# US 9,750,848 B2 · Method of preparing an implantable neuroendoprosthetic system

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- Number: US 9,750,848 B2
- Document ID: US9750848B2
- Status: Granted
- Kind: B2
- Application number: 14/748,183
- Filed: 2015-06-23
- Published: 2015-10-15
- Granted: 2017-09-05
- Earliest claimed: 2008-09-25
- Applicants: Andrey S. Bryukhovetskiy; Viktor I. Sevastianov
- Inventors: Andrey S. Bryukhovetskiy; Viktor I. Sevastianov
- Web page: https://patentgrep.com/patent/US9750848B2

## CPC

- A61L 27/3878: {Nerve tissue, brain, spinal cord, nerves, dura mater}
- A61K 35/30: Nerves; Brain; Eyes; Corneal cells; Cerebrospinal fluid; Neuronal stem cells; Neuronal precursor cells; Glial cells; Oligodendrocytes; Schwann cells; Astroglia; Astrocytes; Choroid plexus; Spinal cord tissue
- A61K 35/44: Vessels; Vascular smooth muscle cells; Endothelial cells; Endothelial progenitor cells
- A61L 27/24: Collagen
- A61L 27/383: {Nerve cells, e.g. dendritic cells, Schwann cells}
- A61L 27/3834: {Cells able to produce different cell types, e.g. hematopoietic stem cells, mesenchymal stem cells, marrow stromal cells, embryonic stem cells}
- A61L 27/3886: {comprising two or more cell types}
- A61L 2430/32: for nerve reconstruction
- A61L: METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES (preservation of bodies or disinfecting characterised by the agents employed A01N; preserving, e.g. sterilising, food or foodstuffs A23; preparations for medical, dental or toiletry purposes A61K)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)

## Abstract

A method of making an implantable neuroendoprosthetic system for transubstantiation of defects of brain, spinal cord and vegetative nervous system in a mammal in reconstructive neurosurgical operations provides a heterogeneous collagen-containing matrix for implantation, a cell preparation of autologous cells of a patient, and the cell preparation is perfused into said matrix to make an elastic cell-biopolymer biologically active mass. The cell preparation comprises placed in a NaCl solution at least one type of cells from a group comprising neural stem cells (NSC), neuroglial ensheathing cells (NGEC), endothelial cells with CD34+ marker (EC), and purified mononuclears (MN). The mass can be additionally subjected to electromagnetic radiation at 1-10 GHz before implanting.

## Claims

### Claim 1

A method of production of an implantable neuroendoprosthetic system for transubstantiation of defects of brain, spinal cord and vegetative nervous system in a mammal in reconstructive neurosurgical operations, comprising the steps of:
providing a heterogeneous collagen-containing matrix for implantation,
providing a cell preparation of autologous cells of a patient, and
perfusing said preparation into said matrix to thereby obtain an elastic cell-biopolymer biologically active mass,
wherein the cell preparation comprises placed in a NaCl solution at least one type of cells from a group comprising neural stem cells (NSC), neuroglial ensheathing cells (NGEC), endothelial cells with CD34+ marker (EC), and purified mononuclears (MN), said cells are in the following ratios (in parts according to numbers of the cells) in the cell preparation: 0.8 to 1.2 of NSC; 1.6 to 2.4 of NGEC; 4 to 6 of EC; 4000 to 6000 of MN.

### Claim 2 (depends on 1)

The method of claim 1, wherein said perfusing includes centrifuging.

### Claim 3 (depends on 2)

The method of claim 2, wherein said centrifuging is carried out within 1.5 to 2.5 minutes at 1500 to 2500 r/m.

### Claim 4 (depends on 1)

The method of claim 1, wherein said cell preparation is prepared by defrosting cryopreserved cell products in a water bath at 37 to 40° C. with subsequent washing them at least twice in a physiological NaCl solution.

### Claim 5 (depends on 1)

The method of claim 1, wherein 0.5 to 1.3% solution of NaCl is used.

### Claim 6 (depends on 1)

The method of claim 1, wherein said NSC and NGEC are obtained from an olfactory sheath of the nose of a patient, and said EC and MN are obtained from either a bone marrow of the patient or a patient mobilized autologous stem cell leukoconcentrate obtained by separating patient's peripheral blood following stimulating the patient with a granulocyte colony-stimulating factor.

### Claim 7 (depends on 1)

The method of claim 1, wherein said cell preparation further comprises stimulators of tissue regeneration, nerve growth factors and vascular growth factors.

### Claim 8 (depends on 1)

The method of claim 1, wherein said production of said implantable neuroendoprosthetic system is carried out in sterile conditions directly in an operation room (ex tempore) or in a culture laboratory.

### Claim 9 (depends on 1)

The method of claim 1, further comprising subjecting the mass to electromagnetic radiation at 1-10 GHz before implanting.