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Cylinder support for repairing spinal cord injury and application method thereof

A spinal cord injury, cylinder technology, applied in the field of biomedical materials, can solve problems such as unsatisfactory clinical effects, achieve ultra-high porosity, promote growth, and promote spinal cord nerve regeneration.

Inactive Publication Date: 2015-01-21
BEIJING LANGJIAYI BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it has not been reported that combined with EGFR antibody to regulate the differentiation of NSCs into neurons
[0006] Studies have shown that the clinical effects of traditional drug administration routes such as injection of factors are not ideal. Therefore, it is urgent to find an ideal method for loading active substances and maintaining effective concentrations in the microenvironment of the spinal cord injury for a certain period of time.

Method used

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  • Cylinder support for repairing spinal cord injury and application method thereof
  • Cylinder support for repairing spinal cord injury and application method thereof
  • Cylinder support for repairing spinal cord injury and application method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1-for the construction of the cylinder support of repairing spinal cord injury

[0027] 1. Construction of Porous Collagen Sponge Cylindrical Scaffolds

[0028] Collagen raw materials were dissolved in 0.5mol / L acetic acid and allowed to stand at 4°C for 8 hours. After mixing evenly, 4mol / L NaOH was added for neutralization. The obtained homogeneous liquid was dialyzed with deionized water for 5 days, and the deionized water was changed every 3 hours during dialysis. ionized water. The dialyzed solution was put into a mold with a diameter of 4 mm, and freeze-dried to obtain a collagen sponge cylindrical scaffold.

[0029] 2. Construction of Porous Collagen Sponge Scaffolds Crosslinked with EGFR Antibody Cetuximab

[0030] Using Traut's Reagent (2-iminohydrothiol chloride) and Sulfo-SMCC (4-(N-maleimidomethyl)cyclohexane-1-carboxylic acid sulfosuccinimidyl ester sodium salt ) two-step coupling reaction to covalently cross-link cetuximab to the collagen scaf...

Embodiment 2

[0032] Example 2 - Characterization of Cylindrical Scaffold Materials for Repairing Spinal Cord Injuries

[0033] 1. Scanning electron microscope (SEM) to observe the morphology and pore size of the collagen scaffold

[0034] The collagen sponge scaffold was sprayed with gold in a dry state, observed and photographed under a scanning electron microscope. Observe the surface morphology, and take 5 fields of view in the material, and count the pore size and range.

[0035] 2. Detection of Cetuximab loaded on the Col-cetuximab sponge scaffold

[0036] After cetuximab was covalently cross-linked or simply adsorbed to the collagen scaffold, the collagen scaffold was blocked with 5% BSA, then excess BSA was removed, washed three times with PBS, and alkaline phosphatase-labeled anti-mouse antibody was added, at 37°C Incubate for 1h. Aspirate the antibody, wash it three times with PBS, and then add 2 mg / mL p-nitrophenyl phosphate disodium salt (p-Npp) for color development, p-Npp i...

Embodiment 3

[0037] Example 3 - The in vitro induction and differentiation of NSCs by cylindrical scaffold materials used for repairing spinal cord injuries

[0038] 1. Extraction of Myelin Proteins

[0039] Prepared by discontinuous sucrose density gradient centrifugation. Spinal cords were obtained from adult rats, homogenized and crushed in 0.3mol / L sucrose solution, and then slowly spread on top of 0.85mol / L sucrose. The samples were centrifuged at 27,000 g for 1 h, and the sheath protein fragments were collected and extracted at the boundary of 0.32 / 0.85 mol / L sucrose. The samples were allowed to stand in deionized water for 1 h, and then collected by centrifugation at 12000 g. The obtained samples were filtered with a 0.22 μm filter membrane to remove large aggregates, sterilized, and stored at -80°C for use.

[0040] 2. Isolation and culture of primary NSCs and inoculation on scaffold materials

[0041] The telencephalon was isolated from SD suckling mice born at 12 hours, cut i...

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Abstract

The invention provides a cylinder support for repairing spinal cord injury. The cylinder support comprises a collagen sponge cylinder support body. An epidermal growth factor receptor antibody cetuximab is crosslinked with the collagen sponge cylinder support body. Neural stem cells are loaded through the collagen sponge cylinder support body crosslinked with the epidermal growth factor receptor antibody cetuximab so as to promote transplanted neural stem cells to be differentiated into neurons, and the cylinder support which facilitates injured spinal nerve regeneration and functional reconstruction is constructed. The collagen sponge cylinder support body has ultrahigh porosity and provides sufficient space for cell growth. More importantly, controlled-release of the antibody cetuximab can relieve effect myelin protein in a microenvironment after spinal cord injury, promotes transplanted neural stem cells to be differentiated into neurons, restrains transplanted neural stem cells from being differentiated into astroglia cells, and also promotes growth of nerve axons so as to facilitate injured spinal nerve regeneration and functional reconstruction. In addition, the invention further provides an application method of the cylinder support for repairing spinal cord injury.

Description

technical field [0001] The invention belongs to the field of biomedical materials, and relates to a cylindrical support for repairing spinal cord injuries and an application method thereof. Background technique [0002] Spinal cord injury is a common severe central nervous system injury clinically, and is currently recognized as one of the diseases with the highest disability rate in the world. Traditional treatment methods, such as injection of high-dose glucocorticoids, removal of constrictors, and removal of connective tissue scars, etc., have relieved symptoms to varying degrees, but they cannot effectively solve the problem of neurological dysfunction. [0003] Stem cell transplant therapy offers hope for repairing spinal cord injuries. Neural stem cells are a group of cells derived from neural tissue with self-renewal and multidirectional differentiation potential, which can be differentiated into neurons and glial cells, and are one of the ideal sources of transplant...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/38A61L27/24A61L27/54A61L27/56
Inventor 李晓然戴建武
Owner BEIJING LANGJIAYI BIOTECH CO LTD
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