阅读说明：本技术 一种双光响应型氧化锌及其制备方法以及具有抗菌成骨性的光敏涂层 (Double-light response type zinc oxide, preparation method thereof and photosensitive coating with antibacterial osteogenesis property ) 是由 廖岚 王小磊 赵思语 于 2020-06-24 设计创作，主要内容包括：本发明涉及外科种植体技术领域,提供了一种双光响应型氧化锌及其制备方法以及具有抗菌成骨性的光敏涂层。本发明在氧化锌制备过程中加入柠檬酸钠和羟丙基甲基纤维素对形貌进行控制,加入光热转换材料使氧化锌具有光热转换能力,加入木质素降低氧化锌的能带间隙；通过微波辐照使冻干后的水热产物碳化,能带间隙进一步降低,使材料对黄光具有响应能力。本发明制备的氧化锌呈银耳状褶皱结构,对黄光和近红外光具有双重响应,具有良好的吸附性、抗菌性和光热稳定性,具有光热转换能力。本发明提供的涂层兼具抗菌性和成骨性,应用于种植体表面能够有效提高种植体的抗菌和成骨能力,且其特殊的光敏性能够实现种植体抗菌和成骨功能的光控起效和按需作用。(The invention relates to the technical field of surgical implants, and provides double-light response type zinc oxide, a preparation method thereof and a photosensitive coating with antibacterial osteogenesis property. In the preparation process of the zinc oxide, sodium citrate and hydroxypropyl methyl cellulose are added to control the appearance, a photo-thermal conversion material is added to enable the zinc oxide to have photo-thermal conversion capability, and lignin is added to reduce the energy band gap of the zinc oxide; the hydrothermal product after freeze-drying is carbonized through microwave irradiation, the energy band gap is further reduced, and the material has response capability to yellow light. The zinc oxide prepared by the invention is of a silver ear-shaped fold structure, has dual response to yellow light and near infrared light, has good adsorbability, antibacterial property and photo-thermal stability, and has photo-thermal conversion capability. The coating provided by the invention has antibacterial property and osteogenesis property, can effectively improve the antibacterial and osteogenesis capacity of the implant when being applied to the surface of the implant, and can realize the light-operated effect and the required effect of the antibacterial and osteogenesis functions of the implant due to the special photosensitivity of the coating.)

1. A preparation method of double-photoresponse zinc oxide comprises the following steps:

(1) mixing soluble zinc salt, hexamethylenetetramine and water to perform a first hydrothermal reaction, and then mixing the reaction feed liquid with sodium citrate, hydroxypropyl methylcellulose, a photothermal conversion material and lignin to perform a second hydrothermal reaction to obtain a hydrothermal product;

(2) and sequentially carrying out freeze drying and microwave irradiation on the hydrothermal product to obtain the double-photoresponse type zinc oxide.

2. The method of claim 1, wherein the soluble zinc salt is zinc nitrate.

3. The production method according to claim 1, wherein the photothermal conversion material is one or more of activated carbon, gold rod, and black phosphorus.

4. The preparation method according to claim 1, 2 or 3, wherein the using ratio of the soluble zinc salt, the hexamethylenetetramine, the water, the sodium citrate, the hydroxypropyl methylcellulose, the photothermal conversion material and the lignin is (1.487-1.488) g, (0.350-0.352) g, (95-110) mL, (0.138-0.142) g, (0.100-0.120) g, (0.025-0.035) g and (0.995-0.115) g.

5. The preparation method according to claim 1, wherein the temperature of the first hydrothermal reaction is 65-67 ℃ and the time is 14-16 min; the temperature of the second hydrothermal reaction is 85-87 ℃, and the time is 10-12 h.

6. The method according to claim 1, wherein the microwave irradiation is performed at a power of 800W or more for a time of 15min or more.

7. A dual-photoresponse zinc oxide prepared by the process of any one of claims 1 to 6 wherein the dual-photoresponse zinc oxide has a microscopic morphology which is in the form of a harpoon-shaped microstructure; the double-photoresponse zinc oxide can respond under the irradiation of yellow light and near infrared light.

8. A photosensitive coating having antibacterial osteogenic properties, prepared from the dual photo-responsive zinc oxide of claim 7.

9. The photosensitive coating of claim 8, wherein the coating is prepared by:

dispersing double-light response type zinc oxide in a solvent, coating the obtained suspension on the surface of a base material, and drying to obtain a photosensitive coating with antibacterial osteogenesis property; the substrate is a surgical implant.

10. The photosensitive coating of claim 8 or 9, wherein the coating is prepared from raw materials further comprising type-collagen powder, and the mass ratio of the double-light response type zinc oxide to the type-collagen powder is 1 (1-5).

Technical Field

The invention relates to the technical field of surgical implants, in particular to double-light response type zinc oxide, a preparation method thereof and a photosensitive coating with antibacterial osteogenesis property.

Background

Oral implant surgery is one of the most common implant operations in humans, and at the same time, the number of patients affected by peri-implant diseases (PIDs) is increasing. Different from the conventional totally-enclosed implantation system, the incidence rate of diseases around the oral implant is as high as about 40%. This is due to exposure of the implant part to the oral microenvironment, which is prone to bacterial invasion and peri-implant mucositis (PIM); further spread of inflammation leads to gradual loss of peri-implantitic supporting bone, which in turn leads to peri-implantitis (PI). The bacterial infection is most closely related to PIDs of the implant, which can cause weak osseointegration around the implant, the implant falls off and finally the implant fails, so that the edentulous patient not only needs more additional operations to treat the PIDs, but also sometimes even needs to go through secondary implant operations to recover the edentulous, and the physical and mental effects of the patient are greatly influenced.

Titanium-based implants are the most widely clinically applied implants at present, but titanium as an inert material has no antibacterial property, is poor in binding property with surrounding bone tissues, and has high implant failure rate.

Disclosure of Invention

In view of the above, the present invention provides a two-photon response zinc oxide, a preparation method thereof, and a photosensitive coating with antibacterial osteogenesis property. The double-light response type zinc oxide provided by the invention has good adsorption capacity, can respond to yellow light and near infrared light, has good antibacterial property and photothermal conversion capacity, and the antibacterial property and osteogenesis property of the implant can be remarkably improved by using the coating prepared by the double-light response type zinc oxide.

In order to achieve the above object, the present invention provides the following technical solutions:

a preparation method of double-photoresponse zinc oxide comprises the following steps:

(1) mixing soluble zinc salt, hexamethylenetetramine and water to perform a first hydrothermal reaction, and then mixing the reaction feed liquid with sodium citrate, hydroxypropyl methylcellulose, a photothermal conversion material and lignin to perform a second hydrothermal reaction to obtain a hydrothermal product;

(2) and sequentially carrying out freeze drying and microwave irradiation on the hydrothermal product to obtain the double-photoresponse type zinc oxide.

Preferably, the soluble zinc salt is zinc nitrate.

Preferably, the photothermal conversion material is one or more of activated carbon, gold rod and black phosphorus.

Preferably, the dosage ratio of the soluble zinc salt, the hexamethylenetetramine, the water, the sodium citrate, the hydroxypropyl methyl cellulose, the photothermal conversion material and the lignin is (1.487-1.488) g, (0.350-0.352) g, (95-110) mL, (0.138-0.142) g, (0.100-0.120) g, (0.025-0.035) g and (0.995-0.115) g.

Preferably, the temperature of the first hydrothermal reaction is 65-67 ℃, and the time is 14-16 min; the temperature of the second hydrothermal reaction is 85-87 ℃, and the time is 10-12 h.

Preferably, the power of the microwave irradiation is more than 800W, and the time is more than 15 min.

The invention provides the double-photoresponse zinc oxide prepared by the method in the scheme, and the microscopic morphology of the double-photoresponse zinc oxide is in a shape of a silver ear; the double-photoresponse zinc oxide can respond under the irradiation of yellow light and near infrared light.

The invention also provides a photosensitive coating with antibacterial osteogenesis property, which is prepared from the double-light response type zinc oxide.

Preferably, the coating is prepared by the following steps:

dispersing double-light response type zinc oxide in a solvent, coating the obtained suspension on the surface of a base material, and drying to obtain a photosensitive coating with antibacterial osteogenesis property; the substrate is a surgical implant.

Preferably, raw materials for preparing the coating further comprise collagen type powder, and the mass ratio of the double-light response type zinc oxide to the collagen type powder is 1 (1-5).

The invention provides a preparation method of double-photoresponse zinc oxide, which comprises the steps of mixing soluble zinc salt, hexamethylenetetramine and water for carrying out a first hydrothermal reaction, and then mixing reaction liquid with sodium citrate, hydroxypropyl methyl cellulose, a photothermal conversion material and lignin for carrying out a second hydrothermal reaction to obtain a hydrothermal product; and then sequentially carrying out freeze drying and microwave irradiation on the hydrothermal product to obtain the double-light response type zinc oxide. In the preparation process, sodium citrate and hydroxypropyl methyl cellulose are added to control the appearance of zinc oxide, a photo-thermal conversion material is added to enable the obtained zinc oxide to have photo-thermal conversion capability, and lignin is added to enable the energy band gap of the zinc oxide to be reduced; after the hydrothermal process is finished, the hydrothermal product is subjected to microwave irradiation after being frozen and dried, and the hydrothermal product is carbonized through the microwave irradiation, so that the energy band gap of the zinc oxide is further reduced, and the material has the response capability to the visible light (yellow light) with long wavelength; meanwhile, the carbonization effect of microwave irradiation can also make the silver ear-shaped fold structure of the zinc oxide more obvious, thereby improving the adsorption capacity of the zinc oxide.

The double-photoresponse zinc oxide prepared by the method is of a silver ear-shaped fold structure, has good adsorbability (can adsorb substances such as pigments, proteins and the like), is good in antibacterial property and photo-thermal stability, has photo-thermal conversion capability, has a photocatalysis effect under yellow light irradiation, can further improve antibacterial capability, has photo-thermal conversion capability under near infrared light irradiation, and is a good photo-thermal treatment material.

The invention also provides a photosensitive coating with antibacterial osteogenesis property, which is prepared from the double-light response type zinc oxide. The coating provided by the invention has antibacterial property and osteogenesis property, can be activated under the irradiation of safe and mild visible light (yellow light), has good antibacterial performance on both gram-positive bacteria and gram-negative bacteria, and particularly has an obvious antibacterial effect on specific oral bacteria. In addition, the coating shows good photothermal conversion capability under Near Infrared (NIR) irradiation, and the appropriate increase of temperature can effectively promote the proliferation and osteogenic gene expression of bone marrow mesenchymal stem cells (BMSCs) and promote the osseointegration of the implant and the surrounding bone tissues; the coating is applied to the surface of the titanium-based implant, so that the antibacterial and osteogenic effects of the titanium-based implant can be effectively improved, and the special photosensitivity of the coating can realize the light-operated effect and the on-demand effect of the antibacterial and osteogenic functions of the implant.

Furthermore, the photosensitive coating provided by the invention also comprises type collagen powder as a raw material for preparing the photosensitive coating, wherein the type collagen powder has the effect of promoting osteogenesis and can further improve the osteogenesis of the coating.

Drawings

FIG. 1 is a scanning electron micrograph of ZnO and ZnO-Col-I with a scale of 1 μm;

FIG. 2 is a graph of energy spectra of ZnO and ZC;

FIG. 3 is an X-ray diffraction pattern of Ti and Ti-ZnO;

FIG. 4 is a standard curve of bovine serum albumin in example 3;

FIG. 5 is a graph showing the results of BSA adsorption at different concentrations of zinc oxide in example 3;

FIG. 6 is a graph of the ultraviolet absorption spectra of the rhodamine B solution at 0h, 12h, 24h, 36h and 48h under dark and light conditions in example 4;

FIG. 7 is a graph showing the change in the degradation rate of rhodamine B in example 4 under dark and light conditions;

FIG. 8 is a graph showing the temperature change under light conditions of different types of zinc oxide dispersions of example 5;

FIG. 9 is a graph of the thermal imaging of differently treated titanium samples of example 5 before and after NIR irradiation;

FIG. 10 is an SEM observation (scale: 1 μm) of the antibacterial ratio against Staphylococcus aureus (S. aureus) and bacteria of the Ti sample in example 6;

fig. 11 is an SEM observation image (scale: 1 μm) of the antibacterial ratio against escherichia coli (e.coli) and bacteria of the Ti sample in example 6;

FIG. 12 is an SEM observation (scale: 1 μm) of the antibacterial ratio and bacteria against Streptococcus mutans (S.mutans) of the Ti sample in example 6;

FIG. 13 is a graph of the temperature change under NIR irradiation of cells treated with different macerating solutions of example 7;

FIG. 14 is an infrared thermography of cells treated with different maceratants in example 7 after 10min of continuous NIR irradiation;

FIG. 15 shows gene expression levels of Runx2 and OCN of cells treated with different maceration solutions in example 8;

FIG. 16 shows the results of ALP staining level measurements (scale: 200 μm) of cells treated with different macerating solutions in example 8.

Detailed Description

The invention provides a preparation method of double-light response type zinc oxide, which comprises the following steps:

(1) mixing soluble zinc salt, hexamethylenetetramine and water to perform a first hydrothermal reaction, and then mixing the reaction feed liquid with sodium citrate, hydroxypropyl methylcellulose, a photothermal conversion material and lignin to perform a second hydrothermal reaction to obtain a hydrothermal product;

(2) and sequentially carrying out freeze drying and microwave irradiation on the hydrothermal product to obtain the double-photoresponse type zinc oxide.

The water used in the present invention is deionized water unless otherwise specified.

According to the invention, soluble zinc salt, Hexamethylenetetramine (HMT) and water are mixed for a first hydrothermal reaction. In the present invention, the soluble zinc salt is preferably zinc nitrate, specifically zinc nitrate hexahydrate; the water is preferably deionized water; the temperature of the first hydrothermal reaction is preferably 65-67 ℃, more preferably 65-66 ℃, and the time is preferably 14-16 min, more preferably 15 min; preferably, soluble zinc salt and hexamethylenetetramine are dissolved in water, the solution is stirred for 10min to completely dissolve the soluble zinc salt and the hexamethylenetetramine, and then a first hydrothermal reaction is carried out; the stirring is preferably carried out under sealed conditions. During the first hydrothermal reaction, the soluble zinc salt provides Zn²⁺HMT slowly releases OH during the reaction^-，Zn²⁺And OH^-Reacting under hydrothermal conditions to form Zn (OH)₂Colloid, and then further reacting to generate zinc oxide.

After the first hydrothermal reaction is finished, the reaction liquid, sodium citrate, hydroxypropyl methylcellulose (HPMC), the photothermal conversion material and lignin are mixed for a second hydrothermal reaction to obtain a hydrothermal product. In the invention, the dosage ratio of the soluble zinc salt, the hexamethylenetetramine, the water, the sodium citrate, the hydroxypropyl methyl cellulose, the photothermal conversion material and the lignin is preferably (1.487-1.488) g, (0.350-0.352) g, (95-110) mL, (0.138-0.142) g, (0.100-0.120) g, (0.025-0.035) g, (0.995-0.115) g, more preferably 1.4875 g: 0.351 g: 100mL of: 0.14 g: 0.1 g: 0.025 g: 0.1 g; the photo-thermal conversion material is preferably one or more of activated carbon, gold rods and black phosphorus, and more preferably activated carbon; the sodium citrate plays a role of a surfactant, the hydroxypropyl methyl cellulose plays a role of forming, the sodium citrate and the hydroxypropyl methyl cellulose can control the appearance of the zinc oxide, the photothermal conversion material can endow the zinc oxide with photothermal conversion capability, and the lignin has good biocompatibility and can reduce the energy band gap of the zinc oxide to a certain degree.

In the invention, the temperature of the second hydrothermal reaction is preferably 85-87 ℃, more preferably 85-86 ℃, and the time is preferably 10-12 h, more preferably 10-11 h. In the second hydrothermal reaction process, HMT in the solution continuously and slowly releases OH^-And reacts with zinc ions to generate more zinc oxide, and the photothermal conversion material, lignin and zinc oxide act in the process, so that the obtained zinc oxide has photothermal conversion capability and the energy band gap is reduced.

After the second hydrothermal reaction is finished, preferably filtering the reaction liquid, and then sequentially washing with absolute ethyl alcohol and water to obtain a hydrothermal product; the number of times of washing by the absolute ethyl alcohol and washing by the water is preferably 2, the number of times of washing by the absolute ethyl alcohol and washing by the water is preferably centrifugal washing, the rotating speed of the centrifugal washing is preferably 7000rpm, and the time of a single washing is preferably 15 min.

After a hydrothermal product is obtained, the hydrothermal product is sequentially subjected to freeze drying and microwave irradiation to obtain the dual-photoresponse zinc oxide. According to the invention, the hydrothermal product is preferably pre-frozen at-80 ℃ and then subjected to vacuum freeze drying; the time of the vacuum freeze drying is preferably 12 hours; the power of the microwave irradiation is preferably more than 800W, more preferably 800-850W, and the time is preferably more than 15min, particularly 15-20 min.

The invention provides the double-photoresponse zinc oxide prepared by the method in the scheme, the microscopic morphology of the double-photoresponse zinc oxide is in a silver ear shape and is formed by stacking folded lamellar structures; the particle size of the double-photoresponse zinc oxide is about 2 mu m, and the energy band gap is 2.125eV (so that the double-photoresponse zinc oxide can respond to yellow light); the double-light response type zinc oxide can respond under the irradiation of yellow light and near infrared light; in the specific embodiment of the invention, the response specifically occurs under 597-577 nm (yellow light) and 808nm (near infrared light); the two-photon response type zinc oxide provided by the invention has good adsorption capacity, can adsorb substances such as protein and pigment, and has good antibacterial property and photothermal conversion capacity.

The invention also provides a photosensitive coating with antibacterial osteogenesis property, which is prepared from the double-light response type zinc oxide. The coating provided by the invention has good antibacterial property, has photocatalytic performance under yellow light, is further improved in antibacterial property, has photo-thermal conversion capability under near infrared light, and is beneficial to promoting osteogenesis due to the increase of temperature. The special photosensitive performance of the coating provided by the invention can realize the light-operated effect and the required effect of the antibacterial and osteogenic functions of the implant, for example, in oral cavity implantation, after the implant is implanted, the part (cuff) contacted with the oral cavity can easily cause the growth of bacteria and further spread to the periodontal, when the surface of the implant contains the coating provided by the invention, a yellow toothbrush can be used, yellow light plays a role in photocatalysis, and the antibacterial property of the implant is improved; further, for example, after implanting the implant provided with the photosensitive coating of the present invention, irradiation with near infrared light (i.e., thermal treatment) is given to promote osseointegration.

In the present invention, the coating is preferably prepared by the following steps:

dispersing the double-light response type zinc oxide in a solvent, coating the obtained suspension on the surface of a base material, and drying to obtain the photosensitive coating with antibacterial osteogenesis property.

In the present invention, the base material is preferably a surgical implant, and the material of the surgical implant may be pure titanium, titanium alloy, titanium-zirconium alloy, or the like.

In the present invention, the solvent is preferably water, physiological saline or PBS buffer; the dispersion is preferably carried out under stirring conditions, the rotation speed of the stirring is preferably 75rpm, and the time is preferably 2 hours; the concentration of the double-light response type zinc oxide in the suspension is preferably 200 mug/mL; the invention has no special requirement on the coating amount of the suspension liquid, and can uniformly and comprehensively cover the surface of the base material; the temperature of the drying is preferably room temperature, and the drying time is not particularly limited in the present invention, and it is preferable to sufficiently dry.

In the invention, the raw material for preparing the coating preferably also comprises collagen type powder (Col-I), and the mass ratio of the double-light response type zinc oxide to the collagen type powder is preferably 1 (1-5), and more preferably 1 (2-3). When the raw material for preparing the coating comprises the type collagen powder, the type collagen powder and the double-light response type zinc oxide are jointly dispersed in the solvent, and other conditions are consistent with the scheme, so that the details are not repeated. Col-I is a main organic matrix of natural bone, participates in the regulation of osteoblast development, differentiation activity and bone remodeling, and plays an important role in tissue repair and regeneration.

The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention.