Implantable liposome embedded matrix composition, uses thereof, and polycaprolactone particles as scaffolds for tissue regeneration

Abstract

In various embodiments, the present invention describes materials and methods for the local reprogramming of cells in a location where the treatment is applied. The invention can be used to replace lost cells or to restore function to tissue damaged due to disease, injury or genetic defect. In various embodiments, the treatment includes a semisolid hydrogel embedded with liposomes. The liposomes can contain an effector molecule or molecules. When phagocytic cells such as monocytes infiltrate the hydrogel, they encounter the liposomes and incorporate the liposomes carrying the effector molecules into the cells. In some embodiments, the effector molecules can be genetic material encoding the expression of specific proteins such as transcription factors, the expression of which can initiate the reprogramming of the cells. In other embodiments, the effector molecules can induce angiogenesis. In other embodiments, the effector molecules are tumor antigens. The matrix can contain other effector molecules designed to attract specific cells to the matrix. The cells can be released from the matrix as the matrix degrades or by active migration from the matrix. The cells can also remain in the matrix and secret molecules such as proteins and hormones that will diffuse through the matrix material to the surrounding tissue.

1 . An implantable, semisolid matrix comprising a hydrogel material and liposomes embedded therein, wherein the liposomes contain one or more bioactive material, and wherein the matrix is permeable to the infiltration of cells from an implantation environment and to products secreted from the cells into the environment. 
     
     
         2 . The matrix of  claim 1 , wherein the bioactive material is selected from the group consisting of a protein, a peptide, a nucleotide, a DNA, a RNA, a siRNA, a cDNA, and a small molecule, a drug or a combinations thereof. 
     
     
         3 . (canceled) 
     
     
         4 . (canceled) 
     
     
         5 . (canceled) 
     
     
         6 . The matrix of  claim 1 , wherein the liposomes carry the bioactive material in the liposomal core, in the liposomal membrane, on the liposome surface, or in a combination thereof. 
     
     
         7 . The matrix of  claim 1 , wherein the hydrogel contains a biological or chemical compound. 
     
     
         8 . The matrix of  claim 7 , wherein the compound is a chemoattractant compound or an agent that promotes differentiation of monocytic cells to dendritic cells. 
     
     
         9 . The matrix of  claim 7 , wherein the compound is selected from the group consisting of monocyte chemotactic protein-1, granulocyte macrophage colony-stimulating factor (GM-CSF), interleukin-4, TNFα, and interleukin-15. 
     
     
         10 . (canceled) 
     
     
         11 . (canceled) 
     
     
         12 . (canceled) 
     
     
         13 . The matrix of  claim 1 , wherein the hydrogel material contains cells. 
     
     
         14 . The matrix of  claim 13 , wherein the cells are monocytes or dendritic cells. 
     
     
         15 . The matrix of  claim 1 , wherein the hydrogel comprises copolymers of two or more polyhydroxy acids, polyorthoesters, polyanhydrides, gelatin, collagen, cellulose, derivatized cellulose, chitosan, alginate, hyaluronan, thiol-modified hyaluronan, and combinations or copolymers thereof. 
     
     
         16 . The matrix of  claim 15 , wherein the polyhydroxy acid is polylactic acid, polyglycolic acid or other polyhydroxy acid. 
     
     
         17 . The matrix of  claim 1 , further comprising a cross-linking agent. 
     
     
         18 . The matrix of  claim 17 , wherein the cross-linking agent is selected from glutaraldehyde, diphenylphosphoryl azide, transglutaminase, dimethyl suberimidate, DMS-treated collagen, dimethyl 3,3′-dithiobispropionimidate, multivalent ions, calcium ions, N,N methylene-bisacrylamide (MBA), acrylamide, allyl methacrylate, ethylene glycol dimethacrylate, tripolyphosphate, and combinations thereof. 
     
     
         19 . The matrix of  claim 1 , wherein the bioactive material contained within the liposomes is a tumor antigen or a nucleic acid molecule encoding a tumor antigen. 
     
     
         20 . (canceled) 
     
     
         21 . (canceled) 
     
     
         22 . (canceled) 
     
     
         23 . The matrix of  claim 1 , wherein the bioactive material contained within the liposomes is an agent that promotes differentiation of monocytic cells to dendritic cells. 
     
     
         24 . The matrix of  claim 1 , wherein the bioactive material is an expression vector encoding a transcription factor. 
     
     
         25 . The matrix of  claim 24 , wherein the transcription factor is capable of differentiating the cell into an insulin producing cell. 
     
     
         26 . The matrix of  claim 24 , wherein the transcription factor is selected from the group consisting of NGN-3, PDX-1, MAFA, Oct4, Sox2, c-Myc, klf-4, Nanog. Polycomb group (PcG) proteins, PAX6, MEIS1, OTX1, ATBF1, DLX5, HAND1, Nuclear factor kappa-B and OENCUT1. 
     
     
         27 . (canceled) 
     
     
         28 . A method of affecting the behavior of a cell in a subject, comprising implanting at a site within the subject an implantable, semisolid matrix comprising a hydrogel material and liposomes embedded therein, wherein the liposomes comprise at least one compound capable of affecting the cell's behavior, such that the cell infiltrates the semisolid matrix comprising the hydrogel material and takes up the liposomes through phagocytosis or through fusion of the liposome membrane with the cell membrane. 
     
     
         29 . The method of  claim 28 , wherein the behavior is selected from the group consisting of an angiogenic activity, an immunosuppressive activity, an inflammatory activity, an antigen presenting activity, cell differentiation, and cell reprogramming. 
     
     
         30 . (canceled) 
     
     
         31 . (canceled) 
     
     
         32 . The method of  claim 28 , wherein the cell is monocytic. 
     
     
         33 . The method of  claim 32 , wherein the monocytic cell secretes vascular growth factors at or near the implantation site. 
     
     
         34 . (canceled) 
     
     
         35 . (canceled) 
     
     
         36 . The method of  claim 28 , wherein the compound is selected from the group consisting of IL-10, IL-4, IL-13, an IL-1 receptor ligand, interleukin-15 PGE2, TGF-β, TNFα, lactic acid, lipoteichoic acid, NADH dehydrogenase 5, subunit 1, poly (adenosine diphosphate-ribose)polymerase, pyruvate, granulocyte macrophage colony stimulating factor (GM-CSF), Colony Stimulating Factor-1, adenosine, an adenosine analogue, NECA, LPS, Pam3CSK4,  E. coli  LPS, R848, imiquimod, non-methylated CpG DNA, ODN2006, thioredoxin peroxidase, Trapidil, TLR2 agonist (e.g., Pam3CysSerLys4, peptidoglycan (Ppg), PamCys), a TLR3 agonist (e.g., IPH 31XX), a TLR4 agonist (e.g., Aminoalkyl glucosaminide phosphates [AGPs], E6020, CRX-675, 5D24.D4, RC-527), a TLR7 agonist (e.g., Imiquimod, 3M-003, Aldara, 852A, R850, R848, CL097), a TLR8 agonist (3M-002), a TLR9 agonist (Flagellin, Vaxlmmune, CpG ODN (AVE0675, HYB2093), CYT005-15 AllQbG10, dSLIM), adenosine A1 agonists (R-PIA, CPA TCPA, CVT-3146, CVT-510, GR 79236, WAG 994), adenosine A2 agonists (CGS 21680, APEC, 2HE-NECA), and/or adenosine A3 agonists (e.g., IB-MECA, CI-IB-MECA, 3′-Aminoadenosine-5′-uronamides), Alendronate, an expression vector and combinations thereof. 
     
     
         37 . A method for delivering a secreted product to a localized site in a subject, comprising
 providing at the localized site the matrix or composition of  claim 1 ; and   maintaining the matrix or composition at the localized site for a period of time sufficient for the secreted product to interact with the localized site.   
     
     
         38 . The method of  claim 28 , wherein the site is within an ischemic tissue. 
     
     
         39 . (canceled) 
     
     
         40 . A method for programming an implantable, semisolid matrix comprising a hydrogel material and liposomes embedded therein to affect the behavior of a cell, comprising formulating within the liposomes a compound capable of affecting the cell's behavior, such that the cell infiltrates the semisolid matrix comprising the hydrogel material and takes up the liposomes through phagocytosis or through fusion of the liposome membrane with the cell membrane. 
     
     
         41 . (canceled) 
     
     
         42 . (canceled) 
     
     
         43 . (canceled) 
     
     
         44 . The method of  claim 28 , wherein the compound is a tumor antigen. 
     
     
         45 . The method of  claim 44 , wherein the tumor antigen is derived from tumor cells or tumor tissue from the subject. 
     
     
         46 . The method of  claim 32 , wherein the monocytic cell displays a tumor antigen for recognition by a T cell at or near the implantation site. 
     
     
         47 . The method of  claim 28 , wherein the liposomes comprise an agent that promotes differentiation of monocytic cells to dendritic cells. 
     
     
         48 . (canceled) 
     
     
         49 . A method for presenting a tumor antigen at a localized site in a subject comprising: providing at the localized site an implantable, semisolid matrix comprising a hydrogel material and liposomes embedded therein, wherein the liposomes comprise the tumor antigen; and maintaining the matrix or composition at the localized site for a period of time sufficient for a phagocytic cell to infiltrate the matrix, phagocytose the antigen and present the antigen to an immune cell at the localized site. 
     
     
         50 . The method of  claim 49 , wherein the localized site is at or near a tumor site, such that an immune response to the tumor is enhanced. 
     
     
         51 . (canceled) 
     
     
         52 . (canceled) 
     
     
         53 . (canceled) 
     
     
         54 . (canceled) 
     
     
         55 . The method of  claim 29 , wherein the behavior is differentiation into insulin producing cells, or dopamine producing cells. 
     
     
         56 . (canceled) 
     
     
         57 . (canceled) 
     
     
         58 . The method of  claim 28 , wherein the cell is a stem cell or stem cell precursor. 
     
     
         59 . (canceled) 
     
     
         60 . (canceled) 
     
     
         61 . The method of  claim 58 , wherein the compound is an expression vector that encodes a transcription factor capable of differentiating the cell into an insulin producing cell. 
     
     
         62 . The method of  claim 61 , wherein the transcription factor is selected from the group consisting of NGN-3, PDX-1, MAFA Oct4, Sox2, c-Myc, klf-4, Nanog, Polycomb group (PcG) proteins, PAX6, MEIS1, OTX1, ATBF1, DLX5, HAND1, Nuclear factor kappa-B and OENCUT1. 
     
     
         63 . (canceled) 
     
     
         64 . A method for promoting insulin secretion at a localized site in a subject comprising, providing at the localized site an implantable, semisolid matrix comprising a hydrogel material and liposomes embedded therein, wherein the liposomes comprise an expression vector encoding a transcription factor capable of differentiating a cell into an insulin producing cell, and maintaining the matrix or composition at the localized site for a period of time sufficient for a cell to infiltrate the matrix and phagocytose the liposomes, such that the transcription factor is expressed in the cell, thereby promoting differentiation of the cell to produce insulin at the localized site. 
     
     
         65 . The method of  claim 64 , wherein the subject is an insulin resistant or insulin deficient subject. 
     
     
         66 - 83 . (canceled)