Prov. App. 61 / 126,344, filed May 2, 2008, the disclosures of which are incorporated herein by reference.FIELD OF THE INVENTION[0002]Disclosed herein are kits, compositions, and methods for the clinical use of erythrocytes in the fields of medical angiography and therapy. The erythrocytes have been pre-loaded with substances for observation of blood flow under physiological conditions to detect circulation abnormalities. The erythrocytes may also be used for delivery of therapeutic substances to localized vascular areas. This technology can be applied to any vasculature that can be directly visualized by an optical means, such as ocular vasculatures.BACKGROUND[0003]Medical angiographic imaging typically involves administration of a detectable substance to a subject (see U.S. Pat. No. 6,915,154). ...

AI Technical Summary

Benefits of technology

[0010](b) reducing the osmolality of the erythrocyte-containing solution in (a) by dialysis against a dialysis buffer having an osmolality less than 300 mOsm / kg to cause pores in the erythrocyte to open and form a dialyzed solution;

[0012](d) combining the dye-containing solution in (c) with a resealing solution having an osmolality of at least 1000 mOsm / kg to increase the osmolality, thereby causing the erythrocyte pores to close and entrap the at least one fluorescent dye within the erythrocytes;

[0020](d) combining the dye-containing solution in (c) with a resealing solution having an osmolality of at least 1000 mOsm / kg to increase the osmolality, thereby causing the erythrocyte pores to close and entrap the at least one fluorescent dye within the erythrocytes;

[0028]In one embodiment, a portion of the erythrocytes contain ICG at a concentration that maximizes fluorescence efficiency, and the remainder contains both the therapeutic agent and ICG at a dye concentration that enhances light absorption, rather than fluorescence.

Problems solved by technology

Then, by returning the solution to normotonicity, the pores close, and the cells return to normal size, trapping the added substances inside; remaining non-entrapped substance can be washed away, leaving substance-loaded osmotically competent erythrocytes; these cells contain reduced amounts of hemoglobin, rendering them incompetent—or at least inefficient—with regard to oxygen delivery.

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