Biological material for promoting optic nerve injury repair and preparation method thereof

Abstract

The invention discloses a biological material for promoting optic nerve injury repair and a preparation method thereof.The biological material comprises a bionic nerve scaffold, a growth factor compound, an anti-inflammatory component and a light-operated neuroelectric signal guiding layer, the bionic nerve scaffold comprises a polycaprolactone-polyethylene glycol block copolymer and self-assembly neuropeptide, the growth factor compound comprises a neurosynaptic growth factor and a vascular endothelial growth factor, the anti-inflammatory component is liposome nanoparticles loaded with interleukin-10, and the light-operated neuroelectric signal guide layer is a polypyrrole film; the oriented nanofiber scaffold of polycaprolactone-polyethylene glycol and self-assembled neuropeptide is prepared by adopting a micro-fluidic spinning technology, a three-dimensional porous scaffold is formed to simulate a natural nerve bundle topological structure and guide oriented regeneration of axons, long-acting slow release is realized through gene activated growth factors, the axon extension efficiency is effectively improved, and the growth rate of the axons is increased. Through the interleukin-10 lipidosome anti-inflammatory component, the inflammation inhibition rate is increased, and secondary injury is relieved.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of optic nerve injury repair, and particularly relates to a biomaterial for promoting optic nerve injury repair and a preparation method thereof. BACKGROUND

[0002] Optic nerve injury repair refers to a process of promoting regeneration and functional reconstruction of damaged optic nerves, reconstructing the integrity and signal transduction function of the optic nerves, and restoring or improving visual function through medical intervention. The core mechanisms include: activating axon regeneration by using neurotrophic factors, reducing secondary damage by using anti-inflammatory drugs or hyperbaric oxygen therapy, bridging nerve defects and providing physical support by using biological materials, and supplementing damaged cells by using gene editing or stem cell transplantation. The repair effect is affected by the type of injury, the timing of treatment and individual differences. Some severe injuries may be irreversible. Current frontier technologies are exploring innovative methods such as optogenetics regulation and intelligent response materials to improve repair efficiency.

[0003] As an important part of the central nervous system, optic nerve injury often leads to irreversible vision loss, with millions of new patients worldwide each year. Traditional treatment methods such as hormone shock, surgical decompression or neurotrophic drugs (such as mecobalamin) can only short-term inhibit secondary damage and cannot achieve axon regeneration and functional reconstruction. The reason is that adult mammalian retinal ganglion cells (RGCs) have very low regenerative capacity, and there are strong inhibitory signals in the local microenvironment after injury. In recent years, multidisciplinary research has provided new ideas for breaking this bottleneck: bioactive materials (such as ciliary neurotrophic factor chitosan) can extend the nutritional support time to 12 weeks by releasing neurotrophic factors, promote optic nerve regeneration distance up to 18 mm, and restore visual electrophysiological function; gene editing technology (such as CRISPR / Cas9 modification of NSF gene) combined with adeno-associated virus vector significantly increases the expression of TrkB receptor and enhances the efficiency of axon extension; stem cell and organoid technology simulates the neural regeneration microenvironment by constructing a three-dimensional retinal sheet. However, single therapy still faces the bottleneck of low regeneration efficiency, immune rejection and insufficient functional integration. Therefore, the present application proposes a biomaterial for promoting optic nerve injury repair and a preparation method thereof to solve the problems existing in the prior art. SUMMARY

[0004] In view of the above problems, the purpose of the present application is to provide a biomaterial for promoting optic nerve injury repair and a preparation method thereof, which prepares a directional nanofiber scaffold of polycaprolactone-polyethylene glycol and self-assembled neuropeptide by using microfluidic spinning technology, forms a three-dimensional porous scaffold to simulate the natural nerve bundle topology, guides the directional regeneration of axons, realizes long-acting release by gene-activated growth factors, effectively improves the efficiency of axon extension, and reduces secondary injury by improving the inflammation inhibition rate through interleukin-10 liposome anti-inflammatory components.

[0005] To achieve the purpose of the present application, the present application realizes the following technical solutions: a biomaterial for promoting optic nerve injury repair and a preparation method thereof, which comprises a biomimetic nerve scaffold, a growth factor compound, an anti-inflammatory component and a light-controlled nerve electrical signal guide layer, the biomimetic nerve scaffold comprises a polycaprolactone-polyethylene glycol block copolymer and a self-assembled neuropeptide, the growth factor compound comprises a neurosynaptic growth factor and a vascular endothelial growth factor, the anti-inflammatory component is an interleukin-10-loaded liposome nanoparticle, and the light-controlled nerve electrical signal guide layer is a polypyrrole film.

[0006] Further improvement lies in that the polycaprolactone-polyethylene glycol block copolymer is combined with the self-assembled neuropeptide to form a three-dimensional porous structure, the porosity is greater than or equal to 80%, and the pore size is 50-200 nm.

[0007] Further improvement lies in that the molecular weight of the polycaprolactone-polyethylene glycol block copolymer is 50000-100000, the mass ratio of polycaprolactone to polyethylene glycol in the polycaprolactone-polyethylene glycol block copolymer is 7:3, and the concentration of the self-assembled neuropeptide is 0.5-2 wt%.

[0008] Further improvement lies in that the nucleic acid sequence of the neurosynaptic growth factor comprises a promoter enhancer element targeting a neurotrophic factor receptor.

[0009] Further improvement lies in that MXene quantum dots are doped in the polypyrrole film, the lateral size of the MXene quantum dots is 5-10 nm, the doping concentration is 0.1-0.5 wt%, the local electric field strength is regulated to 0.1-10 mV / mm by 808 nm near-infrared light, the near-infrared light power density is 1-5 W / cm², and the irradiation time is 10-30 minutes.

[0010] A biomaterial preparation method for promoting optic nerve injury repair, comprising the following steps:

[0011] Step one, dissolve the polycaprolactone-polyethylene glycol block copolymer in tetrahydrofuran, add a self-assembled neuropeptide solution, and form a directional nanofiber scaffold under the condition of pH 7.4 by using a microfluidic chip;

[0012] Step 2: The neural synaptic growth factor gene vector and vascular endothelial growth factor mRNA are encapsulated in lipid nanoparticles and deposited in the pores of the scaffold.

[0013] Step 3: Under low temperature conditions, lipid nanoparticles are combined with polypyrrole film and deposited onto the surface of the biomimetic neural scaffold by electrostatic adsorption to obtain the biomaterial that promotes the repair of optic nerve damage.

[0014] The application method of this biomaterial is as follows: the biomaterial drug is delivered to the damaged site through an implantable microneedle array, and the electrical signal conduction is activated by irradiation with 808nm near-infrared light for 10 minutes.

[0015] A further improvement is that the concentration of the self-assembled neuropeptide solution in step one is 1 mg / mL, the channel width of the microfluidic chip is 50 μm, and the flow rate ratio is tetrahydrofuran phase: peptide solution phase = 4:1.

[0016] The further improvement lies in the fact that the lipid nanoparticles in step two have an encapsulation rate of ≥90% and a release half-life of 72 hours.

[0017] A further improvement is that the implantable microneedle array in step four uses shape memory hydrogel poly-N-isopropylacrylamide, and the microneedle array expands upon contact with body fluid to form a microchannel with a diameter of 200μm.

[0018] The beneficial effects of this invention are as follows: This invention uses microfluidic spinning technology to prepare a directional nanofiber scaffold of polycaprolactone-polyethylene glycol and self-assembled neuropeptides, forming a three-dimensional porous scaffold that simulates the topology of natural nerve bundles, guiding the directional regeneration of axons, achieving long-term sustained release through gene-activated growth factors, effectively improving axon extension efficiency, and enhancing the inflammation suppression rate and reducing secondary damage through the anti-inflammatory component of interleukin-10 liposomes. Attached Figure Description

[0019] Figure 1 This is a flowchart of the steps of the present invention. Detailed Implementation

[0020] To enhance understanding of the present invention, the present invention will be further described in detail below with reference to embodiments. These embodiments are only used to explain the present invention and do not constitute a limitation on the scope of protection of the present invention.

[0021] Unless otherwise specified, the technical means not described in detail in this invention are conventional technical means well known in the art and will not be elaborated here.

[0022] Example 1

[0023] according to Figure 1As shown, this embodiment provides a biomaterial for promoting optic nerve injury repair and its preparation method, including a biomimetic neural scaffold, a growth factor complex, an anti-inflammatory component, and a light-controlled neural electrical signal guiding layer. The biomimetic neural scaffold comprises a polycaprolactone-polyethylene glycol block copolymer and a self-assembled neuropeptide. The polycaprolactone-polyethylene glycol block copolymer and the self-assembled neuropeptide are combined to form a three-dimensional porous structure with a porosity ≥80% and a pore size of 50~200 nm. A multi-dimensional repair mechanism is achieved through the combination of structure guidance, bioactivity, anti-inflammation, and electrical stimulation, significantly improving repair efficiency. The three-dimensional porous structure formed by the polycaprolactone-polyethylene glycol block copolymer and the self-assembled neuropeptide is adapted to cell migration and nutrient transport, providing an ideal microenvironment for nerve regeneration. The molecular weight of the polycaprolactone-polyethylene glycol block copolymer is 50,000 to 100,000, and the mass ratio of polycaprolactone to polyethylene glycol in the polycaprolactone-polyethylene glycol block copolymer is 7:3. By controlling the ratio and molecular weight of the polycaprolactone-polyethylene glycol block copolymer, the mechanical strength and hydrophilicity of the biomimetic neural scaffold are balanced. The concentration of the self-assembled neuropeptide is 0.5 to 2 wt%. Through the synergistic effect of multiple components, four major functions are achieved simultaneously: structural support, nerve regeneration activation, inflammation suppression, and electrical signal regulation, covering the key needs of optic nerve injury repair.

[0024] The growth factor complex includes synaptic growth factor and vascular endothelial growth factor. The nucleic acid sequence of synaptic growth factor contains a promoter enhancement element that targets the neurotrophic factor receptor. Long-term sustained release is achieved through gene-activated synaptic growth factor, avoiding the risk of immune rejection from direct use of exogenous proteins. The mRNA form reduces the risk of genome integration and improves safety.

[0025] The anti-inflammatory component is liposome nanoparticles loaded with interleukin-10, which effectively inhibits the release of pro-inflammatory factors at the site of injury and reduces secondary nerve damage. The liposomes, as a carrier, improve the stability and bioavailability of interleukin-10, prolong its duration of action at the site of injury, and reduce the frequency of administration by utilizing the sustained-release properties, thus reducing the treatment burden on patients.

[0026] The photocontrolled neural signal guiding layer is a polypyrrole film. The polypyrrole film is doped with MXene quantum dots, with a lateral size of 5–10 nm and a doping concentration of 0.1–0.5 wt%. The local electric field intensity is modulated to 0.1–10 mV / mm using 808 nm near-infrared light, with a near-infrared light power density of 1–5 W / cm² and an irradiation time of 10–30 minutes. The MXene quantum dots enhance the conductivity of polypyrrole, and the near-infrared light excites the photothermal effect of MXene, dynamically adjusting the conductivity of polypyrrole to control the electric field intensity.

[0027] A method for preparing a biomaterial that promotes the repair of optic nerve damage includes the following steps:

[0028] Step 1: Dissolve polycaprolactone-polyethylene glycol block copolymer in tetrahydrofuran, add self-assembled neuropeptide solution, and form oriented nanofiber scaffolds under pH 7.4 conditions using a microfluidic chip;

[0029] Self-assembled neuropeptides are a class of short peptide materials that spontaneously form ordered nanofiber structures through intermolecular non-covalent interactions (such as hydrogen bonds and hydrophobic interactions). The concentration of the self-assembled neuropeptide solution is 1 mg / mL, the channel width of the microfluidic chip is 50 μm, and the flow rate ratio is tetrahydrofuran phase: peptide solution phase = 4:1. By precisely controlling the flow rate and direction of the solution through microfluidic technology, the polycaprolactone-polyethylene glycol block copolymer and neuropeptides are uniformly mixed and oriented to form a nanofiber network. This satisfies both the exchange of nutrients and guides axon regeneration along the nerve pathway through the oriented fiber structure.

[0030] Step 2: The neural synaptic growth factor gene vector and vascular endothelial growth factor mRNA are encapsulated and deposited in the pores of the scaffold using lipid nanoparticles; the neural synaptic growth factor gene vector is nerve growth factor (NGF).

[0031] The lipid nanoparticles consist of four lipid components that self-assemble into multilayer or monolayer structures: ionizable cationic lipids, phospholipids, cholesterol, and polyethylene glycol lipids. The lipid nanoparticles have an encapsulation efficiency of ≥90% and a release half-life of 72 hours, enabling long-term sustained release of synaptic growth factor and avoiding the risk of immune rejection associated with direct use of exogenous proteins.

[0032] Step 3: Under low temperature conditions (0~4℃), lipid nanoparticles loaded with interleukin-10 are combined with polypyrrole films and deposited onto the surface of a biomimetic neural scaffold by electrostatic adsorption to obtain a biomaterial that promotes the repair of optic nerve damage. Interleukin-10 inhibits the release of pro-inflammatory factors and reduces secondary nerve damage. Liposomes improve the stability and bioavailability of interleukin-10. Interleukin-10 is mixed using microfluidic mixing technology with Y-type or T-type microfluidic chips to control the flow rate ratio of the lipid phase (ethanol) to the aqueous phase (IL-10 buffer) (e.g., 1:3) to form a laminar shear force driven self-assembly.

[0033] The application method of this biomaterial is as follows: the biomaterial drug is delivered to the damaged site through an implantable microneedle array, and the electrical signal conduction is activated by irradiation with 808nm near-infrared light for 10 minutes.

[0034] The implantable microneedle array uses shape memory hydrogel poly-N-isopropylacrylamide. When the microneedle array expands upon contact with body fluid, it forms a microchannel with a diameter of 200μm. At room temperature, the microneedles are rigid and easy to puncture. After implantation, they expand upon contact with body temperature and are fixed at the site of injury.

[0035] This biomaterial for promoting optic nerve injury repair and its preparation method first employs microfluidic spinning technology to prepare a directional nanofiber scaffold of polycaprolactone-polyethylene glycol and self-assembled neuropeptides, whose three-dimensional porous structure is adapted for cell migration and nutrient transport. Subsequently, lipid nanoparticles encapsulate synaptic growth factor gene carriers and vascular endothelial growth factor mRNA, precisely loading them into the scaffold pores to achieve long-term sustained release of gene-activated growth factors. Next, liposomes loaded with interleukin-10 and a polypyrrole film doped with MXene quantum dots are integrated onto the scaffold surface using electrostatic adsorption to form a photocontrolled electrical signal layer, where the local electric field is modulated by 808 nm near-infrared light. Finally, a shape memory microneedle array is used to encapsulate the biomaterial; upon implantation at the injury site, it expands upon contact with body fluid to form a 200 μm microchannel, achieving minimally invasive delivery and precise electrical stimulation activation. This technology, through the multi-dimensional synergistic effects of structure guidance, gene regulation, anti-inflammatory protection, and photocontrolled electrical signals, significantly improves optic nerve regeneration efficiency, providing an efficient solution for optic nerve injury repair.

[0036] The foregoing has shown and described the basic principles, main features, and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The embodiments and descriptions in the specification are merely illustrative of the principles of the invention. Various changes and modifications can be made to the invention without departing from its spirit and scope, and all such changes and modifications fall within the scope of the present invention as claimed. The scope of protection of the present invention is defined by the appended claims and their equivalents.

Claims

1. A biomaterial for promoting optic nerve injury repair, comprising a biomimetic neural scaffold, a growth factor complex, an anti-inflammatory component, and a light-controlled neural signal guiding layer, wherein the biomimetic neural scaffold comprises a polycaprolactone-polyethylene glycol block copolymer and a self-assembled neuropeptide, the growth factor complex comprises synaptic growth factor and vascular endothelial growth factor, the anti-inflammatory component is liposome nanoparticles loaded with interleukin-10, and the light-controlled neural signal guiding layer is a polypyrrole film.

2. The biomaterial for promoting optic nerve injury repair according to claim 1, characterized in that: The polycaprolactone-polyethylene glycol block copolymer is combined with self-assembled neuropeptides to form a three-dimensional porous structure with a porosity of ≥80% and a pore size of 50~200nm.

3. The biomaterial for promoting optic nerve injury repair according to claim 1, characterized in that: The molecular weight of the polycaprolactone-polyethylene glycol block copolymer is 50,000 to 100,000, the mass ratio of polycaprolactone to polyethylene glycol in the polycaprolactone-polyethylene glycol block copolymer is 7:3, and the concentration of the self-assembled neuropeptide is 0.5 to 2 wt%.

4. The biomaterial for promoting optic nerve injury repair according to claim 1, characterized in that: The nucleic acid sequence of the neurosynaptic growth factor contains a promoter enhancement element that targets the neurotrophic factor receptor.

5. The biomaterial for promoting optic nerve injury repair according to claim 1, characterized in that: The polypyrrole film is doped with MXene quantum dots, the lateral size of which is 5~10 nm and the doping concentration is 0.1~0.5 wt%. The local electric field intensity is modulated to 0.1~10 mV / mm by 808nm near-infrared light, the near-infrared light power density is 1~5 W / cm², and the irradiation time is 10~30 minutes.

6. A method for preparing a biomaterial to promote optic nerve injury repair according to any one of claims 1-5, comprising the following steps: Step 1: Dissolve polycaprolactone-polyethylene glycol block copolymer in tetrahydrofuran, add self-assembled neuropeptide solution, and form oriented nanofiber scaffolds under pH 7.4 conditions using a microfluidic chip; Step 2: The neural synaptic growth factor gene vector and vascular endothelial growth factor mRNA are encapsulated in lipid nanoparticles and deposited in the pores of the scaffold. Step 3: Under low temperature conditions, lipid nanoparticles are combined with polypyrrole film and deposited onto the surface of the biomimetic neural scaffold by electrostatic adsorption to obtain the biomaterial that promotes the repair of optic nerve damage.

7. The method for preparing a biomaterial to promote optic nerve injury repair according to claim 6, characterized in that: The concentration of the self-assembled neuropeptide solution in step one is 1 mg / mL, the channel width of the microfluidic chip is 50 μm, and the flow rate ratio is tetrahydrofuran phase: peptide solution phase = 4:

1.

8. The method for preparing a biomaterial to promote optic nerve injury repair according to claim 6, characterized in that: The lipid nanoparticles in step two have an encapsulation rate of ≥90% and a release half-life of 72 hours.

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