Stem cell active factor capsule for tissue repair and regeneration

Abstract

The invention relates to the technical field of tissue repair and regeneration and regenerative medicine, and discloses a stem cell active factor capsule for tissue repair and regeneration, which comprises a capsule body, and the capsule body has a core-shell-targeting composite structure, and sequentially comprises a load core body, an exosome containing a mesenchymal stem cell source, an exosome containing a mesenchymal stem cell source, and an auxiliary body from inside to outside, at least one recombinant growth factor is loaded on the surface of the exosome; the stem cell exosome is used for carrying growth factors, a human body self-repairing mechanism is synergistically activated, the limitation that only symptoms are relieved traditionally is broken through, and tissue functional regeneration is achieved. Intelligent shell layer targeted delivery is achieved, immunological rejection and systemic side effects are remarkably reduced, and donor shortage is solved. Multi-way administration such as oral administration and intravenous injection is supported, the patient tolerance is good, the utilization degree is improved through enzyme sensitive protection, and the defects that traditional delivery targeting is poor, and absorption is insufficient are effectively overcome.

Description

Technical Field

[0001] This invention relates to the fields of tissue repair and regeneration and regenerative medicine, specifically to a stem cell active factor capsule for tissue repair and regeneration. Background Technology

[0002] Tissue damage and loss of function are major challenges in clinical medicine, involving multiple fields such as trauma, degenerative diseases, and surgical complications. Traditional treatments, such as drug intervention, surgical repair, and organ transplantation, can alleviate some symptoms, but they often have inherent defects such as limited efficacy, large side effects, shortage of donors, immune rejection, and inability to achieve functional regeneration. Therefore, developing new treatment strategies that can actively promote the body's own repair and regeneration capabilities has become the core goal of regenerative medicine.

[0003] Currently, traditional drug and growth factor delivery systems lack targeting and tend to distribute throughout the body, resulting in limited efficacy and potential systemic side effects. While local injections can increase local concentrations, they are invasive, poorly tolerated by patients, and difficult to target deep or diffusely damaged areas. Topical preparations have low transdermal absorption efficiency and are difficult to apply to internal organs. Direct stem cell transplantation faces risks such as immune rejection, ethical controversies, tumorigenesis, and low survival rates. Therefore, we propose a stem cell active factor capsule for tissue repair and regeneration to address these issues. Summary of the Invention

[0004] To address the shortcomings of existing technologies, this invention provides a stem cell active factor capsule for tissue repair and regeneration, which solves the problems of poor targeting and easy systemic side effects of traditional drug and growth factor delivery; strong invasiveness and limited range of action of local injection; difficulty in transdermal absorption and action on internal organs of topical preparations; and the shortcomings of direct stem cell transplantation, such as immune rejection, ethics, tumor formation and low survival rate.

[0005] To achieve the above objectives, the present invention provides the following technical solution: a stem cell active factor capsule for tissue repair and regeneration, comprising a capsule body having a core-shell-targeting composite structure, comprising, from the inside out:

[0006] The payload core comprises exosomes derived from mesenchymal stem cells, the surface of which is loaded with at least one recombinant growth factor;

[0007] A smart shell, which wraps around the outside of the payload core, is composed of a biopolymer cross-linking network containing enzyme-sensitive polypeptide sequences that can be specifically cleaved by matrix metalloproteinases overexpressed at the damage site.

[0008] A targeted functional layer, covering the outside of the smart shell, is surface-modified with specific targeting ligands that can recognize and bind adhesion molecules highly expressed by vascular endothelial cells of damaged tissue.

[0009] Preferably, the recombinant growth factor loaded on the exosomes is selected from at least one of vascular endothelial growth factor, hepatocyte growth factor, and transforming growth factor-β3.

[0010] Preferably, the enzyme-sensitive polypeptide sequence is selected from GPQGIAGQ, PLGLWA or GPLGVRG, and can be specifically cleaved by matrix metalloproteinase-2 or matrix metalloproteinase-9.

[0011] Preferably, the biopolymer of the smart shell is hyaluronic acid or polyethylene glycol, and the enzyme-sensitive polypeptide sequence is covalently linked to the biopolymer chain and cross-linked to form a three-dimensional network structure.

[0012] Preferably, the specific targeting ligand is an anti-ICAM-1 single-chain antibody, an anti-VCAM-1 mimic peptide, or a cyclic RGD polypeptide.

[0013] Preferably, the thickness of the smart shell is 100–300 nanometers, and the thickness of the targeting functional layer is 50–90 nanometers.

[0014] Preferably, the overall hydration kinetic particle size of the capsule is 250–450 nanometers, and the polydispersity index of the particle size distribution is less than 0.12.

[0015] Preferably, after the capsules are incubated in a phosphate buffer solution at pH 7.4 at 37°C for 24 hours, the particle size change rate is less than 10%.

[0016] Preferably, the capsule can be administered via intravenous injection, local injection, or oral administration.

[0017] Beneficial effects

[0018] This invention provides a stem cell active factor capsule for tissue repair and regeneration. Compared with the prior art, it has the following advantages:

[0019] This invention uses mesenchymal stem cell exosomes as carriers to carry recombinant growth factors such as vascular endothelial growth factor, synergistically activating the body's endogenous repair mechanisms to achieve structural and functional reconstruction of damaged tissues. The targeting ligand on the outer layer of the capsule can specifically recognize adhesion molecules highly expressed at the injury site, achieving enrichment of active ingredients in the lesion area. Combined with a low immunogenicity design and biocompatible shell material, it significantly reduces immune rejection and systemic side effects.

[0020] The capsules support intravenous injection, local injection, and oral administration to meet the needs of different injury types and sites. The intelligent shell contains an enzyme-sensitive polypeptide sequence that can be specifically degraded by matrix metalloproteinases overexpressed in the damaged area, achieving targeted and controlled release, effectively protecting the stability of the active ingredients, and improving bioavailability and therapeutic efficiency.

[0021] The capsules have a uniform particle size distribution and are structurally stable in physiological environments. They exhibit good batch consistency and potential for large-scale preparation. The overall design overcomes the problems of poor targeting, single dosage form, high risk of exogenous stem cell transplantation, and easy drug degradation in traditional delivery systems, providing an efficient and safe comprehensive solution for tissue repair and regeneration. Attached Figure Description

[0022] Figure 1 This is a schematic diagram of the overall structure of the present invention.

[0023] In the figure: 1. Capsule body; 101. Load core; 102. Smart shell; 103. Targeting functional layer. Detailed Implementation

[0024] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0025] like Figure 1 As shown:

[0026] A stem cell active factor capsule for tissue repair and regeneration includes a capsule body 1, which has a core-shell-targeting composite structure, comprising, from the inside out:

[0027] The payload core 101 contains exosomes derived from mesenchymal stem cells, and the surface of the exosomes is loaded with at least one recombinant growth factor, wherein the recombinant growth factor loaded on the exosomes is selected from at least one of vascular endothelial growth factor, hepatocyte growth factor, and transforming growth factor-β3.

[0028] The intelligent shell 102 is wrapped around the payload core 101. The intelligent shell 102 is composed of a biopolymer cross-linking network containing enzyme-sensitive polypeptide sequences. The enzyme-sensitive polypeptide sequences can be specifically cleaved by matrix metalloproteinases overexpressed at the damage site. The enzyme-sensitive polypeptide sequences are selected from GPQGIAGQ, PLGLWA or GPLGLVRG and can be specifically cleaved by matrix metalloproteinase-2 or matrix metalloproteinase-9. The biopolymer of the intelligent shell 102 is hyaluronic acid or polyethylene glycol. The enzyme-sensitive polypeptide sequences are covalently linked to the biopolymer chains and cross-linked to form a three-dimensional network structure.

[0029] The targeting functional layer 103 covers the outside of the smart shell 102 and is modified with specific targeting ligands. The ligands can recognize and bind to adhesion molecules highly expressed by vascular endothelial cells of damaged tissue. The specific targeting ligands are anti-ICAM-1 single-chain antibodies, anti-VCAM-1 mimic peptides or cyclic RGD peptides. The thickness of the smart shell 102 is 100–300 nanometers and the thickness of the targeting functional layer 103 is 50–90 nanometers.

[0030] The overall hydration kinetic particle size of capsule 1 is 250–450 nm, and the polydispersity index of the particle size distribution is less than 0.12;

[0031] After incubation in phosphate buffer at pH 7.4 at 37°C for 24 hours, the particle size change rate of capsule 1 was less than 10%.

[0032] Capsule 1 can be administered via intravenous injection, local injection, or oral administration.

[0033] In this implementation plan: Capsule 1 can enter the human body through intravenous injection, local injection or oral administration, adapting to the needs of different injury scenarios;

[0034] Intravenous injection of capsule 1, with its overall hydration dynamics particle size of 250–450 nanometers and polydispersity index of less than 0.12, can smoothly integrate into the blood circulation system, achieve systemic distribution, and accurately reach deep or diffusely damaged areas, solving the problem that traditional local injections cannot cover such areas.

[0035] Local injection: It acts directly on the damaged area, rapidly increasing the local concentration of capsule 1, taking into account both targeting and high efficiency. Compared with the invasiveness of traditional simple local injection, the intelligent structural design of capsule 1 reduces interference with surrounding normal tissues.

[0036] Oral route: Capsule 1 has excellent physiological stability. After incubation at 37°C for 24 hours in phosphate buffer at pH 7.4, the particle size change rate is less than 10%. It can resist the acid and alkali and enzymatic erosion of the gastrointestinal environment and be successfully absorbed into the body through the digestive mucosa, overcoming the limitations of traditional external preparations that cannot act on internal organs and have low transdermal absorption efficiency.

[0037] The outermost targeting functional layer 103 of capsule 1, with a thickness of 50–90 nanometers, plays a core guiding role: the specific targeting ligands modified on the surface of the targeting functional layer 103, such as anti-ICAM-1 single-chain antibody, anti-VCAM-1 mimic peptide or cyclic RGD polypeptide, can specifically recognize adhesion molecules highly expressed by vascular endothelial cells of damaged tissue, such as ICAM-1 and VCAM-1. The ligands bind specifically to the adhesion molecules, so that capsule 1 is precisely anchored to the damaged site, avoiding the systemic distribution problem caused by the lack of targeting in traditional drug and growth factor delivery systems, and greatly reducing the aggregation of capsule 1 in non-targeted sites, laying the foundation for subsequent efficient treatment;

[0038] When capsule 1 accumulates at the injury site, the intelligent shell 102, with a thickness of 100–300 nanometers, initiates the unlocking mechanism. The intelligent shell 102 is composed of a biopolymer cross-linking network containing enzyme-sensitive polypeptide sequences, such as GPQGIAGQ, PLGLWA, or GPLGLVRG. The enzyme-sensitive polypeptide sequences are covalently linked to the biopolymer chains and form a stable three-dimensional network structure through a cross-linking agent. In the blood circulation and normal tissue environment, it can effectively protect the internal load core 101 from degradation. The injury site will overexpress matrix metalloproteinase-2 or matrix metalloproteinase-9. These proteases can specifically cleave the enzyme-sensitive polypeptide sequences in the intelligent shell 102, causing the biopolymer cross-linking network to disintegrate. The disintegration of the intelligent shell 102 opens the structure of capsule 1, realizing the precise release of the load core 101. This avoids the waste and side effects caused by the premature release of active factors at non-injury sites, and solves the pain points of easy degradation, poor targeting, and low bioavailability of active factors in traditional intravenous infusion.

[0039] After the core payload 101 is released, the mesenchymal stem cell-derived exosomes contained therein and the recombinant growth factors loaded on its surface, such as at least one of vascular endothelial growth factor, hepatocyte growth factor, and transforming growth factor-β3, work synergistically. As a functional carrier, the mesenchymal stem cell exosomes have low immunogenicity and good biocompatibility. They can carry recombinant growth factors to penetrate the microenvironment barrier of damaged tissue and reach the vicinity of damaged cells. The recombinant growth factors bind to specific receptors on the surface of cells at the damaged site, initiate intracellular signaling pathways, activate the body's own repair mechanisms, and promote the proliferation, differentiation, and migration of cells at the damaged site. The stem cell-derived active ingredients carried by the exosomes themselves form a synergistic effect with the recombinant growth factors, accelerating the structural repair and functional reconstruction of damaged tissue, and ultimately achieving functional regeneration of damaged tissue, breaking through the limitation of traditional treatments that can only relieve symptoms and cannot achieve regeneration.

[0040] This approach uses mesenchymal stem cell exosomes as carriers to deliver recombinant growth factors such as vascular endothelial growth factor, synergistically activating the body's endogenous repair mechanisms to achieve structural and functional reconstruction of damaged tissues. The targeting ligand on the outer layer of the capsule can specifically recognize adhesion molecules highly expressed at the injury site, achieving enrichment of active ingredients in the lesion area. Combined with a low immunogenicity design and biocompatible shell material, it significantly reduces immune rejection and systemic side effects.

[0041] The capsules support intravenous injection, local injection, and oral administration to meet the needs of different injury types and sites. The intelligent shell contains an enzyme-sensitive polypeptide sequence that can be specifically degraded by matrix metalloproteinases overexpressed in the damaged area, achieving targeted and controlled release, effectively protecting the stability of the active ingredients, and improving bioavailability and therapeutic efficiency.

[0042] The capsules exhibit uniform particle size distribution and structural stability in physiological environments, demonstrating excellent batch-to-batch consistency and potential for scalable production. The overall design overcomes the limitations of traditional delivery systems, such as poor targeting, limited dosage forms, high risks associated with exogenous stem cell transplantation, and easy drug degradation, providing a highly efficient and safe comprehensive solution for tissue repair and regeneration.

[0043] Finally, it should be noted that the above descriptions are merely preferred embodiments of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A capsule containing stem cell active factors for tissue repair and regeneration, characterized in that: Includes a capsule body (1), which has a core-shell-targeting composite structure and comprises, from the inside out: The payload core (101) comprises exosomes derived from mesenchymal stem cells, the surface of which is loaded with at least one recombinant growth factor; A smart shell (102) is wrapped around the outside of the load core (101). The smart shell (102) is composed of a biopolymer cross-linking network containing enzyme-sensitive polypeptide sequences. The enzyme-sensitive polypeptide sequences can be specifically cleaved by matrix metalloproteinases overexpressed at the damage site. A targeting functional layer (103) covers the outside of the smart shell (102) and is surface-modified with a specific targeting ligand that can recognize and bind to adhesion molecules highly expressed by vascular endothelial cells of damaged tissue.

2. The stem cell active factor capsule for tissue repair and regeneration according to claim 1, characterized in that: The recombinant growth factor loaded on the exosomes is selected from at least one of vascular endothelial growth factor, hepatocyte growth factor, and transforming growth factor-β3.

3. The stem cell active factor capsule for tissue repair and regeneration according to claim 1, characterized in that: The enzyme-sensitive polypeptide sequence is selected from GPQGIAGQ, PLGLWA or GPLGVRG, and can be specifically cleaved by matrix metalloproteinase-2 or matrix metalloproteinase-9.

4. The stem cell active factor capsule for tissue repair and regeneration according to claim 1, characterized in that: The biopolymer of the smart shell (102) is hyaluronic acid or polyethylene glycol. The enzyme-sensitive polypeptide sequence is covalently linked to the biopolymer chain and cross-linked to form a three-dimensional network structure.

5. The stem cell active factor capsule for tissue repair and regeneration according to claim 4, characterized in that: The specific targeting ligand is an anti-ICAM-1 single-chain antibody, an anti-VCAM-1 mimic peptide, or a cyclic RGD polypeptide.

6. The stem cell active factor capsule for tissue repair and regeneration according to claim 1, characterized in that: The thickness of the smart shell (102) is 100–300 nanometers, and the thickness of the targeting functional layer (103) is 50–90 nanometers.

7. The stem cell active factor capsule for tissue repair and regeneration according to claim 1, characterized in that: The overall hydration dynamic particle size of the capsule (1) is 250–450 nanometers, and the polydispersity index of the particle size distribution is less than 0.

12.

8. The stem cell active factor capsule for tissue repair and regeneration according to claim 1, characterized in that: After the capsule (1) was incubated in phosphate buffer at pH 7.4 at 37°C for 24 hours, the particle size change rate was less than 10%.

9. The stem cell active factor capsule for tissue repair and regeneration according to claim 1, characterized in that: The capsule (1) can be administered via intravenous injection, local injection or oral administration.

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