An injectable tissue repair material and its preparation method and application

Abstract

The application provides an injectable tissue repair material and a preparation method and application thereof, and the injectable tissue repair material comprises A component and B component; the A component comprises collagen doped with polyphosphate; and the B component comprises an inorganic salt solution.In the application, the injectable tissue repair material can uniformly cover or fill in uneven or cavity-existed wound surfaces, effectively promotes wound healing, can control the release of polyphosphate, has good stability and can be stored for a long time.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of repair materials, and particularly relates to an injectable tissue repair material and a preparation method and application thereof. BACKGROUND

[0002] Chronic wound repair has the characteristics of long treatment course, complex disease condition, multiple lesions and easy recurrence, and is very difficult to treat, resulting in high disability rate and mortality. The occurrence of various chronic wounds is related to poor microcirculation and tissue ischemia, which further causes a series of problems such as impaired tissue nutrient uptake, slow metabolic product excretion, poor tissue nutrition, bacterial infection and loss of tissue regeneration ability. Therefore, providing additional energy and nutrients to the wound microenvironment and promoting blood vessel and tissue regeneration are the key to improving the treatment of chronic wounds.

[0003] In animals and humans, a large amount of energy can be released when adenine nucleotide triphosphate (ATP) is hydrolyzed, which is the most direct source of energy for intracellular metabolism. Amorphous polyphosphate (40-1000 phosphate ester units connected by high-energy phosphoanhydride bonds) has similar functions. Under the action of alkaline phosphatase, the high-energy phosphoanhydride bonds of polyphosphate are hydrolyzed to provide the energy required for biological activity in the extracellular system. In addition, polyphosphate can also increase the expression of skin collagen types I and III, alpha-smooth muscle actin and type I plasminogen activator inhibitor, and improve the epithelialization rate.

[0004] At present, artificial dermis has a relatively wide application in the treatment of chronic wounds. However, the cells and blood vessels of chronic wounds are in a pathological state, and the metabolism is weak. Direct use of artificial dermis has a long blood vesselization cycle and a high risk of infection. Therefore, the existing technology introduces polyphosphate into artificial dermis repair materials to solve the treatment needs of chronic wounds.

[0005] For example, CN114225118A discloses an injectable artificial dermis including collagen-polysaccharide composite hydrogel microspheres loaded with polyphosphate, which supports cell ingrowth, induces cell proliferation and differentiation, can be injected into a specific site, has high viscosity after injection, and has good filling coverage and promotes tissue repair in the lacuna. However, the hydrogel product has no fixed shape, and when coated on a wound surface, the thickness is uneven. CN112402692A discloses an artificial dermis with a structure of a first collagen sponge layer, a hydrogel biological membrane layer, and a second collagen sponge layer, wherein the hydrogel biological membrane layer includes a hydrogel scaffold loaded with a therapeutic agent, which can achieve long-term sustained release, avoid "burst release" or release concentration concentration phenomenon, and better retain and exert the function of the therapeutic agent. However, the artificial dermis cannot better fit the concave part of the wound surface and fill the cavity. CN109529126A discloses an artificial dermis including a collagen sponge layer made of raw materials including collagen, polysaccharide, and functional composite particles, wherein the functional composite particles are amorphous polyphosphate and polylactic acid composite particles. The artificial dermis plays a role in stabilizing and long-term releasing amorphous polyphosphate, but it is not injectable and the release of polyphosphate is slow. At the same time, the technical solutions of CN112402692A and CN109529126A are wet-state heterogeneous products, and polyphosphate may diffuse or hydrolyze during storage, making it difficult to ensure long-term storage effectiveness.

[0006] Therefore, it is an urgent problem in the art to develop an injectable tissue repair material that can effectively control the release of polyphosphate, has good filling coverage, and promotes tissue repair in the lacuna. SUMMARY

[0007] In view of the deficiencies of the prior art, the purpose of the present application is to provide an injectable tissue repair material and its preparation method and application. The injectable tissue repair material can uniformly cover or fill in the uneven or cavity-containing wound surface, effectively promote wound healing, control the release of polyphosphate, have good stability, and can be stored for a long time.

[0008] To achieve this purpose, the present application adopts the following technical solutions:

[0009] In a first aspect, the present application provides an injectable tissue repair material, which includes component A and component B; the component A includes collagen doped with polyphosphate; and the component B includes an inorganic salt solution.

[0010] In the present application, the A component and the B component are independently packaged, which can ensure the stability of the polyphosphate, and realize the long-term effectiveness of the repair material; when used, the A component and the B component are mixed, the polyphosphate is a linear inorganic phosphate with negative charge connected by high-energy phosphate bond; the epsilon-amino group of lysine or the guanidino group of arginine in the collagen is positively charged, therefore, the polyphosphate and the collagen interact by electrostatic interaction, so that the polyphosphate is precipitated around the collagen scaffold, and the addition of the inorganic salt solution with high ionic strength weakens the electrostatic interaction between the polyphosphate and the collagen, thereby promoting the release of the polyphosphate; and the tissue repair material is not in a gel state, and can be uniformly covered or filled in the uneven or cavity-containing wound surface by the injection method, thereby effectively promoting the wound healing.

[0011] Preferably, the polyphosphate includes at least one of calcium polyphosphate, zinc polyphosphate or magnesium polyphosphate.

[0012] Preferably, the molecular chain length of the polyphosphate is 40-120 Pi, for example, can be 40 Pi, 45 Pi, 50 Pi, 55 Pi, 60 Pi, 65 Pi, 70 Pi, 75 Pi, 80 Pi, 85 Pi, 90 Pi, 95 Pi, 100 Pi, 105 Pi, 110 Pi, 115 Pi, 120 Pi, etc.

[0013] Preferably, the particle size of the polyphosphate is 50-500 nm, for example, can be 50 nm, 100 nm, 110 nm, 120 nm, 130 nm, 140 nm, 150 nm, 160 nm, 170 nm, 180 nm, 190 nm, 200 nm, 220 nm, 260 nm, 300 nm, 320 nm, 360 nm, 400 nm, 420 nm, 460 nm, 500 nm, etc.

[0014] Preferably, the collagen exists in the form of particles.

[0015] Preferably, the collagen includes at least one of type I collagen, type II collagen or type III collagen.

[0016] Preferably, the mass ratio of the polyphosphate to the collagen is 1:(15-60), for example, can be 1:15, 1:18, 1:20, 1:22, 1:25, 1:28, 1:30, 1:32, 1:35, 1:38, 1:40, 1:42, 1:45, 1:48, 1:50, 1:52, 1:55, 1:58, 1:60, etc.

[0017] Preferably, the preparation raw material of the A component further includes a crosslinking agent.

[0018] Preferably, the crosslinking agent includes formaldehyde.

[0019] Preferably, the mass ratio of the cross-linking agent to collagen is 1:(100-400), wherein the specific value in (100-400) can be 100, 120, 150, 180, 200, 220, 250, 280, 300, 320, 350, 380, 400, etc.

[0020] Preferably, the A component is prepared by a method comprising:

[0021] The collagen is mixed with a solvent to obtain a collagen solution; after the collagen solution is doped by mixing with a polyphosphate, a cross-linking agent is added for cross-linking, and then dried to obtain the component A.

[0022] In the present application, the drying method comprises freeze-drying; and the method for obtaining the dry collagen particles of the component A comprises a mechanical grinding method.

[0023] Preferably, the mass fraction of the collagen solution is 0.2-0.8%, for example, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, etc.

[0024] Preferably, the inorganic salt solution is at least one of a phosphate buffer (PB) and a phosphate buffered solution (PBS).

[0025] Preferably, the mass ratio of the component A to the component B is 1:(50-80), wherein the specific value in (50-80) can be 50, 52, 54, 55, 56, 57, 58, 59, 60, 61, 62, 64, 65, 66, 68, 68, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, etc.

[0026] Preferably, the degradation period of the tissue repair material is 30-90 days, for example, 30 days, 35 days, 40 days, 45 days, 50 days, 55 days, 60 days, 65 days, 70 days, 75 days, 80 days, 85 days, 90 days, etc.

[0027] In a second aspect, the present application provides a preparation method of the injectable tissue repair material according to the first aspect, the preparation method comprising:

[0028] The component A is mixed with the component B to obtain the injectable tissue repair material.

[0029] Preferably, the mixing method comprises mixing by repeatedly pushing with a syringe.

[0030] In a third aspect, the present application provides a use method of the injectable tissue repair material according to the first aspect, the use method comprising:

[0031] After mixing A component and B component back and forth by using a syringe, a needle is installed, and the wound is filled and covered by injection method; the syringe includes at least two syringes connected by a connector.

[0032] In a fourth aspect, the present application provides an artificial dermis, which comprises the injectable tissue repair material according to the first aspect.

[0033] The numerical range of the present application includes not only the point values listed above, but also any point values between the above numerical ranges that are not listed, and the present application does not exhaustively list the specific point values included in the range for the sake of brevity and simplicity.

[0034] Compared with the prior art, the present application has the following beneficial effects:

[0035] The injectable tissue repair material provided by the present application has the A component and the B component packaged independently, which can ensure the stability of the polyphosphate, realize long-term effectiveness of the repair material; when used, the inorganic salt solution can promote the release of the polyphosphate; and the tissue repair material is not in gel form, and can be uniformly covered or filled in uneven or cavity-containing wound surfaces by injection method, effectively promoting wound healing. BRIEF DESCRIPTION OF DRAWINGS

[0036] Fig. 1 is an SEM image of the injectable tissue repair material provided in Example 1;

[0037] Fig. 2 is an SEM image of the injectable tissue repair material provided in Example 2;

[0038] Fig. 3 is an SEM image of the injectable tissue repair material provided in Example 3; DETAILED DESCRIPTION

[0039] The technical solutions of the present application will be further described below through specific embodiments. Those skilled in the art should understand that the embodiments are only to help understand the present application and should not be regarded as specific limitations of the present application.

[0040] Example 1

[0041] This embodiment provides an injectable tissue repair material, which includes a collagen particle A component doped with polyphosphate and an inorganic salt solution B component. The preparation method of the tissue repair material includes:

[0042] (1) Preparation of 0.20wt% collagen solution; after doping with calcium polyphosphate powder at a mass ratio of polyphosphate to collagen of 1:15, cross-linking is performed by adding formaldehyde solution at a mass ratio of cross-linking agent to collagen of 1:100, and then the cross-linked composite solution is injected into a mold and freeze-dried; the obtained freeze-dried sample is mechanically sheared into granules by a pulverizer, sieved through a 300-mesh sieve, and then 100-1000 μm granules are obtained, which are ready for use by being divided into portions with a syringe, i.e. to obtain component A;

[0043] (2) Preparation of phosphate buffer, the specific steps including: weighing 29.5 g of disodium hydrogen phosphate dodecahydrate and 2.7 g of sodium dihydrogen phosphate dihydrate, dissolving in water, and diluting to 1 L to obtain a phosphate buffer with a concentration of 0.1 M, a pH value of 6.2-7.0, and an osmotic pressure of 281.8 mmol / L, which is ready for use by being divided into portions with a syringe, i.e. to obtain component B;

[0044] (3) After sterilization of components A and B by irradiation, the two components are mixed by being pushed back and forth with a syringe at a ratio of 1:50, to obtain an injectable tissue repair material.

[0045] Example 2

[0046] The present embodiment provides an injectable tissue repair material, which comprises collagen granules doped with polyphosphate A component and an inorganic salt solution B component. The preparation method of the tissue repair material comprises:

[0047] (1) Preparation of 0.65wt% collagen solution; after doping with calcium polyphosphate powder at a mass ratio of polyphosphate to collagen of 1:30, cross-linking is performed by adding formaldehyde solution at a mass ratio of cross-linking agent to collagen of 1:200, and then the cross-linked composite solution is injected into a mold and freeze-dried; the obtained freeze-dried sample is mechanically sheared into granules by a pulverizer, sieved through a 300-mesh sieve, and then 100-1000 μm granules are obtained, which are ready for use by being divided into portions with a syringe, i.e. to obtain component A;

[0048] (2) Preparation of phosphate buffer, the specific steps including: weighing 29.5 g of disodium hydrogen phosphate dodecahydrate and 2.7 g of sodium dihydrogen phosphate dihydrate, dissolving in water, and diluting to 1 L to obtain a phosphate buffer with a concentration of 0.1 M, a pH value of 6.2-7.0, and an osmotic pressure of 281.8 mmol / L, which is ready for use by being divided into portions with a syringe, i.e. to obtain component B;

[0049] (3) After sterilization of components A and B by irradiation, the two components are mixed by being pushed back and forth with a syringe at a ratio of 1:65, to obtain an injectable tissue repair material.

[0050] Example 3

[0051] The present embodiment provides an injectable tissue repair material, which comprises a collagen particle A component doped with polyphosphate and a inorganic salt solution B component. The preparation method of the tissue repair material comprises the following steps:

[0052] (1) A 0.80wt% collagen solution is prepared; after doped with calcium polyphosphate powder at a mass ratio of polyphosphate to collagen of 1:60, a crosslinking agent is added to the solution at a mass ratio of crosslinking agent to collagen of 1:400, followed by crosslinking; then the crosslinked composite solution is perfused into a mold and freeze-dried; the obtained freeze-dried sample is mechanically sheared into particles by a pulverizer, sieved through a 300-mesh sieve to obtain particles of 100-1000 μm, and then the particles are divided into portions by a syringe to obtain the A component;

[0053] (2) A PB buffer is prepared, and the specific steps include: weighing 29.5 g of disodium hydrogen phosphate dodecahydrate and 2.7 g of sodium dihydrogen phosphate dihydrate, dissolving in water, and diluting to 1 L to obtain a PB buffer with a concentration of 0.1 M, a pH value of 6.2-7.0, and an osmotic pressure of 281.8 mmol / L, which is divided into portions by a syringe to obtain the B component;

[0054] (3) After the A component and the B component are sterilized by irradiation, the A component and the B component are mixed by back and forth injection by a syringe at a ratio of 1:80 to obtain an injectable tissue repair material.

[0055] Example 4

[0056] The present embodiment provides an injectable tissue repair material, which is different from the embodiment 1 only in that the total amount of the A component and the B component is unchanged, and the mass ratio is 1:30, and the other components, the amount and the preparation method are the same as those of the embodiment 1.

[0057] Example 5

[0058] The present embodiment provides an injectable tissue repair material, which is different from the embodiment 1 only in that the total amount of the A component and the B component is unchanged, and the mass ratio is 1:100, and the other components, the amount and the preparation method are the same as those of the embodiment 1.

[0059] Example 6

[0060] The present embodiment provides an injectable tissue repair material, which is different from the embodiment 1 only in that the PB buffer is replaced by an aqueous sodium chloride solution with an equal molar amount, and the other components, the amount and the preparation method are the same as those of the embodiment 1.

[0061] The aqueous sodium chloride solution with an equal molar amount has a weak ionic strength and a small influence on electrostatic effect, which is not conducive to the release of polyphosphate, resulting in a low release amount of polyphosphate.

[0062] Test Example 1

[0063] SEM observation was performed on the collagen sponge structure of the unground doped polyphosphate of examples 1-3, respectively as shown in Figs. 1-3 As can be seen from the figures, the collagen sponge doped with polyphosphate has a porous network structure, which is conducive to cell growth.

[0064] Test Example 2

[0065] Macroscopic observation was performed on the properties of the injectable tissue repair material provided in examples 4-5, the product of example 4 had poor moisture and poor adhesion effect, and the materials could not adhere to each other, which would result in the material being unable to fill the wound surface well; the product of example 5 had too strong fluidity and poor adhesion, and the material could not be attached to the wound surface.

[0066] In summary, the injectable tissue repair material provided by the present application comprises A component and B component, and before use, the A component and the B component are independently packaged, which can ensure the stability of the polyphosphate and achieve long-term effectiveness of the repair material; when used, the A component and the B component are mixed, and an inorganic salt solution with high ionic strength is used to promote the release of polyphosphate, avoiding burst release; and the tissue repair material can be evenly covered or filled in the uneven or cavity-containing wound surface through the injection method, effectively promoting wound healing.

[0067] The applicant declares that the above is only a specific embodiment of the present application, but the protection scope of the present application is not limited thereto, and those skilled in the art should understand that any changes or replacements within the technical scope disclosed by the present application can be easily thought of by any person skilled in the art, and all fall within the protection scope and disclosure scope of the present application.

Claims

1. An injectable non-gelatinous tissue repair material, characterized in that, The injectable non-gel tissue repair material is composed of component A and component B; The component A is collagen doped with polyphosphate; the component B is an inorganic salt solution; The mass ratio of the component A to the component B is 1:(50-80); The component A and the component B are independently packaged; The component A is prepared by the following method, which comprises: Mixing collagen with solvent to obtain a collagen solution; After doping the collagen solution with polyphosphate, a crosslinking agent is added for crosslinking, and then drying to obtain component A; The crosslinking agent is formaldehyde; The inorganic salt solution is a phosphate buffer solution.

2. The injectable non-gelatinous tissue repair material of claim 1, wherein, The polyphosphate includes at least one of calcium polyphosphate, zinc polyphosphate or magnesium polyphosphate.

3. The injectable non-gelatinous tissue repair material of claim 1, wherein, The molecular chain length of the polyphosphate is 40-120 Pi.

4. The injectable non-gelatinous tissue repair material of claim 1, wherein, The particle size of the polyphosphate is 50-500 nm.

5. The injectable non-gelatinous tissue repair material of claim 1, wherein, The collagen exists in the form of particles.

6. The injectable non-gelatinous tissue repair material of claim 1, wherein, The collagen includes at least one of type I collagen, type II collagen or type III collagen.

7. The injectable non-gelatinous tissue repair material of claim 1, wherein, The mass ratio of the polyphosphate to the collagen is 1:(15-60).

8. The injectable non-gelatinous tissue repair material of claim 1, wherein, The mass ratio of the crosslinking agent to the collagen is 1:(100-400).

9. The injectable non-gelatinous tissue repair material of claim 1, wherein, The mass fraction of the collagen solution is 0.2-0.8%.

10. The injectable non-gelatinous tissue repair material of claim 1, wherein, The degradation period of the tissue repair material is 30-90 days.

11. A method of preparing an injectable non-gelatinous tissue repair material according to any one of claims 1 to 10, characterized in that, The preparation method comprises: Mixing the component A with the component B to obtain the injectable non-gel tissue repair material.

12. The method of claim 11, wherein, The mixing method comprises using a syringe to push and inject back and forth for mixing.

13. The injectable non-gelatinous tissue repair material according to any one of claims 1 to 10, wherein In use, the component A and the component B are mixed by pushing and injecting back and forth using a syringe, and then a needle is installed, and the wound is filled and covered by injection; The syringe comprises at least two syringes connected by a connector.

14. An artificial dermis, characterized by, The artificial dermis comprises the injectable non-gel tissue repair material according to any one of claims 1-10.

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