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Construction and application of differentially regulated tissue-engineered nerve grafts

Active Publication Date: 2021-09-02
NANTONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention is about creating a special type of nerve that can help repair different types of nerves. This is important because the traditional methods of repairing nerves can cause damage to one type of nerve or the other. The new method uses a special design and construction process to make sure that the nerve can heal both motor nerves and sensory nerves in a way that is separate and independent from each other. This helps to minimize the risk of damaging the nerves and makes the repair more effective.

Problems solved by technology

After nerve injury, if a catheter only made of material is bridged at a local part of the injury, regeneration modes of motor and sensory nerve fibers are disordered and chaotic, and a misconnection rate of the nerve fibers exists.
In addition, even if the nerve fibers eventually find the target organ to be innervated at a later stage, a rate of nerve regeneration has been severely affected.
More seriously, an effector organ at a distal portion of nerve fiber ending cannot fully recover its function due to long-term denervation, for example, a process of atrophy of target muscle is irreversible.
However, during the actual peripheral nerve repair, a motor nerve and a sensory nerve cannot be well distinguished, and the problem of nerve fiber misconnection still exists.

Method used

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  • Construction and application of differentially regulated tissue-engineered nerve grafts
  • Construction and application of differentially regulated tissue-engineered nerve grafts
  • Construction and application of differentially regulated tissue-engineered nerve grafts

Examples

Experimental program
Comparison scheme
Effect test

embodiment 1

Promoting Effects of Neuroligin-1 and Neuroligin-2 on Synapses in Motor and Sensory Neurons

[0038](1) Culture of motor neurons: a SD rat pregnant for 15 d-16 d was dissected under an anatomical lens to remove ventral tunica, dorsal root ganglia and blood vessels of a spinal cord, and a ventral spinal cord of a fetal rat was carefully separated and placed in a petri dish containing ice-cold anatomical fluid. A tissue piece having a size of 0.5 cm3 was cut by a microscissor and then transferred to a 15 mL centrifuge tube. After adding 0.25% trypsin and 200 U / ml DNase, digestion was performed at 37° C. for 30 min, and then digestion was terminated by adding DMEM+10% FBS. Centrifugation was performed at 1000 r / min for 5 min, and supernatant was discarded. 5 mL of Optiprep separation solution was added, and density gradient centrifugation was performed at 3000 r / min for 15 min. Then liquid in the centrifuge tube was divided into 3 layers, wherein the middle layer contained spinal motor ne...

embodiment 2

Promoting Effect of Neurexin-1β in Combination with Neuroligin-1 or Neuroligin-2 on Synapse Formation

[0043]Motor neurons and sensory neurons were cultured for 10 d in vitro and then divided into groups as follows. Neurexin-1β in combination with Neuroligin-1 or Neuroligin-2 was added to motor neuron and sensory neuron media, respectively:

[0044]Motor Neuron Group:

[0045](1) Neurexin-1β (final concentration of 200 ng / mL)+Neuroligin-1 (final concentration of 200 ng / mL);

[0046](2) Neurexin-1β (final concentration of 1 μg / mL)+Neuroligin-1 (final concentration of 200 ng / mL);

[0047](3) Neurexin-1β (final concentration of 1 μg / mL).

[0048]Sensory Neuron Group:

[0049](4) Neurexin-1β (final concentration of 200 ng / mL)+Neuroligin-2 (final concentration of 200 ng / mL);

[0050](5) Neurexin-1β (final concentration of 1 μg / mL)+Neuroligin-2 (final concentration of 200 ng / mL);

[0051](6) Neurexin-1β (final concentration of 1 μg / mL).

[0052]The culture was continued for 24 h. Results of immunocytochemical stainin...

embodiment 3

Construction of a Differential Tissue-Engineered Nerve According to the Present Invention

[0055]1. Preparation of Sustained-Release Microspheres Containing Neuroligin-1, Neuroligin-2 and Neurexin-1β Recombinant Protein

[0056]Polylactic acid-glycolic acid copolymer (PLGA, a molar ratio of lactic acid to ethanol was 53:47, and molecular weight [MW] was 25 kDa). PLGA microspheres were prepared by a water / oil / water multiple emulsion method. 100 mg of PLGA was dissolved in 1 mL of dichloromethane, and then emulsified into 3 mL of a 7% (w / v) poly(vinyl alcohol) (PVA) aqueous solution, and ultrasonicated for 1 min to prepare an emulsion of a first step. Then, the above solution was added to 50 mL of a 1% (w / v) poly(vinyl alcohol) (PVA) aqueous solution (containing 2% isopropanol), and ultrasonicated again for 3 min to form an emulsion of a second step. The emulsion of the second step as mentioned above was slowly stirred overnight at room temperature, and the residual dichloromethane was ful...

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Abstract

A differential tissue-engineered nerve including motor-like nerves and sensory-like nerves. The motor-like nerve and the sensory-like nerve respectively includes a motor-like nerve outer tube and a motor-like nerve fiber in the outer tube as well as a sensory-like nerve outer tube and a sensory-like nerve fiber in the outer tube. Schwann cells and / or fibroblasts derived from motor nerves and sensory nerves are respectively contained in surfaces or pores of the motor-like and sensory-like nerve outer tubes. Transsynaptic signal molecules Neuroligin-1 and Neuroligin-2 are contained in surfaces or pores of the motor-like and sensory-like nerve fibers. Neuroligin-1 is selectively used to specifically promote synaptic remodeling of motor neurons, while Neuroligin-2 is selectively used to specifically promote synaptic remodeling of sensory neurons, so that repair efficiency of motor nerve cells and sensory nerve cells is improved.

Description

BACKGROUNDTechnical Field[0001]The present invention belongs to the field of biomedicine, and particularly relates to a differential tissue-engineered nerve, which can differentially regulate repair of motor nerves and sensory nerves.Related Art[0002]Peripheral nerve injury is a common clinical disease. With social and economic development and aging of a population structure, acute trauma and chronic degenerative diseases resulting from accidents lead to a significant increase trend in peripheral neuropathy. The peripheral nerve injury leads to impaired motor and sensory functions in a corresponding part of a patient. Recovery of the functions after injury depends mainly on precise, selective, and guided growth of regenerative axons for specific motor and sensory pathways. Clinically, nerve repair is mainly performed by direct suture without tension at broken ends, autologous or allogeneic nerve transplantation, tissue-engineered nerve grafts and the like. Because the direct suture ...

Claims

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Application Information

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IPC IPC(8): A61L27/36A61L27/38A61L27/54A61L27/56
CPCA61L27/3675A61L27/383A61L27/54A61L2430/32A61L2300/252A61L2300/412A61L2300/622A61L27/56A61L27/3804A61L27/50A61L2300/62A61L27/3813
Inventor TANG, XINSUN, CHENGCHEN, GANGGU, XIAOSONGYANG, YUMINDING, FEIGONG, YANPEIHE, QIANRUWANG, HONGKUISHEN, MI
Owner NANTONG UNIVERSITY
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