An anti-swelling microgel, its preparation method and application

Abstract

The application discloses anti-swelling microgels, a preparation method and application thereof. The preparation method of the anti-swelling microgels comprises the following steps: preparing the anti-swelling microgels by using a microfluidic technology; wherein the preparation raw materials of the anti-swelling microgels comprise water phase materials, the water phase materials comprise acrylic ester polymerization monomers and degradation-resistant raw materials, the acrylic ester polymerization monomers comprise at least one of polyether F127 diacrylate, polyethylene glycol diacrylate, 4-arm-polyethylene glycol-methacrylate, 8-arm-polyethylene glycol-acrylate or polyethylene glycol-acrylate, and the mass fraction w of the acrylic ester polymerization monomers in the water phase materials satisfies 60% >= w >= 1%. The anti-swelling microgels have the characteristics of good anti-swelling performance and good mechanical properties, and have strong injectability, and have a good application prospect in the fields of biomedical materials, tissue engineering and regenerative medicine technology.

Description

Technical Field

[0001] This invention belongs to the field of biomaterials technology, specifically relating to an anti-swelling microgel, its preparation method, and its application. Background Technology

[0002] Cell therapy, with its advantages of good efficacy, broad therapeutic range, and low toxicity, has become a hot topic in medical research in recent years. Clinically, cell therapy has achieved certain therapeutic effects. However, some drawbacks have emerged during treatment, such as immune rejection, severe and unmanageable cell loss, and low cell survival rates after transplantation. Microgels, due to their high water content, composition similar to the extracellular matrix, and good biocompatibility, can encapsulate cells in hydrogel materials, providing a three-dimensional support, reducing cell loss, and improving cell survival rates after transplantation. The nanopores of hydrogels can also provide natural immune isolation protection for cells. However, traditional microgels are prone to uncontrolled swelling and deformation, thus limiting their application in tissue engineering. Summary of the Invention

[0003] The present invention aims to solve at least one of the technical problems existing in the prior art. To this end, the present invention proposes an anti-swelling microgel with good anti-swelling properties.

[0004] The present invention also proposes a method for preparing anti-swelling microgels.

[0005] This invention also proposes a biomaterial.

[0006] The present invention also proposes a medical product.

[0007] The present invention also proposes the application of the above-mentioned anti-swelling microgel.

[0008] In a first aspect, the present invention provides an anti-swelling microgel, the preparation method of which includes: preparing the anti-swelling microgel using microfluidic technology; wherein the raw materials for preparing the anti-swelling microgel include an aqueous phase material, the aqueous phase material includes acrylate polymeric monomers and degradation-resistant raw materials, the acrylate polymeric monomers include at least one of polyether F127 diacrylate, polyethylene glycol diacrylate, 4-arm-polyethylene glycol-methacrylate, 8-arm-polyethylene glycol-acrylate, or polyethylene glycol-acrylate, and the mass fraction w of the acrylate polymeric monomers in the aqueous phase material satisfies: 60% ≥ w ≥ 1%.

[0009] The anti-swelling microgel according to embodiments of the present invention has at least the following beneficial effects:

[0010] In this invention, aqueous materials are processed using microfluidic technology and undergo monomer polymerization to obtain the anti-swelling microgel. The acrylate monomers used in this invention, such as polyether F127 diacrylate and polyethylene glycol diacrylate, all contain hydrophobic groups in their chains, resulting in an anti-swelling microgel with a high proportion of hydrophobic segments, making it less prone to water absorption. Therefore, the anti-swelling microgel of this invention exhibits excellent anti-swelling properties and good mechanical properties. Furthermore, the addition of degradation-resistant raw materials effectively extends the degradation cycle of the anti-swelling microgel, making it less susceptible to degradation in liquid environments. Therefore, the anti-swelling microgel of this invention can be well applied in biotherapy (such as cell therapy) and tissue engineering. For example, cells can be encapsulated in hydrogel materials, providing not only three-dimensional support but also preventing damage to cells during injection and transplantation. Moreover, the long degradation cycle of the anti-swelling microgel reduces the likelihood of direct cell exposure to the host, leading to transplantation failure and better therapeutic effects. Meanwhile, the microgels in this invention are not prone to uncontrolled swelling and deformation when used in vivo, have strong injectability, long degradation cycle, and their mechanical properties are not easily degraded, and they are not likely to cause damage to surrounding tissues. They have great application prospects in the fields of biomedical materials, tissue engineering, and regenerative medicine.

[0011] In some embodiments of the present invention, the average particle size of the anti-swelling microgel is 50-1200 μm.

[0012] In some embodiments of the present invention, w ≥ 10%.

[0013] Through the above embodiments, the mass fraction w of acrylate polymer monomers in the aqueous phase material is ≥10%, which makes the polymer crosslinking degree of the obtained anti-swelling microgel higher and denser, and can quickly achieve swelling equilibrium, with a lower degree of swelling and better mechanical properties.

[0014] In some embodiments of the present invention, 50% ≥ w ≥ 10%.

[0015] In some embodiments of the present invention, the molecular weight of the polyether F127 diacrylate is 200-20000.

[0016] In some preferred embodiments of the present invention, the molecular weight of the polyether F127 diacrylate is 400-8000.

[0017] In some embodiments of the present invention, the molecular weight of the polyethylene glycol diacrylate is 200-20000.

[0018] In some embodiments of the present invention, the acrylate polymeric monomer includes at least one of polyether F127 diacrylate or polyethylene glycol diacrylate.

[0019] In some embodiments of the present invention, the mass ratio of the polyether F127 diacrylate to the polyethylene glycol diacrylate is (0.1-5):1.

[0020] In some preferred embodiments of the present invention, the mass ratio of the polyether F127 diacrylate to the polyethylene glycol diacrylate is (1-3):1.

[0021] In some embodiments of the present invention, the degradation-resistant raw material includes at least one of sodium carboxymethyl cellulose or modified sodium carboxymethyl cellulose.

[0022] Through the above embodiments, sodium carboxymethyl cellulose and modified sodium carboxymethyl cellulose can effectively improve the degradation cycle of anti-swelling microgels, making them less prone to degradation in liquid environments, which is beneficial for their application in tissue engineering. In cell therapy and related applications, the anti-swelling microgel of the present invention has a long degradation cycle, making it less likely to cause cells to be directly exposed to the host and thus causing transplantation failure, resulting in good therapeutic effects.

[0023] In some preferred embodiments of the present invention, the modified sodium carboxymethyl cellulose comprises acryloyl carboxymethyl cellulose sodium.

[0024] In some preferred embodiments of the present invention, the mass ratio of the acrylate polymer monomer to the degradation-resistant raw material is (2-10):(0.5-1.5).

[0025] In some embodiments of the present invention, the aqueous phase material further includes an initiator.

[0026] In some preferred embodiments of the present invention, the initiator includes a photoinitiator.

[0027] In some preferred embodiments of the present invention, the photoinitiator comprises lithium phenyl (2,4,6-trimethylbenzoyl) phosphate (LAP).

[0028] In some embodiments of the present invention, the aqueous phase material further includes active ingredients.

[0029] In some preferred embodiments of the present invention, the active ingredient includes at least one of cellular, pharmaceutical, or protein-based active factors.

[0030] In some preferred embodiments of the present invention, the cells include at least one of stem cells, progenitor cells, fibroblasts, kidney cells, pancreatic islet cells, tumor cells, or hepatocytes.

[0031] Through the above embodiments, the anti-swelling microgel that encapsulates cells in this invention has superior mechanical properties, good stability, and a long in vivo degradation cycle, which can reduce the apoptosis of encapsulated cells due to human immune rejection and is conducive to achieving long-term treatment.

[0032] In some more preferred embodiments of the present invention, the cell density in the aqueous phase material is 1*102 3 Cells / mL ~ 1*10 9 per mL.

[0033] In some preferred embodiments of the present invention, the drug includes, but is not limited to, ascorbic acid, vitamins, rapamycin, ciprofloxacin, or silymarin.

[0034] In some preferred embodiments of the present invention, the protein-like active factors may include various growth factors such as vascular endothelial growth factor, pancreatic islet development transcription factor regulatory substances, and factors that prevent pancreatic islet cell apoptosis.

[0035] In some preferred embodiments of the present invention, the protein-like active factors include, but are not limited to, factors such as angiogenesis factor or human bone morphogenetic protein-2.

[0036] In some preferred embodiments of the present invention, the mass fraction of the active ingredient in the aqueous phase material is 0.5-30%.

[0037] In a second aspect, the present invention provides a method for preparing an anti-swelling microgel, the method comprising the following steps: taking an aqueous phase material and an oil phase material, and preparing the anti-swelling microgel by employing microfluidic technology.

[0038] In some embodiments of the present invention, the preparation method includes the following steps: taking aqueous phase material and oil phase material, preparing microdroplets using microfluidic technology, polymerizing them, and obtaining the anti-swelling microgel.

[0039] In the above embodiments, the microdroplets are W / O microdroplets.

[0040] In some preferred embodiments of the present invention, the polymerization is a photopolymerization reaction of microdroplets.

[0041] In some embodiments of the present invention, the oil phase material includes surfactants and solvents.

[0042] In some preferred embodiments of the present invention, the surfactant includes a nonionic fluorocarbon surfactant.

[0043] In some preferred embodiments of the present invention, the solvent comprises a fluorinated oil. Preferably, the fluorinated oil is fluorinated oil HFE-7500.

[0044] In some embodiments of the present invention, the preparation method includes the following steps: adding aqueous phase material and oil phase material into a microfluidic device to prepare microdroplets, polymerizing them to obtain the anti-swelling microgel.

[0045] In some preferred embodiments of the present invention, the flow rate ratio of the aqueous phase material to the oil phase material in the microfluidic device is (0.3-5):(8-15).

[0046] In some preferred embodiments of the present invention, the flow rate of the aqueous phase material in the microfluidic device is 0.5-3 mL / h.

[0047] In some preferred embodiments of the present invention, the flow rate of the oil phase material in the microfluidic device is 8-15 mL / h.

[0048] In some embodiments of the present invention, the preparation method includes the following steps:

[0049] S1, Preparation of aqueous phase material, including the following operations: Take a PBS solution containing an initiator, mix it with acrylate polymer monomers and degradation-resistant raw materials to obtain aqueous phase material;

[0050] S2, add aqueous and oily materials into a microfluidic device to prepare microdroplets, and then perform photopolymerization to obtain the anti-swelling microgel.

[0051] In some preferred embodiments of the present invention, in step S1, after mixing the PBS solution containing the initiator with the acrylate polymer monomer and the degradation-resistant raw material, the active ingredient is added to obtain an aqueous phase material.

[0052] In some preferred embodiments of the present invention, in step S1, the PBS solution containing the initiator is mixed with acrylate polymer monomers and degradation-resistant raw materials, sterilized, and then cell resuspension is added to obtain an aqueous phase material.

[0053] In some preferred embodiments of the present invention, in step S1, the initiator and PBS solution are mixed to obtain the PBS solution containing the initiator.

[0054] In some preferred embodiments of the present invention, the initiator in the PBS solution containing the initiator has a mass fraction of 0.05-0.3%.

[0055] In some preferred embodiments of the present invention, step S1, preparing the aqueous phase material, specifically includes the following operations: mixing the initiator and PBS solution to obtain the PBS solution containing the initiator, and mixing it with acrylate polymer monomers and degradation-resistant raw materials to obtain the aqueous phase material.

[0056] In some more preferred embodiments of the present invention, in step S1, the ratio of the amount of acrylate polymer monomer, the degradation-resistant raw material and the PBS solution is (2-10)g:(0.5-1.5)g:(9-11)mL.

[0057] In some preferred embodiments of the present invention, step S1 further includes preparing an oil phase material, including the following operation: mixing a surfactant and a solvent to obtain an oil phase material.

[0058] In some more preferred embodiments of the present invention, step S1, preparing the oil phase material, specifically includes the following operation: mixing a nonionic fluorocarbon surfactant and fluorinated oil HFE-7500 to obtain the oil phase material.

[0059] In some preferred embodiments of the present invention, in step S2, during photopolymerization, the light wavelength is 405-420 nm and the light exposure time is 1-5 min.

[0060] In some preferred embodiments of the present invention, in step S2, the obtained anti-swelling microgel is washed with fluorinated oil HFE7500, then washed three times with PBS solution, and then transferred to PBS solution for storage.

[0061] A third aspect of the present invention provides a biomaterial comprising the above-described anti-swelling microgel.

[0062] In a fourth aspect, the present invention provides a medical product comprising at least one of the above-described anti-swelling microgels or the above-described biomaterials.

[0063] In a fifth aspect, the present invention proposes the application of the above-mentioned anti-swelling microgel in the preparation of medical materials or biomaterials. Attached Figure Description

[0064] The present invention will be further described below with reference to the accompanying drawings and embodiments, wherein:

[0065] Figure 1 This is an optical image of the anti-swelling microgel in Example 1 of the present invention;

[0066] Figure 2 This is an optical image of the anti-swelling microgel in Example 2 of the present invention;

[0067] Figure 3 This is an optical image of the anti-swelling microgel in Example 3 of the present invention;

[0068] Figure 4 This is an optical image of the anti-swelling microgel in Example 4 of the present invention;

[0069] Figure 5This is a graph showing the swelling performance test results of the anti-swelling microgel in Example 5 of the present invention;

[0070] Figure 6 The figure shows the swelling performance test results of the microgel in Comparative Example 1.

[0071] Figure 7 The figure shows the swelling performance test results of the microgel in Comparative Example 2;

[0072] Figure 8 This is a diagram showing the cell viability and mortality characterization results in the anti-swelling microgel in Example 5 of the present invention. Detailed Implementation

[0073] The following will describe the concept and technical effects of the present invention clearly and completely with reference to embodiments, so as to fully understand the purpose, features and effects of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are all within the scope of protection of the present invention.

[0074] Unless otherwise specified, the experimental methods used in the following examples were generally performed under conventional conditions in the art or as recommended by the manufacturer. Unless otherwise specified, all raw materials and reagents used are commercially available from the general market. The cells (or cell populations) used in the examples were all commercially available (or may be obtained through self-differentiation).

[0075] Example 1

[0076] This embodiment discloses an anti-swelling microgel, the preparation process of which includes:

[0077] (I) Preparation of aqueous precursor solution (aqueous material):

[0078] 0.2 g of lithium phenyl (2,4,6-trimethylbenzoyl) phosphate (LAP) was dissolved in 100 mL of PBS solution to obtain a 0.2 wt% LAP solution. 2 g of polyether F127 diacrylate (F127DA), 2 g of Ficoll (density = 1.084 g / mL), 1 g of polyethylene glycol diacrylate (PEGDA), and 0.5 g of sodium carboxymethyl cellulose (CMC-Na) were added to a sampling cup, followed by 10 mL of the aforementioned 0.2 wt% LAP solution. The mixture was stirred until the solids were completely dissolved to obtain an aqueous precursor solution. The sum of the mass fractions of F127DA and PEGDA in the aqueous precursor solution was 16.6%. F127DA was purchased from EFL, lot number EFL-PEGDA-600, with a molecular weight of 600; sodium carboxymethyl cellulose had a molecular weight of 240 kDa; and PEGDA had a molecular weight of 700. In some other embodiments of the present invention, the molecular weight of F127DA can be 400-8000; the molecular weight of sodium carboxymethyl cellulose can be 90-500 kDa; and the molecular weight of PEGDA can be 200-20000.

[0079] (II) Preparation of oil phase materials:

[0080] Take 1 mL of a 10% (g / mL) nonionic fluorocarbon surfactant (wherein the solvent is fluorinated oil HFE-7500 and the solute is a nonionic fluorocarbon surfactant), add 9 mL of fluorinated oil HFE-7500 and dilute to a 1% (g / mL) concentration, stir well to obtain the oil phase material.

[0081] (III) Preparation of anti-swelling microgels:

[0082] Aqueous and oil-phase materials were added to a microfluidic device. The microfluidic chip used in the experiment contained a T-shaped flow channel with a rectangular cross-section, the dimensions of which were 300 μm * 300 μm (width * height). W / O microdroplets were prepared using microfluidic technology, with the flow rates of the aqueous and oil phases set at 1 mL / h and 15 mL / h, respectively. The microdroplets generated in the microfluidic chip were collected and irradiated under blue light (405-420 nm) for 2 min to crosslink them into microgel spheres. After washing with fluorinated oil HFE7500, the microgels were washed three times with PBS solution and then transferred back to PBS solution for storage, yielding anti-swelling microgels.

[0083] This embodiment provides a biomaterial, including the anti-swelling microgel prepared by the above operations.

[0084] This embodiment provides a medical product comprising the anti-swelling microgel prepared by the above operations.

[0085] Example 2

[0086] This embodiment discloses an anti-swelling microgel, which differs from Example 1 in that the same mass of sodium carboxymethyl cellulose methacrylamide is used in this embodiment instead of sodium carboxymethyl cellulose in Example 1; and in step (III) preparation of the anti-swelling microgel, the flow rates of the aqueous phase material and the oil phase material are set to 1 mL / h and 10 mL / h, respectively.

[0087] This embodiment provides a biomaterial, including the anti-swelling microgel prepared by the above operations.

[0088] This embodiment provides a medical product comprising the anti-swelling microgel prepared by the above operations.

[0089] Example 3

[0090] This embodiment discloses an anti-swelling microgel, which is a cell-encapsulating anti-swelling microgel. The only difference between this embodiment and Embodiment 1 is that the aqueous phase material in this embodiment also includes MSC cells (mesenchymal stem cells); the microfluidic chip selected in this embodiment contains a T-shaped flow channel, and the flow channel cross-section is rectangular, with the rectangle having dimensions of 100μm*100μm (width*height); and in step (III) preparation of the anti-swelling microgel, the flow rates of the aqueous phase material and the oil phase material are set to 1mL / h and 8mL / h, respectively.

[0091] Specifically, the preparation of the aqueous phase material in this embodiment includes the following steps:

[0092] 0.2 g of lithium phenyl (2,4,6-trimethylbenzoyl) phosphate (LAP) was dissolved in 100 mL of PBS solution to obtain a 0.2 wt% LAP solution. 2 g of polyether F127 diacrylate (F127DA), 2 g of Ficoll (density = 1.084 g / mL), 1 g of polyethylene glycol diacrylate (PEGDA), and 0.5 g of sodium carboxymethyl cellulose (CMC-Na) were added to a sampling cup, along with 10 mL of the aforementioned 0.2 wt% LAP solution. The mixture was stirred, filtered for sterilization, and a cell-free aqueous phase was obtained. 200 μL of this aqueous phase was then added to 200 μL of MSC cell resuspension (cell density 1*10⁻⁶). 6 Cells / mL, cell culture medium: 1640 medium), to obtain an aqueous precursor solution. The sum of the mass fractions of F127DA and PEGDA in the aqueous precursor solution is 10%. In some other embodiments of the present invention, the volume ratio of the aqueous liquid to the cell resuspension is not limited to 1:1.

[0093] This embodiment provides a biomaterial, including the anti-swelling microgel prepared by the above operations.

[0094] This embodiment provides a medical product comprising the anti-swelling microgel prepared by the above operations.

[0095] Example 4

[0096] This embodiment discloses an anti-swelling microgel, which is a cell-encapsulating anti-swelling microgel. The only difference between this embodiment and Embodiment 1 is that the aqueous phase material in this embodiment also includes NPC cells (renal progenitor cells), the microfluidic chip selected in this embodiment contains a T-shaped flow channel, the flow channel cross-section is rectangular, and the size of the rectangle is 100μm*100*μm (width*height); and in the preparation of the anti-swelling microgel in step (Ⅲ), the flow rates of the aqueous phase material and the oil phase material in this embodiment are set to 1mL / h and 6mL / h, respectively.

[0097] Specifically, the preparation of the aqueous phase material in this embodiment includes the following steps:

[0098] 0.2 g of lithium phenyl (2,4,6-trimethylbenzoyl) phosphate (LAP) was dissolved in 100 mL of PBS solution to obtain a 0.2 wt% LAP solution. 2 g of polyether F127 diacrylate (F127DA), 2 g of Ficoll (density = 1.084 g / mL), 1 g of polyethylene glycol diacrylate (PEGDA), and 0.5 g of sodium carboxymethyl cellulose (CMC-Na) were added to a sampling cup, along with 10 mL of the aforementioned 0.2 wt% LAP solution. After stirring and sterilization by filtration, a cell-free aqueous phase was obtained. 200 μL of this aqueous phase was then added, along with 200 μL of NPC cell resuspension (cell density 1*10⁻⁶). 6 Cells / mL, cell culture medium: 1640 medium), to obtain an aqueous precursor solution. The sum of the mass fractions of F127DA and PEGDA in the aqueous precursor solution was 10%.

[0099] This embodiment provides a biomaterial, including the anti-swelling microgel prepared by the above operations.

[0100] This embodiment provides a medical product comprising the anti-swelling microgel prepared by the above operations.

[0101] Example 5

[0102] This embodiment discloses an anti-swelling microgel, which is a cell-encapsulating anti-swelling microgel. The only difference between this embodiment and Embodiment 1 is that the aqueous phase material in this embodiment also includes pancreatic islet cells; and in step (III) preparation of the anti-swelling microgel, the flow rates of the aqueous phase material and the oil phase material in this embodiment are set to 1 mL / h and 10 mL / h, respectively.

[0103] Specifically, the preparation of the aqueous phase material in this embodiment includes the following steps:

[0104] 0.2 g of lithium phenyl (2,4,6-trimethylbenzoyl) phosphate (LAP) was dissolved in 100 mL of PBS solution to obtain a 0.2 wt% LAP solution. 2 g of polyether F127 diacrylate (F127DA), 2 g of Ficoll (density = 1.084 g / mL), 1 g of polyethylene glycol diacrylate (PEGDA), and 0.5 g of sodium carboxymethyl cellulose (CMC-Na) were added to a sampling cup, along with 10 mL of the aforementioned 0.2 wt% LAP solution. After stirring and sterilization by filtration, a cell-free aqueous phase was obtained. 200 μL of this aqueous phase was then added to 200 μL of a resuspension of pancreatic islet cells (cell density 1*10⁻⁶). 6 Cells / mL, cell culture medium: 1640 medium), to obtain an aqueous precursor solution. The sum of the mass fractions of F127DA and PEGDA in the aqueous precursor solution was 10%.

[0105] This embodiment provides a biomaterial, including the anti-swelling microgel prepared by the above operations.

[0106] This embodiment provides a medical product comprising the anti-swelling microgel prepared by the above operations.

[0107] Example 6

[0108] This embodiment discloses an anti-swelling microgel, which differs from Example 1 in that: in this embodiment, the same mass of 4-arm-polyethylene glycol-methacrylate is used instead of F127DA in Example 1.

[0109] This embodiment provides a biomaterial, including the anti-swelling microgel prepared by the above operations.

[0110] This embodiment provides a medical product comprising the anti-swelling microgel prepared by the above operations.

[0111] Example 7

[0112] This embodiment discloses an anti-swelling microgel, which differs from Example 1 in that: in this embodiment, the same mass of 8-arm polyethylene glycol-acrylate is used instead of F127DA in Example 1, and the same mass of polyethylene glycol-acrylate is used instead of PEGDA in Example 1.

[0113] This embodiment provides a biomaterial, including the anti-swelling microgel prepared by the above operations.

[0114] This embodiment provides a medical product comprising the anti-swelling microgel prepared by the above operations.

[0115] In other embodiments of the present invention, at least one of 8-arm polyethylene glycol-acrylate or polyethylene glycol-acrylate of the same mass may be used instead of F127DA or PEGDA in Example 1.

[0116] Example 8

[0117] This embodiment discloses an anti-swelling microgel, which is a cell-encapsulating anti-swelling microgel. The difference between this embodiment and Example 5 is that the same mass of polyethylene glycol-acrylate is used instead of PEGDA in Example 5.

[0118] This embodiment provides a biomaterial, including the anti-swelling microgel prepared by the above operations.

[0119] This embodiment provides a medical product comprising the anti-swelling microgel prepared by the above operations.

[0120] Example 9

[0121] This embodiment discloses an anti-swelling microgel, which is a cell-encapsulating anti-swelling microgel. The difference between this embodiment and Example 5 is that the same mass of 4-arm polyethylene glycol-methacrylate is used instead of F127DA in Example 5, and the same mass of polyethylene glycol-acrylate is used instead of PEGDA in Example 1.

[0122] This embodiment provides a biomaterial, including the anti-swelling microgel prepared by the above operations.

[0123] This embodiment provides a medical product comprising the anti-swelling microgel prepared by the above operations.

[0124] Comparative Example 1

[0125] This comparative example discloses a microgel, which differs from Example 1 only in that: in step (I) of this comparative example, the polymerizable monomer in the aqueous phase material is methacryloyl gelatin (GelMA) instead of F127DA and PEGDA in Example 1, while the remaining raw materials and preparation steps are the same as in Example 1. The microgel prepared in this comparative example is a GelMA-CMC microgel.

[0126] Comparative Example 2

[0127] This comparative example discloses a microgel, which differs from Example 1 in that: in step (I) of this comparative example, the aqueous phase material does not include PEGDA, and the prepared microgel is a PEGDA-free microgel.

[0128] The sodium methacrylamide carboxymethyl cellulose used in the above embodiments was prepared by the following method, specifically including the following steps:

[0129] 2.00 g of sodium carboxymethyl cellulose was dissolved in 100 mL of water to obtain a CMC-Na solution. Throughout the dissolution and reaction process, the pH of the CMC-Na solution was adjusted to 8.0 using a 3 mol / L sodium hydroxide aqueous solution. After cooling the solution at 4 °C, 4 mL of methacrylic anhydride (MA) was added dropwise to the pH-adjusted CMC-Na solution, and the reaction was continued at 4 °C for 24 hours. The resulting mixture was precipitated in ethanol and dialyzed for 3 days to remove unreacted methacrylic acid and methacrylic anhydride, yielding methacrylated carboxymethyl cellulose sodium. After dialyzing, the methacrylated carboxymethyl cellulose sodium was freeze-dried for two days for later use.

[0130] The addition of Ficoll in the various embodiments and comparative examples can be used to adjust the viscosity of the aqueous precursor solution, which is beneficial for microgel formation. When the aqueous precursor solution contains cells, the addition of Ficoll can effectively prevent cell sedimentation.

[0131] Test case

[0132] This experimental example tested the performance of the microgels obtained in the examples and comparative examples, specifically as follows:

[0133] (1) The optical images of the anti-swelling microgels prepared in Examples 1-4 are as follows: Figure 1-4 As shown. By Figure 1-4 It can be seen that the preparation method of the present invention successfully prepared anti-swelling microgels with uniform size distribution and good quality. These anti-swelling microgels can be used to encapsulate different cells.

[0134] The average particle sizes of the anti-swelling microgels prepared in Examples 1-5 were measured to be 360 ​​μm, 420 μm, 80 μm, 60 μm, and 420 μm, respectively.

[0135] (2) The swelling properties of the microgels prepared in Example 5 and Comparative Examples 1-2 were tested, and the test results are as follows: Figure 5-7 As shown, the test steps for swelling performance testing include: soaking the prepared microgel in 1640 culture medium for 30 days, and observing and measuring the size of the microgel before and after swelling using a microscope.

[0136] Depend on Figure 5 It can be seen that the anti-swelling microgel prepared by the method of the present invention does not show significant changes in size before and after swelling, and can still maintain its original size well after 30 days of swelling, indicating that the anti-swelling microgel has good anti-swelling properties.

[0137] from Figure 6-7 It can be seen that, in Comparative Example 1, the microgels were 480 μm in size before swelling. After swelling for 30 days, the size of the microgels increased to 1210 μm. Figure 6Comparative Example 2 yielded microgels with a size of 560 μm before swelling. After 30 days of swelling, the microgel size increased to 920 μm. Figure 7 Furthermore, cracks appeared in the swollen microgel, indicating that the mechanical properties of the swollen microgel deteriorated and it was difficult to support the original structure.

[0138] (3) The survival status of cells in the anti-swelling microgel (encapsulating islet cell tumors) prepared in Example 5 was characterized, and the test results are as follows: Figure 8 As shown:

[0139] The specific steps include the following: Biological cells (L929) were cultured in the anti-swelling microgel prepared in Example 5 for one day, then 1-2 mL of Calcein AM / PI detection working solution was added, and the cells were incubated at 37°C in the dark for 30 min. After incubation, the cells were thoroughly washed with PBS, and the staining effect was observed under a fluorescence microscope (Calcein AM is green fluorescence, Ex / Em = 494 / 517 nm; PI is red fluorescence, Ex / Em = 535 / 617 nm). Figure (a) shows a 40x microscope image; Figures (b)-(d) show fluorescence images after live and dead cell staining, with Calcein AM (green, live cells) and PI (red, dead cells): (b) live and dead cell co-staining; (c) live cell staining; (d) dead cell staining.

[0140] Depend on Figure 8 It can be seen that a large number of green fluorescent signals were observed after staining, indicating that the cells are in good condition. This anti-swelling microgel has excellent biocompatibility and a microenvironment suitable for cell survival.

[0141] This invention proposes a long-lasting, stable, and anti-swelling microgel and its preparation method. The prepared anti-swelling microgel has the characteristics of injectability, high mechanical strength, poor degradation, and good anti-swelling performance.

[0142] It should be noted that, unless otherwise specified, the "molecular weight" of organic compounds in this article refers to the "weight-average molecular weight". The word "approximately" in numerical values ​​used in this article means an error of ±2%.

[0143] The embodiments of the present invention have been described in detail above with reference to the accompanying drawings. However, the present invention is not limited to the above embodiments, and various changes can be made within the scope of knowledge possessed by those skilled in the art without departing from the spirit of the present invention. Furthermore, the embodiments of the present invention and the features thereof can be combined with each other unless otherwise specified.

Claims

1. An anti-swell microgel, characterized in that, The preparation method comprises: using a microfluidic technology to prepare the anti-swelling microgel; wherein the raw material for preparing the anti-swelling microgel comprises a water phase material, the water phase material comprises acrylic ester polymerization monomers and a degradation-resistant raw material, the acrylic ester polymerization monomers comprise polyether F127 diacrylate and polyethylene glycol diacrylate, and the mass fraction w of the acrylic ester polymerization monomers in the water phase material satisfies 60% ≥ w ≥ 10%; and the degradation-resistant raw material comprises at least one of sodium carboxymethyl cellulose and methacrylated sodium carboxymethyl cellulose.

2. The anti-swell microgel according to claim 1, characterized in that, The mass ratio of the polyether F127 diacrylate and the polyethylene glycol diacrylate is (0.1-5):

1.

3. The anti-swell microgel according to claim 1, characterized in that, The mass ratio of the acrylic ester polymerization monomers and the degradation-resistant raw material is (2-10):(0.5-1.5).

4. The anti-swell microgel according to claim 1, characterized in that, The water phase material further comprises an active ingredient.

5. The anti-swell microgel according to claim 4, characterized in that, The active ingredient comprises at least one of cells, drugs or protein active factors.

6. The anti-swell microgel according to claim 5, characterized in that, The cells comprise at least one of stem cells, progenitor cells, fibroblasts, kidney cells, islet cells, tumor cells or liver cells.

7. A process for the preparation of the anti-swellen microgel according to claim 1, characterized in that, The method comprises the following steps: taking the water phase material and an oil phase material, and preparing the anti-swelling microgel by using a microfluidic technology.

8. A biomaterial, characterized by, The anti-swelling microgel as claimed in any one of claims 1-6 or prepared by the method as claimed in claim 7.

9. A medical product, characterized in that, The anti-swelling microgel as claimed in any one of claims 1-6 or prepared by the method as claimed in claim 7 or the biomaterial as claimed in claim 8.

10. The anti-swelling microgel as claimed in any one of claims 1-6 or prepared by the method as claimed in claim 7 in the preparation of a medical material or a biomaterial.

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