A gelatin-based hydrogel membrane, its preparation method and application
By reacting gelatin with bovine serum albumin and the cross-linking agent genipin, a non-toxic, biodegradable hydrogel membrane was prepared, solving the problem of poor biocompatibility in existing technologies and realizing the application of high-performance biomedical dressings.
Patent Information
- Application Number
- CN202310113992.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2023-02-15
- Publication Date
- 2025-10-31
- Estimated Expiration
- 2043-02-15
AI Technical Summary
Existing gelatin-based hydrogel membrane materials use aldehyde-based crosslinking agents and organic solvents in their preparation process, which affects biocompatibility and leaves behind free aldehyde groups.
Hydrogels were prepared by using gelatin, bovine serum albumin and crosslinking agent genipin via Schiff base reaction or nucleophilic substitution reaction. Bovine serum albumin was added to improve flexibility, forming a high-performance gel material that combines rigidity and flexibility.
The prepared hydrogel membrane has good biocompatibility and mechanical strength, adapts to skin movement, is low in cost, easy to mass-produce, and is non-toxic and biodegradable.
Smart Images
Figure CN116284902B_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of biopolymer materials technology, specifically relating to a gelatin-based hydrogel membrane, its preparation method, and its application. Background Technology
[0002] Hydrogels are polymeric network systems with a hydrophilic three-dimensional cross-linked structure. Hydrogels are soft, can maintain their shape, can absorb large amounts of water, and exhibit good biocompatibility and biodegradability, making them suitable for applications in tissue engineering, drug delivery systems, medical dressings, and cell culture.
[0003] There are many types of polymers used to construct hydrogels, among which gelatin (GEL) is the most widely used. It is a hydrolyzed product of collagen and possesses the prominent characteristics of collagen, such as biodegradability, good biocompatibility, non-immunogenicity, and the presence of RGD (R: arginine; G: glycine; D: aspartic acid) peptide sequences. Therefore, it can serve as a multifunctional platform for applications in various fields, such as wound repair, tissue engineering, bone repair, and cartilage repair. An ideal hydrogel scaffold should possess the following characteristics: appropriate mechanical properties, good water retention, anti-infection ability, injectability, and good cell biocompatibility.
[0004] Existing technologies include the use of gelatin to construct hydrogel materials. For example, Pensak Jantrawut et al. reported a low-methoxyl pectin (LMP) / gelatin / carboxymethyl cellulose (CMC) crosslinked hydrogel film material for use in wound dressings (Jantrawut, P.; Bunrueangtha, J.; Suerthong, J.; Kantrong, N. Fabrication and Characterization of Low Methoxyl Pectin / Gelatin / Carboxymethyl Cellulose Absorbent Hydrogel Film for Wound Dressing Applications. Materials 2019, 12, 1628.). Tze-Wen Chung et al. reported the use of gelatin combined with PLGA or PLGA-NH2 to produce drug carrier hydrogel films for delivering hydrophobic drugs. The hydrogel films were found to have high loading of indomethacin (IDM), low explosive release, and sustained release of IDM for up to 96 hours (Chung TW, Chou TH, Wu K Y. Gelatin / PLGA hydrogel films and their delivery of hydrophobic drugs[J]. Journal of the Taiwan Institute of Chemical Engineers, 2016, 60:8-14.). Patricia Hubner et al. reported a gelatin / PVA hydrogel film prepared by casting and UV crosslinking. The optimized hydrogel film has good swelling (65%), elasticity (146%) and good mechanical properties (3.40 MPa) (Hubner P, Marcilio NR, Tessaro I C. Gelatin / poly(vinyl alcohol) based hydrogel film – A potential biomaterial for wound dressing: Experimental design and optimization followed by rotatable central composite design[J]. Journal of Biomaterials Applications, 2021, 36(4):682-700.).Satish Patel et al. prepared a chitosan-gelatin hydrogel (LCGH) membrane with glutaraldehyde crosslinking agent. The test results showed that the glycerol-plasticized membrane significantly changed the properties, strength, and thickness of the hydrogel, and exhibited high swelling capacity and drug controlled release for up to 24 hours (Patel S, Srivastava S, Singh MR, et al. Preparation and optimization of chitosan-gelatin films for sustained delivery of lupeol for wound healing[J]. International Journal of Biological Macromolecules, 2018, 107:1888-1897.).
[0005] However, most of the hydrogel membrane materials prepared from gelatin use aldehyde-containing crosslinking agents and organic solvents to prepare hydrogels, which have problems such as free aldehyde groups and incomplete evaporation of organic solvents, and seriously affect the biocompatibility of the materials. Summary of the Invention
[0006] To address the aforementioned technical problems, this invention provides a gelatin-based hydrogel membrane, which is prepared from gelatin, bovine serum albumin (BSA), a crosslinking agent, and water. The mass ratio of gelatin, bovine serum albumin, and crosslinking agent is 2-5:0.5-5:0.05-1, and the mass ratio of the total mass of bovine serum albumin and crosslinking agent to water is 2-11:89-98.
[0007] Gelatin can undergo Schiff base reaction or nucleophilic substitution reaction at the above temperature under the action of crosslinking agent and initiator. At the same time, due to the addition of bovine serum albumin with helical fold conformation and random coil structure, high-performance gel materials with both rigidity and flexibility can be prepared, resulting in gelatin-based hydrogel membranes.
[0008] Preferably, the mass ratio of the gelatin, bovine serum albumin and crosslinking agent is 2-4:1-4:0.05-0.5.
[0009] Furthermore, the mass ratio of the gelatin, bovine serum albumin, and cross-linking agent is 2-4:1-3.5:0.05-0.1.
[0010] Preferably, the gelatin is type A gelatin and / or type B gelatin.
[0011] Furthermore, the adhesive strength of the type A gelatin is 100-500g Bloom.
[0012] Preferably, the crosslinking agent is genipin.
[0013] The present invention also provides a method for preparing the above-mentioned gelatin-based hydrogel membrane, comprising the following steps: dissolving gelatin in water, adding bovine serum albumin and a crosslinking agent, reacting, and drying to obtain the gelatin-based hydrogel membrane.
[0014] Preferably, the reaction temperature is 10-40°C.
[0015] Preferably, the reaction time is 2-10 hours.
[0016] Preferably, the drying time is 12-48 hours. This step is intended to remove excess moisture and obtain a pure hydrogel membrane after drying.
[0017] Preferably, the reaction is carried out in a polytetrafluoroethylene mold.
[0018] The present invention also provides a biomedical dressing comprising the above-mentioned gelatin-based hydrogel film.
[0019] The technical solution of the present invention has the following advantages compared with the prior art:
[0020] This invention uses type A gelatin and / or type B gelatin as raw materials. Under the action of a cross-linking agent, they can undergo a Schiff base reaction or a nucleophilic substitution reaction in solution, forming a hydrogel that is non-toxic, biodegradable, and has good biocompatibility. Simultaneously, bovine serum albumin is added as a flexible chain to prepare a composite gel, improving the flexibility of the hydrogel membrane. This invention adds bovine serum albumin, which has a helical folding conformation and random coil structure, to gelatin molecular chains that exhibit a certain degree of rigidity and poor molecular chain flexibility, to prepare a high-performance gel material that combines rigidity and flexibility. Moreover, the hydrogel membrane of this invention, as a biomedical hydrogel dressing, can adapt to skin movement and has good mechanical strength and relative stability.
[0021] Furthermore, the hydrogel membrane of the present invention has low raw material cost, simple preparation method, and is easy to scale up production and application. Attached Figure Description
[0022] Figure 1 This is a graph showing the tensile properties of the hydrogel film prepared using Comparative Example 1;
[0023] Figure 2 The graph shows the compression properties of the hydrogel membrane prepared using Comparative Example 1.
[0024] Figure 3 This is a graph showing the tensile properties of the hydrogel film prepared using Example 1;
[0025] Figure 4 The graph shows the compression properties of the hydrogel membrane prepared using Example 1.
[0026] Figure 5 This is a graph showing the tensile properties of the hydrogel film prepared using Example 2;
[0027] Figure 6 The graph shows the compression properties of the hydrogel membrane prepared using Example 2.
[0028] Figure 7 The diagram shows the adhesion performance of the hydrogel film prepared using Example 2;
[0029] Figure 8 This is a comparison chart of the tensile strength of hydrogel films;
[0030] Figure 9 This is a comparison chart of compressive stress in hydrogel films. Detailed Implementation
[0031] The present invention will be further described in detail below with reference to specific embodiments. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a thorough and complete understanding of the disclosure of the present invention.
[0032] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein in the specification of this invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.
[0033] the term
[0034] Unless otherwise stated or in case of contradiction, the terms or phrases used herein shall have the following meanings:
[0035] As used herein, the terms “and / or,” “or / and,” and “and / or” may include any one of two or more of the related listed items, as well as any and all combinations of the related listed items, including any two related listed items, any more related listed items, or a combination of all the related listed items.
[0036] In this article, "one or several" refers to any one, two, or more of the listed items. Among them, "several" refers to any two or more.
[0037] In this document, "preferred" is only used to describe a better implementation method or embodiment, and should be understood as not constituting a limitation on the scope of protection of this invention.
[0038] In this invention, the technical features described in an open-ended manner include both closed-ended technical solutions composed of the listed features and open-ended technical solutions that include the listed features.
[0039] In this invention, numerical ranges are involved, and unless otherwise specified, they include the two endpoints of the numerical range.
[0040] Unless otherwise specified, the percentage content involved in this invention refers to mass percentage for solid-liquid mixtures and solid-phase-solid mixtures, and volume percentage for liquid-phase-liquid mixtures.
[0041] Unless otherwise specified, all percentage concentrations mentioned in this invention refer to the final concentration. The final concentration refers to the proportion of the added component in the system after the addition of that component.
[0042] Unless otherwise specified, the temperature parameters in this invention can be either constant temperature processing or processing within a certain temperature range. The constant temperature processing allows temperature fluctuations within the precision range controlled by the instrument.
[0043] The following detailed description is provided in conjunction with specific embodiments. Unless otherwise specified, all raw materials and instruments used in the following specific embodiments are commercially available.
[0044] Example 1
[0045] First, prepare 0.3g of gelatin, 0.3g of bovine serum albumin, 0.08g of cross-linking agent Genipin, and water; the gelatin is 0.15g of type A gelatin and 0.15g of type B gelatin.
[0046] Preparation of gelatin solution: Add gelatin to cold water, heat for 10 minutes, and then heat at a temperature not exceeding 70°C. Stir until the gelatin dissolves to obtain a gelatin solution.
[0047] Bovine serum albumin and genipin were added to a gelatin solution to obtain a mixed solution. The mixed solution was then added to a polytetrafluoroethylene mold. The gelatin and bovine serum albumin were cross-linked at 26°C for 4 hours under the action of the cross-linking agent genipin to obtain a gelatin / bovine serum albumin composite hydrogel.
[0048] The gelatin / bovine serum albumin composite hydrogel was dried in a fume hood for 24 hours to obtain a pure gelatin-based hydrogel membrane.
[0049] Example 2
[0050] First, prepare 0.3g of gelatin, 0.15g of bovine serum albumin, 0.08g of cross-linking agent Genipin, and water; the gelatin is 0.15g of type A gelatin and 0.15g of type B gelatin.
[0051] Preparation of gelatin solution: Add gelatin to cold water, heat for 10 minutes, and then heat at a temperature not exceeding 70°C. Stir until the gelatin dissolves to obtain a gelatin solution.
[0052] Bovine serum albumin and genipin were added to a gelatin solution to obtain a mixed solution. The mixed solution was then added to a polytetrafluoroethylene mold. The gelatin and bovine serum albumin were cross-linked at 26°C for 4 hours under the action of the cross-linking agent genipin to obtain a gelatin / bovine serum albumin composite hydrogel.
[0053] The gelatin / bovine serum albumin composite hydrogel was dried in a fume hood for 24 hours to obtain a pure gelatin-based hydrogel membrane.
[0054] Example 3
[0055] First, prepare 0.3g of gelatin, 0.3g of bovine serum albumin, 0.08g of cross-linking agent Genipin, and water; the gelatin is 0.15g of type A gelatin and 0.15g of type B gelatin.
[0056] Preparation of gelatin solution: Add gelatin to cold water, heat for 5 minutes, and then heat at a temperature not exceeding 70°C. Stir until the gelatin dissolves to obtain a gelatin solution.
[0057] Bovine serum albumin and genipin were added to a gelatin solution to obtain a mixed solution. The mixed solution was then added to a polytetrafluoroethylene mold. The gelatin and bovine serum albumin were cross-linked at 10°C for 2 hours under the action of the cross-linking agent genipin to obtain a gelatin / bovine serum albumin composite hydrogel.
[0058] The gelatin / bovine serum albumin composite hydrogel was dried in a fume hood for 12 hours to obtain a pure gelatin-based hydrogel membrane.
[0059] Example 4
[0060] First, prepare 0.3g of gelatin, 0.3g of bovine serum albumin, 0.08g of cross-linking agent Genipin, and water; the gelatin is 0.15g of type A gelatin and 0.15g of type B gelatin.
[0061] Preparation of gelatin solution: Add gelatin to cold water, heat for 20 minutes, and then heat at a temperature not exceeding 70°C. Stir until the gelatin dissolves to obtain a gelatin solution.
[0062] Bovine serum albumin and genipin were added to a gelatin solution to obtain a mixed solution. The mixed solution was then added to a polytetrafluoroethylene mold. The gelatin and bovine serum albumin were cross-linked at 40°C for 10 hours under the action of the cross-linking agent genipin to obtain a gelatin / bovine serum albumin composite hydrogel.
[0063] The gelatin / bovine serum albumin composite hydrogel was dried in a fume hood for 48 hours to obtain a pure gelatin-based hydrogel membrane.
[0064] Example 5
[0065] First, prepare 0.3g of gelatin, 0.3g of bovine serum albumin, 0.08g of cross-linking agent Genipin, and water; the gelatin is 0.15g of type A gelatin and 0.15g of type B gelatin.
[0066] Preparation of gelatin solution: Add gelatin to cold water, heat for 10 minutes, and then heat at a temperature not exceeding 70°C. Stir until the gelatin dissolves to obtain a gelatin solution.
[0067] Bovine serum albumin and genipin were added to a gelatin solution to obtain a mixed solution. The mixed solution was then added to a polytetrafluoroethylene mold. The gelatin and bovine serum albumin were cross-linked at 30°C for 2 hours under the action of the cross-linking agent genipin to obtain a gelatin / bovine serum albumin composite hydrogel.
[0068] The gelatin / bovine serum albumin composite hydrogel was dried in a fume hood for 36 hours to obtain a pure gelatin-based hydrogel membrane.
[0069] Example 6
[0070] First, prepare 0.3g of gelatin, 0.3g of bovine serum albumin, 0.08g of cross-linking agent Genipin, and water; the gelatin is 0.15g of type A gelatin and 0.15g of type B gelatin.
[0071] Preparation of gelatin solution: Add gelatin to cold water, heat for 18 minutes, and then heat at a temperature not exceeding 70°C. Stir until the gelatin dissolves to obtain a gelatin solution.
[0072] Bovine serum albumin and genipin were added to a gelatin solution to obtain a mixed solution. The mixed solution was then added to a polytetrafluoroethylene mold. The gelatin and bovine serum albumin were cross-linked at 30°C for 4 hours under the action of the cross-linking agent genipin to obtain a gelatin / bovine serum albumin composite hydrogel.
[0073] The gelatin / bovine serum albumin composite hydrogel was dried in a fume hood for 36 hours to obtain a pure gelatin-based hydrogel membrane.
[0074] Example 7
[0075] First, prepare 0.2g of gelatin, 0.05g of bovine serum albumin, 0.005g of cross-linking agent Genipin, and water; the gelatin is 0.1g of type A gelatin and 0.1g of type B gelatin.
[0076] Preparation of gelatin solution: Add gelatin to cold water, heat for 10 minutes, and then heat at a temperature not exceeding 70°C. Stir until the gelatin dissolves to obtain a gelatin solution.
[0077] Bovine serum albumin and genipin were added to a gelatin solution to obtain a mixed solution. The mixed solution was then added to a polytetrafluoroethylene mold. The gelatin and bovine serum albumin were cross-linked at 26°C for 4 hours under the action of the cross-linking agent genipin to obtain a gelatin / bovine serum albumin composite hydrogel.
[0078] The gelatin / bovine serum albumin composite hydrogel was dried in a fume hood for 24 hours to obtain a pure gelatin-based hydrogel membrane.
[0079] Example 8
[0080] First, prepare 0.5g of gelatin, 0.5g of bovine serum albumin, 0.1g of cross-linking agent Genipin, and water; the gelatin is 0.25g of type A gelatin and 0.25g of type B gelatin.
[0081] Preparation of gelatin solution: Add gelatin to cold water, heat for 10 minutes, and then heat at a temperature not exceeding 70°C. Stir until the gelatin dissolves to obtain a gelatin solution.
[0082] Bovine serum albumin and genipin were added to a gelatin solution to obtain a mixed solution. The mixed solution was then added to a polytetrafluoroethylene mold. The gelatin and bovine serum albumin were cross-linked at 26°C for 4 hours under the action of the cross-linking agent genipin to obtain a gelatin / bovine serum albumin composite hydrogel.
[0083] The gelatin / bovine serum albumin composite hydrogel was dried in a fume hood for 24 hours to obtain a pure gelatin-based hydrogel membrane.
[0084] Example 9
[0085] First, prepare 0.2g of gelatin, 0.5g of bovine serum albumin, 0.005g of cross-linking agent Genipin, and water; the gelatin is 0.2g of type A gelatin.
[0086] Preparation of gelatin solution: Add gelatin to cold water, heat for 10 minutes, and then heat at a temperature not exceeding 70°C. Stir until the gelatin dissolves to obtain a gelatin solution.
[0087] Bovine serum albumin and genipin were added to a gelatin solution to obtain a mixed solution. The mixed solution was then added to a polytetrafluoroethylene mold. The gelatin and bovine serum albumin were cross-linked at 10°C for 4 hours under the action of the cross-linking agent genipin to obtain a gelatin / bovine serum albumin composite hydrogel.
[0088] The gelatin / bovine serum albumin composite hydrogel was dried in a fume hood for 24 hours to obtain a pure gelatin-based hydrogel membrane.
[0089] Example 10
[0090] First, prepare 0.4g of gelatin, 0.5g of bovine serum albumin, 0.05g of cross-linking agent Genipin, and water; the gelatin is 0.4g of type B gelatin.
[0091] Preparation of gelatin solution: Add gelatin to cold water, heat for 10 minutes, and then heat at a temperature not exceeding 70°C. Stir until the gelatin dissolves to obtain a gelatin solution.
[0092] Bovine serum albumin and genipin were added to a gelatin solution to obtain a mixed solution. The mixed solution was then added to a polytetrafluoroethylene mold. The gelatin and bovine serum albumin were cross-linked at 15°C for 8 hours under the action of the cross-linking agent genipin to obtain a gelatin / bovine serum albumin composite hydrogel.
[0093] The gelatin / bovine serum albumin composite hydrogel was dried in a fume hood for 36 hours to obtain a pure gelatin-based hydrogel membrane.
[0094] Comparative Example 1
[0095] First, prepare 0.3g of gelatin, 0.08g of crosslinking agent Genipin, and water; the gelatin is 0.15g of type A gelatin and 0.15g of type B gelatin.
[0096] Preparation of gelatin solution: Add gelatin to cold water, heat for 10 minutes, and then heat at a temperature not exceeding 70°C. Stir until the gelatin dissolves to obtain a gelatin solution.
[0097] Genipin was added to the gelatin solution to obtain a mixed solution. The mixed solution was then added to a polytetrafluoroethylene mold. Under the action of the crosslinking agent genipin, the gelatin underwent a crosslinking reaction at 26°C for 4 hours to obtain a gelatin hydrogel.
[0098] The gelatin hydrogel was dried in a fume hood for 24 hours, and a pure gelatin-based hydrogel membrane was obtained after drying.
[0099] Effect Evaluation 1
[0100] The hydrogels prepared in Examples 1, 2, and 1 (Comparative Example 1) were used as test materials, and their mechanical properties were tested using a universal mechanical analyzer. The obtained mechanical data are as follows: Figure 1-6 As shown, Figure 1 The tensile properties of the hydrogel film prepared using Comparative Example 1 are shown. Figure 2 The display shows the compressibility of the hydrogel membrane prepared using Comparative Example 1; Figure 3 The tensile properties of the hydrogel film prepared using Example 1 are shown. Figure 4 The compression properties of the hydrogel membrane prepared using Example 1 are shown; Figure 5 The tensile properties of the hydrogel film prepared using Example 1 are shown. Figure 6The compressibility of the hydrogel membrane prepared using Example 2 is shown; Figure 7 The adhesion of the hydrogel film prepared using Example 2 is shown.
[0101] Depend on Figures 1 to 7 It can be seen that Examples 1, 2, and Comparative Example 1 exhibit good adhesion to the skin surface, can adapt to skin movement, and possess certain mechanical strength and relative stability. The mechanical strength of the hydrogels was characterized using a universal tensile testing machine, and the results are shown in Table 1.
[0102] Table 1 Test Table of Mechanical Properties of Hydrogels
[0103]
[0104] The tensile strength, elongation at break, tensile stress at break, and compressive stress at failure of the hydrogels in Examples 1 and 2 were all higher than those of the hydrogel in Comparative Example 1. Notably, the hydrogel in Example 2 exhibited higher mechanical properties than both the hydrogel in Comparative Example 1 and the hydrogel in Example 1. This suggests that adding BSA can increase the mechanical properties of gelatin hydrogels in their initial state, while excessive BSA may reduce the chemical crosslinking of the hydrogel.
[0105] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.
[0106] Obviously, the above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations here. However, obvious variations or modifications derived therefrom are still within the scope of protection of this invention.
Claims
1. A biomedical dressing, characterized in that, The invention includes a gelatin-based hydrogel membrane, which is prepared from gelatin, bovine serum albumin, a crosslinking agent, and water. The mass ratio of gelatin, bovine serum albumin, and crosslinking agent is 2-5:0.5-5:0.05-1, and the mass ratio of the total mass of gelatin, bovine serum albumin, and crosslinking agent to water is 2-11:89-98. The crosslinking agent is selected from genipin.
2. The biomedical dressing as described in claim 1, characterized in that, The gelatin is type A gelatin and / or type B gelatin.
3. The biomedical dressing as described in claim 2, characterized in that, The adhesive strength of the type A gelatin is 100-500 gBloom.
4. The biomedical dressing as described in claim 1, characterized in that, The preparation method of the gelatin-based hydrogel membrane includes the following steps: dissolving gelatin in water, adding bovine serum albumin and a cross-linking agent, reacting, and drying to obtain the gelatin-based hydrogel membrane.
5. The biomedical dressing as described in claim 4, characterized in that, The reaction temperature is 10-40℃.
6. The biomedical dressing as described in claim 4, characterized in that, The reaction time is 2-10 h.
7. The biomedical dressing as described in claim 4, characterized in that, The drying time is 12-48 h.
8. The biomedical dressing as described in claim 4, characterized in that, The reaction is carried out in a polytetrafluoroethylene mold.
Citation Information
Patent Citations
Water-insoluble silk fibroin-gelatin blended film and preparation method thereof
CN104530456A