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Functional biomaterial for inducing tissue regeneration after craniocerebral injury and application thereof

A technology for tissue regeneration and functional materials, applied in the fields of applications, biochemical equipment and methods, growth factors/growth regulators, etc., which can solve the problems of difficulty in maintaining VEGF, maintaining effective concentration, and rapid diffusion of soluble VEGF with a short half-life.

Pending Publication Date: 2018-07-31
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main problem in the application of exogenous VEGF is the short half-life of soluble VEGF and the rapid diffusion into the cerebrospinal fluid.
These problems make it difficult for VEGF in the brain to maintain an effective concentration in the injured area

Method used

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  • Functional biomaterial for inducing tissue regeneration after craniocerebral injury and application thereof
  • Functional biomaterial for inducing tissue regeneration after craniocerebral injury and application thereof
  • Functional biomaterial for inducing tissue regeneration after craniocerebral injury and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0098] Embodiment 1, the preparation of collagen gel

[0099] 1. Take the isolated bovine tendon tissue from which fat and muscle have been removed.

[0100] 2. Take the tissue obtained in step 1 and follow the steps below:

[0101] (1) Soak in 1M NaCl aqueous solution for 24 hours, then wash with deionized water 5 times, 3 hours each time.

[0102] (2) Soak in PBS buffer solution containing 50U / ml DNaseI for 3 hours, then wash with deionized water 5 times, 3 hours each time.

[0103] (3) Soak in 1M KOH aqueous solution for 20 minutes, then wash with deionized water 10 times, 30 minutes each time.

[0104] (4) Dissolve in 0.5M acetic acid aqueous solution for 48 hours, centrifuge at 12000 g for 20 minutes to remove the precipitate, and take the supernatant.

[0105] (5) The supernatant was dialyzed with a dialysis bag, and the external liquid was deionized water, and dialyzed to neutrality to obtain a collagen gel.

[0106] Scanning electron micrographs of collagen gels as...

Embodiment 2

[0107] Embodiment 2, growth factor preparation

[0108] 1. The small fragment between the XbaI and XhoI restriction sites of the pET-28a(+) vector was replaced with the DNA molecule shown in the 1st-552th position of Sequence 1 in the sequence listing to obtain the recombinant plasmid pET28a-CBD-VEGF. The exogenously inserted DNA molecule is fused with part of the nucleotides on the carrier backbone to form a fusion gene shown in sequence 1 of the sequence listing, which encodes a fusion protein shown in sequence 2 of the sequence listing.

[0109] In sequence 2 of the sequence listing, amino acid residues 1-166 from the N-terminal are VEGF segments, amino acid residues 178-184 are CBD segments, and amino acid residues 187-192 are histidine tags.

[0110] In Sequence 1 of the sequence listing, nucleotides 1-498 from the 5' end are the coding gene of the VEGF segment, nucleotides 532-552 are the gene coding for the CBD segment, and nucleotides 559-576 are Acid is the coding se...

Embodiment 3

[0130] Embodiment 3, the preparation of collagen gel repair material

[0131] Prepare the following collagen gel repair materials:

[0132] 1, 125 I-NAT-VEGF collagen gel repair material: 10 μL of Na prepared in Example 2 125 I-labeled NAT-VEGF protein solution (protein content: 0.5 nmol) was mixed with 190 μL of the collagen gel prepared in Example 1.

[0133] 2, 125 I-CBD-VEGF collagen gel repair material: 10 μL of Na prepared in Example 2 125 The I-labeled CBD-VEGF protein solution (protein content: 0.5 nmol) was mixed with 190 μL of the collagen gel prepared in Example 1.

[0134] 3. NAT-VEGF collagen gel repair material: 10 μL of the unlabeled NAT-VEGF protein solution (protein content: 0.5 nmol) prepared in Example 2 was mixed with 190 μL of the collagen gel prepared in Example 1.

[0135] 4. CBD-VEGF collagen gel repair material: Mix 10 μL of the unlabeled CBD-VEGF protein solution (protein content: 0.5 nmol) prepared in Example 2 with 190 μL of the collagen gel pr...

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Abstract

The invention discloses a functional biomaterial for inducing tissue regeneration after craniocerebral injury and application thereof. A preparation method for the functional biomaterial comprises a step of mixing collagen gel with a vascular endothelial growth factor with a collagen binding zone, wherein a ratio of the vascular endothelial growth factor with the collagen binding zone to the collagen gel is 0.5 nmol: 190 [mu]L. Through gene modification of the vascular endothelial growth factor, the vascular endothelial growth factor can specifically bind to collagen, and then the obtained mixture is loaded on a collagen support material so as to form a functional repair system capable of effectively releasing the vascular endothelial growth factor in a long time; so repair of cerebral injury can be effectively promoted.

Description

technical field [0001] The invention relates to a functional biological material for inducing tissue regeneration of craniocerebral injury and its application. Background technique [0002] Traumatic brain injury (Traumatic brain iniury, TBI), which can cause brain structural damage, is an important cause of death and disability, especially in children and young adults. About 75,000 to 100,000 people die from TBI every year in the world, and about 70,000 to 90,000 TBI survivors suffer from long-term or permanent disabilities. At present, there are almost no effective therapeutic methods that can prevent nerve injury after TBI or effectively promote the regeneration of nerve tissue. [0003] After brain injury, biomaterials can provide a scaffold for the repair of damaged tissue and guide the migration of cells. However, the use of biomaterials alone often has no activity-inducing function, and the therapeutic effect is not good. [0004] On the other hand, angiogenesis is...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/24A61L27/52A61L27/54C12N15/63C07K14/71C12N15/12
Inventor 戴建武陈冰赵燕南关健侯祥林肖志峰
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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