Engineered cell intraocular transplantation device based on hyaluronic acid scaffold

Abstract

The invention discloses an engineered cell intraocular transplantation device based on a hyaluronic acid scaffold, the engineered cell intraocular transplantation device comprises a vitreous body and a cell intraocular transplantation device fixed in the vitreous body, the cell intraocular transplantation device comprises engineered cells, a hyaluronic acid hydrogel scaffold and a suture sclera; according to the engineered cell storage container of the capsule-shaped structure, a semi-permeable polymer film is adopted, the semi-permeable film is provided with gaps of 15 nm, engineered cells wrapped by hyaluronic acid hydrogel are arranged in the semi-permeable film, the bottom of the semi-permeable film is a sealed end, and a detachable sealing end cover is arranged at the top end of the semi-permeable film; the sealing end is used for preventing invasion of an eye immune system; and the sealing end cover is sealed by using a medical-grade sealant, so that the cells are prevented from being invaded by host immune cells. The invention provides a new technical solution for the field of protection and transplantation of engineered cells, and shows remarkable economical efficiency, practicability, safety, operation convenience and stability.

Description

Technical Field

[0001] The invention relates to the technical field of medical engineered cell encapsulation, and in particular to an engineered cell intraocular transplantation device based on a hyaluronic acid scaffold. Background Art

[0002] In recent years, ophthalmic diseases have grown rapidly, especially those affecting the posterior segment of the eye, such as age-related macular degeneration, retinitis pigmentosa, diabetic retinopathy, and glaucomatous optic neuropathy, which are the main causes of blindness. However, the limited permeability of the ocular barrier poses a challenge to the development of effective therapeutic interventions. Current treatments include local administration, systemic administration, intravitreal injection, and gene therapy. Whether it is local administration, systemic administration, or intravitreal injection, they all face problems such as low drug concentration reaching the lesion, short half-life in the body, and poor stability. Therefore, a large number of multiple injections are required to achieve the desired therapeutic effect, resulting in poor patient compliance. Although gene therapy can achieve the sustained expression of a given protein, the dosage of the therapeutic protein is difficult to control due to the lack of a reliable method to regulate the exact expression level of the transgene. Once the gene is integrated into the host cell and transcription begins, the treatment process will be irreversible.

[0003] Engineered cells are a process of modifying cells using biotechnology to give them specific functions or properties. Typically, this involves modifying or regulating the morphology and function of cells through gene editing technology, gene modification, cell reprogramming, synthetic biology and other methods to achieve the desired biological effects. Transplanting such specially edited cells into the eye and releasing freshly synthesized protein drugs or specific cytokines in situ can achieve long-term effective delivery and presence of drugs in the eye. Therefore, only relatively small amounts of protein are needed to achieve a therapeutic effect, and stable endogenous protein secretion not only ensures the continuous availability of proteins at the target site, but also for a long time, thereby eliminating the need for frequent injections and related side effects, and improving patient compliance. Compared with gene therapy, the target gene is integrated into allogeneic cells rather than the host cell genome, avoiding unexpected gene mutations or affecting the normal function of host cells, thereby eliminating potential safety hazards.

[0004] Since the editing steps of autologous engineered cells are cumbersome and the safety verification time is too long, most of the existing engineered cells are allogeneic or xenogeneic cells. Patients may have a strong immune response to them, which will not only destroy the transplanted engineered cells and weaken the therapeutic effect, but also cause damage to the patient's normal tissues and aggravate the condition. The existing encapsulation devices have poor mobility, and trauma or strenuous activities can easily cause eye discomfort and damage to normal eye tissues in patients. In addition, the cell storage systems in the existing devices are mostly artificial synthetic fiber spinning. Due to their low biocompatibility, they cannot effectively interact with transplanted cells, and their metabolites may be toxic or accumulate in the body to cause side effects. Therefore, finding a safe and effective device for encapsulating engineered cells, especially one that can protect engineered cells from the host's immune response, while ensuring the free exchange of small-sized substances such as drugs, oxygen, nutrients, metabolites, etc., so as to maintain the various biological activities and normal functions of cells, has become an urgent problem to be solved in the current intraocular engineered cell transplantation.

[0005] To address the above problems, we proposed an engineered cell intraocular transplantation device based on a hyaluronic acid scaffold. Summary of the invention

[0006] The purpose of the present invention is to provide an engineered cell intraocular transplantation device based on a hyaluronic acid scaffold, which solves the existing problems.

[0007] To achieve the above object, the present invention provides the following technical solutions:

[0008] An engineered cell intraocular transplantation device based on a hyaluronic acid scaffold, comprising:

[0009] Vitreous body, and a cell intraocular transplantation device for fixation in the vitreous body, the cell intraocular transplantation device comprising engineered cells, a hyaluronic acid hydrogel scaffold and a sutured sclera;

[0010] The engineered cell storage container with a capsule-like structure adopts a semipermeable polymer membrane with a 15nm gap, a sealed end at the bottom, and a detachable sealed end cap at the top;

[0011] A sealed end to prevent invasion of the eye's immune system;

[0012] Sealed end caps, sealed with medical-grade sealants, protect cells from invasion by host immune cells;

[0013] Engineered cell survival support system, using an extracellular matrix-like network formed based on hyaluronic acid hydrogel, and using photoactivated methacryloyl hyaluronic acid, activated by photoinitiator to form gel, as a support scaffold for cell adhesion;

[0014] An anchoring suture clip, used for suturing the sclera during transplantation surgery and fixing the cell intraocular transplantation device in the vitreous body;

[0015] A spring-like link connects the capsule-shaped engineered cell storage container and the anchoring suture clip, allowing the engineered cell storage container to move in a small range to avoid damaging normal eye tissues;

[0016] Preferably, the sealing end cap adopts a threaded connection or a plug-in structure to prevent the invasion of the eye's immune system.

[0017] Preferably, the spring-like links are used to increase the mobility of the engineered cell storage container with a capsule-like structure through the spring-like structure of the spring-like links.

[0018] Preferably, the normal angle between the two clamp arms of the anchoring suture clamp is V-shaped.

[0019] Preferably, the semipermeable polymer membrane is made of natural or synthetic polymer materials, including alginate and polyvinyl chloride.

[0020] Preferably, the shape of the engineered cell storage container is not limited to a capsule, and can have different shapes, including a disc, a sphere, and a cylinder.

[0021] Preferably, the engineered cells encapsulated into the engineered cell survival support system are endowed with specific functions or properties through a series of biotechnological means, and the drugs released by the encapsulated engineered cells include cytokines and proteins.

[0022] Preferably, the photocrosslinking agent used to form the hydrogel scaffold in the engineered cell survival support system includes one or more of Irgacure 2959, LAP, Eosin Y and Rose Bengal.

[0023] Compared with the prior art, the present invention has the following beneficial effects:

[0024] The invention adopts an economical and practical design, is easy to manufacture and use, and greatly improves its applicability and potential for industrial application.

[0025] The present invention significantly improves the living environment of engineered cells, effectively prolongs the sustained release time of protein drugs, and improves patient compliance by utilizing the selective permeability of semipermeable polymer membranes, the supporting effect of hyaluronic acid hydrogels, and the small-range activity allowed by spring-like links.

[0026] The engineered cell intraocular transplantation device of the present invention also takes into account the convenience of user operation, so that medical workers can easily assemble the device and get started without complicated training, which greatly improves operational efficiency and convenience.

[0027] According to the method provided by the present invention, the effect of intraocular transplantation of engineered cells is more stable and reliable, with good repeatability, and provides a solid foundation for the successful treatment of eye diseases.

[0028] The present invention brings a new technical solution to the field of protection and transplantation of engineered cells, showing its remarkable practicality, safety, ease of operation and stability. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 It is a schematic diagram of the mechanism of action of the present invention;

[0030] Figure 2 It is a structural schematic diagram of the present invention;

[0031] Figure 3 is a cross-sectional view of the present invention;

[0032] Figure 4 is an operation flow chart of the present invention;

[0033] Figure 5 It is a specific operation flow chart of the present invention.

[0034] Figure Number:

[0035] 1. Engineered cell storage container; 2. Sealed end; 3. Sealed end cap; 4. Engineered cell survival support system; 5. Anchored suture clip; 6. Spring-like link; 7. Engineered cells; 8. Suturing sclera; 9. Immune cells; 10. Drugs. DETAILED DESCRIPTION

[0036] The technical solutions in the embodiments of the present invention will be described clearly and completely below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments.

[0037] Embodiment 1:

[0038] like Figure 1-3 As shown, an engineered cell intraocular transplantation device based on a hyaluronic acid scaffold comprises:

[0039] A vitreous body, and a cell intraocular transplantation device for fixing inside the vitreous body, the cell intraocular transplantation device comprising engineered cells 7, a hyaluronic acid hydrogel scaffold and a sutured sclera 8;

[0040] The engineered cell storage container 1, the shape of the engineered cell storage container 1 is not limited to a capsule shape, and can have different shapes, including a disc shape, a sphere shape, and a cylinder shape;

[0041] The engineered cell storage container 1 of capsule structure adopts a semipermeable polymer membrane, which is made of natural or synthetic polymer materials, including alginate and polyvinyl chloride. The semipermeable membrane has a gap of 15nm, which allows the target drug 10 and oxygen and nutrient factors to pass through. The bottom is a sealed end 2, which is used to prevent the invasion of the eye immune system. The top is provided with a detachable sealed end cap 3 to facilitate the placement and removal of the engineered cells 7.

[0042] The engineered cell storage container 1 and the sealing end cap are connected by a threaded connection or a plug-in structure, that is, the two form a capsule-like structure and the threaded connection or the plug-in structure makes it easy to disassemble and assemble, and the internal structure and the engineered cells can be taken out more conveniently and quickly;

[0043] The sealing end cap 3 and the engineered cell storage container 1 are connected by a threaded connection or a plug-in structure to ensure the sealing of the container, and are sealed with a medical-grade sealant to protect the cells from the invasion of the host immune cells 9.

[0044] Embodiment 2:

[0045] like Figure 1-3 As shown, the engineered cell survival support system 4 adopts an extracellular matrix-like network formed based on hyaluronic acid hydrogel, and adopts methacryloyl hyaluronic acid, which can be activated into gel by a photoinitiator. By adjusting the illumination time and intensity, the cross-linking density of the hydrogel can be adjusted, thereby regulating its mechanical properties and degradation rate, thereby controlling the action time of the engineered cells in the eye. As a supporting scaffold for cell adhesion, the formula of this system usually contains a variety of nutrients, electrolytes, buffers and necessary antibiotics. These ingredients work together to simulate the natural environment in the human eye to support the basic life activities of the engineered cell.

[0046] The photocrosslinking agent used to form the hydrogel scaffold in the engineered cell survival support system 4 includes one or more of Irgacure 2959, LAP, Eosin Y and Rose Bengal.

[0047] The anchoring suture clip 5 is a suture clip commonly used in ophthalmic surgery. The normal angle between the two clamp arms of the anchoring suture clip 5 is V-shaped. During the transplantation operation, the sclera 8 is sutured and the cell intraocular transplantation device is fixed in the vitreous body.

[0048] The spring-like link 6 connects the engineered cell storage container 1 with a capsule-like structure and the anchoring suture clip 5. The spring-like link 6 is used to increase the mobility of the engineered cell storage container 1 with a capsule-like structure through the spring-like structure of the spring-like link 6, allowing the engineered cell storage container to move in a small range to avoid eye discomfort to the patient and damage to normal eye tissue caused by trauma or strenuous activities.

[0049] Embodiment 3:

[0050] like Figure 1-3 As shown, the engineered cells 7 encapsulated into the engineered cell survival support system 4 are endowed with specific functions or properties through a series of biotechnology means (including gene editing technology, gene modification, cell reprogramming, synthetic biology and other methods), and the drugs 10 released by the encapsulated engineered cells 7 include cytokines and proteins.

[0051] Working principle:

[0052] First, the edited engineered cells 7 capable of producing specific drugs are placed in an engineered cell survival support system 4, which uses an extracellular matrix-like network formed based on hyaluronic acid hydrogel to simulate the human eye environment and has good biocompatibility;

[0053] The support system containing the engineered cells is then encapsulated with an engineered cell storage container 1, which employs a semipermeable polymer membrane having 15 nm pores to allow oxygen and other nutrients 11 to nourish the engineered cells 7, while providing diffusion control when the target drug 10 leaves the capsule and enters the surrounding vasculature or tissue, and prevents cell rejection by constructing an immunoisolation membrane, thereby eliminating destructive elements of the host immune system 9 from entering the cell-containing cavity;

[0054] Then, the sealing end cap 3 is covered and sealed with a medical-grade sealant to prevent the transplanted cells from leaking out and the eye's immune system from invading;

[0055] Among them, during the transplantation operation, the engineered cell intraocular transplantation device is fixed in the vitreous cavity using an anchoring suture clamp 5, and a spring-like link 6 is used between the engineered cell storage container 1 and the anchoring suture clamp 5 to allow the engineered cell storage container 1 to move in a small range to avoid eye discomfort and damage to normal eye tissue caused by trauma or strenuous activities. This design ensures the stability and safety of the engineered cell before and after transplantation, effectively prolongs the sustained release time of protein drugs, and improves patient compliance.

[0056] like Figure 4 and Figure 5 As shown, the following is a detailed method for using the engineered cell intraocular transplantation device to ensure that the engineered cells can obtain the greatest degree of protection before and after transplantation:

[0057] Step S1: Preparation of engineered cell storage container:

[0058] Check the basic container to ensure that the container has good selective filtration to prevent leakage and microbial contamination of the engineered cell survival support system, and thoroughly clean the basic container and sterilize it when necessary to ensure that it is free of bacteria and other contaminants;

[0059] Step S2: Preparation of engineered cell survival support system:

[0060] Add methacrylic anhydride and hyaluronic acid in a ratio of 1:1 to PBS, stir the solution and adjust the pH value to 8.0-8.5, and allow the reaction to proceed at room temperature for 4-6 hours to form methacryloyl hyaluronic acid. Add an appropriate amount of photoinitiator (recommended 0.05-0.1% w / v) to the PBS solution containing the desired concentration of methacryloyl hyaluronic acid, stir until the photoinitiator is completely dissolved, and add an appropriate amount of nutrients, electrolytes, buffers, and necessary antibiotics, and then inoculate the engineered cells. Ensure that the engineered cells have a suitable living environment before transplantation;

[0061] Step S3: Inoculation of engineered cells:

[0062] Under sterile conditions, the edited engineered cells 7 capable of producing specific drugs are added to a hyaluronic acid solution containing a photoinitiator, and gently stirred to evenly disperse the cells in the survival support system to avoid contact inhibition that may damage the normal function of the engineered cells;

[0063] Step S4, assembly of the device:

[0064] The support system containing the engineered cells is placed in the engineered cell storage container 1, and the solution is irradiated with a light source of appropriate wavelength. The irradiation time varies according to the type and concentration of the photoinitiator, and is generally 1-5 minutes. During the irradiation process, the mixed solution will form a gel, and then the sealing end cap 3 is covered and sealed with a medical grade sealant;

[0065] Step S5: Fixation of the device during surgery:

[0066] The engineered cell intraocular transplant device is inserted through sclerotomy, and the sclera 8 is sutured using an anchoring suture clip 5 to fix the engineered cell intraocular transplant device in the vitreous cavity;

[0067] Step S6, evaluation of transplantation effect:

[0068] Patients undergoing transplantation are examined regularly, and the status of the engineered cell intraocular transplant device located in the lens cavity is examined with a slit lamp after the pupil is dilated.

[0069] In the description of this article, it should be noted that relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. The terms "installed", "connected", and "connected" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection, a direct connection, or an indirect connection through an intermediate medium, or it can be the internal connection of two elements. Moreover, the terms "include", "comprise", or any other variant thereof are intended to cover non-exclusive inclusion, so that a process, method, article, or device that includes a series of elements includes not only those elements, but also other elements that are not explicitly listed, or also includes elements inherent to such process, method, article, or device.

[0070] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. An engineered cell intraocular transplantation device based on a hyaluronic acid scaffold, It is characterized in that include: Vitreous body, and a cell intraocular transplantation device for fixing inside the vitreous body, the cell intraocular transplantation device comprising engineered cells (7), a hyaluronic acid hydrogel scaffold and a sutured sclera (8); An engineered cell storage container (1) with a capsule-like structure adopts a semipermeable polymer membrane with a 15 nm gap, a sealed end (2) at the bottom, and a detachable sealed end cap (3) at the top; A sealed end (2) for preventing invasion of the ocular immune system; A sealed end cap (3) is sealed using a medical grade sealant to protect the cells from invasion by host immune cells (9); Engineered cell survival support system (4), using an extracellular matrix-like network formed based on hyaluronic acid hydrogel, and using photoactivated methacryloyl hyaluronic acid to activate the gel through a photoinitiator as a support scaffold for cell adhesion; An anchoring suture clip (5) is used to suture the sclera (8) during the transplantation operation and fix the cell intraocular transplantation device in the vitreous body; The spring-like link (6) connects the capsule-shaped engineered cell storage container (1) and the anchoring suture clip (5), allowing the engineered cell storage container to move in a small range to avoid damaging normal eye tissue.

2. The engineered cell intraocular transplantation device according to claim 1, It is characterized in that The sealing end cover (3) adopts a threaded connection or a plug-in structure to prevent the invasion of the eye's immune system.

3. The engineered cell intraocular transplantation device according to claim 1, It is characterized in that The spring-like link (6) is used to increase the mobility of the engineered cell storage container (1) with a capsule-like structure through the spring-like structure of the spring-like link (6).

4. The engineered cell intraocular transplantation device according to claim 1, It is characterized in that The normal included angle of the two clamp arms of the anchoring suture clamp (5) is V-shaped.

5. The engineered cell intraocular transplantation device according to claim 1, It is characterized in that The semipermeable polymer membrane is made of natural or artificially synthesized polymer materials, including alginate and polyvinyl chloride.

6. The engineered cell intraocular transplantation device according to claim 1, It is characterized in that The shape of the engineered cell storage container (1) is not limited to a capsule shape, and can have different shapes, including a disc shape, a sphere shape, and a cylinder shape.

7. The engineered cell intraocular transplantation device according to claim 1, It is characterized in that The engineered cells (7) encapsulated in the engineered cell survival support system (4) are endowed with specific functions or properties through a series of biotechnological means, and the drugs (10) released by the encapsulated engineered cells (7) include cytokines and proteins.

8. The engineered cell intraocular transplantation device according to claim 1, It is characterized in that The photocrosslinking agent used to form the hydrogel scaffold in the engineered cell survival support system (4) includes one or more of Irgacure 2959, LAP, Eosin Y and RoseBengal.

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