CN-121971602-A - Injectable bioactive matrix hydrogels with anti-angiogenic and anti-neurogenic microenvironment and uses thereof

Abstract

The invention discloses an injectable bioactive matrix hydrogel with anti-angiogenesis and anti-neurogenesis microenvironment and application thereof. The injectable bioactive matrix hydrogel comprises bevacizumab, concentrated growth factors and methacrylic chondroitin sulfate. The hydrogels prepared by the invention not only exhibit excellent injectability and biocompatibility, but also inhibit pathological vascularization and neurogenesis, polarize macrophages to promote regeneration of the M2 phenotype to reduce inflammation, and directly enhance deposition of cartilage-specific ECM components (such as type II collagen and aggrecan). By utilizing the coordinated multi-mechanism strategy, the comprehensive structure and function repair of the degenerated intervertebral disc is realized.

Inventors

• TIAN BAOXING
• HUANG MEILIN
• LI CHENYANG
• HOU MENGJIE
• Bai Baoshuai
• WANG BIN
• LIN XIAOXI
• LEI DONG
• HU BAOYANG
• LUO JIE
• XIE SHANHONG

Assignees

• 上海交通大学医学院附属第九人民医院

Dates

Publication Date

20260505

Application Date

20260402

Claims (10)

1. 1. An injectable bioactive matrix hydrogel having anti-angiogenic and anti-neurogenic microenvironment, wherein the composition of the injectable bioactive matrix hydrogel comprises bevacizumab, concentrated growth factor, and methacryated chondroitin sulfate.
2. 2. The injectable bioactive matrix hydrogel having anti-angiogenic and anti-neurogenic microenvironment according to claim 1, wherein the concentration of bevacizumab in the injectable bioactive matrix hydrogel is 10ug/mL and the concentration of concentrated growth factor is 10mg/mL.
3. 3. The injectable bioactive matrix hydrogel with anti-angiogenesis and anti-neurogenesis microenvironment according to claim 1 or 2, wherein the preparation method of the methacrylic acid chondroitin sulfate comprises the steps of dissolving the chondroitin sulfate in deionized water to form a chondroitin sulfate solution, then dropwise adding methacrylic anhydride into the chondroitin sulfate solution, stirring at a low temperature for reaction, purifying the obtained product, and freeze-drying to obtain the methacrylic acid chondroitin sulfate.
4. 4. The injectable bioactive matrix hydrogel with anti-angiogenic and anti-neurogenic microenvironment according to claim 1 or 2, wherein the preparation method of the concentrated growth factor comprises the steps of centrifuging collected blood at speeds of 2700 rpm, 2400 rpm, 2700 rpm and 3000 rpm for 2 minutes, 4 minutes and 3 minutes in sequence, standing for delamination, extracting an intermediate layer, and subjecting the extracted intermediate layer to pre-cooling, freeze-drying and powdering treatment to obtain the concentrated growth factor.
5. 5. A method of preparing an injectable bioactive matrix hydrogel having anti-angiogenic and anti-neurogenic microenvironment according to any one of claims 1 to 4, comprising the steps of: S1, dissolving methacrylic acid chondroitin sulfate in a solvent containing a photoinitiator to obtain a precursor solution a; s2, adding bevacizumab and concentrated growth factors into the precursor solution a, and uniformly stirring to obtain a precursor solution b; and S3, inducing photo-crosslinking of the precursor solution b under the condition of UV illumination to obtain the injectable bioactive matrix hydrogel.
6. 6. The method of preparing an injectable bioactive matrix hydrogel having anti-angiogenic and anti-neurogenic microenvironment according to claim 5, wherein in step a, the concentration of the methacryiated chondroitin sulfate in the precursor solution a is 3-7%.
7. 7. The method of preparing an injectable bioactive matrix hydrogel having anti-angiogenic and anti-neurogenic microenvironment according to claim 6, wherein the concentration of the methacrying chondroitin sulfate in the precursor solution a is 5%.
8. 8. The method of preparing an injectable bioactive matrix hydrogel having anti-angiogenic and anti-neurogenic microenvironment according to claim 5, wherein in step a, the photoinitiator is selected from the group consisting of phenyl-2, 4, 6-trimethylbenzoyl lithium phosphonate; in the solvent containing the photoinitiator, the concentration of the photoinitiator is 0.2-0.5%.
9. 9. The method of claim 5, wherein in step C, the UV light is at a wavelength of 320-400nm, an illumination intensity of 8-12 mW/cm2, and an illumination time of 3-8 seconds.
10. 10. Use of an injectable bioactive matrix hydrogel according to any one of claims 1-4 or an injectable bioactive matrix hydrogel prepared according to the method of any one of claims 5-9 in the preparation of a product for repair of disc degeneration and/or promotion of cartilage regeneration.

Description

Injectable bioactive matrix hydrogels with anti-angiogenic and anti-neurogenic microenvironment and uses thereof Technical Field The invention relates to the technical field of bone regeneration, in particular to injectable bioactive matrix hydrogel with anti-angiogenesis and anti-neurogenesis microenvironment and application thereof. Background Lower Back Pain (LBP) is a huge global health challenge, leading to years of global disability. It places a heavy burden on individuals and medical systems. The number of people disabled by LBP has increased by 54% over the last three decades, and this trend is expected to continue. This rising trend, along with a decrease in the quality of life of the patient and an increase in medical expenditure, together put a tremendous strain on social resources. Among them, disc degeneration (intervertebral disc degeneration, IDD) is the main pathological basis. The pathogenesis of IDD is multifactorial and complex, characterized by a deleterious cascade of reactions involving apoptosis of the nucleus pulposus (nucleus pulposus, NP), impaired extracellular matrix (extracellular matrix, ECM) metabolism, and a chronic inflammatory microenvironment. Thus, in addition to palliative symptom management, there is an urgent need to develop therapeutic strategies that can effectively stop disc degeneration and promote recovery of disc function. Disclosure of Invention In view of the shortcomings of the prior art, it is an object of the present invention to provide an injectable bioactive matrix hydrogel with anti-angiogenic and anti-neurogenic microenvironment and uses thereof. The invention aims at realizing the following technical scheme: In a first aspect, the invention provides an injectable bioactive matrix hydrogel having anti-angiogenic and anti-neurogenic microenvironment, the composition of the injectable bioactive matrix hydrogel comprising bevacizumab, concentrated growth factor (concentrated growth factors, CGF) and methacryated chondroitin sulfate (METHACRYLATED CHONDROITIN SULFATE, CSMA). Preferably, in the injectable bioactive matrix hydrogel, the concentration of bevacizumab is 10ug/mL, and the concentration of the concentrated growth factor is 10mg/mL. The preparation method of the methacrylic acid chondroitin sulfate comprises the following steps of dissolving the chondroitin sulfate in deionized water to form a chondroitin sulfate solution, then dropwise adding methacrylic anhydride into the chondroitin sulfate solution, stirring at a low temperature for reaction, purifying the obtained product, and then freeze-drying to obtain the methacrylic acid chondroitin sulfate. The preparation method of the concentrated growth factor comprises the following steps of centrifuging collected blood at speeds of 2700 rpm, 2400 rpm, 2700 rpm and 3000 rpm for 2 minutes, 4 minutes and 3 minutes in sequence, standing for layering, extracting an intermediate layer, and pre-cooling, freeze-drying and powdering the extracted intermediate layer to obtain the concentrated growth factor. In a second aspect, the present invention provides a method for preparing an injectable bioactive matrix hydrogel having anti-angiogenic and anti-neurogenic microenvironment, comprising the steps of: S1, dissolving methacrylic acid chondroitin sulfate in a solvent containing a photoinitiator to obtain a precursor solution a; s2, adding bevacizumab and concentrated growth factors into the precursor solution a, and uniformly stirring to obtain a precursor solution b; and S3, inducing photo-crosslinking of the precursor solution b under the condition of UV illumination to obtain the injectable bioactive matrix hydrogel. Preferably, in the step a, the concentration of the methacryiated chondroitin sulfate in the precursor solution a is 3-7%. Preferably, the concentration of the methacryized chondroitin sulfate in the precursor solution a is 5%. Preferably, in the step A, the photoinitiator is selected from phenyl-2, 4, 6-trimethyl benzoyl lithium phosphonate, and the solvent is selected from PBS solution. Preferably, the concentration of the photoinitiator in the solvent containing the photoinitiator is 0.2-0.5%. In the preferred scheme, in the step C, the UV illumination condition is that the wavelength of UV light is 320-400nm, the illumination intensity is 8-12 mW/cm < 2 >, and the illumination time is 3-8 seconds. In a third aspect, the present invention provides the use of an injectable bioactive matrix hydrogel according to the foregoing or an injectable bioactive matrix hydrogel prepared according to the foregoing method for the preparation of a product for repair of disc degeneration and/or promotion of cartilage regeneration. Compared with the prior art, the invention has the following beneficial effects: The invention develops a 'one-stone three-bird' strategy for intervertebral disc degeneration, and the strategy has not been reported before by constructing a novel injectable bioactive matrix hydrog