A room temperature collagen mineralization material and its preparation method and application

Abstract

This invention relates to a room-temperature collagen mineralization material, its preparation method, and its application, comprising the following steps: pretreating fish maw material to obtain treated fish maw; immersing the treated fish maw in a mineralization solution for mineralization treatment; and drying to obtain the room-temperature collagen mineralization material. This invention can also use bio-adhesive as a binder to laminate and press the room-temperature collagen mineralization material to obtain a block-shaped mineralized adhesive raw material. This invention uses fish maw as the adhesive raw material for mineralization, which is readily available, inexpensive, and has a relatively large material size that facilitates processing. The collagen fibers are finely arranged, and the mineralized material exhibits good mechanical properties. Simultaneously, the lamination and pressing process produces a block-shaped mineralized adhesive raw material with good mechanical properties.

Description

Technical Field

[0001] This invention relates to the field of biomimetic mineralization materials, specifically to a room-temperature collagen mineralization material, its preparation method, and its application. Background Technology

[0002] The production of building materials (ceramics, cement, etc.) consumes a large amount of energy, resulting in significant carbon emissions. A reliable direction for reducing carbon emissions in manufacturing processes is to learn from nature. Over tens of thousands of years of evolution, many organisms have developed various structures with specific functions. Furthermore, organisms often undergo a series of biochemical reactions at room temperature or even lower temperatures, forming multi-level hard materials ranging from microscopic to macroscopic. This low-consumption, high-efficiency biological structure synthesis process provides a new approach to optimizing current high-carbon-emission synthesis technologies. In recent years, Academician Fu Zhengyi of Wuhan University of Technology has been the first in the world to propose learning from the structural formation processes in nature to gain inspiration for guiding the synthesis of artificial materials—a concept known as "bioprocess-inspired material preparation technology."

[0003] Biomineralization is the process by which most minerals are synthesized within living organisms. Many high-performance hard materials, such as bone and tooth enamel, are formed through biomineralization. Taking lightweight, high-strength bone as an example, mineralized collagen fibers are its basic building blocks. Nano-hydroxyapatite grows within the confined space of collagen fibers, along their long axis. The assembly of inorganic and organic phases in bone from the molecular to the macroscopic scale gives it excellent mechanical properties. Furthermore, researchers at Pinghang have confirmed that the mineralization process of collagen fibers generates megapascal-level prestress. These mineralized fibers, on the one hand, provide mineral content to the bone, and on the other hand, act like "prestressed steel bars," giving the entire bone prestress and greatly improving its toughness.

[0004] By simulating the bone biomineralization process and preparing suitable mineralization solutions, in vitro mineralization of collagen raw materials can be achieved. However, currently, the collagen raw materials used for mineralization are mostly one-dimensional collagen fibers and two-dimensional collagen films. These materials have low collagen concentrations, making it impossible to prepare bulk (three-dimensional) materials. The thickness of collagen films is often only a few hundred nanometers, making it difficult to conduct mechanical property tests, such as common tensile tests. Furthermore, the arrangement of collagen fibers in these materials is relatively disordered, which cannot be compared with the precision of collagen fiber arrangement in bone. In addition, these collagen raw materials are difficult to obtain and expensive, which does not meet the requirements of practical industrial production.

[0005] Therefore, there is currently no method for preparing bulk materials at room temperature using inexpensive, finely structured adhesive raw materials through mineralization, and no basic process flow and parameters have been found. Summary of the Invention

[0006] The purpose of this invention is to overcome the above-mentioned technical deficiencies and provide a room temperature collagen mineralization material, its preparation method and application, thereby solving the technical problems of high cost of raw materials for mineralization and inability to prepare bulk materials in the prior art.

[0007] To achieve the above-mentioned technical objectives, the technical solution provided by this invention is as follows:

[0008] In a first aspect, the present invention provides a method for preparing room temperature collagen mineralization material, comprising the following steps: S1, pretreating fish bladder material to obtain treated fish bladder; S2, immersing the treated fish bladder in a mineralization solution for mineralization treatment, and drying to obtain room temperature collagen mineralization material.

[0009] Secondly, the present invention provides a room-temperature collagen mineralized material prepared by the above preparation method.

[0010] Thirdly, the present invention provides a bulk mineralized adhesive raw material, comprising the above-mentioned room temperature collagen mineralization material stacked in layers, wherein an adhesive layer is provided between each layer of the room temperature collagen mineralization material.

[0011] Fourthly, the present invention provides a method for preparing a bulk mineralized adhesive raw material, comprising the following steps: using bio-adhesive as a binder, layering and pressing room temperature collagen mineralization materials to obtain a bulk mineralized adhesive raw material.

[0012] Compared with the prior art, the beneficial effects of the present invention include:

[0013] 1. This invention uses fish bladders as the raw material for mineralization, which is not only easy to obtain and inexpensive, but also has a large material size that is conducive to processing; in addition, the collagen fibers in the fish bladder material have a high degree of fineness in arrangement, comparable to bone tissue, and the material has good performance after mineralization.

[0014] 2. This invention simulates the process of collagen mineralization in bone tissue, prepares a mineralization solution in vitro, and realizes the directional mineralization and growth of nanoscale minerals along the long axis of fish bladder collagen fibers. It makes full use of the interaction between organic and inorganic materials, further improving the mineral content and mechanical properties of the composite material. The resulting mineralized fish bladder material has good mechanical properties.

[0015] 3. This invention utilizes bio-adhesive to laminate and bond multiple sheets of room-temperature collagen mineralized material into a single unit, thereby improving the material's dimensions; and employs compression molding to enhance the interlayer bonding strength. Through a rapid two-step process, the size and interfacial bonding of the material are improved at room temperature, resulting in a bulk mineralized adhesive raw material with good mechanical properties.

[0016] Therefore, this invention can prepare large-sized bulk collagen mineralized materials at room temperature with excellent mechanical properties. The fish maw used in this invention is inexpensive, readily available, and utilizes waste materials; the mineralization process can be carried out at room temperature and pressure, demonstrating strong potential for industrial mass production. Attached Figure Description

[0017] Figure 1 The images show the morphology of the materials prepared in Example 1 and Comparative Example 1; where A is an optical image of the unmineralized fish bubble prepared in Comparative Example 1, B is an optical image of the mineralized fish bubble prepared in Example 1, C is an actual image of the mineralized fish bubble prepared in Example 1, D is a cross-sectional scanning electron microscope image of the unmineralized fish bubble obtained in Comparative Example 1, and E is a cross-sectional scanning electron microscope image of the mineralized fish bubble in Example 1.

[0018] Figure 2 Transmission electron microscopy (TEM) images of fish bladders after mineralization in Example 1 and unmineralized fish bladders in Comparative Example 1; where A is Comparative Example 1 (low magnification), B is Comparative Example 1 (high magnification); C is Example 1 (low magnification), and D is Example 1 (high magnification).

[0019] Figure 3 The image shows a transmission electron microscope (TEM) image of a fish bladder slice after mineralization in Example 1; where A represents the mineralized collagen fibers within the fish bladder; B is a high-resolution image of the white rectangular portion of A, and the inset is an enlarged view of the dashed rectangular portion of B; C is a selected area electron diffraction pattern of the white rectangular portion of A.

[0020] Figure 4 Tensile tests were performed on the fish bladders after mineralization in Example 1 and the unmineralized fish bladders in Comparative Example 1; where A is the tensile test curve, B is the elastic modulus comparison graph, C is the ultimate tensile strength comparison graph, and D is the elongation at break comparison graph.

[0021] Figure 5 Thermogravimetric analysis (TGA) results of the mineralized fish bladder in Example 1;

[0022] Figure 6 This is a photograph of the block-shaped mineralized adhesive raw material prepared in Example 1; A is the angle of being laid flat, and B is the angle of being held in hand;

[0023] Figure 7 The three-point flexural strength test curves of the blocky mineralized adhesive raw material in Example 1;

[0024] Figure 8 The cut, unmineralized block material obtained in Comparative Example 2;

[0025] Figure 9 This is the displacement-load curve of the three-point bending test of the unmineralized blocky material in Comparative Example 2. Detailed Implementation

[0026] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.

[0027] The purpose of this invention is to provide a room-temperature collagen mineralization material, its preparation method, and its application. By selecting readily available and inexpensive fish maw as the raw material, the mineralization process of fish maw is simulated to achieve mineralization of the material at room temperature. Multiple mineralized fish maw sheets are bonded together using chitosan bio-adhesive, improving the overall size of the material. Cold isostatic pressing further enhances the bonding between the layers.

[0028] In a first aspect, the present invention provides a method for preparing room temperature collagen mineralization materials, comprising the following steps:

[0029] S1, pre-treat the fish maw material to obtain the treated fish maw;

[0030] S2, the treated fish bladder is soaked in a mineralization solution for mineralization treatment, and then dried to obtain room temperature collagen mineralized material.

[0031] Preferably, in step S1, the pretreatment includes delipidation, decalcification, and removal of impurities and proteins.

[0032] Further preferred conditions include: immersion in a 0.1–1 mol / L NaOH solution for 12–48 h; immersion in a 0.1–1 mol / L HCl solution for 12–48 h; and removal of impurities and proteins by ultrasonic cleaning in a 0.1–1 mol / L NaCl solution for 10–240 min.

[0033] In an even better manner, the fish maw material is cleaned with deionized water before and after degreasing and decalcification.

[0034] It is understandable that the processed fish maw can be cut into different shapes as needed, and the operation is a standard procedure, so no specific restrictions are made here.

[0035] Preferably, in step S2, after the treated fish bladder is fixed on the clamp (flattened), it is then immersed in the mineralization solution for mineralization treatment. This invention, by fixing the fish bladder on the clamp to maintain flatness and ensuring complete immersion in the mineralization solution, facilitates better mineralization and avoids wrinkles and bends that could affect subsequent layering.

[0036] Further preferably, the clamp is made of materials such as polypropylene and polytetrafluoroethylene; which do not react with the components in the mineralization solution.

[0037] Preferably, in step S2, the mineralization solution is prepared by mixing a calcium source, a phosphorus source, a buffer, sodium chloride, polyacrylic acid, and a solvent in a mass ratio of 1:(1-3):(4-8):(20-24):(0.1-0.5):(2200-2500). This invention uses calcium and phosphorus sources as raw materials to form a hydroxyapatite mineralization solution under the action of a buffer, sodium chloride, and polyacrylic acid; through this hydroxyapatite mineralization solution, the internal mineralization of fish maw collagen fibers is achieved.

[0038] Further preferably, the calcium source includes calcium chloride dihydrate; the phosphorus source includes dipotassium hydrogen phosphate trihydrate; the buffer includes hydroxyethylpiperazine ethanethiolic acid; and the solvent is water.

[0039] Preferably, in step S2, the mineralization conditions include: a mineralization temperature of 25–40 °C, replacement of the mineralization solution every 12–48 hours, and a total mineralization time of 3–7 days. This invention increases the mineral content within the fish maw material by periodically replacing the mineralization solution and extending the mineralization reaction time. Furthermore, increasing the mineralization temperature accelerates the mineralization reaction; extending the mineralization cycle increases the mineral content within the fish maw. Increased mineral content within the fish maw improves its mechanical properties.

[0040] However, it's important to note that the mineral content cannot increase indefinitely. First, the collagen fibers within the fish maw are finite; once all mineralizable fibers have been mineralized, extending the mineralization time or temperature is meaningless. Second, the mineralization process is influenced by already formed minerals. These minerals affect the diffusion of amorphous precursors, leading to a lower or even halted mineralization rate in later stages. Therefore, the mineralization of fish maw materials can only reach a relatively stable level. In other words, the mineral content within the fish maw increases very slowly or not at all after reaching a certain value, i.e., relatively complete mineralization. The parameters that enable relatively complete mineralization of fish maw materials are quite broad. This invention, by controlling the mineralization treatment conditions within the aforementioned parameter range, can achieve the mineralization of fish maw materials. Theoretically, the performance of fish maws that have achieved relatively complete mineralization is not significantly different.

[0041] Preferably, in step S2, the drying conditions include a temperature of 25–40 °C and a time of 12–48 h. This invention allows the mineralized fish maw to be naturally dried for subsequent use.

[0042] Secondly, the present invention provides a room-temperature collagen mineralized material prepared by the above preparation method.

[0043] Thirdly, the present invention provides a bulk mineralized adhesive raw material, comprising the above-mentioned room temperature collagen mineralization material stacked in layers, wherein an adhesive layer is provided between each layer of the room temperature collagen mineralization material.

[0044] Preferably, the number of layers in the room temperature collagen mineralization material is 2 to 10. The number of bonding layers affects the final block size and some mechanical properties, and can be selected according to the actual application.

[0045] Fourthly, the present invention provides a method for preparing a bulk mineralized adhesive raw material, comprising the following steps:

[0046] Using bio-adhesive as a binder, room temperature collagen mineralization materials are laminated, bonded, and pressed into shape to obtain block-shaped mineralized adhesive raw materials.

[0047] Preferably, the bio-adhesive is a mixture of chitosan, acetic acid, and solvent in a mass ratio of 1g:(5-10)mL:(30-50)mL; the amount of bio-adhesive used is 0.03-1.3mL / cm³. 2 This invention allows for spin-coating of chitosan solution to bond multiple layers of room-temperature collagen mineralized material together.

[0048] A further preferred solvent is water.

[0049] Preferably, the compression molding process involves placing the laminated and bonded materials into a vacuum bag and evacuating it. A cold isostatic press (LDJ320 / 700-400, Sichuan Aviation Industry West Machinery Factory) is then used to improve the interlayer bonding strength and enhance the interfacial bonding effect. The working pressure of the cold isostatic press is 100 MPa to 400 MPa, and the holding time is 5 to 30 minutes.

[0050] This invention opens up new applications for collagen waste, enabling the preparation of large-sized materials at room temperature through biomineralization, thus promoting waste utilization and integrated green production. Furthermore, the invented large-sized, bulk collagen raw materials possess excellent mechanical properties and hold great promise for future applications.

[0051] The invention will be further described in detail below through specific embodiments. The fish maw must be kept fresh, without obvious discoloration or off-odors.

[0052] Example 1

[0053] A method for preparing a bulk mineralized adhesive raw material includes the following steps:

[0054] S1. Pretreatment and shaping of fish maw material: Fresh fish maw was washed to remove blood, then soaked in 0.1 M NaOH solution for 24 h to remove fat; subsequently, the fish maw was washed with deionized water. The fish maw was then soaked in 0.1 M HCl solution for 24 h to remove any possible calcium; subsequently, the fish maw was washed again with deionized water. Finally, the fish maw was immersed in 0.1 M NaCl solution and ultrasonically cleaned for 10 min to remove impurities and proteins. The treated fish maw material was then cut into circles with a diameter of 2 cm using a circular mold.

[0055] S2, Mineralization of Collagen Fibers in Fish Bulb Material: A hydroxyapatite mineralization solution was prepared, with the following mass values ​​for calcium chloride dihydrate, potassium dihydrogen phosphate trihydrate, hydroxyethylpiperazine ethanethiolic acid, sodium chloride, polyacrylic acid, and water: 0.04 g, 0.062 g, 0.238 g, 0.876 g, 0.01 g, and 100 g, respectively. The treated fish bubble material was fixed on a polypropylene clamp and immersed in the mineralization solution. The mineralization solution was replaced every 12 hours, with a total mineralization reaction time of 7 days. The mineralized fish bubble was removed from the clamp, rinsed with deionized water, and then dried at 37 °C for 24 hours to obtain room temperature collagen mineralized material (mineralized fish bubble).

[0056] S3. Using a layer-by-layer stacking technique combined with cold isostatic pressing, a bulk mineralized adhesive raw material is prepared: The room-temperature collagen mineralized material obtained in step S2 (mineralized fish-eye discs, 2 cm in diameter) is each coated with 1 mL of chitosan bio-adhesive solution, which is composed of 0.6 g chitosan, 5 mL glacial acetic acid, and 24.4 mL water. Eight mineralized fish-eye discs are bonded together as a single unit, placed in a vacuum bag, and vacuum-sealed. A cold isostatic press is then used to enhance the interlayer adhesion. The working pressure of the cold isostatic press is 200 MPa, and the holding time is 5 min.

[0057] Comparative Example 1

[0058] To study the morphology, structure, and properties of fish bladders before hydroxyapatite mineralization, and to demonstrate that mineralization solutions can achieve the mineralization of fish bladder materials, fresh fish bladders were pretreated and then dried directly without mineralization to obtain single unmineralized fish bladder pieces.

[0059] The difference between Comparative Example 1 and Example 1 is that the mineralization treatment in step S2 and the multi-layer superposition treatment in step S3 are not performed; the fish bladder material obtained after step S1 of Example 1 is used directly.

[0060] Comparative Example 2

[0061] To study the effect of mineralization on the properties of bulk materials, fresh fish maw was pretreated and then dried directly without mineralization to obtain unmineralized fish maw. The unmineralized fish maw was then stacked to prepare bulk materials.

[0062] The difference between Comparative Example 2 and Example 1 is that the mineralization treatment in step S2 is not performed. Instead, the fish bladder material obtained in step S1 is directly subjected to the multi-layer stacking treatment in step S3 to prepare a block material.

[0063] Performance testing

[0064] 1. The mineralized fish-like bubbles obtained in step S2 of Example 1 and the unmineralized fish-like bubbles obtained in Comparative Example 1 were characterized by optical microscopy. The results are as follows: Figure 1 As shown.

[0065] Depend on Figure 1 As can be seen, the surface of the fish bubble after mineralization in Example 1 is smooth, and the fish bubble becomes opaque due to the growth of hydroxyapatite inside it. Figure 1 B). The unmineralized fish maw obtained in Comparative Example 1 was transparent after drying. Figure 1 A).

[0066] In Example 1, by cutting the fish maw into a suitable shape, a circular sheet of mineralized fish maw material can be prepared for use in subsequent processes. Figure 1 C). The monolithically mineralized fish bladder prepared in Example 1 and the unmineralized fish bladder obtained in Comparative Example 1 were characterized by scanning electron microscopy, and the results are as follows: Figure 1 D and Figure 1 E. It can be found that, in Figure 1 In D, the unmineralized fish bladder obtained in Comparative Example 1 contained many adjacent collagen lamellae, thus it can be determined that the fish bladder material was formed by the continuous stacking of many collagen lamellae. Figure 1 In E, the scan image of the mineralized fish maw prepared in Example 1 still shows adjacent collagen plates, but the growth of many hydroxyapatite can be seen inside, indicating that the fish maw material has been mineralized from the inside out, but the basic structure has not changed.

[0067] 2. The mineralized fish bladders prepared in Example 1 and the unmineralized fish bladders in Comparative Example 1 were subjected to fixation, tissue dehydration, infiltration, embedding and polymerization, sectioning, and staining to produce ultrathin sections, allowing electrons to penetrate the samples for observation using a transmission electron microscope (Talos F200S). The results are as follows: Figure 2 As shown.

[0068] It can be seen that, Figure 2 A and Figure 2 In Comparative Example 1, shown in B, the unmineralized fish-bubble structure contains [data / images / data]. Figure 1 The collagen microfiber lamellar layers in D) are in close contact with each other. Within the same collagen lamellar layer, the collagen microfibers are aligned in the same direction. There are differences in the angle of arrangement of collagen microfibers between adjacent layers, and the size of the collagen microfibers is approximately 50 nm. Figure 2 As can be seen from C, the internal fibers of the mineralized fish maw fibers are darkened in color due to the filling of hydroxyapatite, as observed under high magnification. Figure 2As can be seen from D, the mineralized hydroxyapatite exists in the form of needles inside the collagen fibers, and there are no hydroxyapatite minerals between the fibers, indicating that the present invention has achieved internal mineralization of fish bladder collagen fibers.

[0069] 3. The monolithically mineralized fish maw prepared in Example 1 was prepared into an ultrathin section, and the arrangement of hydroxyapatite inside was observed using a high-resolution transmission electron microscope. The results are as follows: Figure 3 As shown.

[0070] Figure 3 B is a resolved transmission electron microscope (TEM) image of the mineralized fish maw collagen fibers. The interplanar spacing of 0.34 nm is found to perfectly match the interplanar spacing of the (002) crystal plane of the HAP crystal, and the lattice fringes are neatly arranged and aligned along the direction of the crystal. Figure 3 The fact that the collagen fibers in A are arranged along their long axis indicates that hydroxyapatite grows along the axial direction of the fish bladder collagen fibers. Figure 3 C represents the selected area electron diffraction (SED) analysis of the fish-bubble mineralized collagen fibers. The SED pattern of the single-crystal-like structure, after calculation, analysis, and comparison, confirmed that the crystal composition was hydroxyapatite. Furthermore, the symmetry directions of the arc-shaped diffraction planes (002) and (004) coincided with the long axis of the collagen microfibers, indicating that the HAP crystals grew in an oriented manner within the fibers, and that the growth direction was along the long axis (c-axis) of the collagen microfibers. This demonstrates that Example 1 achieved the directional growth of hydroxyapatite within the fish-bubble collagen fibers.

[0071] 4. The mineralized fish maw prepared in Example 1 (denoted as Mineralized) and the unmineralized fish maw from Comparative Example 1 (denoted as Original) were cut into strips and tested using a universal testing machine (Instron 5967) to compare the differences in mechanical properties. The results are as follows: Figure 4 .

[0072] Figure 4 A is a tensile curve of the fish maw after mineralization in Example 1 and the unmineralized fish maw in Comparative Example 1. As shown in the figure, the fish maw after mineralization in Example 1 exhibits the properties of a brittle material compared to the unmineralized fish maw in Comparative Example 1. Figure 4 As shown in B, the elastic modulus of the mineralized fish maw in Example 1 is 1213.4 ± 140.1 MPa, while the elastic modulus of the unmineralized fish maw in Comparative Example 1 is 430.6 ± 162.1 MPa. The elastic modulus of the mineralized fish maw in Example 1 is 2.8 times higher than that of the unmineralized fish maw in Comparative Example 1. Figure 4 As shown in C, the ultimate tensile strength of the mineralized fish maw in Example 1 is 27.4 ± 3.7 MPa, while the ultimate tensile strength of the unmineralized fish maw in Comparative Example 1 is 21.2 ± 2.2 MPa. The ultimate tensile strength of the mineralized fish maw in Example 1 is 1.29 times higher than that of the unmineralized fish maw in Comparative Example 1. Figure 4 As shown in D, the elongation at break of the mineralized fish maw in Example 1 is approximately 5%, while the elongation at break of the unmineralized fish maw in Comparative Example 1 is approximately 15%. The mineralized fish maw in Example 1 has a shorter elongation at break than the unmineralized fish maw in Comparative Example 1, exhibiting characteristics of a brittle material. In summary, the mechanical properties of the mineralized fish maw in Example 1 are significantly improved compared to the unmineralized fish maw in Comparative Example 1, and the elongation at break is shorter, exhibiting characteristics of a brittle material.

[0073] 5. Thermogravimetric analysis was performed on the single mineralized fish maw prepared in Example 1. The results are as follows: Figure 5 It can be seen that after seven days of mineralization, the hydroxyapatite mineral content in the fish bubble is around 60%.

[0074] 6. The bulk mineralized adhesive raw material obtained in Example 1 through lamination and cold isostatic pressing, such as... Figure 6 As shown.

[0075] As can be seen, the bulk mineralized adhesive raw material prepared by the three steps of this invention has a diameter of 20 mm and also has a certain thickness, indicating that this invention can prepare macroscopic large-sized materials at room temperature. The bulk material appears white due to the internal growth of hydroxyapatite.

[0076] 7. The block-shaped mineralized adhesive raw material prepared in Example 1 was cut into strips 20 mm long and 2 mm wide, and subjected to a three-point bending test using a universal testing machine. The results are as follows: Figure 7 As shown, the flexural strength and elastic modulus are both average values ​​obtained from three tests.

[0077] Depend on Figure 7 It can be calculated that the flexural strength of the block prepared by the present invention is 51.89 MPa ± 9.54 MPa, and the elastic modulus is 3.00 GPa ± 1.15 GPa.

[0078] 8. Cut the unmineralized bulk material prepared in Comparative Example 2 into the same way, such as... Figure 8 As shown; then a three-point bending test was performed, and the results are as follows. Figure 9 As shown.

[0079] Depend on Figure 8 and Figure 9 The results show that the unmineralized material appears yellow and is relatively transparent. During the sample testing process, the curve did not decrease before reaching the instrument's maximum range, indicating very poor bending resistance. This demonstrates that the unmineralized sample exhibits typical characteristics of organic materials. Although it possesses toughness, it lacks the characteristics of brittle materials; it can be bent with very little force, thus limiting its application scenarios.

[0080] This invention fills the gap in the preparation method of room-temperature bulk collagen mineralized materials, solving problems related to the size, mechanical properties, and economic efficiency of existing collagen mineralized materials. This invention achieves the preparation of room-temperature bulk collagen mineralized materials by using fish bladder material—a waste product from industrial production—as a matrix. By simulating the bone mineralization process in vivo, the fish bladder material undergoes internal mineralization, and then a layering process is used to prepare large-size materials. The preparation method of this invention is convenient, simple, environmentally friendly, and highly economical, suitable for mass production. Furthermore, the produced material exhibits good mechanical properties and shows promise for use as a biological scaffold material, tissue regeneration material, and tissue replacement material.

[0081] The specific embodiments of the present invention described above do not constitute a limitation on the scope of protection of the present invention. Any other corresponding changes and modifications made in accordance with the technical concept of the present invention should be included within the scope of protection of the claims of the present invention.

Claims

1. A method for preparing room temperature collagen mineralized material, characterized in that, The following steps are involved: S1, pre-treat the fish maw material to obtain the treated fish maw; S2, the treated fish bladder is soaked in a mineralization solution for mineralization treatment and dried to obtain room temperature collagen mineralization material; In step S1, the pretreatment includes delipidation, decalcification, and removal of impurities and proteins; The degreasing conditions include: soaking in a NaOH solution with a concentration of 0.1–1 mol / L for 12–48 h; The decalcification conditions include: soaking in an HCl solution with a concentration of 0.1–1 mol / L for 12–48 h; The conditions for removing impurities include: ultrasonic cleaning with a NaCl solution of concentration of 0.1–1 mol / L for 10–240 min.

2. The method for preparing room temperature collagen mineralized material according to claim 1, characterized in that, In step S2, the treated fish maw is fixed and stretched flat on the clamp, and then soaked in the mineralization solution for mineralization treatment.

3. The method for preparing room temperature collagen mineralized material according to claim 1, characterized in that, In step S2, the mineralization solution is a mixture of calcium source, phosphorus source, buffer, sodium chloride, polyacrylic acid and solvent in a mass ratio of 1:(1~3):(4~8):(20~24):(0.1~0.5):(2200~2500).

4. The method for preparing room temperature collagen mineralized material according to claim 3, characterized in that, The calcium source includes calcium chloride dihydrate; the phosphorus source includes dipotassium hydrogen phosphate trihydrate; the buffer includes hydroxyethylpiperazine ethanethiolic acid; and the solvent is water.

5. The method for preparing room temperature collagen mineralized material according to claim 1, characterized in that, In step S2, the mineralization conditions include: a mineralization temperature of 25–40 °C, replacement of the mineralization solution every 12–48 h, and a total mineralization time of 3–7 days.

6. Room temperature collagen mineralization material prepared by the preparation method according to any one of claims 1-5.

7. A block-shaped mineralized adhesive raw material, characterized in that, The room temperature collagen mineralization material as described in claim 6 is provided in a laminated configuration, wherein each layer of the room temperature collagen mineralization material is provided with an adhesive layer.

8. The method for preparing the bulk mineralized adhesive raw material as described in claim 7, characterized in that, The following steps are involved: Using bio-adhesive as a binder, room temperature collagen mineralization materials are laminated, bonded, and pressed into shape to obtain block-shaped mineralized adhesive raw materials.

9. The method for preparing the bulk mineralized adhesive raw material according to claim 8, characterized in that, The bio-adhesive is composed of chitosan, acetic acid, and solvent mixed in a mass ratio of 1g:(5-10)mL:(30-50)mL; the amount of bio-adhesive used is 0.03-1.3mL / cm³. 2 ; The pressing process involves placing the laminated and bonded materials into a vacuum bag, evacuating the vacuum, and then performing cold isostatic pressing. The pressure for cold isostatic pressing is 100 MPa to 400 MPa, and the holding time is 5 to 30 min.

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