Protein biological cement and preparation method thereof

Abstract

The invention discloses protein biological cement and a preparation method thereof, and belongs to the technical field of chemical materials, the raw materials comprise anionic surfactant dispersed corn protein colloid, a primary amine polymer, hydroxyapatite and genipin. Protein of the protein biological cement is natural corn protein, and is low in cost, renewable and wide in source, and amino acid composition is diversified and rich in variety; according to the invention, a concentrated soft substance is obtained by utilizing electrostatic interaction between the protein and the primary amine polymer, and then bulk phase cohesion and interface adhesion are increased through synergistic crosslinking between genipin and hydroxyapatite and between the protein and the primary amine polymer. The high bonding strength and Young modulus of the cement are proved through underwater apparent bonding, stretching and other experiments on various substrates. And the adhesive can be used for underwater bonding of different substrates. And the brand-new protein biological cement is prepared by combining the good biocompatibility of the protein biological cement.

Description

Technical Field

[0001] The invention relates to the technical field of chemical materials, in particular to a protein biocement composed of natural protein, primary amine polymer, hydroxyapatite and genipin and a preparation method thereof. Background Art

[0002] High-performance bioadhesives have a wide range of applications in the medical field, such as bone bonding, bone defect repair, connection between medical devices and human tissues, wound closure, tooth repair and eye surgery (R. Yang, et al., Tunable backbone-degradable robust tissue adhesives via in situ radical ring-opening polymerization. Nature communications, 2023, 14, 6063.). However, most reported bioadhesives have greatly reduced bonding strength due to the hydration layer on the tissue surface, and their underwater bonding stability is also reduced (J. Yu, et al., Molecular architecture regulation for the design of instant and robust underwater adhesives. Science advances, 2023, 9, 4031.). Especially in clinical treatment, the bonding of bones and implants with high Young's modulus faces important challenges.

[0003] Current bioadhesives mainly include two categories: synthetic polymers and protein adhesives. Synthetic polymer adhesives are adhesive groups covalently grafted to polymer side chains, and super strong underwater adhesion is achieved through entanglement and non-covalent cross-linking of polymer chains. However, synthetic polymer adhesives have problems such as poor biocompatibility and difficult degradation (S.Lee, et al., Surface-patterned gallol pressure-sensitive adhesives for strong underwater adhesion. Materials Design, 2023, 236, 112505.). In addition, people have prepared protein adhesives containing catechol by genetic recombination technology (S.Huang, et al., Programmable adhesion and morphing of protein hydrogels for underwater robots. Nature communications, 2024, 15, 195.). Although recombinant protein adhesives have the advantage of good biosafety, their bonding strength is generally low, which seriously limits their biological applications. Natural proteins have the characteristics of abundant sources, renewable, low cost and good biocompatibility, and have become one of the ideal materials for preparing underwater adhesives / medical adhesives.

[0004] Therefore, providing a protein biocement prepared using natural protein and a preparation method thereof is a technical problem that technical personnel in the field urgently need to solve. Summary of the invention

[0005] In view of this, the present invention provides a protein biocement with high bonding strength and good biocompatibility and a preparation method thereof. The biocement uses natural corn protein, primary amine polymer, hydroxyapatite and genipin as raw materials, and the prepared biocement has high bonding strength and Young's modulus, and has good biocompatibility.

[0006] To achieve the above object, the present invention adopts the following technical solution:

[0007] A protein biocement is composed of the following raw materials: zein colloid dispersed by anionic surfactant, primary amine polymer, hydroxyapatite and genipin.

[0008] Preferably, it is composed of the following raw materials in percentage by mass: 45-60 wt.% of zein colloid dispersed by anionic surfactant, 15-20 wt.% of primary amine polymer, 5-10 wt.% of hydroxyapatite and 15-30 wt.% of genipin.

[0009] The beneficial effect of adopting the above scheme is that the above defined scheme of the present invention is conducive to the formation of concentrated soft material hydroxyapatite as a physical crosslinker and genipin as a chemical crosslinker by anionic surfactant dispersed corn protein colloid and primary amine polymer through electrostatic action, which is conducive to increasing the crosslinking density, improving the cohesion, and obtaining protein biocement. In the present invention, the primary amine polymer is an ideal covalent / non-covalent crosslinker; hydroxyapatite is a natural bioactive material that can act as a physical crosslinker; genipin is a product obtained from the hydrolysis of gardenia jasminoides by β-glucosidase, and a chemical crosslinker that specifically reacts with primary amines.

[0010] Furthermore, the primary amine polymer is any one of α-polylysine, ε-polylysine, polyethyleneimine and polyethylenediamine.

[0011] Furthermore, the molecular weight of the primary amine polymer is 3,000-70,000.

[0012] The present invention also provides a method for preparing the above protein biocement, comprising the following steps:

[0013] (1) weighing the raw materials according to the above mass percentages, and then dissolving the zein colloid dispersed by the anionic surfactant and the primary amine polymer in water to form corresponding aqueous solutions;

[0014] (2) adding the primary amine polymer aqueous solution dropwise to the zein colloidal aqueous solution dispersed by an anionic surfactant, shaking at room temperature to obtain a concentrated soft substance, and then freeze-drying it into a powder;

[0015] (3) Mix the freeze-dried powder obtained in step (2) with hydroxyapatite and genipin, and then grind for 5-10 minutes. Place the ground powder in 10-50 μL of double distilled water, stir at room temperature for 3-5 minutes, and then place at 20-65° C. for 24-48 hours to obtain protein biocement.

[0016] The mixed grinding in the above scheme of the present invention is conducive to uniform mixing of poorly water-soluble genipin with the above freeze-dried powder and hydroxyapatite, which is convenient for subsequent covalent cross-linking in water.

[0017] Furthermore, the preparation method of the anionic surfactant-dispersed zein colloid is as follows: zein powder is mixed with an aqueous solution of anionic surfactant to obtain a mixed solution, and the mixed solution is ultrasonicated at 90-100W for 5-30min to obtain the anionic surfactant-dispersed zein colloid.

[0018] Furthermore, the mass concentration of zein in the mixed solution is 40-90 mg / mL, and the mass concentration of the anionic surfactant is 20-60 mg / mL.

[0019] Furthermore, the anionic surfactant is any one of sodium dodecyl sulfate, sodium dodecylbenzene sulfonate, lauric acid, sodium lauroyl sarcosinate, sodium lauroyl glutamate, sodium lauroyl glycinate, dodecyl phosphate, and dipotassium cetyl phosphatidic acid.

[0020] Furthermore, in step (2), the shaking reaction is to first adjust the pH of the solution to 4-9, and then shake the reaction at 25-45° C. for 3-5 minutes;

[0021] The freeze drying is performed in a freeze dryer at -80 to -70°C for 24 to 48 hours.

[0022] The beneficial effect of adopting the above further scheme is that the pH range adopted in the above scheme of the present invention is conducive to the electrostatic compounding of the zein colloid dispersed by the anionic surfactant and the primary amine polymer to form a concentrated soft material. The shaking is for a full reaction. The freeze-drying is to obtain a powder sample for further physical grinding.

[0023] The beneficial effects of the present invention are as follows: the protein of the protein biocement of the present invention is natural corn protein, which is low-cost, renewable, and widely available, and has a diverse amino acid composition and rich variety; the present invention utilizes the electrostatic interaction between protein and primary amine polymers to obtain concentrated soft matter, and then increases the bulk cohesion and interface adhesion through the synergistic crosslinking between genipin and hydroxyapatite and protein and primary amine polymers. The high bonding strength and Young's modulus of this cement are also confirmed by underwater apparent bonding and stretching experiments on various substrates. It can be used for underwater bonding of different substrates. Combined with its good biocompatibility, a new protein biocement is prepared. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 Optical images of concentrated soft matter and freeze-dried powder formed by zein-SDS composite colloid and ε-polylysine (molecular weight: 3-4k) provided by the present invention (SDS concentration is 44 mg / mL, zein concentration is 76 mg / mL, ε-polylysine (molecular weight: 3-4k) concentration: 40 mg / mL);

[0025] Figure 2 The invention provides a protein biocement formed by physically mixing zein-SDS composite colloid, ε-polylysine (molecular weight: 3-4k), hydroxyapatite and genipin and spontaneously cross-linking and curing.

[0026] Figure 3Qualitative bonding diagram of the zein-SDS composite colloid-polylysine-hydroxyapatite-genipin biocement provided by the present invention and different substrates (including polypropylene, polyetheretherketone, polycarbonate, metal titanium, stainless steel, glass, stone, wood, shell) in an underwater polymethyl methacrylate apparatus.

[0027] Figure 4 The data graph of underwater bonding strength between the zein-SDS composite colloid-polylysine-hydroxyapatite-genipin biocement provided by the present invention and different substrates (wood, polyetheretherketone, stainless steel, cattle bone, pig skin).

[0028] Figure 5 The invention provides a stress-strain curve diagram and Young's modulus of the zein-SDS composite colloid-polylysine-hydroxyapatite-genipin biocement.

[0029] Figure 6 This is a graph showing the cytotoxicity results of the zein-SDS composite colloid-polylysine-hydroxyapatite-genipin biocement provided by the present invention. DETAILED DESCRIPTION

[0030] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0031] Example 1 A protein biocement

[0032] (1) Dissolve 76 mg of zein in 1 mL of 44 mg / mL SDS aqueous solution, and disperse the zein in the SDS aqueous solution under ultrasound for 20 min to form a zein-SDS composite colloidal solution with a negatively charged surface. Adjust the pH value of the solution to 7.4 with NaOH. Dissolve 40 mg of ε-polylysine (molecular weight: 3-4k) in 1 mL of distilled water, and adjust the pH value to 7.4 with NaOH. Subsequently, mix the zein-SDS composite colloidal solution with the ε-polylysine (molecular weight: 3-4k) aqueous solution, and shake at 25°C for 3 min to form a concentrated soft material. Remove the supernatant by decantation, and freeze-dry the concentrated soft material in a freeze dryer for 48 h to obtain zein-SDS / polylysine powder ( Figure 1 ).

[0033] (2) Weigh 80 mg of zein-SDS / polylysine powder, 5 mg of hydroxyapatite powder, and 15 mg of genipin powder in a mortar and grind at 25°C for 5 min. Then, the ground powder was placed in 0.02 mL of double distilled water and stirred for 3 min to form a yellow adhesive soft material. The yellow adhesive soft material was allowed to stand at 25°C for 24 h, and the genipin encapsulated in the adhesive soft material was solidified by covalent cross-linking with polylysine and zein, ultimately forming a hard blue-black biocement ( Figure 2 ).

[0034] Test Example 1

[0035] The bonding performance (qualitative and quantitative underwater bonding performance) was tested using the following methods:

[0036] The yellow adhesive was used to bond different substrates (including polypropylene, polyetheretherketone, polycarbonate, titanium, stainless steel, glass, stone, wood, shell) to the inner wall of a polymethyl methacrylate container filled with water, and the mixture was allowed to stand at 25°C for 24 hours. The blue-black biocement obtained was able to firmly bond different substrates to the inner wall of the polymethyl methacrylate container, which showed that the present invention was a good underwater bonding agent for protein biocement ( Figure 3 ).

[0037] The obtained yellow adhesive was applied to the surface of two solid substrates (wood, polyetheretherketone, stainless steel, cattle bones, pig skin) with a length of 12mm and a width of 8mm, and bonded together in an overlapping manner with an overlap length of 1.2cm. The bonded solid substrates were placed in a container filled with water at 25°C for 24 hours, and then the two ends of the solid substrates were vertically fixed on an electronic universal material testing machine and longitudinally stretched at a speed of 30mm / min. The maximum load force required when the solid substrates were pulled apart was measured, and the shear tensile strength per unit area was calculated based on the overlap area. At 25°C, the underwater bonding strength of the protein biocement on wood, polyetheretherketone, stainless steel, cattle bones and pig skin were 4.34, 0.76, 0.94, 2.15 and 0.2MPa, respectively. ( Figure 4 ).

[0038] Test Example 2 The Young's modulus was tested using the following method:

[0039] The yellow adhesive was filled into a 25 mm, 9 mm wide, 1 mm thick polytetrafluoroethylene rectangular mold and placed at 25°C for 24 h to obtain a rectangular blue-black biocement specimen with a length of 25 mm, a width of 9 mm, and a thickness of 1 mm. Subsequently, at 25°C, the rectangular blue-black biocement specimen was placed on the lap shear Instron 5944 universal material testing machine equipped with a 2KN sensor and tested at a test speed of 30 mm min-1 to obtain the stress-strain curve. The stress-strain formula was used to calculate the stress-strain curve. The Young's modulus is 373MPa. Figure 5 ).

[0040] Test Example 3

[0041] To test the biocompatibility of the biocement, NIH-3T3 cells were co-cultured with zein-SDS composite colloid-polylysine-hydroxyapatite-genipin in 500 μL of complete culture medium, and the culture medium was replaced with a complete culture medium containing 10% CCK-8 solution after 24 hours. Incubate at 37°C for 2 hours, measure the absorbance at 450nm, and calculate the cell viability value: [ODsample / ODcontrol]×100%. The survival rate of NIH-3T3 cells co-cultured with the protein biocement provided by the present invention remained above 85% within 24 hours, showing excellent biocompatibility ( Figure 6 ).

[0042] Example 2

[0043] ε-polylysine (molecular weight: 3-4k) was replaced with ε-polylysine (molecular weight: 7-15k), and other conditions were consistent with Example 1. The shear tensile strength of the obtained hard blue-black biocement on wood, polyetheretherketone, stainless steel, cattle bone and pig skin was 4.15, 0.71, 0.88, 2.02 and 0.18 MPa, respectively, and the Young's modulus was 227 MPa.

[0044] Example 3

[0045] ε-polylysine (molecular weight: 3-4k) was replaced with ε-polylysine (molecular weight: 10-30k), and other conditions were consistent with Example 1. The shear tensile strength of the obtained hard blue-black biocement on wood, polyetheretherketone, stainless steel, cattle bone and pig skin was 3.99, 0.68, 0.81, 1.98 and 0.18 MPa, respectively, and the Young's modulus was 220 MPa.

[0046] Example 4

[0047] ε-polylysine (molecular weight: 3-4k) was replaced with ε-polylysine (molecular weight: 30-70k), and other conditions were consistent with Example 1. The shear tensile strength of the obtained hard blue-black biocement on wood, polyetheretherketone, stainless steel, cattle bone and pig skin was 387, 0.65, 0.79, 1.95 and 0.16 MPa, respectively, and the Young's modulus was 221 MPa.

[0048] Example 5

[0049] 80 mg zein-SDS / polylysine powder, 5 mg hydroxyapatite powder and 15 mg genipin powder were replaced with 76 mg zein-SDS / polylysine powder, 6 mg hydroxyapatite powder and 18 mg genipin powder, and other conditions were consistent with Example 1. The shear tensile strength of the obtained hard blue-black biocement on wood, polyetheretherketone, stainless steel, cattle bone and pig skin was 4.39, 0.75, 0.90, 2.14 and 0.17 MPa, respectively, and the Young's modulus was 360 MPa.

[0050] Example 6

[0051] 80 mg zein-SDS / polylysine powder, 5 mg hydroxyapatite powder and 15 mg genipin powder were replaced with 68 mg zein-SDS / polylysine powder, 8 mg hydroxyapatite powder and 24 mg genipin powder, and other conditions were consistent with Example 1. The shear tensile strength of the obtained hard blue-black biocement on wood, polyetheretherketone, stainless steel, cattle bone and pig skin was 4.08, 0.70, 0.91, 2.01 and 0.18 MPa, respectively, and the Young's modulus was 355 MPa.

[0052] Example 7

[0053] 80 mg zein-SDS / polylysine powder, 5 mg hydroxyapatite powder and 15 mg genipin powder were replaced with 60 mg zein-SDS / polylysine powder, 10 mg hydroxyapatite powder and 30 mg genipin powder, and other conditions were consistent with Example 1. The shear tensile strength of the obtained hard blue-black biocement on wood, polyetheretherketone, stainless steel, cattle bone and pig skin was 4.25, 0.76, 0.83, 1.96 and 0.17 MPa, respectively, and the Young's modulus was 340 MPa.

[0054] Example 8

[0055] ε-polylysine (molecular weight: 3-4k) was replaced with polyethyleneimine (molecular weight: 10k), and other conditions were consistent with Example 1. The shear tensile strength of the obtained hard blue-black biocement on wood, polyetheretherketone, stainless steel, cattle bone and pig skin was 4.51, 0.52, 0.98, 2.34 and 0.16 MPa, respectively, and the Young's modulus was 226 MPa.

[0056] Example 9

[0057] ε-polylysine (molecular weight: 3-4k) was replaced with polyethyleneimine (molecular weight: 25k), and other conditions were consistent with Example 1. The shear tensile strength of the obtained hard blue-black biocement on wood, polyetheretherketone, stainless steel, cattle bone and pig skin was 4.45, 0.50, 0.96, 2.29 and 0.15 MPa, respectively, and the Young's modulus was 240 MPa.

[0058] Example 10

[0059] ε-polylysine (molecular weight: 3-4k) was replaced with polyethyleneimine (molecular weight: 70k), and other conditions were consistent with Example 1. The shear tensile strength of the obtained hard blue-black biocement on wood, polyetheretherketone, stainless steel, cattle bone and pig skin were 4.61, 0.56, 0.78, 2.31 and 0.16 MPa, respectively, and the Young's modulus was 235 MPa.

[0060] Embodiment 11

[0061] ε-polylysine (molecular weight: 3-4k) was replaced with polyethylenediamine, and other conditions were consistent with Example 1. The shear tensile strength of the obtained hard blue-black biocement on wood, polyetheretherketone, stainless steel, cattle bone and pig skin was 4.51, 0.82, 0.94, 2.18 and 0.15 MPa, respectively, and the Young's modulus was 212 MPa.

[0062] Example 12

[0063] ε-polylysine (molecular weight: 3-4k) was replaced with α-polylysine, and other conditions were consistent with Example 1. The shear tensile strength of the obtained hard blue-black biocement on wood, polyetheretherketone, stainless steel, cattle bone and pig skin was 3.86, 0.63, 0.74, 1.86 and 0.14 MPa, respectively, and the Young's modulus was 252 MPa.

[0064] Example 13

[0065] The pH was adjusted to 5, and the other conditions were consistent with Example 1. The shear tensile strength of the obtained hard blue-black biocement on wood, polyetheretherketone, stainless steel, cattle bone and pig skin were 4.29, 0.73, 0.97, 2.21 and 0.19 MPa, respectively, and the Young's modulus was 226 MPa.

[0066] Embodiment 14

[0067] The pH was adjusted to 6, and the other conditions were consistent with Example 1. The shear tensile strength of the obtained hard blue-black biocement on wood, polyetheretherketone, stainless steel, cattle bone and pig skin were 4.27, 0.75, 0.94, 2.28 and 0.19 MPa, respectively, and the Young's modulus was 221 MPa.

[0068] Embodiment 15

[0069] The pH was adjusted to 8, and the other conditions were consistent with Example 1. The shear tensile strength of the obtained hard blue-black biocement on wood, polyetheretherketone, stainless steel, cattle bone and pig skin were 4.21, 0.74, 0.91, 2.23 and 0.18 MPa, respectively, and the Young's modulus was 229 MPa.

[0070] Example 16

[0071] The pH was adjusted to 9, and the other conditions were consistent with those in Example 1. The shear tensile strength of the obtained hard blue-black biocement on wood, polyetheretherketone, stainless steel, cattle bone and pig skin were 4.12, 0.66, 0.84, 2.11 and 0.15 MPa, respectively, and the Young's modulus was 289 MPa.

[0072] Embodiment 17

[0073] The grinding time of the powder sample was extended to 10 min, and the other conditions were consistent with Example 1. The shear tensile strength of the obtained hard blue-black biocement on wood, polyetheretherketone, stainless steel, cattle bone and pig skin were 4.31, 0.77, 0.99, 2.34 and 0.18 MPa, respectively, and the Young's modulus was 251 MPa.

[0074] Embodiment 18

[0075] The curing time at 25°C was extended to 48 h, and the other conditions were consistent with Example 1. The shear tensile strength of the obtained hard blue-black biocement on wood, polyetheretherketone, stainless steel, cattle bone and pig skin were 4.34, 0.79, 0.93, 2.28 and 0.19 MPa, respectively, and the Young's modulus was 236 MPa.

[0076] Embodiment 19

[0077] The curing temperature was changed to 65, and the other conditions were consistent with Example 1. The shear tensile strength of the obtained hard blue-black biocement on wood, polyetheretherketone, stainless steel, cattle bone and pig skin were 8.29, 1.73, 1.97, 3.21 and 0.2 MPa, respectively, and the Young's modulus was 212 MPa.

[0078] Embodiment 20

[0079] SDS was replaced by lauric acid, and other conditions were the same as those in Example 1. The shear tensile strength of the obtained hard blue-black biocement on wood, polyetheretherketone, stainless steel, cattle bone and pig skin were 4.18, 0.69, 0.89, 2.15 and 0.17 MPa, respectively, and the Young's modulus was 207 MPa.

[0080] Embodiment 21

[0081] SDS was replaced with sodium lauroyl sarcosinate, and other conditions were consistent with Example 1. The shear tensile strength of the obtained hard blue-black biocement on wood, polyetheretherketone, stainless steel, cattle bone and pig skin were 4.32, 0.83, 0.86, 2.11 and 0.17 MPa, respectively, and the Young's modulus was 212 MPa.

[0082] Embodiment 22

[0083] SDS was replaced with sodium lauroyl glutamate, and other conditions were consistent with Example 1. The shear tensile strength of the obtained hard blue-black biocement on wood, polyetheretherketone, stainless steel, cattle bone and pig skin were 4.26, 0.65, 0.73, 2.21 and 0.14 MPa, respectively, and the Young's modulus was 215 MPa.

[0084] Embodiment 23

[0085] SDS was replaced by sodium lauroyl glycinate, and other conditions were consistent with Example 1. The shear tensile strength of the obtained hard blue-black biocement on wood, polyetheretherketone, stainless steel, cattle bone and pig skin were 4.16, 0.73, 0.93, 2.20 and 0.16 MPa, respectively, and the Young's modulus was 236 MPa.

[0086] Embodiment 24

[0087] SDS was replaced by sodium dodecylbenzene sulfonate, and other conditions were consistent with Example 1. The shear tensile strength of the obtained hard blue-black biocement on wood, polyetheretherketone, stainless steel, cattle bone and pig skin were 4.31, 0.70, 0.78, 1.92 and 0.15 MPa, respectively, and the Young's modulus was 252 MPa.

[0088] Embodiment 24

[0089] SDS was replaced with dodecyl phosphate, and other conditions were the same as those in Example 1. The shear tensile strength of the obtained hard blue-black biocement on wood, polyetheretherketone, stainless steel, cattle bone and pig skin were 4.32, 0.71, 0.79, 1.95 and 0.16 MPa, respectively, and the Young's modulus was 280 MPa.

[0090] Embodiment 24

[0091] SDS was replaced with dipotassium cetyl phosphate, and other conditions were consistent with Example 1. The shear tensile strength of the obtained hard blue-black biocement on wood, polyetheretherketone, stainless steel, cattle bone and pig skin were 4.25, 0.76, 0.93, 1.99 and 0.15 MPa, respectively, and the Young's modulus was 282 MPa.

[0092] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and vary the above embodiments within the scope of the present invention.

Claims

1. A protein biocement, characterized in that: The invention is composed of the following raw materials: zein colloid dispersed by anionic surfactant, primary amine polymer, hydroxyapatite and genipin.

2. A protein biocement according to claim 1, characterized in that: The invention is composed of the following raw materials in percentage by mass: 45-60 wt.% of corn protein colloid dispersed by anionic surfactant, 15-20 wt.% of primary amine polymer, 5-10 wt.% of hydroxyapatite and 15-30 wt.% of genipin.

3. A protein biocement according to claim 1 or 2, characterized in that: The primary amine polymer is any one of α-polylysine, ε-polylysine, polyethyleneimine and polyethylenediamine.

4. A protein biocement according to claim 3, characterized in that: The molecular weight of the primary amine polymer is 3,000-70,000.

5. A method for preparing protein biocement, characterized in that: The following steps are involved: (1) weighing the raw materials according to the mass percentages described in any one of claims 1 to 4, and then dissolving the zein colloid dispersed by the anionic surfactant and the primary amine polymer in water to form corresponding aqueous solutions; (2) adding the primary amine polymer aqueous solution dropwise to the zein colloidal aqueous solution dispersed by an anionic surfactant, shaking at room temperature to obtain a concentrated soft substance, and then freeze-drying it into a powder; (3) Mix the freeze-dried powder obtained in step (2) with hydroxyapatite and genipin, and then grind for 5-10 minutes. Place the ground powder in 10-50 μL of double distilled water, stir at room temperature for 3-5 minutes, and then place at 20-65° C. for 24-48 hours to obtain protein biocement.

6. The method for preparing protein biocement according to claim 5, characterized in that: The preparation method of the anionic surfactant-dispersed zein colloid is as follows: zein powder is mixed with an anionic surfactant aqueous solution to obtain a mixed solution, and the mixed solution is ultrasonicated at 90-100W for 5-30min to obtain the anionic surfactant-dispersed zein colloid.

7. The method for preparing protein biocement according to claim 6, characterized in that: The mass concentration of corn protein in the mixed solution is 40-90 mg / mL, and the mass concentration of anionic surfactant is 20-60 mg / mL.

8. The method for preparing protein biocement according to claim 7, characterized in that: The anionic surfactant is any one of sodium dodecyl sulfate, sodium dodecylbenzene sulfonate, lauric acid, sodium lauroyl sarcosinate, sodium lauroyl glutamate, sodium lauroyl glycinate, dodecyl phosphate, and dipotassium cetyl phosphatidic acid.

9. The method for preparing protein biocement according to claim 5, characterized in that: The shaking reaction in step (2) is to first adjust the pH of the solution to 4-9, and then shake the solution at 25-45° C. for 3-5 minutes; The freeze drying is performed in a freeze dryer at -80 to -70°C for 24 to 48 hours.

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