Bone adhesive as well as preparation method and application thereof

Abstract

The invention provides a bone adhesive as well as a preparation method and application thereof, and belongs to the technical field of medical bone adhesives. The bone adhesive provided by the invention comprises isinglass, polyphenol and dialdehyde starch, the mass ratio of the isinglass to the polyphenol to the dialdehyde starch is (1-5): (0.2-2): (0.1-10). The bone adhesive provided by the invention is formed by crosslinking the isinglass, the polyphenol substance and the dialdehyde starch, has the advantages of good bone adhesiveness, high strength, good biocompatibility and degradability, can adhere fracture fragments, and provides a new strategy for fracture repair.

Description

Technical Field

[0001] This invention belongs to the field of medical bone adhesive technology, specifically relating to a bone adhesive, its preparation method, and its application. Background Technology

[0002] Current fracture treatment primarily involves the reduction and fixation of bone fragments to restore their original position and achieve close apposition, thereby promoting bone healing through endogenous osteogenesis. Tissue adhesives are materials that can seal soft tissues such as skin and muscles through their surface adhesion and adhesive properties; examples include soft tissue adhesives and bone adhesives. Bone adhesives exhibit rapid and strong adhesion to bone tissue while promoting bone repair. Compared to internal fixation methods, they offer advantages in convenience, safety, efficiency, and cost-effectiveness.

[0003] Several bone adhesives have undergone clinical trials. Fibrin glue, for example, exhibits excellent bioactivity and has shown potential in accelerating the repair of non-weight-bearing fractures, such as cartilage fractures, but its adhesive strength is limited. Cyanoacrylate adhesives have unavoidable toxicity issues and offer poorer fracture fixation compared to metal plates and screws. Polyurethane bone adhesives offer high adhesive strength, but still suffer from complications and long curing times. Similarly, "Os-tamer" has been clinically tested but found to have poor adhesive properties, leading to complications such as infection and tissue necrosis. Polymethyl methacrylate (PMMA) and CaP bone cement have been tested in clinical settings, but both show unsatisfactory adhesion at the bone interface, lacking the adhesive strength required for bone adhesives. Therefore, a bone adhesive product that combines both adhesive strength and safety is urgently needed. Summary of the Invention

[0004] The purpose of this invention is to provide a bone adhesive, its preparation method, and its application. The bone adhesive provided by this invention has excellent adhesive strength, excellent cell compatibility and blood compatibility, and good safety.

[0005] To achieve the objectives of this invention, the following technical solutions are provided: A bone adhesive includes fish maw glue, polyphenols, and dialdehyde starch; the mass ratio of the fish maw glue, polyphenols, and dialdehyde starch is 1~5:0.2~2:0.1~10; the water content of the bone adhesive is 10~50%.

[0006] Preferably, the polyphenol is a flavonoid polyphenol.

[0007] Preferably, the flavonoid polyphenol is tannic acid.

[0008] Preferably, the number-average molecular weight of the tannic acid is 500~20000 Da.

[0009] Preferably, the dialdehyde starch is obtained by modifying starch with sodium periodate.

[0010] The present invention also provides a method for preparing the bone adhesive described in the above technical solution, comprising the following steps: The bone adhesive is obtained by mixing fish maw glue, polyphenols, dialdehyde starch and water.

[0011] Preferably, the mixing temperature is 60~70℃ and the mixing time is 20~60min.

[0012] Preferably, the mixing is carried out under stirring conditions; the stirring rate is 50~100 rpm.

[0013] The present invention also provides the application of the bone adhesive described in the above technical solution or the bone adhesive prepared by the preparation method described in the above technical solution in the preparation of products with the function of promoting tissue healing.

[0014] This invention provides a bone adhesive comprising fish maw glue, polyphenols, and dialdehyde starch; the mass ratio of the fish maw glue, polyphenols, and dialdehyde starch is 1-5:0.2-2:0.1-10; the water content of the bone adhesive is 10-50%. The fish maw glue in this invention is not only environmentally friendly and has strong adhesion, but also does not damage bone tissue structure. Fish maw is composed of collagen, elastin, and polysaccharides, among which collagen and elastin constitute collagen fibers and elastic fibers, respectively; the polysaccharides are mainly chondroitin sulfate (95%, type A / C and type B, in a ratio of 1.4:1) and hyaluronic acid (HA, 5%); chondroitin sulfate plays an important role in promoting wound healing and can be used for cartilage repair and tissue engineering. Furthermore, fish maw is rich in inorganic salts and vitamins, especially calcium, followed by potassium and magnesium, as well as trace elements such as zinc, copper, iron, and selenium. These trace elements can maintain normal physiological functions and health in the human body. In this invention, the catechol groups in polyphenols can undergo cross-linking reactions with fish maw glue, while the aldehyde groups in dialdehyde starch also undergo cross-linking reactions with the groups in fish maw glue, making the internal structure of fish maw glue more compact and giving it better interfacial adhesion. Attached Figure Description

[0015] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0016] Figure 1 This is a schematic diagram of adhesive adhesion. Figure 2The adhesion curves of the bone adhesive, 502 glue, fibrin glue and fish maw glue obtained in Examples 1 and 2 on the surface of wet cancellous bone fragments are shown. Figure 3 The bar chart shows the adhesion strength of the bone adhesive, 502 glue, fibrin glue and fish maw glue obtained in Examples 1 and 2 on the surface of wet cancellous bone fragments. Figure 4 The adhesion curves of the bone adhesive, 502 glue, fibrin glue and fish maw glue obtained in Examples 1 and 2 on the wet titanium sheet surface are shown. Figure 5 The bonding strength curves of the bone adhesive, 502 glue, fibrin glue and fish maw glue obtained in Examples 1 and 2 on the surface of a wet glass slide are shown. Figure 6 This is a schematic diagram illustrating the actual adhesion effect of the bone adhesive obtained in Example 1 to bovine bone; Figure 7 Bar chart showing the cell viability of MC3T3-E1 cells for the bone adhesives obtained in Examples 1 and 2; Figure 8 The coagulation effect of the bone adhesives obtained in Examples 1 and 2; Figure 9 The results of blood compatibility testing of the bone adhesives obtained in Examples 1 and 2 are shown. Figure 10 The degradation effect of the bone adhesive obtained in Example 1 in PBS. Detailed Implementation

[0017] The present invention provides a bone adhesive comprising fish maw glue, polyphenols and dialdehyde starch; the mass ratio of the fish maw glue, polyphenols and dialdehyde starch is 1~5:0.2~2:0.1~10; the water content of the bone adhesive is 10~50%.

[0018] In this invention, unless otherwise specified, all raw materials used in the preparation are commercially available products well known to those skilled in the art.

[0019] In this invention, the fish maw glue can be from marine fish of the Sciaenidae family, specifically including large yellow croaker, yellow croaker, red-lipped croaker, white croaker, as well as sturgeon, cod, beluga, and croaker. This invention does not impose any particular limitation on the preparation method of the fish maw glue; any method well-known to those skilled in the art can be used. In a specific embodiment, the fish maw glue is prepared from fish maw through steaming, repeated pounding, and drying.

[0020] In this invention, the polyphenol is a flavonoid polyphenol, specifically tannic acid; the tannic acid can be hydrolyzed tannin or condensed tannic acid; preferably, the number average molecular weight of the tannic acid is 500~20000 Da, and in specific embodiments it can be 1000, 1500, 2500, 5000 or 10000 Da.

[0021] In this invention, the dialdehyde starch is obtained by modifying starch with sodium periodate; in this invention, the dialdehyde starch is prepared by conventional preparation methods well known to those skilled in the art, and for details, please refer to "Preparation and Characteristic Study of Dialdehyde Waxy Corn Starch" (Hu Lei, Gao Qunyu, Food Industry Technology, 2012, (03): 195-198.DOI: 10.13386 / j.issn1002-0306.2012.03.067).

[0022] The present invention also provides a method for preparing the bone adhesive described in the above technical solution, comprising the following steps: The bone adhesive is obtained by mixing fish maw glue, polyphenols, modified starch and water.

[0023] In this invention, the mixing method of fish maw glue, polyphenols, dialdehyde starch and water can be as follows: Fish maw glue and water are first mixed to obtain a precursor solution; the volume ratio of fish maw glue to water is 0.5~2:1; the temperature of the first mixing is 65~70℃, the rotation speed is 50~100 rpm, and the time is 30~60 min; Polyphenols and water are mixed in a second step to obtain a polyphenol solution; the water is sterile water; the temperature of the second mixing step is room temperature; the concentration of polyphenols in the polyphenol solution is 0.04~0.24 g / mL, and in specific embodiments it can be 0.06, 0.08, 0.1 or 0.18 g / mL; The dialdehyde starch and water are mixed in a third step to obtain a dialdehyde starch solution; the temperature of the third mixing is 60~70℃; or the dialdehyde starch is dissolved in water at a temperature of 60~70℃; the concentration of dialdehyde starch in the dialdehyde starch solution is 0.01~0.08 g / mL, and in specific embodiments, it can be 0.02, 0.04 or 0.06 g / mL.

[0024] The precursor solution and polyphenol solution are mixed in a fourth step, and then a dialdehyde starch solution is added and mixed in a fifth step to obtain the bone adhesive. The volume ratio of the precursor solution and polyphenol solution to the dialdehyde starch solution is 3:0.2~1:0.1~0.6, and in a specific embodiment, it can be 3:0.3:0.3 or 3:0.5:0.5. The fourth and fifth mixing steps are performed at a temperature of 60~70℃, a rotation speed of 50~150 rpm, and a time of 10~15 min.

[0025] The present invention also provides the application of the bone adhesive described in the above technical solution or the bone adhesive prepared by the preparation method described in the above technical solution in the preparation of products with the function of promoting tissue healing.

[0026] To further illustrate the present invention, the bone adhesive, its preparation method, and its application provided by the present invention will be described in detail below with reference to the accompanying drawings and embodiments, but these should not be construed as limiting the scope of protection of the present invention.

[0027] Example 1 Fish swim bladder gelatin and water were mixed in a ratio of 0.5:1 and mixed at 100 rpm and 65°C for 30 min to obtain a precursor solution. Prepare a tannic acid solution with a concentration of 0.12 g / mL by dissolving tannic acid (molecular weight 500 Da) in sterile water at room temperature. Dissolve the dialdehyde starch in water at 70℃ to prepare a dialdehyde starch solution with a concentration of 0.02 g / mL; The precursor solution, tannic acid solution, and dialdehyde starch solution were mixed in a magnetically heated stirrer at a volume ratio of 3:0.5:0.5. The temperature was 60-70℃ and the speed was 50-100 rpm. After stirring for 15 minutes, the dialdehyde starch solution was added and the mixture was stirred for another 10 minutes to obtain the bone binder.

[0028] Example 2 Fish swim bladder gelatin and water were mixed in a ratio of 0.5:1 and mixed at 100 rpm and 65°C for 30 min to obtain a precursor solution. Dissolve tannic acid (molecular weight 1500 Da) in sterile water at room temperature to prepare a tannic acid solution with a concentration of 0.24 g / mL. Dissolve the dialdehyde starch in water at 70℃ to prepare a dialdehyde starch solution with a concentration of 0.04 g / mL; The precursor solution, tannic acid solution, and dialdehyde starch solution were mixed in a magnetically heated stirrer at a volume ratio of 3:0.5:0.5. The temperature was 60-70℃ and the speed was 50-100 rpm. After stirring for 15 minutes, the dialdehyde starch solution was added and the mixture was stirred for another 10 minutes to obtain the bone binder.

[0029] Example 3 Fish maw glue and water were mixed in a 1:1 ratio and mixed at 100 rpm and 65°C for 30 min to obtain a precursor solution. Dissolve tannic acid (molecular weight 2000 Da) in sterile water at room temperature to prepare a tannic acid solution with a concentration of 0.08 g / mL. Dissolve the dialdehyde starch in water at 70℃ to prepare a dialdehyde starch solution with a concentration of 0.06 g / mL; The precursor solution, tannic acid solution, and dialdehyde starch solution were mixed in a magnetically heated stirrer at a volume ratio of 3:0.5:0.5. The temperature was 60-70℃ and the speed was 50-100 rpm. After stirring for 15 minutes, the dialdehyde starch solution was added and the mixture was stirred for another 10 minutes to obtain the bone binder.

[0030] Example 4 Fish maw glue and water were mixed in a 1:1 ratio and mixed at 100 rpm and 65°C for 30 min to obtain a precursor solution. Dissolve tannic acid (molecular weight 5000 Da) in sterile water at room temperature to prepare a tannic acid solution with a concentration of 0.1 g / mL; Dissolve the dialdehyde starch in water at 70℃ to prepare a dialdehyde starch solution with a concentration of 0.06 g / mL; The precursor solution, tannic acid solution, and dialdehyde starch solution were mixed in a magnetically heated stirrer at a volume ratio of 3:0.3:0.3. The temperature was 60-70℃ and the speed was 50-100 rpm. After stirring for 15 minutes, the dialdehyde starch solution was added and the mixture was stirred for another 10 minutes to obtain the bone binder.

[0031] Example 5 Fish maw glue and water were mixed in a 1:1 ratio and mixed at 100 rpm and 65°C for 30 min to obtain a precursor solution. Dissolve tannic acid (molecular weight 10000 Da) in sterile water at room temperature to prepare a tannic acid solution with a concentration of 0.1 g / mL; Dissolve the dialdehyde starch in water at 70℃ to prepare a dialdehyde starch solution with a concentration of 0.08 g / mL; The precursor solution, tannic acid solution, and dialdehyde starch solution were mixed in a magnetically heated stirrer at a volume ratio of 3:0.2:0.2. The temperature was 60-70℃ and the speed was 50-100 rpm. After stirring for 15 minutes, the dialdehyde starch solution was added and the mixture was stirred for another 10 minutes to obtain the bone binder.

[0032] Test Example 1 The wet adhesion properties of bone tissue adhesives on different material surfaces were tested using the following methods: Wet cancellous bone fragments, wet titanium sheets, and wet glass slides were selected as test substrates. The cancellous bone fragments were taken from fresh bovine bones, cut and polished, and then soaked in physiological saline to keep them moist. Their porous structure and organic components can simulate the complex interface of human bone tissue. The medical-grade titanium sheets were polished and ultrasonically cleaned, and then sprayed with deionized water to simulate a humid environment, representing commonly used metal implant materials in orthopedics.

[0033] according to Figure 1The two substrates were adhered together using an overlapping shear test method, with the bone tissue to be tested evenly applied between them. Simultaneously, commercially available medical 502 adhesive and fibrin glue samples were prepared in the same manner. After the adhesives had completely cured, the samples were fixed on an electronic universal testing machine, and shear tests were performed at a rate of 1 mm / min. The shear force versus displacement curve was recorded in real time. The results are shown below. Figures 2-5 As shown.

[0034] Note: Figures 2-5 FBG is a fish maw gelatin made by mixing fish maw and water in a 0.5:1 ratio; FBG / DS / TA 0.12 and FBG / DS / TA 0.24 The bone tissue adhesives obtained in Examples 1 and 2 are respectively. (From...) Figures 2-5 The results show that the bone tissue adhesive provided by the present invention has excellent adhesion properties.

[0035] Test Example 2 The bone tissue adhesive obtained in Example 1 was applied to the cut surface of fresh pig bone. After bonding, it was placed in a constant temperature and humidity chamber at 37°C and 100% humidity. The adhesion process and adhesion effect are as follows. Figure 6 As shown. By Figure 6 The results showed that the glued pig bones could support a 65kg adult standing up.

[0036] Test Example 3 The biocompatibility of the bone tissue adhesives obtained in Examples 1 and 2 was tested using an in vitro cell culture method. Adhesive extracts with different TA (tannic acid) contents were added to 96-well plates containing osteoblasts, with TA-free adhesive extracts and blank culture medium serving as controls. Cell viability was assessed using the CCK-8 assay. Figure 7 The results showed that the cell survival rate of the low TA content group was over 90% after 1 and 3 days of culture, which was significantly higher than that of the high TA content group. The low TA content adhesive showed good cell compatibility and promoted osteoblast proliferation. At the same time, it had good coagulation properties, which is beneficial for hemostasis and sealing of wound sites.

[0037] Test Example 4 First, the bone tissue adhesive obtained in Example 1 was equilibrated in physiological saline at 37°C. Then, the saline was discarded, and fresh anticoagulated whole blood and 0.2M calcium chloride solution were sequentially added to the surface of the bone tissue adhesive to initiate coagulation, and the mixture was immediately incubated at a constant temperature of 37°C. After 2 hours, ultrapure water was added to the system to terminate the reaction and lyse uncoagulated red blood cells. After gentle mixing, the liquid was aspirated and centrifuged, and the absorbance of the supernatant was measured at 540 nm using a microplate reader. Finally, the procoagulant properties of the hydrogel were evaluated by calculating the coagulation index or coagulation rate at each time point; a lower absorbance value indicated a better coagulation effect.

[0038] Figure 8 The figure shows the coagulation effect of the bone adhesives obtained in Examples 1 and 2. The results show that the bone adhesives prepared in this invention have a good coagulation effect.

[0039] Test Example 5 The blood compatibility of the bone tissue adhesives obtained in Examples 1 and 2 was determined by measuring the hemolysis rate. The adhesives were co-incubated with a red blood cell suspension, and the absorbance of the supernatant was measured. The results are as follows: Figure 9 As shown, Figure 9 In the diagram, ① represents the FBG group, and ② represents FBG / DS / TA. 0.12 Group ③ is FBG / DS / TA 0.24 Groups ④ and ⑤ are PBS and deionized water groups, respectively. The calculated hemolysis rates were only 1.21% and 3.63%, respectively, far below the safety standard of 5%, indicating that the bone tissue adhesive provided by this invention has no significant destructive effect on blood components, has good blood compatibility, and provides a safety guarantee for clinical application.

[0040] Test Example 6 The bone adhesive obtained in Example 1 was soaked in PBS buffer and placed in an environment of 37°C. The material was taken out at regular intervals, the surface moisture was absorbed with filter paper, and the material was weighed to test the degradation performance of the bone adhesive obtained in Example 1.

[0041] Figure 10 The figure shows the degradation effect of the bone adhesive obtained in Example 1 in PBS. The results show that the bone adhesive prepared in this invention can be completely degraded in PBS within 35 days.

[0042] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A bone adhesive, characterized by, The bone adhesive comprises swim bladder glue, polyphenol and dialdehyde starch; the mass ratio of the swim bladder glue, polyphenol and dialdehyde starch is 1-5:0.2-2:0.1-10; the water content of the bone adhesive is 10-50%.

2. The bone adhesive of claim 1, wherein, The polyphenol is flavonoid polyphenol.

3. The bone adhesive of claim 2, wherein, The flavonoid polyphenol is tannic acid.

4. The bone cement according to claim 3, characterized in that The number average molecular weight of the tannic acid is 500-20000 Da.

5. The bone adhesive of claim 1, wherein, The dialdehyde starch is obtained by modifying starch with sodium periodate.

6. Process for the preparation of the bone cement according to any one of claims 1 to 5, characterized in that, The method comprises the following steps: The swim bladder glue, polyphenol, dialdehyde starch and water are mixed to obtain the bone adhesive.

7. The production method according to claim 6, wherein The mixing temperature is 60-70 DEG C, and the mixing time is 20-60 min.

8. The production method according to claim 6 or 7, characterized by, The mixing is carried out under stirring; the stirring rate is 50-100 rpm.

9. Use of the bone adhesive of any one of claims 1-5 or the bone adhesive prepared by the preparation method of any one of claims 6-8 in the preparation of a product with the efficacy of promoting tissue healing.

Images