Matrix bound nanovesicles and their use
AssigneeUniversity of Pittsburgh—Of the Commonwealth System of Higher Education
InventorsStephen Francis Badylak, Luai Huleihel, George S. Hussey, Juan Diego Naranjo Gutierrez
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Abstract
A composition is disclosed herein that includes isolated ECM-derived nanovesicles and a pharmaceutically acceptable carrier. Methods are producing the ECM-derived nanovesicles are also disclosed. These ECM-derived nanovesicles can be included in pharmaceutical compositions, bioscaffolds, and devices. Methods for using these ECM-derived nanovesicles are provided.
Claims31
A composition comprising isolated nanovesicles derived from an extracellular matrix and a pharmaceutically acceptable carrier.
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A method of isolating nanovesicles from an extracellular matrix, wherein the nanovesicles do not express CD63 or CD81, or are CD63loCD81lo, comprising:
- digesting the extracellular matrix with an enzyme to produce digested extracellular matrix;
- (b) centrifuging the digested extracellular matrix to remove collagen fibril remnants and thus to produce a fibril-free supernatant;
- (c) centrifuging the fibril-free supernatant to isolate solid materials;
- and
- (d) suspending the solid materials in a carrier,
- thereby isolating nanovesicles from the extracellular matrix.
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A method of altering cell proliferation, migration and/or differentiation on an extracellular matrix of interest, comprising introducing isolated nanovesicles derived from a second extracellular matrix into the extracellular matrix of interest wherein the nanovesicles do not express CD63 or CD81, or are CD63loCD81lo;
- thereby altering cell proliferation, migration and/or differentiation on the matrix.
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A method of isolating nanovesicles from an extracellular matrix, wherein the nanovesicles do not express CD63 or CD81, or are CD63loCD81lo, comprising:
- (a) incubating an extracellular matrix (ECM) at a salt concentration of greater than about 0.1 M to produce an ECM-salt solution;
- (b) centrifuging the ECM-salt solution to remove collagen fibril remnants, and isolating the supernatant;
- (c) centrifuging the supernatant to isolate solid materials;
- and
- (d) suspending the solid materials in a carrier, thereby isolating nanovesicles from the extracellular matrix.
A method of isolating nanovesicles from an extracellular matrix, wherein the nanovesicles do not express CD63 or CD81, or are CD63loCD81lo, comprising:
- (a) suspending the extracellular matrix in isotonic buffered saltsaline solution to form a suspension;
- and optionally centrifuging the suspension to remove collagen fibrils to produce a collagen fibril-free supernatant;
- (b) performing ultrafiltration to isolate particles from the suspension or the collagen fibril-free supernatant of between about 10 nm and about 10,000 nm in diameter;
- thereby isolating nanovesicles from the extracellular matrix.
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Dependencies are inferred from the claim text.