Biocomposite for regeneration of injured tissue and organs, a kit for making the biocomposite, a method of making the biocomposite and a method of treating injuries

• US 9,730,959 B2
• Filed: 12/24/2012
• Issued: 08/15/2017
• Est. Priority Date: 12/29/2011
• Status: Active Grant

• Current Assignee: NextGen Company Limited (Human Stem Cells Institute OJSC)
• Original Assignee: NextGen Company Limited (Human Stem Cells Institute OJSC)
• Inventors: Bozo, Il'ya Yadigerovich, Filonenko, Elena Sergeevna, Deev, Roman Vadimovich, Isaev, Artur Aleksandrovich, Kiselev, Sergej L'vovich
• Primary Examiner(s): EPPS -SMITH, JANET L
• Application Number:
  US14/369,008
• Publication Number:
  US 20140341870A1
• Time in Patent Office: 1,695 Days
• Field of Search: 424 9321
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61K 47/6901 Conjugates being cells, cell fragments, viruses, ghosts, red blood cells or viral vectors
• CPC Class Codes: A61K 47/6923 the form being an inorganic particle, e.g. ceramic particles, silica particles, ferrite or synsorb
• CPC Class Codes: A61K 48/0041 the non-active part being polymeric
• CPC Class Codes: A61L 2300/258 Genetic materials, DNA, RNA, genes, vectors, e.g. plasmids
• CPC Class Codes: A61L 2300/64 Animal cells
• CPC Class Codes: A61L 2400/06 Flowable or injectable implant compositions
• CPC Class Codes: A61L 2430/02 for reconstruction of bones; weight-bearing implants
• CPC Class Codes: A61L 27/12 Phosphorus-containing materials, e.g. apatite
• CPC Class Codes: A61L 27/24 Collagen
• CPC Class Codes: A61L 27/3804 characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
• CPC Class Codes: A61L 27/46 with phosphorus-containing inorganic fillers
• CPC Class Codes: A61L 27/54 Biologically active materials, e.g. therapeutic substances A61L27/227 takes precedence
• CPC Class Codes: A61L 27/58 Materials at least partially resorbable by the body
• CPC Class Codes: C08L 89/06 derived from leather or skin , e.g. gelatin

Abstract

Provided is a biocomposite including cells, a genetic construction, and a scaffold and a method for repairing tissue and organs in mammalians with the biocomposite. The interaction of the components of the biocomposite provides a complex effect on reparative regeneration processes. Also provided is a method for administration of gene-cellular therapeutic constructions to a recipient which can be used in medicine and veterinary to provide reparative processes. After administering the biocomposite to a recipient, the scaffold structure releases the nucleic acids which enter into the cells of a recipient bed and cells of the transplanted product. The nucleic acids are expressed, which results in the increased concentration of a target product responsible for reparative processes.

Claims

1. A biocomposite suitable for providing reparative processes in injured mammalians consisting of:

a three-dimensional solid material matrix as a scaffold consisting of at least one calcium phosphate,

at least one nucleic acid encoding vascular endothelial growth factor (VEGF), stromal cell-derived factor (SDF), or VEGF and SDF, and

cells which provide reparative regeneration, wherein the cells are selected from the group consisting of multipotent mesenchymal stromal cells, fibroblasts, periostal osteogenic cells, fresh blood with cells, and stromal vascular cells fraction.

2. The biocomposite according to claim 1, wherein the at least one nucleic acid is a DNA which is included in a vector molecule, wherein the vector is a plasmid, a virus, an episome, or a transposone.

3. The biocomposite according to claim 1, which comprises autogenous and/or allogenous cells.

4. The biocomposite according to claim 1, wherein the cells are derived from mammalians immediately prior to adding the cells to the biocomposite, or cells which were preliminarily exposed to laboratory technologies of cellular processing.

10. A method for healing injury comprising administering the biocomposite according to claim 1 to a mammalian in need thereof.

14. A method of delivery of at least one nucleic acid comprised in the biocomposite according claim 1 to a mammalian in need thereof, the method comprises applying the biocomposite to the mammalian.

15. A kit for preparation of the biococomposite according to claim 1, comprising:

a first vessel comprising a complex of a three-dimensional solid material matrix as a scaffold and at least one nucleic acid encoding vascular endothelial growth factor (VEGF), stromal cell-derived factor (SDF), or VEGF and SDF, wherein the scaffold consists of at least one calcium phosphate, and

a second separate vessel for combining the complex with a biological fluid or cultural medium with cells which provide reparative regeneration, wherein the cells are selected from the group consisting of multipotent mesenchymal stromal cells, fibroblasts, periostal osteogenic cells, fresh blood with cells, and stromal vascular cells fraction.

17. The biocomposite of claim 1, wherein the scaffold and the at least one nucleic acid create a complex.

5. A method of making a biocomposite, the method comprising:

combining at least one nucleic acid encoding vascular endothelial growth factor (VEGF), stromal cell-derived factor (SDF), or VEGF and SDF with a three-dimensional solid material matrix as a scaffold, thereby creating a complex, and

adding cells which provide histotypical reparative regeneration to the created complex of the scaffold and the at least one nucleic acid,

wherein the scaffold consists of at least one calcium phosphate, and

wherein the cells are selected from the group consisting of multipotent mesenchymal stromal cells, fibroblasts, periostal osteogenic cells, fresh blood with cells, and stromal vascular cells fraction.

6. The method according to claim 5, further comprising deriving the cells from mammalians immediately prior to the adding the cells or preliminarily exposing the cells to laboratory technologies of cellular processing.

7. A method of making a biocomposite, the method comprising:

combining at least one nucleic acid encoding vascular endothelial growth factor (VEGF), stromal cell-derived factor (SDF), or VEGF and SDF with a three-dimensional solid material matrix as a scaffold, thereby creating a complex, and

adding cells to the created complex, wherein the cells comprise

(i) cells which were preliminarily transfected with at least one nucleic acid, which can be the same or different from the at least one nucleic acid combined with the scaffold, and

(ii) cells which were not preliminary transfected,

wherein the cells (i) and (ii) provide histotypical reparative regeneration,

wherein the scaffold consists of at least one calcium phosphate, and

wherein the cells are selected from the group consisting of multipotent mesenchymal stromal cells, fibroblasts, periostal osteogenic cells, fresh blood with cells, and stromal vascular cells fraction.

11. The method according to claim 7, further comprising administering the biocomposite directly to a damaged tissue area as a part of a surgical operation or manipulation.

12. The method according to claim 7, further comprising administering the biocomposite in an intravenous, intra-arterial, intramuscular, intradermic, subcutaneous, intraosteal, endolumbar, subdural, intraarticular, or an orendobulbar injection.

13. The method according to claim 7, further comprising topically applying the biocomposite.

8. A method of making a biocomposite, the method comprising:

combining at least one nucleic acid encoding vascular endothelial growth factor (VEGF), stromal cell-derived factor (SDF), or VEGF and SDF with a three-dimensional solid material matrix as a scaffold, thereby creating a complex,

preliminarily transfecting cells with at least one nucleic acid, which can be the same or different from the at least one nucleic acid combined with the scaffold, wherein the cells provide histotypical reparative regeneration, and

adding the transfected cells to the created complex,

wherein the scaffold consists of at least one calcium phosphate, and

wherein the cells are selected from the group consisting of multipotent mesenchymal stromal cells, fibroblasts, periostal osteogenic cells, fresh blood with cells, and stromal vascular cells fraction.

9. A method of making the biocomposite, the method comprising:

preliminary transfecting cells, which provide histotypical reparative regeneration, with at least one nucleic acid encoding vascular endothelial growth factor (VEGF), stromal cell-derived factor (SDF), or VEGF and SDF, and

combining the transfected cells with a three-dimensional solid material matrix as a scaffold,

wherein the scaffold consists of at least one calcium phosphate, and

wherein the cells are selected from the group consisting of multipotent mesenchymal stromal cells, fibroblasts, periostal osteogenic cells, fresh blood with cells, and stromal vascular cells fraction.

16. A biocomposite suitable for providing reparative processes in injured mammalians comprising:

a three-dimensional solid material matrix as a scaffold,

at least one nucleic acid encoding vascular endothelial growth factor (VEGF), stromal cell-derived factor (SDF), or VEGF and SDF, and

cells which provide reparative regeneration,

wherein the scaffold consists of at least one of calcium phosphate, and

wherein the cells are selected from the group consisting of multipotent mesenchymal stromal cells, a stromal vascular cells fraction, and cells of fresh blood.