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Tissue engineering nerve complex as well as preparation method and application thereof

A tissue engineering and complex technology, applied in the field of biomedical materials, can solve problems such as unfavorable standardized operation, leakage, abnormal shape of blood vessels, etc., to improve the local microenvironment of injury, reduce immune rejection, and improve local microcirculation. Effect

Active Publication Date: 2022-04-08
NANTONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The disadvantage of using vascular endothelial cells (or endothelial cells derived from stem cell-induced differentiation) to build a vascular network in vitro and then transplant it in vivo to anastomose with the host's vascular network is that to form a stable, long-term, and effective capillary network, it is necessary to form a blood vessel wall. Other cells such as pericytes, smooth muscle cells, etc.
Even though a variety of cells can be used to construct a vascular network in vitro, the process involved is complicated, and the operability and controllability are poor, which is not conducive to the later standardized operation
The disadvantage of vascularization of engineered grafts based on growth factors or cytokines is that these factors are highly unstable in vivo
The transient large release of growth factors in the body cannot effectively promote the occurrence of vascularization, but will cause abnormal morphology and leakage of blood vessels due to excessive concentration of local short-term factors

Method used

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  • Tissue engineering nerve complex as well as preparation method and application thereof
  • Tissue engineering nerve complex as well as preparation method and application thereof
  • Tissue engineering nerve complex as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] 1. Construction of VEGF-iSTB sustained release system

[0048] Prepare 18% w / v gelatin solution (type A, G1890, Sigma, CA, USA) and 10% w / v SNPs solution (Laponite XLG, BYK Additives Ltd, TX, USA) with Milli-Q water respectively; Composite hydrogel iSTB of proportional gelatin and SNPs.

[0049] Biocompatibility, angiogenic effect, and cell migration experiments were carried out on iSTB prepared by different ratios of gelatin and SNPs, human umbilical vein endothelial cells (HUVECs) and human skin fibroblasts (HDF) in vitro, and the construction of VEGF was determined according to the experimental results. - Optimal ratio of gelatin and SNPs in iSTB sustained release system.

[0050] In order to test the angiogenic effect of VEGF-iSTB slow-release system on vascular endothelial cells, human umbilical vein endothelial cells (HUVECs) were cultured in well plates coated with different VEGF-iSTB slow-release hydrogels, and photographed at different time points Recorded, a...

Embodiment 2

[0059] Construction of prevascularized tissue-engineered neural complexes

[0060] VEGF-iSTB tissue-engineered nerve grafts consisting of chitosan nerve guides, built-in silk fibroin scaffolds, and iSTB containing different concentrations of VEGF were pre-implanted subcutaneously on the back of rats, and the gross and histological observations were made at different time points after transplantation Inflammatory immune response and prevascularization of tissue engineered neural complexes.

[0061] VEGF-iSTB was injected into the nerve conduit through a syringe to completely fill the cavity space. Tissue-engineered nerve grafts composed of iSTB containing 1 μg / mL VEGF were pre-implanted subcutaneously on the back of rats, and samples were taken 4 days, 7 days, and 14 days after transplantation for direct observation under a stereomicroscope and immunohistochemical staining (CD34) to observe the tissue Prevascularization of engineered neural complexes.

[0062] Such as Figur...

Embodiment 3

[0065] 1. Evaluation of the effect of prevascularized tissue engineered nerve complex pontogenesis on angiogenesis

[0066]Establish rat sciatic nerve defect model, after intraperitoneal injection of anesthesia, routine skin preparation, incision of the skin and subcutaneous tissue with traces of gluteus gluteus superficialis and biceps femoris, blunt dissection of the oblongus muscle space to expose sciatic nerve, experimental group, autologous group and bracket The rats in group 1 were excised about 10mm of sciatic nerve at the middle part of the femur. The 10 mm defect of rat sciatic nerve was repaired by bridging the prevascularized tissue engineered nerve complex, and the non-prevascularized tissue engineered nerve graft repaired the rat sciatic nerve defect was set up as the negative control group. The tissue-engineered nerve complex and the epineurium were sutured with microscopic sutures respectively, the positions of the nerves of each group in the intermuscular space...

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Abstract

The invention discloses a tissue engineering nerve complex as well as a preparation method and application thereof, and belongs to the field of biomedical materials. The tissue engineering nerve complex comprises a nerve conduit, a fiber scaffold and an injectable shear thinning biological material, wherein the nerve conduit is filled with the fiber scaffold and the injectable shear thinning biological material; the injectable shear thinning biological material (VEGF-iSTB) is prepared from gelatin, nano silicate SNPs, a vascular endothelial growth factor and water. The tissue engineering nerve complex containing VEGF-iSTB is prepared by utilizing a VEGF-iSTB slow release system and is transplanted to the subcutaneous part of a host, so that the pre-vascularized tissue engineering nerve complex is constructed; then, the pre-vascularized tissue engineering nerve complex is used for transplanting and repairing long-distance defects of sciatic nerves of rats, fusion with a host vascular system after transplantation can be achieved, and then the nerve injury repairing effect is improved.

Description

technical field [0001] The invention belongs to the field of biomedical materials, and in particular relates to a tissue engineering nerve complex and its preparation method and application. Background technique [0002] Vascularization is one of the biggest challenges hindering the clinical application of tissue engineering products and technologies. The degree of vascularization of tissue engineered nerve grafts in vivo is closely related to the effect of peripheral nerve repair. Therefore, research on tissue-engineered nerve grafts is far from enough to be limited to biomaterials, seed cells, cytokines, and other active ingredients. How to promote early vascularization of tissue-engineered nerve grafts is particularly important. At present, simulating the complex natural vascular system in vitro is still facing great challenges, and further research on biomaterials and engineering technologies is urgently needed. Establishing an efficient and easy pre-vascularization met...

Claims

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Application Information

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IPC IPC(8): A61L27/54A61L27/50A61L27/22A61L27/02A61L27/20
Inventor 薛成斌王鸿奎朱慧顾晓松杨宇民
Owner NANTONG UNIVERSITY
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