MATRIX BOUND NANOVESICLES AND THEIR USE
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.
1 . A composition comprising isolated nanovesicles derived from an extracellular matrix and a pharmaceutically acceptable carrier.
2 . The composition of claim 1 , wherein the nanovesicles do not express CD63 or CD81, or are CD63 lo CD81 lo .
3 . The composition of claim 1 , wherein the extracellular matrix is a mammalian extracellular matrix.
4 . The composition of claim 2 , wherein the mammalian extracellular matrix is a human extracellular matrix.
5 . The composition of claim 3 , wherein the extracellular matrix is from esophageal tissue, urinary bladder, small intestinal submucosa, dermis, umbilical cord, pericardium, cardiac tissue, tumor tissue, or skeletal muscle.
6 . The composition of claim 3 , wherein the nanovesicles comprise miR-145 and/or miR-181.
7 . The composition of claim 3 , wherein the extracellular matrix is digested with an enzyme.
8 - 9 . (canceled)
10 . The composition of claim 3 , further comprising an exogenous therapeutic agent.
11 - 12 . (canceled)
13 . A method of isolating nanovesicles from an extracellular matrix, comprising:
digesting the extracellular matrix with an enzyme to produce digested extracellular matrix;
centrifuging the digested extracellular matrix to remove collagen fibril remnants and thus to produce a fibril-free supernatant;
centrifuging the fibril-free supernatant to isolate the solid materials; and
suspending the solid materials in a carrier,
thereby isolating nanovesicles from the extracellular matrix.
14 - 22 . (canceled)
23 . 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;
thereby altering cell proliferation, migration and/or differentiation on the matrix.
24 - 34 . (canceled)
35 . A bioscaffold comprising nanovesicles derived from an extracellular matrix and a heterologous extracellular matrix.
36 . The bioscaffold of claim 35 , wherein the nanovesicles are derived from a mammalian extracellular matrix.
37 . The bioscaffold of claim 36 , wherein the mammalian extracellular matrix is a human or a porcine extracellular matrix.
38 . The bioscaffold of claim 36 , wherein the extracellular matrix is from esophageal cells, urinary bladder cells, a small intestinal submucosa, or a dermis
39 . The bioscaffold of claim 36 , wherein the heterologous extracellular matrix is derived from a different tissue than the nanovesicles.
40 . The bioscaffold of claim 36 , wherein the heterologous extracellular matrix is from a different species than the nanovesicles.
41 . A medical device comprising the composition of claim 1 .
42 . The medical device of claim 41 , wherein the device is a surgical mesh, a stent, a pacemaker, a catheter, heart valve, biosensor, a drug delivery device, or an orthopedic implant.
43 . A method of isolating nanovesicles from an extracellular matrix, comprising
a) incubating an extracellular matrix at a salt concentration of greater than about 0.1 M;
b) centrifuging the digested extracellular matrix to remove collagen fibril remnants, and isolating the supernatant;
c) centrifuging the supernatant to isolate the solid materials; and
d) suspending the solid materials in a carrier, thereby isolating nanovesicles from the extracellular matrix.
44 . (canceled)
45 . A method of isolating nanovesicles from an extracellular matrix, comprising:
a) suspending the extracellular matrix in isotonic buffered saline solution to form a suspension; and
b) performing ultrafiltration to isolate particles from the suspension of between about 10 nm and about 10,000 nm in diameter;
thereby isolating nanovesicles from the extracellular matrix.
46 - 49 . (canceled)
50 . A method for reducing the proliferation of a tumor cell, increasing apoptosis of a tumor cell, and/or decreasing migration of a tumor cell, comprising
contacting the tumor cell with an effective amount of the composition of claim 3 , thereby reducing the proliferation of the tumor cell, increasing apoptosis of the tumor cell, and/or decreasing migration of the tumor cell.
51 - 54 . (canceled)
55 . A method of treating a subject with a tumor, comprising administering to the subject a therapeutically effective amount of the composition of claim 3 , thereby treating the tumor in the subject.
56 - 57 . (canceled)
58 . A method of increasing M2 macrophages in a subject, comprising
administering to a subject a therapeutically effect amount of the composition of claim 1 , thereby increasing M2 macrophages in the subject.
59 - 60 . (canceled)