CN-121513197-B - Piezoelectric hydrogel loaded with response polypeptide and application thereof

Abstract

The invention relates to the field of polypeptide medicines, in particular to the technical field of ultrasonic physical field regulation biological signals and bone tissue repair, and in particular relates to a piezoelectric hydrogel loaded with response polypeptides and application thereof, wherein the application is a biological signal regulation method based on ultrasonic induction response polypeptides LLPS and application for promoting bone differentiation, and space-time specificity activation of biological signal paths is realized through accurate regulation and control mediated by physical fields.

Inventors

• WANG XIAOGANG
• Gong Zunlei
• Han Jien

Assignees

• 南方医科大学第三附属医院（广东省骨科研究院）

Dates

Publication Date

20260505

Application Date

20260114

Claims (6)

1. 1. A piezoelectric hydrogel loaded with a responsive polypeptide, the method of preparation comprising the steps of: 1) Mixing 5-40-wt% of methacrylic acid-acylated gelatin GelMA solution and 5-20-wt% of oxidized beta-cyclodextrin solution to obtain a clear solution, and cooling to room temperature; 2) Adding response polypeptide into the clarified solution until the concentration reaches 25-50 mug/mL, and fully dissolving; 3) Adding barium titanate microparticles into the solution until the concentration reaches 5-20 mg/mL, stirring uniformly, injecting into a mould, and solidifying to obtain the piezoelectric hydrogel loaded with the response polypeptide; the response polypeptide is subjected to liquid-liquid phase separation under ultrasonic conditions; the amino acid sequence of the response polypeptide is shown as SEQ ID NO. 3.
2. 2. The polypeptide-loaded piezoelectric hydrogel according to claim 1, wherein the barium titanate is modified with APTES ethanol solution.
3. 3. The piezoelectric hydrogel loaded with a response polypeptide according to claim 1, wherein the concentration of the methacryloylated gelatin GelMA is 8-12 wt%, the concentration of the oxidized β -cyclodextrin is 8-12 wt%, the concentration of the response polypeptide is 25-35 μg/mL, and the concentration of the barium titanate microparticles is 8-12 mg/mL.
4. 4. The piezoelectric hydrogel loaded with a response polypeptide according to claim 3, wherein the concentration of the methacryloylated gelatin GelMA is 10 wt%, the concentration of the oxidized β -cyclodextrin is 10 wt%, the concentration of the response polypeptide is 30 μg/mL, and the concentration of the barium titanate microparticles is 10 mg/mL.
5. 5. A pharmaceutical composition comprising the piezoelectric hydrogel of the load-responsive polypeptide of any one of claims 1-4 and a pharmaceutically acceptable excipient.
6. 6. Use of a piezoelectric hydrogel loaded with a responsive polypeptide according to any one of claims 1-4 or a pharmaceutical composition according to claim 5 for the preparation of a medicament for the treatment of bone damage.

Description

Piezoelectric hydrogel loaded with response polypeptide and application thereof Technical Field The invention belongs to the technical field of ultrasonic physical field regulation biological signals and bone tissue repair, and particularly relates to a piezoelectric hydrogel loaded with response polypeptides, a biological signal regulation method based on ultrasonic induction response polypeptides LLPS and application of promoting bone differentiation, which realize space-time specific activation of biological signal paths through accurate regulation and control mediated by physical fields. Background Systematic bottleneck of existing biological signal accurate regulation technology The field of tissue regeneration, in particular osteoporosis bone repair, is highly dependent on precise regulation of key biological signals (e.g., growth factors, gene expression products) in time, space and dose. However, the prior art systems are faced with significant bottlenecks in terms of gene delivery, in that widely used viral vectors (such as adeno-associated viral AAV) generally have higher transfection efficiency, but have potential genome integration risks, which may raise long-term safety concerns, and non-viral vectors are relatively improved in safety, but often suffer from low transfection efficiency, delayed expression of the target gene and unstable duration, which makes it difficult to meet the requirements of rapid initiation repair. In the delivery level of bioactive molecules, slow-release scaffolds (such as hydrogels and microspheres) based on growth factors often cause local concentration sudden rise due to the excessively fast initial release rate, are easy to induce side effects such as ectopic ossification of non-target areas, and the release behavior of the slow-release scaffolds is difficult to dynamically regulate according to the repair process. In the aspect of physical regulation and control tools, although the emerging optogenetic technology can realize space-time regulation with higher precision, the tissue penetration capability of excitation light is limited (especially in compact structures such as bone tissues) and is difficult to effectively act on deep regions, and a light source device is generally required to be implanted invasively, so that the complexity and risk of clinical application are increased. These bottlenecks together limit the safety and effectiveness of biosignal modulation therapies. Core contradiction and clinical requirements in the field of bone repair Focusing on bone repair, the technical bottleneck is concentrated and presented as a core contradiction to be solved, namely, firstly, when local injection is used, medicines or growth factors are easy to diffuse and flow away from a defect part, and effective treatment concentration is difficult to maintain, so that the curative effect is reduced, and the medicine or growth factors are key factors for inducing ectopic ossification. Secondly, although gene therapy strategies have long-acting regulatory potential, delay often exists in the effect appearance, and urgent needs of a scene requiring rapid bone formation (such as acute injury repair or large-segment bone defect bridging) are difficult to meet. It is particularly critical that the prior art platforms generally have difficulty in achieving three key performance metrics, namely, effective action of deep tissue (e.g., centimeter level bone defect centers), rapid response (the speed required to simulate dynamic changes in physiological signals), and fully non-invasive operation at the same time. The dilemma that the depth-response speed-noninvasive performance is difficult to be compatible is a fundamental obstacle for restricting the development of the next-generation intelligent bone repair technology, and is a core challenge for clinically realizing efficient, controllable and safe bone regeneration. Potential and conversion bottleneck of emerging technologies—phase separation and piezoelectric materials To seek breakthrough, leading-edge research explored the combination of LLPS phenomenon with deep penetration physical field-ultrasound. LLPS is formed by biomolecular aggregates, can enhance the specificity and efficiency of local signal transduction, and provides possibility for constructing bionic regulation nodes. However, its practical application faces challenges of poor space-time controllability of spontaneous nucleation processes of endogenous phase separation, and concomitant risk of cytotoxicity of exogenous chemical inducers. More critical, there is currently a lack of physical regulatory means that can efficiently and noninvasively trigger LLPS in deep tissues. Meanwhile, the ultrasonic mechanical energy induces the piezoelectric material to generate local electric signals or chemical micro-environment changes, and is considered as a potential way for realizing deep noninvasive regulation. However, biomedical transformation has bottlenecks that o