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CN117442743A - Preparation method and drug release application of composite hollow multi-shell microspheres - Google Patents

Preparation method and drug release application of composite hollow multi-shell microspheres Download PDF

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CN117442743A
CN117442743A CN202210847310.1A CN202210847310A CN117442743A CN 117442743 A CN117442743 A CN 117442743A CN 202210847310 A CN202210847310 A CN 202210847310A CN 117442743 A CN117442743 A CN 117442743A
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hollow multi
polyethylene glycol
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赵德偲
王丹
杨乃亮
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Institute of Process Engineering of CAS
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Abstract

The invention relates to an organic-inorganic composite heterogeneous hollow multi-shell material, and provides a preparation method of a composite hollow multi-shell microsphere, which comprises the following steps: 1) Carrying out hydrothermal reaction on a carbon source to obtain a carbon microsphere template; 2) Dispersing the carbon sphere template obtained in the step 1) in a metal salt solution to obtain a solid precursor; 3) Roasting the precursor obtained in the step 2) to obtain hollow multi-shell microspheres; 4) And 3) contacting the hollow multi-shell microsphere obtained in the step 3) with a solution containing a polymer or a monomer to obtain the composite hollow multi-shell microsphere. The drug-loaded microsphere prepared by the invention can effectively prolong the slow release of the drug and has the effect of pH response release, thereby realizing excellent antibacterial performance.

Description

一种复合中空多壳层微球的制备方法及药物释放应用Preparation method and drug release application of composite hollow multi-shell microspheres

技术领域Technical field

本发明涉及有机无机复合的异质中空多壳层材料，更具体而言，本发明涉及复合中空多壳层微球的制备方法及其作为抗菌药物缓释体系的载体的用途。The present invention relates to organic-inorganic composite heterogeneous hollow multi-shell materials. More specifically, the present invention relates to a preparation method of composite hollow multi-shell microspheres and their use as carriers of antibacterial drug sustained-release systems.

背景技术Background technique

人的生活环境里存在着大量细菌、真菌等微生物。从服装到电器，从塑料到建材，微生物无处不在。它们滋生在仪器、设备表面，导致物质的腐蚀、变质、发霉以及伤口化脓等，甚至威胁着人类的生命安全。因此，抗菌剂广泛应用于生产生活中，而抗菌材料的研发成为当今新材料的研究热点之一。传统的有机抗菌剂寿命较短，稳定性较差，长期使用对环境具有一定的危害，并且容易对人类的健康造成损害。而无机抗菌材料主要是金属离子和光催化型抗菌剂，其使用环境受到限制。相比之下，释放型抗菌剂在环境友好性、长效性、灵活性方面均具有较大优势，因此成为近年来的研究热点。There are a large number of bacteria, fungi and other microorganisms in the human living environment. From clothing to appliances, from plastics to building materials, microorganisms are everywhere. They breed on the surface of instruments and equipment, causing material corrosion, deterioration, mold, wound suppuration, etc., and even threatening human life. Therefore, antibacterial agents are widely used in production and life, and the research and development of antibacterial materials has become one of the research hotspots of new materials today. Traditional organic antibacterial agents have a short lifespan and poor stability. Long-term use is harmful to the environment and can easily cause damage to human health. Inorganic antibacterial materials are mainly metal ions and photocatalytic antibacterial agents, and their use environment is restricted. In contrast, releasing antibacterial agents have great advantages in environmental friendliness, long-lasting effect, and flexibility, and therefore have become a research hotspot in recent years.

近年来，中空多壳层结构在分子递送领域引起了广泛关注。多壳层结构为分子的负载提供了不同的表面和内部空间。例如，壳层的外表面和内表面都允许分子的吸附和解吸，而由于物理屏障和局部浓度梯度，每个壳层的吸附和解吸速率不同。同样，每个壳层内的孔隙和相邻壳层之间的空隙也为分子的储存提供了空间，而药物分子的扩散速率受到物理屏障和浓度梯度的影响。因此，中空多壳层结构作为物质储库，可以通过一系列次序的阶段实现持续的释放。研究已经证明，中空多壳层作为一类药物载体，具有时空顺序性。然而，精确地控制药物分子对释放速率，对于多壳层载体而言仍然存在着巨大的挑战。In recent years, hollow multi-shell structures have attracted widespread attention in the field of molecular delivery. The multi-shell structure provides different surfaces and internal spaces for the loading of molecules. For example, both the outer and inner surfaces of the shell allow adsorption and desorption of molecules, and each shell has different rates of adsorption and desorption due to physical barriers and local concentration gradients. Likewise, the pores within each shell and the gaps between adjacent shells also provide space for the storage of molecules, while the diffusion rate of drug molecules is affected by physical barriers and concentration gradients. Therefore, the hollow multi-shell structure serves as a material reservoir that can achieve sustained release through a series of sequential stages. Research has proven that hollow multi-shells, as a type of drug carrier, have spatiotemporal order. However, precisely controlling the release rate of drug molecules remains a huge challenge for multi-shell carriers.

聚合物材料已被开发为各种小分子药物的载体。它们的聚合物外壳能够增加所负载的药物的稳定性，并且可以通过调控不同的交联度、表面电性等来调控载药性能，如降解性、pH响应性、环境相容性等。利用pH、温度等变化，可以开发对环境敏感的载体。通过亲水/疏水相互作用、共价键、范德华相互作用和静电相互作用，使用各种功能性高分子或门控分子将其接枝到中空多壳层材料的表面或孔道上，从而优化材料的物理化学特征。将有机聚合物与无机微纳材料相结合作为载体，是一种新的研究趋势。Polymeric materials have been developed as carriers for various small molecule drugs. Their polymer shells can increase the stability of the loaded drugs, and can regulate drug-loading properties, such as degradability, pH responsiveness, environmental compatibility, etc., by adjusting different cross-linking degrees, surface electrical properties, etc. Using changes in pH, temperature, etc., carriers that are sensitive to the environment can be developed. Various functional polymers or gated molecules are grafted onto the surface or pores of hollow multi-shell materials through hydrophilic/hydrophobic interactions, covalent bonds, van der Waals interactions and electrostatic interactions to optimize the material. physical and chemical characteristics. Combining organic polymers with inorganic micro-nano materials as carriers is a new research trend.

发明内容Contents of the invention

本发明的目的在于克服上述问题，提供一种复合的异质中空多壳层结构抗菌剂载体。其明显的优势在于：(1)长效抗菌能力强；(2)具有环境响应性；(3)灵活控制释放速率。本发明所制得的载药微球能有效延长药物的缓释，并具有pH响应释放的效果，从而实现优异的抗菌性能。The purpose of the present invention is to overcome the above problems and provide a composite heterogeneous hollow multi-shell structure antibacterial agent carrier. Its obvious advantages are: (1) strong long-lasting antibacterial ability; (2) environmental responsiveness; (3) flexible control of release rate. The drug-loaded microspheres prepared by the present invention can effectively prolong the sustained release of drugs and have a pH-responsive release effect, thereby achieving excellent antibacterial properties.

为达到上述目的，本发明提供了制备复合中空多壳层金属氧化物微球的方法，首先制备出中空多壳层微球；在其表面包覆聚合物，随后加入药物分子，以得到缓释体系。所述方法具体包括如下步骤：In order to achieve the above purpose, the present invention provides a method for preparing composite hollow multi-shell metal oxide microspheres. First, hollow multi-shell microspheres are prepared; polymers are coated on their surfaces, and then drug molecules are added to obtain sustained release. system. The method specifically includes the following steps:

1)对碳源进行水热反应，获得碳微球模板；1) Perform a hydrothermal reaction on the carbon source to obtain the carbon microsphere template;

2)将步骤1)得到的碳球模板分散于金属盐溶液中，得到固体前驱体；2) Disperse the carbon sphere template obtained in step 1) into the metal salt solution to obtain a solid precursor;

3)将步骤2)得到的前驱体进行焙烧处理，得到中空多壳层微球；3) Calculate the precursor obtained in step 2) to obtain hollow multi-shell microspheres;

4)将步骤3)得到的中空多壳层微球与含有聚合物或单体的溶液接触，以便将聚合物包覆至所述中空多壳层微球表面，以得到复合中空多壳层微球，即，中空多壳层二氧化钛@聚合物复合结构；4) Contact the hollow multi-shell microspheres obtained in step 3) with a solution containing polymer or monomer, so as to coat the polymer on the surface of the hollow multi-shell microspheres to obtain composite hollow multi-shell microspheres. Sphere, i.e., hollow multi-shell titanium dioxide@polymer composite structure;

5)将药物水溶液加入到步骤4)所制备复合中空多壳层微球的材料中，搅拌以完成对药物的负载。5) Add the drug aqueous solution to the material of the composite hollow multi-shell microsphere prepared in step 4), and stir to complete the loading of the drug.

在一些实施方案中，步骤2)中所述金属盐溶液包括1-3mol/L四氯化钛的水溶液或0.1-0.5mol/L四氯化钛的丙酮溶液。In some embodiments, the metal salt solution in step 2) includes an aqueous solution of 1-3 mol/L titanium tetrachloride or an acetone solution of 0.1-0.5 mol/L titanium tetrachloride.

在一些实施方案中，步骤3)中所述中空多壳层微球为通过次序模板法制得的单壳层、双壳层和三壳层的二氧化钛微球。In some embodiments, the hollow multi-shell microspheres described in step 3) are single-shell, double-shell and triple-shell titanium dioxide microspheres prepared by sequential template method.

在一些实施方案中，步骤4)中所述的聚合物包括聚乳酸-羟基乙酸共聚物，聚乙二醇，单体包括盐酸多巴胺。进一步优选为，所述的聚合物为聚乙二醇，聚乙二醇作为一种生物安全的聚合物，广泛地应用于无机药物载体的修饰，以提高材料的生物相容性。结合聚乙二醇的pH敏感性和中空多壳层结构的多级相互作用，精确响应环境变化，控制载体的释放速率。In some embodiments, the polymer described in step 4) includes polylactic acid-co-glycolic acid, polyethylene glycol, and the monomer includes dopamine hydrochloride. It is further preferred that the polymer is polyethylene glycol. As a biosafe polymer, polyethylene glycol is widely used in the modification of inorganic drug carriers to improve the biocompatibility of materials. Combining the pH sensitivity of polyethylene glycol and the multi-level interaction of the hollow multi-shell structure, it accurately responds to environmental changes and controls the release rate of the carrier.

在一些实施方案中，步骤5)中所述药物水溶液的药物包括抗菌剂卡松，抗生素布洛芬、多西环素、降糖药物二甲双胍和抗肿瘤药物阿霉素中的一种或两种以上，其浓度为1mg/mL-100mg/mL。In some embodiments, the drug in the drug aqueous solution in step 5) includes one or two of the antibacterial agent kason, the antibiotic ibuprofen, doxycycline, the hypoglycemic drug metformin and the anti-tumor drug doxorubicin Above, its concentration is 1mg/mL-100mg/mL.

在一些实施方案中，步骤4)中所述使用的聚乙二醇，包括聚乙二醇4000，聚乙二醇6000，聚乙二醇200000和聚乙二醇100000中的一种或两种以上。In some embodiments, the polyethylene glycol used in step 4) includes one or two of polyethylene glycol 4000, polyethylene glycol 6000, polyethylene glycol 200000 and polyethylene glycol 100000. above.

在一些实施方案中，步骤4)中所述使中空多壳层微球与含有聚乙二醇的水溶液接触，包括在室温下搅拌所述混合液2-48小时。所述中空多壳层微球与含有聚乳酸-羟基乙酸的二氯甲烷溶液接触，超声乳化，逐滴加入到聚乙烯醇水溶液中，在室温下搅拌2-10小时；所述中空多壳层微球与含有盐酸多巴胺的水溶液接触，在室温下搅拌5-24小时。In some embodiments, contacting the hollow multi-shell microspheres with an aqueous solution containing polyethylene glycol as described in step 4) includes stirring the mixture at room temperature for 2-48 hours. The hollow multi-shell microspheres are contacted with a methylene chloride solution containing polylactic acid-glycolic acid, ultrasonic emulsified, added dropwise to the polyvinyl alcohol aqueous solution, and stirred at room temperature for 2-10 hours; the hollow multi-shell microspheres are The microspheres are contacted with an aqueous solution containing dopamine hydrochloride and stirred at room temperature for 5-24 hours.

在一些实施方案中，步骤4)中所述的聚乙二醇水溶液浓度包括为10-500mg/mL，优选为10mg/mL，100mg/mL和500mg/mL。所述的聚乳酸-羟基乙酸共聚物的二氯甲烷溶液的浓度为1-50mg/mL，优选为20mg/mL。聚乙烯醇水溶液浓度为0.1％-2％所述的盐酸多巴胺水溶液的浓度为0.1-5mg/mL，pH为8.0-8.5。In some embodiments, the concentration of the polyethylene glycol aqueous solution described in step 4) includes 10-500 mg/mL, preferably 10 mg/mL, 100 mg/mL and 500 mg/mL. The concentration of the dichloromethane solution of the polylactic acid-glycolic acid copolymer is 1-50 mg/mL, preferably 20 mg/mL. The concentration of the polyvinyl alcohol aqueous solution is 0.1%-2%, the concentration of the dopamine hydrochloride aqueous solution is 0.1-5mg/mL, and the pH is 8.0-8.5.

在一些实施方案中，聚合物复合异质中空多壳层药物微球缓释体系，在酸性条件的刺激下，能够加快释放所负载药物，且酸性强弱不同，释放速率不同。In some embodiments, the polymer composite heterogeneous hollow multi-shell drug microsphere sustained-release system can accelerate the release of loaded drugs under the stimulation of acidic conditions, and the release rates are different depending on the acidity.

在一些实施方案中，聚合物复合异质中空多壳层微球药物载体，其酸性条件为pH＜5；其药物是卡松或多西环素。In some embodiments, the acidic condition of the polymer composite heterogeneous hollow multi-shell microsphere drug carrier is pH<5; the drug is cassone or doxycycline.

在一些实施方案中，中空多壳层药物缓释体系，其中所述的中空多壳层微球为三壳层的。In some embodiments, the hollow multi-shell drug sustained release system, wherein the hollow multi-shell microspheres are three-shelled.

与现有技术相比，本发明的优势在于：Compared with the existing technology, the advantages of the present invention are:

(1)本发明采用聚合物包覆的中空多壳层氧化物微球作为抗菌剂载体，与普通中空多壳层相比，除具有中空结构容纳大量药物，多壳层为药物释放提供物理阻隔等优点，更重要的是，功能化的壳层间的多级次相互作用使材料具有开关与调速功能，能够根据环境pH调控药物释放速率。(1) The present invention uses polymer-coated hollow multi-shell oxide microspheres as antibacterial agent carriers. Compared with ordinary hollow multi-shell layers, in addition to having a hollow structure to accommodate a large amount of drugs, the multi-shell layers provide physical barriers for drug release. More importantly, the multi-level interaction between the functionalized shell layers enables the material to have switching and speed-regulating functions, and can regulate the drug release rate according to the environmental pH.

(2)本发明所采用的技术实现了温和条件下的聚合物修饰与药物装载，并便于调节高分子包覆量与药物装载量。修饰与装载同时进行，能够增加药物吸附深度，提高中空多壳层结构的空间利用率。(2) The technology used in the present invention realizes polymer modification and drug loading under mild conditions, and facilitates adjustment of the polymer coating amount and drug loading amount. Modification and loading are carried out simultaneously, which can increase the drug adsorption depth and improve the space utilization of the hollow multi-shell structure.

(3)本发明所用的载体具有pH开关的特点，该特点使得在无菌的中性环境下，开关关闭，大部分药物储存在载体中；在细菌或霉菌大量生存的低pH值的环境中，开关打开，且载体能够依据pH的不同调控释放速率。(3) The carrier used in the present invention has the characteristics of a pH switch, which allows the switch to be closed in a sterile neutral environment and most of the drugs are stored in the carrier; in a low pH environment where bacteria or molds survive in large numbers , the switch is turned on, and the carrier can regulate the release rate according to different pH values.

为便于理解本发明，本发明列举实施例如下。本领域技术人员应该明了，所述实施例仅仅是帮助理解本发明，不应视为对本发明的具体限制。In order to facilitate understanding of the present invention, the following examples are enumerated. Those skilled in the art should understand that the embodiments are only to help understand the present invention and should not be regarded as specific limitations of the present invention.

附图说明Description of the drawings

图1(a)为本发明制备的单层TiO2空心球@聚乙二醇的透射电镜照片；Figure 1(a) is a transmission electron microscope photo of the single-layer TiO 2 hollow sphere@polyethylene glycol prepared by the present invention;

图1(b)为本发明制备的双壳层TiO2空心球@聚乙二醇的透射电镜照片；Figure 1(b) is a transmission electron microscope photo of the double-shell TiO 2 hollow sphere@polyethylene glycol prepared by the present invention;

图1(c)为本发明制备的三壳层TiO2空心球@聚乙二醇的透射电镜照片；Figure 1(c) is a transmission electron microscope photo of the three-shell TiO 2 hollow sphere@polyethylene glycol prepared by the present invention;

图2为本发明制备的三壳层TiO2空心球@聚乙二醇的激光共聚焦显微镜照片；Figure 2 is a laser confocal microscope photo of the three-shell TiO 2 hollow sphere@polyethylene glycol prepared in the present invention;

图3为以制备的单、双、三壳层TiO2空心球@聚乙二醇为载体装载有机抗菌剂卡松后的缓释性能曲线，图3(a)为实际-释放量曲线，3(b)为时间-释放率曲线；Figure 3 shows the sustained-release performance curve after loading the organic antibacterial agent Casson using the prepared single-, double- and triple-shell TiO 2 hollow spheres@polyethylene glycol as carriers. Figure 3(a) is the actual-release curve. 3 (b) is the time-release rate curve;

图4为以制备的三壳层TiO2空心球@聚乙二醇为载体装载有机抗菌剂卡松后的pH响应释放性能曲线；Figure 4 shows the pH response release performance curve after loading the organic antibacterial agent kason using the prepared three-shell TiO 2 hollow spheres@polyethylene glycol as a carrier;

图5为以制备的三壳层TiO2空心球@聚乙二醇为载体装载抗生素多西环素后的pH响应释放性能曲线；Figure 5 shows the pH response release performance curve after loading the antibiotic doxycycline using the prepared three-shell TiO 2 hollow spheres@polyethylene glycol as a carrier;

图6显示了一些细菌荧光照片，为抗菌剂卡松、实施例1-3制备的装载卡松的单壳层、双壳层以及三壳层TiO2@聚乙二醇的抗菌性能测试结果；图中第一行显示绿的荧光的为被异硫氰酸荧光素染色的全部细菌(死和活)；第二行显示红色荧光的为被碘化吡啶染色的，为死菌；第三行为前两列的叠加，黄色为死菌，绿色为活菌；Figure 6 shows some bacterial fluorescence photos, which are the antibacterial performance test results of the antibacterial agent kason, and the kason-loaded single-shell, double-shell and triple-shell TiO 2 @polyethylene glycol prepared in Examples 1-3; The first row of the figure shows all the bacteria (dead and alive) stained by fluorescein isothiocyanate; the second row shows red fluorescence, which is stained by pyridinium iodide, and is dead bacteria; the third row shows In the superposition of the first two columns, yellow represents dead bacteria and green represents live bacteria;

图7为实施例4制备的三壳层TiO2@聚多巴胺的透射电镜照片；Figure 7 is a transmission electron microscope photo of the three-shell TiO 2 @polydopamine prepared in Example 4;

图8为实施例5制备的三壳层TiO2@聚乳酸-羟基乙酸共聚物的透射电镜照片；Figure 8 is a transmission electron microscope photo of the three-shell TiO 2 @polylactic acid-glycolic acid copolymer prepared in Example 5;

图9为以制备的三壳层TiO2空心球@聚乙二醇为载体装载二甲双胍后的缓释性能曲线。Figure 9 shows the sustained release performance curve of metformin loaded with the prepared three-shell TiO2 hollow spheres@polyethylene glycol as a carrier.

具体实施方式Detailed ways

本发明中，术语“中空多壳层微球”或“中空微球”均指具有空腔与多层结构、主要成分为金属氧化物的微球。换言之，金属氧化物形成该微球的壳体，而壳体内部大部分为非致密结构，能够容纳其他分子。“中空微球”的尺寸可以在200-1000nm，优选范围为500-800nm。In the present invention, the terms "hollow multi-shell microspheres" or "hollow microspheres" refer to microspheres with cavities and multi-layer structures and whose main components are metal oxides. In other words, the metal oxide forms the shell of the microsphere, and the interior of the shell is mostly non-dense and can accommodate other molecules. The size of "hollow microspheres" can be in the range of 200-1000nm, and the preferred range is 500-800nm.

下面以具体实施示例对本发明的技术方案做进一步说明，但是不能以此限制本发明的范围。The technical solution of the present invention is further described below with specific implementation examples, but the scope of the present invention cannot be limited in this way.

实施例1Example 1

单壳层TiO2中空微球-卡松的制备及其缓释与抗菌性能研究，包括如下步骤：The preparation of single-shell TiO 2 hollow microsphere-casone and the study of its sustained release and antibacterial properties include the following steps:

(1)仪器与试剂(1)Instruments and reagents

本发明实施例所用的烘箱是上海一恒DHG-9240A恒温干燥箱，所用的天平是天马衡基仪器的FA1204电子天平，所用搅拌器为IKA RCT Basic加热磁力搅拌器，所用马弗炉为中环实验公司箱式电阻炉SX2-5-12，所用紫外可见分光光度计为岛津仪器UV-1780。The oven used in the embodiment of the present invention is Shanghai Yiheng DHG-9240A constant temperature drying oven, the balance used is the FA1204 electronic balance of Tianma Hengji Instruments, the stirrer used is IKA RCT Basic heating magnetic stirrer, and the muffle furnace used is from Zhonghuan Experimental Company The box-type resistance furnace SX2-5-12 was used, and the UV-visible spectrophotometer used was Shimadzu Instruments UV-1780.

所用的试剂为阿拉丁试剂公司的四氯化钛分析纯；所用卡松抗菌剂为郑州三三日化公司14％的卡松。The reagent used was analytically pure titanium tetrachloride from Aladdin Reagent Company; the kason antibacterial agent used was 14% kason from Zhengzhou Sansan Daily Chemical Company.

(2)单壳层TiO2微球的制备(2) Preparation of single-shell TiO 2 microspheres

单壳层TiO2微球的制备：将0.6g碳球分散到30mL 3M的四氯化钛溶液中，室温搅拌3小时后抽滤，用去离子水清洗3遍，置于70℃烘箱中干燥12小时，将所得的固体粉末置于马弗炉中，以2℃/min的速率升温到500℃，保温3h，自然冷却后得到单壳层TiO2空心球。Preparation of single-shell TiO 2 microspheres: Disperse 0.6g of carbon spheres into 30 mL of 3M titanium tetrachloride solution, stir at room temperature for 3 hours, then filter with suction, wash with deionized water 3 times, and dry in a 70°C oven For 12 hours, the obtained solid powder was placed in a muffle furnace, heated to 500°C at a rate of 2°C/min, kept for 3 hours, and single-shell TiO 2 hollow spheres were obtained after natural cooling.

(3)单壳层TiO2微球@PEG-卡松的制备(3) Preparation of single-shell TiO 2 microspheres@PEG-casone

将上述制备的6mg单壳层TiO2空心球加入到1mL去离子水中，分散均匀后，向其中加入1mL聚乙二醇水溶液，在室温下搅拌2小时之后，加入2mL 14wt％的卡松水溶液，室温下避光搅拌24小时。离心分离去上清，将得到的固体于35℃真空干燥12小时，备用。TEM照片显示于图1a。Add 6 mg of single-shell TiO2 hollow spheres prepared above to 1 mL of deionized water. After uniform dispersion, add 1 mL of polyethylene glycol aqueous solution to it. After stirring at room temperature for 2 hours, add 2 mL of 14 wt% Casson aqueous solution. Stir for 24 hours at room temperature in the dark. The supernatant was removed by centrifugation, and the obtained solid was vacuum dried at 35°C for 12 hours and set aside. The TEM photo is shown in Figure 1a.

(4)缓释性能测试(4) Sustained release performance test

将5mg载有抗菌剂的聚乙二醇包覆的单壳层微球(即上述经真空干燥处理的固体物)室温分散于1mL去离子水中，接着将混合液转移至透析袋中(截留分子量：14000Da)。将透析袋置于100mL水中，每隔一段时间取上清液，测量在273nm下紫外可见光谱吸收度。根据紫外可见光谱标准曲线求得实际溶液中药物浓度及含量，与热重实验得到的药物装载量对比，得到释放率，进而得出释放率-时间对应曲线(图3中最上方的曲线)。5 mg of polyethylene glycol-coated single-shell microspheres loaded with antibacterial agents (i.e., the above-mentioned vacuum-dried solids) were dispersed in 1 mL of deionized water at room temperature, and then the mixture was transferred to a dialysis bag (molecular weight cutoff :14000Da). Place the dialysis bag in 100 mL of water, take the supernatant at regular intervals, and measure the UV-visible spectrum absorbance at 273 nm. The concentration and content of the drug in the actual solution were obtained based on the UV-visible spectrum standard curve, and compared with the drug loading obtained by the thermogravimetric experiment, the release rate was obtained, and then the release rate-time corresponding curve was obtained (the top curve in Figure 3).

(5)抗菌性能测试(5) Antibacterial performance test

载有0.2mg卡松的微球与5mL LB琼脂培养基混合，每隔24小时向体系中加入200μL大肠杆菌溶液，使得细菌浓度为106CFU/mL。每隔一段时间取出200μL混合液体，使用异硫氰酸荧光素和碘化丙啶染色，并在第17天时于荧光电子显微镜下观察细菌生长情况。通过对比视野中死亡细菌的含量评估抗菌性能。荧光显微镜图显示在图5中第2列，其死亡细菌的比例为56％。Microspheres loaded with 0.2 mg of Casson were mixed with 5 mL of LB agar medium, and 200 μL of E. coli solution was added to the system every 24 hours to achieve a bacterial concentration of 10 6 CFU/mL. At regular intervals, 200 μL of the mixed liquid was taken out, stained with fluorescein isothiocyanate and propidium iodide, and bacterial growth was observed under a fluorescence electron microscope on the 17th day. Antimicrobial performance was evaluated by comparing the content of dead bacteria in the field of view. The fluorescence microscopy picture is shown in column 2 of Figure 5, and the proportion of dead bacteria is 56%.

实施例2Example 2

双壳层TiO2中空微球-卡松的制备及其缓释与抗菌性能研究，包括如下步骤：The preparation of double-shell TiO 2 hollow microsphere-casone and the study of its sustained release and antibacterial properties include the following steps:

(1)仪器与试剂(1)Instruments and reagents

本发明实施例所用的烘箱是上海一恒DHG-9240A恒温干燥箱，所用的天平是天马衡基仪器的FA1204电子天平，所用搅拌器为IKA RCT Basic加热磁力搅拌器，所用马弗炉为中环实验公司箱式电阻炉SX2-5-12，所用紫外可见分光光度计为岛津仪器UV-1780。The oven used in the embodiment of the present invention is Shanghai Yiheng DHG-9240A constant temperature drying oven, the balance used is the FA1204 electronic balance of Tianma Hengji Instruments, the stirrer used is IKA RCT Basic heating magnetic stirrer, and the muffle furnace used is from Zhonghuan Experimental Company The box-type resistance furnace SX2-5-12 was used, and the UV-visible spectrophotometer used was Shimadzu Instruments UV-1780.

所用的试剂为阿拉丁试剂公司的四氯化钛分析纯；所用卡松抗菌剂为郑州三三日化公司14％的卡松。The reagent used was analytically pure titanium tetrachloride from Aladdin Reagent Company; the kason antibacterial agent used was 14% kason from Zhengzhou Sansan Daily Chemical Company.

(2)双壳层TiO2中空微球的制备(2) Preparation of double-shell TiO 2 hollow microspheres

双壳层TiO2中空微球的制备：将0.6g碳球分散到30mL 3M的四氯化钛溶液中，40℃搅拌5小时后抽滤，用去离子水清洗3遍，置于70℃烘箱中干燥12小时，将所得的固体粉末置于马弗炉中，以2℃/min的速率升温到500℃，保温3h，自然冷却后得到单壳层TiO2空心球。Preparation of double-shell TiO 2 hollow microspheres: Disperse 0.6g carbon spheres into 30 mL of 3M titanium tetrachloride solution, stir at 40°C for 5 hours, then filter, wash 3 times with deionized water, and place in a 70°C oven Medium dry for 12 hours, place the obtained solid powder in a muffle furnace, raise the temperature to 500°C at a rate of 2°C/min, keep it for 3 hours, and obtain single-shell TiO2 hollow spheres after natural cooling.

(3)双壳层TiO2中空微球@PEG-卡松的制备(3) Preparation of double-shell TiO 2 hollow microspheres @PEG-casson

将上述制备的6mg双壳层TiO2空心球加入到1mL去离子水中，分散均匀后，向其中加入1mL聚乙二醇水溶液，在室温下搅拌1小时之后，加入2mL 14wt％的卡松水溶液，室温下避光搅拌24小时。离心分离去上清，将得到的固体于35℃真空干燥12小时，备用。TEM照片显示于图1b。Add 6 mg of the double-shell TiO2 hollow spheres prepared above to 1 mL of deionized water. After uniform dispersion, add 1 mL of polyethylene glycol aqueous solution to it. After stirring at room temperature for 1 hour, add 2 mL of 14 wt% Casson aqueous solution. Stir for 24 hours at room temperature in the dark. The supernatant was removed by centrifugation, and the obtained solid was vacuum dried at 35°C for 12 hours and set aside. The TEM picture is shown in Figure 1b.

(4)缓释性能测试(4) Sustained release performance test

将5mg载有抗菌剂的聚乙二醇包覆的双壳层微球(即上述经真空干燥处理的固体物)室温分散于1mL去离子水中，接着将混合液转移至透析袋中(截留分子量：14000Da)。将透析袋置于100mL水中，每隔一段时间取上清液，测量在273nm下紫外可见光谱吸收度。根据紫外可见光谱标准曲线求得实际溶液中药物浓度及含量，与热重实验得到的药物装载量对比，得到释放率，进而得出释放率-时间对应曲线(图3中第二条曲线)。5 mg of polyethylene glycol-coated double-shell microspheres loaded with antibacterial agents (i.e., the above-mentioned vacuum-dried solids) were dispersed in 1 mL of deionized water at room temperature, and then the mixture was transferred to a dialysis bag (molecular weight cutoff :14000Da). Place the dialysis bag in 100 mL of water, take the supernatant at regular intervals, and measure the UV-visible spectrum absorbance at 273 nm. The concentration and content of the drug in the actual solution were obtained based on the UV-visible spectrum standard curve, and compared with the drug loading obtained by the thermogravimetric experiment, the release rate was obtained, and then the release rate-time corresponding curve was obtained (the second curve in Figure 3).

(5)抗菌性能测试(5) Antibacterial performance test

载有0.2mg卡松的微球与5mL LB琼脂培养基混合，每隔24小时向体系中加入200μL大肠杆菌溶液，使得细菌浓度为106CFU/mL。每隔一段时间取出200μL混合液体，使用异硫氰酸荧光素和碘化丙啶染色，并在第17天时于荧光电子显微镜下观察细菌生长情况。通过对比视野中死亡细菌的含量评估抗菌性能。荧光显微镜图显示在图5中第3列，其死亡细菌的比例为84％。Microspheres loaded with 0.2 mg of Casson were mixed with 5 mL of LB agar medium, and 200 μL of E. coli solution was added to the system every 24 hours to achieve a bacterial concentration of 10 6 CFU/mL. At regular intervals, 200 μL of the mixed liquid was taken out, stained with fluorescein isothiocyanate and propidium iodide, and bacterial growth was observed under a fluorescence electron microscope on the 17th day. Antimicrobial performance was evaluated by comparing the content of dead bacteria in the field of view. The fluorescence microscopy picture is shown in column 3 of Figure 5, and the proportion of dead bacteria is 84%.

实施例3Example 3

三壳层TiO2中空微球-卡松的制备及其缓释与抗菌性能研究，包括如下步骤：The preparation of three-shell TiO 2 hollow microspheres-casone and the study of its sustained release and antibacterial properties include the following steps:

(1)仪器与试剂(1)Instruments and reagents

本发明实施例所用的烘箱是上海一恒DHG-9240A恒温干燥箱，所用的天平是天马衡基仪器的FA1204电子天平，所用搅拌器为IKA RCT Basic加热磁力搅拌器，所用马弗炉为中环实验公司箱式电阻炉SX2-5-12，所用紫外可见分光光度计为岛津仪器UV-1780。The oven used in the embodiment of the present invention is Shanghai Yiheng DHG-9240A constant temperature drying oven, the balance used is the FA1204 electronic balance of Tianma Hengji Instruments, the stirrer used is IKA RCT Basic heating magnetic stirrer, and the muffle furnace used is from Zhonghuan Experimental Company The box-type resistance furnace SX2-5-12 was used, and the UV-visible spectrophotometer used was Shimadzu Instruments UV-1780.

所用的试剂为阿拉丁试剂公司的四氯化钛分析纯；所用卡松抗菌剂为郑州三三日化公司14％的卡松。The reagent used was analytically pure titanium tetrachloride from Aladdin Reagent Company; the kason antibacterial agent used was 14% kason from Zhengzhou Sansan Daily Chemical Company.

(2)三壳层TiO2中空微球的制备(2) Preparation of three-shell TiO 2 hollow microspheres

3壳层TiO2微球的制备：将0.6g碳球分散到30mL 3M的四氯化钛溶液中，40℃搅拌5小时后抽滤，用去离子水清洗3遍，置于70℃烘箱中干燥12小时，将所得的固体粉末置于马弗炉中，以2℃/min的速率升温到500℃，保温3h，自然冷却后得到单壳层TiO2空心球。Preparation of 3-shell TiO 2 microspheres: Disperse 0.6g carbon spheres into 30 mL 3M titanium tetrachloride solution, stir at 40°C for 5 hours, then filter with suction, wash 3 times with deionized water, and place in a 70°C oven Dry for 12 hours, place the obtained solid powder in a muffle furnace, raise the temperature to 500°C at a rate of 2°C/min, keep it for 3 hours, and obtain single-shell TiO2 hollow spheres after natural cooling.

(3)三壳层TiO2中空微球@PEG-卡松的制备(3) Preparation of three-shell TiO 2 hollow microspheres @PEG-casone

将上述制备的6mg单壳层TiO2空心球加入到1mL去离子水中，分散均匀后，向其中加入1mL聚乙二醇水溶液，在室温下搅拌1小时之后，加入2mL 14wt％的卡松水溶液，室温下避光搅拌24小时。离心分离去上清，将得到的固体于35℃真空干燥12小时，备用。TEM照片显示于图1c，激光共聚焦显微镜照片显示于图2。Add 6 mg of single-shell TiO2 hollow spheres prepared above to 1 mL of deionized water. After uniform dispersion, add 1 mL of polyethylene glycol aqueous solution to it. After stirring at room temperature for 1 hour, add 2 mL of 14 wt% Casson aqueous solution. Stir for 24 hours at room temperature in the dark. The supernatant was removed by centrifugation, and the obtained solid was vacuum dried at 35°C for 12 hours and set aside. The TEM photo is shown in Figure 1c and the laser confocal microscopy photo is shown in Figure 2.

(4)缓释性能测试(4) Sustained release performance test

将5mg载有抗菌剂的聚乙二醇包覆的双壳层微球(即上述经真空干燥处理的固体物)室温分散于1mL去离子水中，接着将混合液转移至透析袋中(截留分子量：14000Da)。将透析袋置于100mL水中，每隔一段时间取上清液，测量在273nm下紫外可见光谱吸收度。根据紫外可见光谱标准曲线求得实际溶液中药物浓度及含量，与热重实验得到的药物装载量对比，得到释放率，进而得出释放率-时间对应曲线(图3中最下方的曲线)。5 mg of polyethylene glycol-coated double-shell microspheres loaded with antibacterial agents (i.e., the above-mentioned vacuum-dried solids) were dispersed in 1 mL of deionized water at room temperature, and then the mixture was transferred to a dialysis bag (molecular weight cutoff :14000Da). Place the dialysis bag in 100 mL of water, take the supernatant at regular intervals, and measure the UV-visible spectrum absorbance at 273 nm. The concentration and content of the drug in the actual solution were obtained based on the UV-visible spectrum standard curve, and compared with the drug loading obtained by the thermogravimetric experiment, the release rate was obtained, and then the release rate-time corresponding curve was obtained (the bottom curve in Figure 3).

(5)pH响应释放性能测试(5) pH response release performance test

将1mg载有抗菌剂的聚乙二醇包覆的双壳层微球室温分散于1mL去离子水中，每隔一段时间离心分离，取出上清液，测量在273nm下紫外可见光谱吸收度。当释放达到平衡后，用HCl将溶液pH调至4，每隔一段时间离心分离，取出上清液，测量在273nm下紫外可见光谱吸收度，观察释放趋势。释放曲线显示在图4，结果表明在pH＝4的环境下，药物释放量增加，因此该载体具有pH响应释放的性能。Disperse 1 mg of polyethylene glycol-coated double-shell microspheres loaded with antibacterial agents in 1 mL of deionized water at room temperature, centrifuge at regular intervals, take out the supernatant, and measure the UV-visible spectrum absorbance at 273 nm. When the release reaches equilibrium, use HCl to adjust the pH of the solution to 4, centrifuge at regular intervals, take out the supernatant, measure the UV-visible spectrum absorbance at 273nm, and observe the release trend. The release curve is shown in Figure 4. The results show that in an environment of pH=4, the amount of drug release increases, so the carrier has pH-responsive release properties.

抗菌性能测试：载有0.2mg卡松的微球与5mL LB琼脂培养基混合，每隔24小时向体系中加入200μL大肠杆菌溶液，使得细菌浓度为106CFU/mL。每隔一段时间取出200μL混合液体，使用异硫氰酸荧光素和碘化丙啶染色，并在第17天时于荧光电子显微镜下观察细菌生长情况。通过对比视野中死亡细菌的含量评估抗菌性能。荧光显微镜图显示在图5中第4列，其死亡细菌的比例为100％。Antibacterial performance test: Microspheres loaded with 0.2 mg Casson were mixed with 5 mL LB agar medium, and 200 μL E. coli solution was added to the system every 24 hours so that the bacterial concentration was 10 6 CFU/mL. At regular intervals, 200 μL of the mixed liquid was taken out, stained with fluorescein isothiocyanate and propidium iodide, and bacterial growth was observed under a fluorescence electron microscope on the 17th day. Antimicrobial performance was evaluated by comparing the content of dead bacteria in the field of view. The fluorescence microscopy picture is shown in column 4 of Figure 5, and the proportion of dead bacteria is 100%.

(6)防霉性能测试(6) Anti-mildew performance test

地毯抗菌防霉性能按照国标GB/T 24346-2009执行。将聚酯纤维地毯裁剪为4cm×4cm的正方形试片，然后采用高压蒸汽法进行灭菌处理，灭菌温度为121℃，时间为20min。用负载了0.2mg卡松的三壳层TiO2中空微球@PEG对地毯进行处理，并将处理后的地毯片置于琼脂-葡萄糖培养基上。用滴管在地毯表明滴1mL配置好的黑曲霉菌孢子悬浮液，置于恒温恒湿箱中观察。28天后，地毯表面无霉菌生长，达到了0级抗菌标准。The antibacterial and mildew-proof performance of carpets is implemented in accordance with the national standard GB/T 24346-2009. The polyester fiber carpet was cut into square test pieces of 4cm × 4cm, and then sterilized using high-pressure steam method. The sterilization temperature was 121°C and the time was 20 minutes. The carpet was treated with three-shell TiO 2 hollow microspheres @PEG loaded with 0.2 mg of Casson, and the treated carpet pieces were placed on agar-glucose medium. Use a dropper to drop 1 mL of the prepared Aspergillus niger spore suspension on the surface of the carpet, and place it in a constant temperature and humidity box for observation. After 28 days, there was no mold growth on the carpet surface, reaching level 0 antibacterial standards.

实施例4Example 4

三壳层TiO2中空微球@聚多巴胺的制备，包括如下步骤：The preparation of three-shell TiO2 hollow microspheres@polydopamine includes the following steps:

3壳层TiO2微球的制备：将0.6g碳球分散到30mL 3M的四氯化钛溶液中，40℃搅拌8小时后抽滤，用去离子水清洗3遍，置于70℃烘箱中干燥12小时，将所得的固体粉末置于马弗炉中，以2℃/min的速率升温到500℃，保温3h，自然冷却后得到三壳层TiO2空心球。Preparation of 3-shell TiO 2 microspheres: Disperse 0.6g carbon spheres into 30 mL of 3M titanium tetrachloride solution, stir at 40°C for 8 hours, then filter with suction, wash 3 times with deionized water, and place in a 70°C oven Dry for 12 hours, place the obtained solid powder in a muffle furnace, raise the temperature to 500°C at a rate of 2°C/min, keep it for 3 hours, and obtain three-shell TiO2 hollow spheres after natural cooling.

将上述制备的10mg三壳层TiO2空心球加入到20mL pH＝8.5的缓冲液中，然后加入25mg盐酸多巴胺。室温搅拌6h，离心分离，40℃真空干燥过夜。三壳层TiO2空心球@聚多巴胺的透射电镜照片如图6所示。Add 10 mg of the three-shell TiO2 hollow spheres prepared above into 20 mL of pH=8.5 buffer, and then add 25 mg of dopamine hydrochloride. Stir at room temperature for 6 hours, centrifuge, and vacuum dry at 40°C overnight. The transmission electron microscope photo of three-shell TiO2 hollow spheres@polydopamine is shown in Figure 6.

实施例5Example 5

3壳层TiO2中空微球@聚乳酸-羟基乙酸共聚物(TiO2中空微球@PLGA)的制备，包括如下步骤：The preparation of 3-shell TiO 2 hollow microspheres@polylactic acid-glycolic acid copolymer (TiO 2 hollow microspheres@PLGA) includes the following steps:

3壳层TiO2微球的制备：将0.6g碳球分散到30mL 3M的四氯化钛溶液中，40℃搅拌8小时后抽滤，用去离子水清洗3遍，置于70℃烘箱中干燥12小时，将所得的固体粉末置于马弗炉中，以2℃/min的速率升温到500℃，保温3h，自然冷却后得到三壳层TiO2空心球。Preparation of 3-shell TiO 2 microspheres: Disperse 0.6g carbon spheres into 30 mL of 3M titanium tetrachloride solution, stir at 40°C for 8 hours, then filter with suction, wash 3 times with deionized water, and place in a 70°C oven Dry for 12 hours, place the obtained solid powder in a muffle furnace, raise the temperature to 500°C at a rate of 2°C/min, keep it for 3 hours, and obtain three-shell TiO2 hollow spheres after natural cooling.

将聚乳酸-羟基乙酸共聚物溶解在2mL乙酸：二氯甲烷1:1的溶剂中，将上述制备的5mg三壳层TiO2空心球分散到1mL去离子水中。分散均匀后，将二者混合，涡旋1min，超声2min，形成微乳液。然后将其逐滴加入2％的40mL聚乙烯醇(PVA)水溶液中，超声10min，在室温下搅拌5小时。将样品进行多次离心，并用乙醇清洗。随后，对离心所得的产物进行冷冻干燥。三壳层TiO2空心球@PLGA的透射电镜照片如图7所示。The polylactic acid-glycolic acid copolymer was dissolved in 2 mL of acetic acid:dichloromethane 1:1 solvent, and 5 mg of the three-shell TiO2 hollow spheres prepared above were dispersed into 1 mL of deionized water. After evenly dispersed, mix the two, vortex for 1 minute, and sonicate for 2 minutes to form a microemulsion. Then it was added dropwise to 2% 40 mL polyvinyl alcohol (PVA) aqueous solution, sonicated for 10 min, and stirred at room temperature for 5 hours. The samples were centrifuged multiple times and washed with ethanol. Subsequently, the centrifuged product was freeze-dried. The transmission electron microscope photo of three-shell TiO2 hollow sphere@PLGA is shown in Figure 7.

实施例6Example 6

3壳层TiO2中空微球@PEG-布洛芬缓释平台的构建，包括如下步骤：The construction of 3-shell TiO2 hollow microspheres@PEG-ibuprofen sustained-release platform includes the following steps:

3壳层TiO2微球的制备：将0.6g碳球分散到30mL 3M的四氯化钛溶液中，40℃搅拌8小时后抽滤，用去离子水清洗3遍，置于70℃烘箱中干燥12小时，将所得的固体粉末置于马弗炉中，以2℃/min的速率升温到500℃，保温3h，自然冷却后得到单壳层TiO2空心球。Preparation of 3-shell TiO 2 microspheres: Disperse 0.6g carbon spheres into 30 mL of 3M titanium tetrachloride solution, stir at 40°C for 8 hours, then filter with suction, wash 3 times with deionized water, and place in a 70°C oven Dry for 12 hours, place the obtained solid powder in a muffle furnace, raise the temperature to 500°C at a rate of 2°C/min, keep it for 3 hours, and obtain single-shell TiO2 hollow spheres after natural cooling.

将上述制备的6mg单壳层TiO2空心球加入到1mL去离子水中，分散均匀后，向其中加入1mL聚乙二醇水溶液，在室温下搅拌1小时之后，加入2mL 200mg/mL的布洛芬溶液，室温下避光搅拌24小时。离心分离去上清，将得到的固体于35℃真空干燥12小时，备用。Add 6 mg of single-shell TiO2 hollow spheres prepared above to 1 mL of deionized water. After evenly dispersed, add 1 mL of polyethylene glycol aqueous solution to it. After stirring at room temperature for 1 hour, add 2 mL of 200 mg/mL ibuprofen. The solution was stirred at room temperature in the dark for 24 hours. The supernatant was removed by centrifugation, and the obtained solid was vacuum dried at 35°C for 12 hours and set aside.

缓释性能测试：将5mg载有布洛芬的聚乙二醇包覆的三壳层微球(即上述经真空干燥处理的固体物)室温分散于1mL去离子水中，接着将混合液转移至透析袋中(截留分子量：14000Da)。将透析袋置于100mL水中，每隔一段时间取上清液，测量在220nm下紫外可见光谱吸收度。根据紫外可见光谱标准曲线求得实际溶液中药物浓度及含量，与热重实验得到的药物装载量对比，得到释放率，进而得出释放率-时间对应曲线。结果如图8所示，表明聚乙二醇包覆的三壳层TiO2微球对布洛芬具有缓释性能。Sustained release performance test: 5 mg of ibuprofen-loaded polyethylene glycol-coated three-shell microspheres (i.e., the above-mentioned vacuum-dried solid matter) were dispersed in 1 mL of deionized water at room temperature, and then the mixture was transferred to In dialysis bag (molecular weight cutoff: 14000Da). Place the dialysis bag in 100 mL of water, take the supernatant at regular intervals, and measure the UV-visible spectrum absorbance at 220 nm. The concentration and content of the drug in the actual solution were obtained based on the UV-visible spectrum standard curve, and compared with the drug loading obtained by the thermogravimetric experiment, the release rate was obtained, and then the release rate-time corresponding curve was obtained. The results are shown in Figure 8, indicating that polyethylene glycol-coated three-shell TiO2 microspheres have sustained-release properties for ibuprofen.

实施例7Example 7

3壳层TiO2中空微球@PEG-二甲双胍缓释平台的构建，包括如下步骤：The construction of 3-shell TiO2 hollow microspheres@PEG-metformin sustained-release platform includes the following steps:

3壳层TiO2微球的制备：将0.6g碳球分散到30mL 3M的四氯化钛溶液中，40℃搅拌8小时后抽滤，用去离子水清洗3遍，置于70℃烘箱中干燥12小时，将所得的固体粉末置于马弗炉中，以2℃/min的速率升温到500℃，保温3h，自然冷却后得到单壳层TiO2空心球。Preparation of 3-shell TiO 2 microspheres: Disperse 0.6g carbon spheres into 30 mL of 3M titanium tetrachloride solution, stir at 40°C for 8 hours, then filter with suction, wash 3 times with deionized water, and place in a 70°C oven Dry for 12 hours, place the obtained solid powder in a muffle furnace, raise the temperature to 500°C at a rate of 2°C/min, keep it for 3 hours, and obtain single-shell TiO2 hollow spheres after natural cooling.

将上述制备的6mg三壳层TiO2空心球加入到1mL去离子水中，分散均匀后，向其中加入1mL聚乙二醇水溶液，在室温下搅拌1小时之后，加入2mL二甲双胍水溶液，室温下避光搅拌24小时。离心分离去上清，将得到的固体于35℃真空干燥12小时，备用。Add 6 mg of three-shell TiO2 hollow spheres prepared above to 1 mL of deionized water. After evenly dispersed, add 1 mL of polyethylene glycol aqueous solution to it. After stirring at room temperature for 1 hour, add 2 mL of metformin aqueous solution and keep away from light at room temperature. Stir for 24 hours. The supernatant was removed by centrifugation, and the obtained solid was vacuum dried at 35°C for 12 hours and set aside.

缓释性能测试：将5mg载有二甲双胍的聚乙二醇包覆的双壳层微球(即上述经真空干燥处理的固体物)室温分散于1mL去离子水中，接着将混合液转移至透析袋中(截留分子量：14000Da)。将透析袋置于100mL水中，每隔一段时间取上清液，测量在230nm下紫外可见光谱吸收度。根据紫外可见光谱标准曲线求得实际溶液中药物浓度及含量，与热重实验得到的药物装载量对比，得到释放率，进而得出释放率-时间对应曲线。结果如图9所示。当pH由7调整为4时，释放率增加，且经过多个循环，释放率达到100％。表明TiO2@聚乙二醇中空多壳层载体对二甲双胍的释放具有pH响应的性能。Sustained release performance test: 5 mg of metformin-loaded polyethylene glycol-coated double-shell microspheres (i.e., the above-mentioned vacuum-dried solids) were dispersed in 1 mL of deionized water at room temperature, and then the mixture was transferred to a dialysis bag. Medium (molecular weight cutoff: 14000Da). Place the dialysis bag in 100 mL of water, take the supernatant at regular intervals, and measure the UV-visible spectrum absorbance at 230 nm. The concentration and content of the drug in the actual solution were obtained based on the UV-visible spectrum standard curve, and compared with the drug loading obtained by the thermogravimetric experiment, the release rate was obtained, and then the release rate-time corresponding curve was obtained. The results are shown in Figure 9. When the pH was adjusted from 7 to 4, the release rate increased, and after multiple cycles, the release rate reached 100%. It shows that the TiO 2 @polyethylene glycol hollow multi-shell carrier has pH-responsive properties for the release of metformin.

本发明未详细说明的内容均可采用本领域的常规技术知识。Contents not described in detail in the present invention may adopt conventional technical knowledge in the field.

最后所应说明的是，以上实施例仅用以说明本发明的技术方案而非限制。尽管参照实施例对本发明进行了详细说明，本领域的普通技术人员应当理解，对本发明的技术方案进行修改或者等同替换，都不脱离本发明技术方案的精神和范围，其均应涵盖在本发明的权利要求范围当中。Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not limiting. Although the present invention has been described in detail with reference to the embodiments, those of ordinary skill in the art will understand that modifications or equivalent substitutions may be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and they shall all be covered by the scope of the present invention. within the scope of the claims.

Claims (10)

1. A preparation method of a composite hollow multi-shell microsphere comprises the following steps:
1) Carrying out hydrothermal reaction on a carbon source to obtain a carbon microsphere template;
2) Dispersing the carbon sphere template obtained in the step 1) in a metal salt solution to obtain a solid precursor;
3) Roasting the precursor obtained in the step 2) to obtain hollow multi-shell microspheres;
4) And 3) contacting the hollow multi-shell microsphere obtained in the step 3) with a solution containing a polymer or a monomer to obtain the composite hollow multi-shell microsphere.
2. The method of claim 1, wherein the metal salt solution comprises an aqueous solution of titanium tetrachloride at a concentration of 1-3mol/L or an acetone solution of titanium tetrachloride at a concentration of 0.1-0.5 mol/L.
3. The method according to claim 2, characterized in that: and 3) preparing the titanium dioxide hollow multi-shell microspheres with single shell, double shell and three shells by using the hollow multi-shell microspheres in the step 3) through a sequential template method.
4. The method according to claim 1, characterized in that: the polymer in the step 4) comprises polylactic acid-glycolic acid copolymer or polyethylene glycol, and the monomer is dopamine hydrochloride.
5. The method according to claim 4, wherein: the polyethylene glycol comprises one or more than two of polyethylene glycol 4000, polyethylene glycol 6000, polyethylene glycol 200000 and polyethylene glycol 100000.
6. The method according to claim 4, wherein: the hollow multi-shell microspheres are contacted with an aqueous solution containing polyethylene glycol, and the mixed solution is stirred for 2-48 hours at room temperature; the concentration of the polyethylene glycol aqueous solution is 10-500mg/mL;
the hollow multi-shell microsphere is contacted with dichloromethane solution containing polylactic acid-glycolic acid, is subjected to ultrasonic emulsification, is added into polyvinyl alcohol water solution dropwise, and is stirred for 2-10 hours at room temperature; the concentration of the methylene dichloride solution of the polylactic acid-glycolic acid copolymer is 1-50mg/mL, and the concentration of the polyvinyl alcohol aqueous solution is 0.1% -2%;
the hollow multi-shell microspheres are contacted with an aqueous solution containing dopamine hydrochloride, and are stirred at room temperature for 5-24 hours, wherein the water concentration of the dopamine hydrochloride is 0.1-5mg/mL, and the pH is 8.0-8.5.
7. The use of the composite hollow multi-shell microsphere prepared by the method of any one of claims 1 to 6 in a drug sustained release system, characterized in that: and (3) adding the aqueous solution of the drug into the composite hollow multi-shell microsphere prepared in the step (4) to finish the drug loading.
8. The use according to claim 7, characterized in that: the medicine comprises one or more than two of an antibacterial agent of pinacolin, an antibiotic of ibuprofen, doxycycline, a hypoglycemic medicine of metformin and an antitumor medicine of doxorubicin.
9. The use according to claim 7, characterized in that: under the stimulation of acidic conditions, the composite hollow multi-shell microsphere drug carrier can accelerate the release of the loaded drug;
wherein the acidic condition is that the pH is less than 5; the medicine is one of kathon and doxycycline.
10. The use according to claim 7, characterized in that: the composite hollow multi-shell microsphere is three-shell.
CN202210847310.1A 2022-07-19 2022-07-19 Preparation method and drug release application of composite hollow multi-shell microspheres Pending CN117442743A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN118766857A (en) * 2024-09-12 2024-10-15 潍坊大耀新材料有限公司 A hollow multi-shell structure oral drug carrier with controllable internal microenvironment and its preparation method and application

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN118766857A (en) * 2024-09-12 2024-10-15 潍坊大耀新材料有限公司 A hollow multi-shell structure oral drug carrier with controllable internal microenvironment and its preparation method and application
CN118766857B (en) * 2024-09-12 2025-03-11 潍坊大耀新材料有限公司 Hollow multi-shell structure oral drug carrier with controllable internal microenvironment and preparation method and application thereof

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