A hydrogel for preventing post-glaucoma filtration surgery scarring reaction and its preparation method and application

Abstract

The application discloses a hydrogel for resisting post-glaucoma filtration surgery cicatrization reaction and a preparation method and application thereof, and ingredients of the hydrogel include 2-methacrylate hydroxyethyl and 2-((2-hydroxy-3-(methacryloxy)propyl)dimethylamino) acetate. The hydrogel has good optical performance, mechanical performance and biological antifouling performance. The hydrogel is used for glaucoma filtration surgery of rabbit eyes, can more effectively reduce intraocular pressure of the rabbit eyes and maintain filtration blebs, and significantly reduces the cicatrization reaction of scleral flaps of the rabbit eyes. The hydrogel can also be used as a drug carrier platform, loads anti-cicatrization drugs and hormone drugs, and provides a new strategy for clinical application of anti-cicatrization after glaucoma filtration surgery.

Description

Technical Field

[0001] This invention belongs to the field of biomedicine and relates to a novel hydrogel for preventing scarring after glaucoma filtration surgery (GFS), its preparation method, and its application. Background Technology

[0002] Glaucoma surgery is widely considered a highly effective treatment for patients whose intraocular pressure cannot be effectively controlled by anti-glaucoma drugs or laser therapy. GFS (Glassy Foaming Surgery), a common surgical method for glaucoma, relies heavily on the efficiency of external drainage of the aqueous humor, and its efficacy depends on incomplete healing of the surgical wound. However, GFS damage can stimulate fibroblast proliferation and differentiation, as well as excessive deposition of extracellular matrix (ECM) in the filtration area. Scarring in the surgical area is a significant factor limiting the success rate of GFS. Uncontrolled intraocular pressure after GFS can lead to further deterioration of optic disc cupping and subsequent visual field loss. Clinically, antimetabolites such as mitomycin C (MMC) and 5-fluorouracil (5-FU) are often used to inhibit scar formation. However, the use of these drugs must be strictly controlled within an appropriate dosage range; overdose or inappropriate use can lead to a series of complications, including persistent low intraocular pressure, corneal toxicity, filtration bleb leakage, and endophthalmitis, which can potentially cause permanent visual impairment. Therefore, developing novel anti-scarring strategies with greater anti-fibrotic effects and lower cytotoxicity is crucial for improving the safety and effectiveness of GFS surgery.

[0003] 2-((2-hydroxy-3-(methacryloyloxy)propyl)dimethylammonium)acetate (CBOH) is renowned for its excellent biofouling-resistant properties. As an amphoteric ion, CBOH can interact with water molecules through ionic solvation, forming a hydrated layer on the material surface and endowing it with superhydrophilicity. This effectively reduces the adsorption of non-specific proteins and cell adhesion, opening up new possibilities for anti-scarring treatments. However, CBOH lacks toughness. Improving its mechanical properties through modification would better meet the suture requirements of GFS surgery, thereby enhancing its applicability in clinical applications.

[0004] Hydroxyethyl methacrylate (HEMA) has gained attention for its excellent mechanical properties and biocompatibility. Its side-chain structure is similar to that of CBOH monomer, making it easier to bind with CBOH. This invention combines HEMA and CBOH monomers, effectively solving the problems of insufficient lubrication and potential foreign body reactions caused by long-term implantation. It also improves the flexibility of the hydrogel, meeting the material suture requirements in GFS surgery. Therefore, the development of P(HEMA-CBOH) hydrogel provides an important approach for the innovation of novel anti-scarring treatment methods in GFS surgery. Summary of the Invention

[0005] The purpose of this invention is to solve the problem of scarring after glaucoma filtration surgery, and to provide a novel hydrogel that reduces scarring after glaucoma filtration surgery, as well as its preparation method and application.

[0006] This invention successfully prepared a P(HEMA-CBOH) zwitterionic hydrogel. The P(HEMA-CBOH) hydrogel comprises HEMA monomers and CBOH monomers. The CBOH polymer PCBOH exhibits excellent biofouling resistance; its superhydrophilicity effectively reduces fibroblast adhesion and adsorption, minimizing postoperative inflammatory reactions and fibrous capsule formation. The HEMA polymer (PHEMA) possesses good mechanical properties and biocompatibility, meeting the material suture requirements of GFS surgery and ensuring convenient surgical procedures and stable postoperative recovery. The results of this invention demonstrate that the P(HEMA-CBOH) hydrogel possesses excellent optical, mechanical, and biofouling resistance properties, effectively maintaining postoperative GFS outcomes and reducing scarring after rabbit eye GFS surgery. This method provides a new strategy for promoting the clinical application of anti-scarring after GFS surgery. Furthermore, this hydrogel can serve as a drug carrier platform, loading anti-scarring and hormone-based drugs, laying a solid foundation for further clinical application and promotion.

[0007] The technical solution of this invention is:

[0008] A method for preparing a hydrogel that resists scarring after glaucoma filtration surgery includes the following steps:

[0009] 1) The CBOH monomer is heated and dissolved in the HEMA monomer solution to form a homogeneous solution;

[0010] 2) Add 2-hydroxy-2-methyl-1-phenyl-1-propanone (photoinitiator 1173) and chemical crosslinking agent triethylene glycol dimethacrylate (TEGDMA) to the solution obtained in step 1), mix evenly at room temperature to obtain HEMA-CBOH solution;

[0011] 3) Thoroughly mix the HEMA-CBOH solution obtained in step 2) and transfer it into a mold. Irradiate it under 365 nm UV light (predetermined light intensity) for 1 hour to obtain P(HEMA-CBOH) hydrogel.

[0012] 4) Soak the P(HEMA-CBOH) hydrogel obtained in step 3) in 75% ethanol overnight, and dialyze it in PBS to remove residual small molecules for at least 4 days to obtain a sterile hydrogel.

[0013] In this invention, the mass percentage of the CBOH monomer is 0.1-15 wt% (relative to the total mass), more preferably 5-13 wt% (relative to the total mass), and most preferably 12 wt% (relative to the total mass).

[0014] In this invention, the heating temperature in step 1) is 80-100 degrees Celsius, and the heating time is 2-5 minutes, with the most preferred temperature being 80 degrees Celsius for 3 minutes.

[0015] In this invention, the photoinitiator 1173 is present in a mass ratio of 0.1-5 wt% relative to the total mass of the solution, more preferably 0.5-3 wt%, and most preferably 1 wt%. The chemical crosslinking agent TEGDMA is present in a mass ratio of 0.1-1 wt% relative to the total mass, more preferably 0.2-0.6 wt%, and most preferably 0.5 wt%.

[0016] In this invention, the thickness of the P(HEMA-CBOH) hydrogel is 100-1000 μm, more preferably 100-500 μm, and most preferably 100 μm.

[0017] In this invention, a novel hydrogel for resisting scarring after glaucoma filtration surgery is disclosed, which can be loaded with anti-scarring drugs, hormones, etc.

[0018] All raw materials used in this invention are commercially available products. Unless otherwise specified, all methods used in this invention are conventional methods in the field.

[0019] The technical effects of this invention are:

[0020] CBOH is renowned for its excellent biofouling-resistant properties. As an amphoteric ion, CBOH can interact with water molecules through ionic solvation, forming a hydrated layer on the material surface and endowing it with superhydrophilicity. This effectively reduces the adsorption of non-specific proteins and cell adhesion, opening up new possibilities for anti-scarring treatment. However, CBOH lacks flexibility, necessitating improved mechanical properties to meet the sutureability requirements of GFS (Growth Forecasting). To overcome this limitation, this invention introduces HEMA, which has a similar side-chain structure to CBOH monomers. HEMA is valued for its good mechanical properties and biocompatibility, but using HEMA hydrogels alone still has some drawbacks, such as low water content, high modulus leading to severe foreign body sensation, insufficient lubrication, and the potential for severe foreign body reactions from long-term implantation. This invention combines HEMA monomers with CBOH monomers to successfully synthesize a novel amphoteric hydrogel, P (HEMA-CBOH). This hydrogel not only inherits the excellent mechanical properties of PHEMA but also incorporates the biofouling-resistant properties of PCBOH, effectively solving the problems of insufficient lubrication and potential foreign body reactions from long-term implantation. Simultaneously, it also improves the flexibility of the hydrogel, meeting the material suture requirements in GFS surgery. Therefore, the development of P(HEMA-CBOH) hydrogel provides important insights for the innovation of novel anti-scarring treatment methods in GFS surgery.

[0021] The mass ratio of CBOH monomers in the P(HEMA-CBOH) hydrogel directly affects the water content of the hydrogel. In this invention, the mass ratio of CBOH monomers is 0.1-15 wt% (relative to the total mass), more preferably 5-13 wt% (relative to the total mass), and most preferably 12 wt% (relative to the total mass). As the zwitterionic CBOH content increases, the water content of the gel increases, leading to a decrease in modulus, effectively reducing the possible foreign body sensation after surgery and improving comfort. 1173 and TEGDMA, as photoinitiator and crosslinking agent respectively, affect the mechanical properties and chemical stability of the hydrogel in their mass ratio. In this invention, the mass ratio of photoinitiator 1173 is 0.1-5 wt% relative to the total mass of the solution, more preferably 0.5-3 wt%, and most preferably 1 wt%. The mass ratio of chemical crosslinking agent TEGDMA is 0.1-1 wt% relative to the total mass, more preferably 0.2-0.6 wt%, and most preferably 0.5 wt%.

[0022] This invention utilizes rabbit eyes for GFS (Glass Fibrosis Follicle). P(HEMA-CBOH) hydrogel can effectively reduce intraocular pressure, maintain the stability of the filtering bleb, significantly reduce foreign body reactions such as inflammation and fibrous capsule formation, reduce scleral collagen deposition, alleviate GFS surgical scar formation, and improve the success rate of GFS. Attached Figure Description

[0023] Figure 1 This is a schematic diagram of the synthesis of P(HEMA-CBOH) hydrogel.

[0024] Figure 2 The transmittance (A) and equilibrium water content (B) of P(HEMA-CBOH) hydrogel in the wavelength range of 400-800 nm are given.

[0025] Figure 3 The tensile strength and modulus of P(HEMA-CBOH) hydrogel are given.

[0026] Figure 4 Crack insensitivity test for P(HEMA-CBOH) hydrogel.

[0027] Figure 5 Data for protein adsorption characterization of P(HEMA-CBOH) hydrogel (A), relative cell viability (B), and anti-cell adhesion ability (C).

[0028] Figure 6 This is a schematic diagram of the application of P(HEMA-CBOH) hydrogel in the eye.

[0029] Figure 7 The changes in intraocular pressure in rabbits before and 3, 7, 14 and 28 days after GFS surgery were studied.

[0030] Figure 8 UBM examinations were performed on rabbits 7, 14, and 28 days after GFS surgery.

[0031] Figure 9 HE staining of rabbit eyeballs was performed 7, 14 and 28 days after GFS surgery.

[0032] Figure 10 Masson staining of rabbit eyeballs was performed 7, 14 and 28 days after GFS surgery.

[0033] Figure 11 The expression of α-SMA in rabbits was observed 7, 14, and 28 days after GFS surgery. Detailed Implementation

[0034] Example 1: Preparation of P(HEMA-CBOH) hydrogel

[0035] Reagents and their sources:

[0036] 2-Hydroxyethyl methacrylate (HEMA) and 2-hydroxy-2-methylpropenone (IRGACURE 1173, 97%) were purchased from Tianjin Xien Biochemical Technology Co., Ltd., and tetraethylene glycol diacrylate (TEGDMA) was purchased from Shanghai Mairui Co., Ltd.

[0037] Preparation steps:

[0038] At 80 °C, different masses of CBOH monomer and HEMA monomer solution were mixed and stirred for 3 min to form homogeneous transparent solutions with CBOH monomer mass ratios of 0, 3, 6, or 12 wt% (relative to total mass). Photoinitiator 1173 (1 wt% relative to total mass) and chemical crosslinking agent TEGDMA (0.5 wt% relative to total mass) were added to the above solutions and stirred evenly at room temperature. Irradiation under 365 nm UV light (predetermined intensity) for 1 hour formed PHEMA, P(HEMA-CBOH)-3, P(HEMA-CBOH)-6, and P(HEMA-CBOH)-12 hydrogels. The resulting hydrogels were soaked in 75% ethanol overnight, dialyzed in PBS to remove residual small molecules for at least 4 days, and then soaked in sterile PBS to obtain sterile hydrogels for later use.

[0039] The preparation process of P(HEMA-CBOH) hydrogel is as follows: Figure 1 As shown.

[0040] Example 2: Preparation of P(HEMA-CBOH) hydrogel

[0041] Unlike Example 1, at 100 °C, different masses of CBOH monomers were mixed with HEMA monomer solutions and stirred for 2 min to form homogeneous transparent solutions with CBOH monomer mass ratios of 5 and 15 wt% (relative to total mass). Photoinitiator 1173 (3 wt% relative to total mass) and chemical crosslinking agent TEGDMA (0.1 wt% relative to total mass) were added to the above solutions and stirred evenly at room temperature. The solutions were irradiated with 365 nm UV light (predetermined intensity) for 1 hour to form hydrogels. The resulting P(HEMA-CBOH)-5 and P(HEMA-CBOH)-10 hydrogels were soaked in 75% ethanol overnight, dialyzed in PBS to remove residual small molecules for at least 4 days, and then soaked in sterile PBS to obtain sterile hydrogels for later use.

[0042] Example 3: Preparation of P(HEMA-CBOH) hydrogel

[0043] Unlike Examples 1 and 2, different masses of CBOH monomers were mixed with HEMA monomer solutions and stirred for 5 min at 90 °C to form homogeneous, transparent solutions with CBOH monomer mass ratios of 0.1, 1, or 10 wt% (relative to the total mass). Photoinitiator 1173 (0.1 wt% relative to the total mass) and chemical crosslinking agent TEGDMA (1 wt% relative to the total mass) were added to the above solutions and stirred until homogeneous at room temperature. The solutions were irradiated with 365 nm UV light (predetermined intensity) for 1 hour to form hydrogels. The resulting P(HEMA-CBOH)-0.1, P(HEMA-CBOH)-1, and P(HEMA-CBOH)-10 hydrogels were soaked in 75% ethanol overnight, dialyzed in PBS to remove residual small molecules for at least 4 days, and then soaked in sterile PBS to obtain sterile hydrogels for later use.

[0044] Example 4: Evaluation of optical properties and measurement of equilibrium water content of P(HEMA-CBOH) hydrogel

[0045] Test methods: UV-Vis spectrophotometry and gravimetric method

[0046] Testing tool: UV-Vis spectrophotometer (GENESYS 180, ThermoFisher Scientific)

[0047] Experimental steps:

[0048] Evaluation of optical properties: At room temperature, using PBS as a blank, the transmittance of the hydrogels was measured in the wavelength range of 400-800 nm using a UV-Vis spectrophotometer. PHEMA, P(HEMA-CBOH)-3, P(HEMA-CBOH)-6, and P(HEMA-CBOH)-12 hydrogels were cut into strips, and their transmittance was measured in a quartz dish of a UV-Vis spectrophotometer.

[0049] Equilibrium water content measurement: The equilibrium water content (EWCs) of the hydrogel was determined by gravimetric method. The prepared hydrogel was placed in PBS for 4 days to allow it to reach swelling equilibrium. Then, the hydrogel was removed and weighed to obtain m. wet Then, it was placed in a freeze dryer to remove moisture, and the weight was measured to obtain m. dry Three parallel samples were made for each scale sample.

[0050] The EWCs of the hydrogel are:

[0051]

[0052] like Figure 2As shown, the prepared hydrogel possesses excellent optical properties. Furthermore, its high equilibrium water content helps reduce the modulus, thereby significantly reducing the foreign body sensation during use and improving comfort.

[0053] Example 5: Tensile strength and modulus of P(HEMA-CBOH) hydrogel

[0054] Test method: Uniaxial tensile test

[0055] Testing tool: Electronic universal testing machine (Legend 2344, Instron, USA)

[0056] Experimental steps:

[0057] For tensile testing, dumbbell-shaped samples (35 × 2.1 × 1 mm) of PHEMA, P(HEMA-CBOH)-3, P(HEMA-CBOH)-6, and P(HEMA-CBOH)-12 hydrogels were prepared according to ASTM standards. 3 All tests were conducted at a constant rate of 50 mm / min. Young's modulus was calculated from the initial slope of the stress-strain curve, with a strain range of 3% to 8%, and toughness was defined as the area under the stress-strain curve.

[0058] Excessive tensile strength and modulus can lead to severe foreign body sensation after implantation in the eye; therefore, the hydrogel used in GFS needs to be as soft as possible to meet suture requirements. Figure 3 As shown, the performance indicators of each group of hydrogels were relatively low, which can effectively reduce the foreign body sensation after implantation. Furthermore, with the increase of zwitterionic components, the hydrogels became more flexible, facilitating suturing.

[0059] Example 6: Crack Insensitivity Test of P(HEMA-CBOH) Hydrogel

[0060] Test method: Crack notch tensile cycle test

[0061] Testing tool: Electronic universal testing machine (Legend 2344, Instron, USA)

[0062] Experimental steps:

[0063] like Figure 4As shown in Figure A, this invention evaluates the crack insensitivity of each group of hydrogels through a crack-notch tensile cyclic test. In the load-unload tensile test, the notched specimens of P(HEMA-CBOH)-3, P(HEMA-CBOH)-6, and P(HEMA-CBOH)-12 hydrogels were first stretched to a predetermined strain (10%) at a tensile rate of 50 mm / min, and then immediately restored to the initial state at the same rate. The notch length of the notched specimen was 15% of the specimen width, and its stability was observed.

[0064] like Figure 4 As shown in B, after 10 cycles of testing, this notched hydrogel exhibited excellent stability with no further crack propagation, fully demonstrating that the hydrogel is stable under tensile strain, ensuring intraoperative suturing capability and suitability for glaucoma filtration surgery.

[0065] Example 7: Evaluation of the biological properties of P(HEMA-CBOH) hydrogel

[0066] Test method: BCA protein assay kit (Boster Biotechnology), CCK8 method (Dojindo, Japan)

[0067] Testing tools: Tecan Spark multi-functional microplate reader, inverted fluorescence microscope (Eclipse Ti-SR, Nikon, Japan)

[0068] Experimental steps:

[0069] 1. Measurement of nonspecific protein adsorption: PHEMA, P(HEMA-CBOH)-3, P(HEMA-CBOH)-6, and P(HEMA-CBOH)-12 hydrogels in swelling equilibrium were immersed in 1 mg / mL BSA and incubated at 37 °C for 1 day. The hydrogels were then rinsed three times with physiological saline to remove loosely adsorbed proteins from the surface. The proteins adsorbed in the hydrogels were then sonicated in 1% sodium dodecyl sulfate (SDS) aqueous solution for 10 minutes and analyzed using a BCA protein assay kit and a microplate reader. The average values ​​were recorded.

[0070] 2. Cell viability assay: Mouse fibroblasts (L929) were used, and cell compatibility was evaluated using the CCK-8 assay. L929 cells were seeded in 96-well plates at a density of 1 × 10⁶ cells / well. 4Cells per well were inoculated at 37 °C under a 5% CO2 atmosphere for 24 h. The culture medium was removed, and hydrogel extracts of PHEMA, P(HEMA-CBOH)-3, P(HEMA-CBOH)-6, and P(HEMA-CBOH)-12 containing the culture medium were added, with untreated cells serving as a control. After 24 h of incubation, the cells were analyzed using a CCK-8 assay kit and a microplate reader, and the data were recorded.

[0071] 3. Anti-cell adhesion assay: PHEMA, P(HEMA-CBOH)-3, P(HEMA-CBOH)-6, and P(HEMA-CBOH)-12 hydrogels were placed in 48-well plates and incubated at 37 °C for 1 h. L929 cells were then sputtered at a concentration of 1 × 10⁻⁶ cells / well. 5 L929 cells were seeded at a density of 10 cells / well and incubated at 37°C in 5% CO2 for 48 h. The cells were then gently washed with sterile PBS to remove suspension. The L929 cells adhering to the hydrogel surface were stained with calcein AM and observed under an inverted fluorescence microscope.

[0072] To determine the antifouling ability of P(HEMA-CBOH)-3, P(HEMA-CBOH)-6, and P(HEMA-CBOH)-12 hydrogels, this invention evaluated their non-specific protein adsorption properties. For example... Figure 5 As shown in Figure A, compared to pHEMA hydrogels, the resistance of each group of P(HEMA-CBOH) hydrogels to non-specific protein adsorption is significantly improved.

[0073] like Figure 5 B showed that the cell viability of each group exceeded 98%, indicating that the P(HEMA-CBOH) hydrogel has excellent biocompatibility.

[0074] like Figure 5 As shown in Figure C, after 48 hours of cell incubation, a large number of L929 cells were observed adhering to the surface of the cell culture plate (TCPS). Similarly, a small number of L929 cells were also observed on the surface of the hydrophilic PHEMA hydrogel, while almost no cells adhered to the P(HEMA-CBOH) hydrogels, indicating that they have excellent anti-cell adhesion ability.

[0075] Example 8: Rabbit Eye GFS Surgery

[0076] Animals and their source: New Zealand white rabbits (2.0-2.5 kg), purchased from Spiford (Beijing) Biotechnology Co., Ltd.

[0077] Experimental equipment: Ophthalmic surgical microscope (YZ20P5, Suzhou Liuliu)

[0078] Experimental steps:

[0079] 1. New Zealand white rabbits were randomly divided into a normal control group, a trabecular group, and a hydrogel group, with 6 rabbits in each group.

[0080] 2. Surgical procedure for the trabecular meshwork group: Rabbits were anesthetized with thiamethoxam hydrochloride (approximately 0.5 mL). The eyeball was exposed using a speculum and disinfected with povidone-iodine. Obuprocaine hydrochloride was used for local anesthesia of the ocular surface. The conjunctiva and subconjunctival tissue were carefully dissected. Then, a 3*4 mm incision was made to a depth of two-thirds the thickness of the sclera. 2 A rectangular scleral flap was created, soaked in 0.2 mg / mL MMC, covered with a sponge for 2 minutes, and then rinsed for 30 seconds. Trabeculectomy was then performed, followed by intermittent suturing of the scleral flap to the conjunctiva using 10-0 nylon sutures. Postoperatively, TobraDex ointment and levofloxacin were used to reduce inflammation.

[0081] 3. Surgical procedure for the hydrogel group: Rabbits were anesthetized with thiamethoxam hydrochloride (approximately 0.5 mL). The eyeball was exposed using a speculum and disinfected with povidone-iodine. Obuprocaine hydrochloride was used for local anesthesia of the ocular surface. The conjunctiva and subconjunctival tissue were carefully dissected. Then, a 3*4 mm incision was made to a depth of two-thirds the thickness of the sclera. 2 A rectangular scleral flap was created, soaked in 0.2 mg / mL MMC, covered with a sponge for 2 minutes, and then rinsed for 30 seconds. Trabeculectomy was then performed, and hydrogel was placed under the scleral flap. The scleral flap was then intermittently sutured to the conjunctiva with 10-0 nylon sutures. Postoperatively, TobraDex ointment and levofloxacin were used to reduce inflammation.

[0082] A schematic diagram of the hydrogel group surgical method is shown below. Figure 6 As shown.

[0083] Example 9: Postoperative intraocular pressure measurement after GFS

[0084] Animals and their source: New Zealand white rabbits (2.0-2.5 kg), purchased from Spiford (Beijing) Biotechnology Co., Ltd.

[0085] Experimental equipment: TonoLab, ICare tonometer

[0086] Experimental steps:

[0087] 1. New Zealand white rabbits were randomly divided into a normal control group, a trabecular group, and a hydrogel group, with 6 rabbits in each group.

[0088] 2. Intraocular pressure (IOP) was measured in rabbits using a tonometer through the center of the cornea before and after surgery. To ensure accuracy, measurements were taken at 2 PM. IOP values ​​were measured before surgery and at 3, 7, 14, and 28 days post-surgery.

[0089] like Figure 7As shown, on postoperative day 3, intraocular pressure (IOP) in both the trabecular group and the material group decreased compared to the normal control group. Subsequently, on postoperative day 7, the hydrogel group showed a more significant IOP-lowering effect than the trabecular group. On postoperative day 28, the IOP in the trabecular group returned to normal levels, while the hydrogel group continued to maintain its IOP-lowering effect, indicating that P(HEMA-CBOH) hydrogel can effectively reduce IOP.

[0090] Example 10: Postoperative UBM measurement after GFS

[0091] Animals and their source: New Zealand white rabbits (2.0-2.5 kg), purchased from Spiford (Beijing) Biotechnology Co., Ltd.

[0092] Experimental equipment: Ultrasonic biological microscope (UBM) (MD-300L, MEDA Co., Ltd, Tianjin, China)

[0093] Experimental steps:

[0094] 1. New Zealand white rabbits were randomly divided into a normal control group, a trabecular group, and a hydrogel group, with 6 rabbits in each group.

[0095] 2. Rabbits were first anesthetized, followed by local anesthesia of the eyes with oxybuprocaine hydrochloride to carefully expose the follicles for clear imaging. The follicles were examined using an ultrasonic biomicroscope and a covered sterile probe (QUANTEL MEDICAL, Clermont-Frand, France).

[0096] Filtration vesicles are structures formed during GFS (Gastrostomy for Failure to Filter Follicles) and are one of the key indicators of surgical success. For example... Figure 8 As shown, the red arrows mark the locations of the filtration vesicles. The morphology of the vesicles can be clearly observed using UBM. The filtration vesicles in the hydrogel group were still present after 28 days, while the vesicles in the trabecular group were only maintained for 14 days, indicating that the P(HEMA-CBOH) hydrogel can better maintain the presence of vesicles.

[0097] Example 11: HE staining after GFS surgery

[0098] Animals and their source: New Zealand white rabbits (2.0-2.5 kg), purchased from Spiford (Beijing) Biotechnology Co., Ltd.

[0099] Test method: Hematoxylin and eosin (H&E) staining of paraffin sections.

[0100] Testing tool: BX51 microscope (Olympus Optical Co., Ltd., Tokyo, Japan)

[0101] Experimental steps:

[0102] 1. New Zealand white rabbits were randomly divided into a normal control group, a trabecular group, and a hydrogel group, with 6 rabbits in each group.

[0103] 2. Rabbits were euthanized on postoperative days 7, 14, and 28, and the eyeballs were separated. The eye tissue was then fixed in 10% formalin to preserve its cellular structure and embedded in paraffin. Serial sections with a thickness of 3 μm were cut.

[0104] 3. Stain with hematoxylin and eosin (H&E) and observe under a microscope.

[0105] like Figure 9 As shown, histopathological examinations were performed on postoperative days 7, 14, and 28 to observe changes in ocular tissue. The results clearly showed that in the trabecular group, significant aggregation of blue-stained cell nuclei (marked with red arrows) was observed in the sclera at all observation time points. This phenomenon was significantly more prevalent than in the normal control group and the hydrogel group, suggesting a stronger inflammatory response. Furthermore, the presence of P(HEMA-CBOH) hydrogel was still observed at all time points, fully demonstrating its excellent biological stability.

[0106] Example 12: Masson staining after GFS surgery

[0107] Animals and their source: New Zealand white rabbits (2.0-2.5 kg), purchased from Spiford (Beijing) Biotechnology Co., Ltd.

[0108] Test method: Masson staining of paraffin sections

[0109] Testing tool: BX51 microscope (Olympus Optical Co., Ltd., Tokyo, Japan)

[0110] Experimental steps:

[0111] 1. New Zealand white rabbits were randomly divided into a normal control group, a trabecular group, and a hydrogel group, with 6 rabbits in each group.

[0112] 2. Rabbits were euthanized on postoperative days 7, 14, and 28, and the eyeballs were separated. The eye tissue was then fixed in 10% formalin to preserve its cellular structure and embedded in paraffin. Serial sections with a thickness of 3 μm were cut.

[0113] 3. All sections were stained with Masson's solution and observed under a microscope.

[0114] Collagen deposition is a crucial step in the healing process of GFS (Gastrostomy Fibrillation); however, excessive collagen formation may adversely affect surgical outcomes. Figure 10As shown, at various postoperative observation time points, the trabecular group showed significantly deeper staining in the scleral region than the hydrogel group, revealing excessive collagen deposition. When tissue is damaged or inflammatory, the spaces between collagen fibers expand, resulting in a loose and irregular fiber arrangement. Through comparative observation, this invention found that the collagen fibers in the trabecular group were more sparsely arranged compared to the other two groups. These results indicate that P(HEMA-CBOH) hydrogel can more effectively reduce collagen deposition and inflammatory responses.

[0115] Example 13: Immunohistochemical staining after GFS surgery

[0116] Animals and their source: New Zealand white rabbits (2.0-2.5 kg), purchased from Spiford (Beijing) Biotechnology Co., Ltd.

[0117] Test method: Immunohistochemical staining of paraffin sections (primary antibody: α-SMA, secondary antibody: HRP)

[0118] Testing tool: BX51 microscope (Olympus Optical Co., Ltd., Tokyo, Japan)

[0119] Experimental steps:

[0120] 1. New Zealand white rabbits were randomly divided into a normal control group, a trabecular group, and a hydrogel group, with 6 rabbits in each group.

[0121] 2. Rabbits were euthanized on postoperative days 7, 14, and 28, and the eyeballs were separated. The eye tissue was then fixed in 10% formalin to preserve its cellular structure and embedded in paraffin. Serial sections with a thickness of 3 μm were cut.

[0122] 3. Primary antibody was labeled with α-SMA (Proteintech, 1:200), and secondary antibody was labeled with horseradish peroxidase HRP (Thermo, 1:1000). After staining, the samples were observed under a microscope.

[0123] like Figure 11 As shown, this invention detected the expression level of α-SMA in rabbit eyes at different time points. α-SMA is commonly used as a marker protein for smooth muscle cells or myofibroblasts. Myofibroblasts are cells that emerge during wound healing and tissue fibrosis. Compared to the normal group and the trabecular group, the hydrogel group showed lower α-SMA expression. This indicates that P(HEMA-CBOH) hydrogel has a significant effect on preventing tissue fibrosis after GFS surgery.

[0124] Based on the above description, this invention, through optimization of reactants and processes, yields a hydrogel that can effectively reduce intraocular pressure and alleviate scarring after glaucoma filtration surgery. This invention fully utilizes the excellent mechanical properties and biocompatibility of PHEMA, combining it with CBOH monomers to solve the problems of low water content, high modulus leading to severe foreign body sensation, insufficient abrasion resistance, and inadequate lubrication that still exist in PHEMA-based hydrogels. Through experimental verification, the optimal addition amounts of CBOH monomer, initiator 1173, and chemical crosslinking agent TEGDMA were determined to be 0.1-15 wt%, 0.1-5 wt%, and 0.1-1 wt% of total mass, respectively, resulting in the hydrogel of this invention. Furthermore, the optimal addition amounts of the three components were determined to be 5-13 wt%, 0.5-3 wt%, and 0.2-0.6 wt%, respectively, achieving the desired technical effects for the hydrogel. Even further, this invention determined the addition amounts of the three components to be 12 wt%, 1 wt%, and 0.5 wt% of total mass, resulting in outstanding technical effects for the hydrogel product.

[0125] It should be further noted that the above embodiments are only used for understanding the technical solution of the present invention and are not intended to limit the scope of protection of the present invention. Any obvious adjustments and modifications made to the technical solution of the present invention that are part of the technical concept of the present invention should also be within the scope of protection of the present invention.

Claims

1. A method for preparing a hydrogel that resists scarring after glaucoma filtration surgery, characterized in that... The steps are as follows: 1) The CBOH monomer is heated and dissolved in the HEMA solution to form a homogeneous solution; 2) Add the photoinitiator 2-hydroxy-2-methyl-1-phenyl-1-propanone and the chemical crosslinking agent triethylene glycol dimethacrylate to the solution obtained in step 1), react at room temperature for 20 min, and then mix thoroughly to obtain a HEMA-CBOH solution; 3) In a UV crosslinking chamber, the HEMA-CBOH solution obtained in step 2) is irradiated under UV light for 1 hour to obtain P(HEMA-CBOH) hydrogel; 4) Soak the P(HEMA-CBOH) hydrogel obtained in step 3) in 75% ethanol overnight, and then dialyze it in sterile PBS for 4-8 days to remove residual small molecules. After solubilization and equilibrium, a sterile hydrogel is obtained. The resulting sterile hydrogel can be used for suturing; In step 1), the mass ratio of the added CBOH monomer is 0.1-15 wt% relative to the total mass, the heating temperature is 80-100 degrees Celsius, and the heating time is 2-5 minutes; In step 2), the photoinitiator 2-hydroxy-2-methyl-1-phenyl-1-propanone has a mass ratio of 0.1-5 wt% relative to the total mass, and the chemical crosslinking agent triethylene glycol dimethacrylate has a mass ratio of 0.1-1 wt% relative to the total mass. Step 3) The UV light wavelength used is 365nm; After solubilization equilibrium in step 4), the final thickness of the hydrogel is 100-1000 micrometers.

2. The method for preparing the hydrogel for anti-scarring reaction after glaucoma filtration surgery as described in claim 1, characterized in that, The amount of CBOH monomer added is 5-13 wt% of the total mass.

3. The method for preparing the hydrogel for anti-scarring reaction after glaucoma filtration surgery as described in claim 1, characterized in that, The photoinitiator 2-hydroxy-2-methyl-1-phenyl-1-propanone is present in a mass ratio of 0.5-5 wt% relative to the total mass, and the chemical crosslinking agent triethylene glycol dimethacrylate is present in a mass ratio of 0.2-0.6 wt% relative to the total mass.

4. An anti-scarring drug carrier, characterized in that, The hydrogel obtained by any one of claims 1-3 is used to load anti-scarring and hormonal drugs.

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