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A kind of tissue engineered nerve graft and preparation method thereof

A nerve graft and tissue engineering technology, applied in the field of tissue engineered nerve grafts, can solve the problems of lack of biocompatibility, three-dimensional ideal permeable scaffold, difficulty in obtaining neuron cells, etc., so as to avoid gene integration and mutation. Potential safety hazards, promotion of nerve regeneration and functional recovery, minimal trauma

Active Publication Date: 2021-09-28
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the technical problem that needs to be solved urgently is the difficulty in obtaining neuron cells, and the lack of an ideal transparent scaffold with good biocompatibility and three-dimensional communication.

Method used

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  • A kind of tissue engineered nerve graft and preparation method thereof
  • A kind of tissue engineered nerve graft and preparation method thereof
  • A kind of tissue engineered nerve graft and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Preparation of tissue-engineered nerve grafts

[0045] according to figure 1 Schematic diagram of the schematic diagram, first construct the silk protein scaffold, extract the seed cell fibroblasts; then seed the fibroblasts on the silk protein scaffold, expand and culture; finally induce the fibroblasts in the scaffold to transdifferentiate into neurons by small molecule compounds, and The connection between the axons of neurons is established to form a neural network, thereby obtaining a tissue-engineered nerve graft.

[0046] 1) Construction of porous scaffolds of silk protein micro-nanofibers

[0047] After the silk is degummed by boiling at 0.05% (w / v), it is dispersed in 98% formic acid containing 0.1% calcium chloride (w / v) (silk:solution ratio 1:50), and the silk dispersion is placed at -20°C Freeze for 24 hours, then thaw in 75% ethanol for 1 hour, then fully wash with deionized water, and freeze-dry the obtained wet silk protein scaffold to obtain a three-di...

Embodiment 2

[0058] Preparation of tissue-engineered nerve grafts

[0059] according to figure 1 Schematic diagram of the schematic diagram, first construct silk protein scaffolds, extract seed cells human skin fibroblasts; then seed fibroblasts on silk protein scaffolds, expand and culture; finally induce the fibroblasts in the scaffolds to transdifferentiate into neurons by small molecule compounds , and establish the connection between the axons of neurons to form a neural network, thereby obtaining a tissue engineered nerve graft.

[0060] 1) Construction of porous scaffolds of silk protein micro-nanofibers

[0061] After the silk is degummed by boiling at 0.05% (w / v), it is dispersed in 98% formic acid containing 0.1% calcium chloride (w / v) (silk:solution ratio 1:50), and the silk dispersion is placed at -20°C Freeze for 24 hours, then thaw in 75% ethanol for 1 hour, then fully wash with deionized water, and freeze-dry the obtained wet silk protein scaffold to obtain a three-dimensi...

Embodiment 3

[0069] Transplantation of tissue-engineered neural grafts in the treatment of spinal cord injuries

[0070] 1) Establishment of spinal cord transection model

[0071] The rat spinal cord transection model was constructed, and the spinal cord tissue with a length of 2 mm at the T10 level was excised.

[0072] 2) Transplant tissue engineered neural grafts

[0073] Filling the spinal cord defect with tissue engineered nerve grafts, such as Figure 7 shown.

[0074] 3) Behavioral observation and scoring

[0075] Put the animal into an open pot, tap the pot wall to make it crawl, and observe the animal's buttocks, knees, ankles, trunk movement and coordination.

[0076] Such as Figure 8 with 9 As shown, it can be seen from the BBB score that all mice were completely paralyzed 1 day after the operation, indicating that the modeling was successful. After stent implantation, the BBB score increased gradually, and there were significant differences among the groups within 1 wee...

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Abstract

The invention provides a preparation method of a tissue engineered nerve graft, comprising the following steps: constructing a three-dimensional silk protein fiber porous scaffold; inoculating fibroblasts on the scaffold, and culturing and expanding the cells; adding a small molecule compound combination CFLSSVY to induce fibrogenesis Cells differentiate into neurons and form neuronal networks. The preparation method of the invention is simple and efficient, and the silk protein material used has a wide source, the source of adult cells is convenient, and the preparation is easy to be standardized. The present invention provides a bionic tissue engineered nerve graft in the true sense, which not only contains a permeable silk protein fiber scaffold that is beneficial to the growth of cells and tissues, but also contains a neuron network formed by seed cell neurons, which is a peripheral It is an ideal graft for the repair of central nervous system injury, and has broad application prospects and value.

Description

technical field [0001] The invention relates to the field of nerve injury transplantation of tissue engineering, in particular to a tissue engineered nerve graft used for repairing long-distance peripheral nerve defects and central nerve damage. Background technique [0002] There are millions of patients with organ or tissue damage and functional loss due to diseases and accidents every year. In the United States alone, more than 8 million operations are needed to treat such patients every year, and the economic cost is more than 400 billion US dollars. With the development of modern medicine and surgical techniques, tissue or organ transplantation to repair functional loss has been widely accepted, however, it faces a huge donor gap. The formation of tissues or organs in vivo or in vitro by means of regenerative medicine provides a new treatment option for the repair of damaged functions. Among them, the construction of tissue engineered grafts has become one of the keys ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/22A61L27/38A61L27/56
CPCA61L27/227A61L27/3804A61L27/56A61L2430/32C08L89/00
Inventor 张锋胡雅楠张焕相范志海秦建忠金志高沈影超杨敏何其圣
Owner SUZHOU UNIV
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