CN102430151A - Tissue engineering bone cartilage composite bracket and integrated photocuringable forming method thereof - Google Patents

Abstract

Translated fromChinese

组织工程骨软骨复合支架及其一体化光固化成型方法，将作为骨修复部分、表面富有孔隙结构的陶瓷支架固定于光固化快速成型机托板上，利用CAD模型驱动激光扫描路径，激光将含陶瓷成分的水凝胶以及纯水凝胶支架先后直接曝光固化于陶瓷支架上，形成了三层复合支架，实现了类骨软骨组织形态的三维复合支架的快速、精细制备，避免传统复合支架制备过程中的结构不可控以及手工参与的低效率，提高制备效率并能够在陶瓷支架表面依靠铆定及两相材料粘接复合具有精细结构的类钙化软骨层支架及水凝胶软骨支架。

Tissue-engineered osteochondral composite scaffold and its integrated photocuring molding method, fix the ceramic scaffold with porous structure on the surface of the photocuring rapid prototyping machine as the bone repair part, use the CAD model to drive the laser scanning path, and the laser will contain The hydrogel of ceramic components and the pure hydrogel scaffold were directly exposed and cured on the ceramic scaffold, forming a three-layer composite scaffold, which realized the rapid and fine preparation of the three-dimensional composite scaffold with osteocartilage-like tissue morphology, avoiding the preparation of traditional composite scaffolds. The structure in the process is uncontrollable and the low efficiency of manual participation improves the preparation efficiency and can rely on riveting and two-phase material bonding on the surface of the ceramic scaffold to compound calcified cartilage-like layer scaffolds and hydrogel cartilage scaffolds with fine structures.

Description

Translated fromChinese

组织工程骨软骨复合支架及其一体化光固化成型方法Tissue engineered osteochondral composite scaffold and its integrated photocuring molding method

技术领域technical field

本发明涉及生物制造技术领域，特别涉及组织工程骨软骨复合支架及其一体化光固化成型方法。The invention relates to the technical field of biomanufacturing, in particular to a tissue engineering osteochondral composite scaffold and an integrated photocuring molding method thereof.

背景技术Background technique

由于成年人关节软骨的自修复能力有限，临床上针对关节软骨及骨的缺损，通常需要手术干预，包括关节镜下清理术、镜下微骨折术、骨软骨自体移植等。上述手段存在着修复效果差，受限于患者年龄及缺损面积，移植物来源有限等缺陷。通过近些年对人工骨软骨缺损修复的研究，骨软骨组织工程已经有了较深入的发展，并且显示出在临床上应用的巨大潜能。Due to the limited self-repair ability of adult articular cartilage, surgical intervention is usually required for articular cartilage and bone defects in clinical practice, including arthroscopic debridement, microscopic microfracture, and osteochondral autografting. The above methods have disadvantages such as poor repair effect, limited by the patient's age and defect area, and limited sources of grafts. Through the research on the repair of artificial osteochondral defects in recent years, osteochondral tissue engineering has been developed deeply, and has shown great potential in clinical application.

骨软骨组织工程支架包含有单纯骨(软骨)支架，骨软骨复合支架以及包含钙化软骨层的骨软骨复合支架等种类，目前大多数骨软骨复合支架主要依靠手工缝合、粘接等方式制备，操作过程复杂，自动化及定制化程度低，并且造成复合支架的结构不稳定，在支架植入后容易产生脱层，剥落等情况，无法为新生组织的长入及支架与周围组织的融合提供一个良好的力学支撑环境。此外，自然骨软骨之间具有一层钙化软骨层，它起到支持软骨压力承载以及阻止血管入侵的作用，传统支架的制备并没有考虑到制备钙化软骨层支架这一过渡结构，而采用三维打印的方法制备的分层结构支架又存在着结合力较差，支架植入后容易塌陷等问题。Osteochondral tissue engineering scaffolds include simple bone (cartilage) scaffolds, osteochondral composite scaffolds, and osteochondral composite scaffolds containing calcified cartilage layers. At present, most osteochondral composite scaffolds are mainly prepared by manual suturing and bonding. The process is complicated, the degree of automation and customization is low, and the structure of the composite stent is unstable. After the stent is implanted, delamination and peeling are easy to occur, and it cannot provide a good environment for the growth of new tissue and the fusion of the stent and the surrounding tissue. mechanical support environment. In addition, there is a layer of calcified cartilage between the natural osteocartilage, which plays a role in supporting the pressure bearing of cartilage and preventing blood vessel invasion. The preparation of traditional scaffolds does not take into account the transitional structure of preparing calcified cartilage layer scaffolds, and the use of 3D printing The layered structure scaffold prepared by the method has the problems of poor binding force and easy collapse after implantation.

发明内容Contents of the invention

为了克服上述现有技术的缺陷，本发明的目的在于提供组织工程骨软骨复合支架及其一体化光固化成型方法，能够快速制备出定制化外形的骨软骨组织工程多层复合支架，复合支架具有类似钙化软骨层功能的钙化层，依托陶瓷骨表面的孔隙及胶体在表面的粘接作用，能够有效提高复合支架之间的粘接强度，增强支架植入后的稳定性，为细胞和组织的长入提供一个良好的力学支撑环境，该制备方法具有精度高、操作简单、周期短、可重复性高的优点。In order to overcome the above-mentioned defects in the prior art, the object of the present invention is to provide a tissue-engineered osteochondral composite scaffold and its integrated light-curing molding method, which can quickly prepare a customized multi-layer composite scaffold for osteochondral tissue engineering. The composite scaffold has The calcified layer, which is similar to the function of the calcified cartilage layer, relies on the pores on the surface of the ceramic bone and the bonding effect of the colloid on the surface, which can effectively improve the bonding strength between the composite scaffolds, enhance the stability of the scaffold after implantation, and provide support for cells and tissues. The ingrowth provides a good mechanical support environment, and the preparation method has the advantages of high precision, simple operation, short cycle and high repeatability.

为了达到上述目的，本发明的技术方案是这样实现的：In order to achieve the above object, the technical solution of the present invention is achieved in that:

一种组织工程骨软骨复合支架的一体化光固化成型方法，包括以下步骤：An integrated light-curing molding method for a tissue-engineered osteochondral composite scaffold, comprising the following steps:

步骤一，采用凝胶注模或光固化成型技术或其他方法制备多孔陶瓷骨支架1，多孔陶瓷骨支架1表面均匀分布孔径不小于200μm的孔隙；Step 1, using gel injection molding or photo-curing molding technology or other methods to prepare a porousceramic bone scaffold 1, the surface of the porousceramic bone scaffold 1 is evenly distributed with pores with a diameter of not less than 200 μm;

步骤二，根据陶瓷骨支架尺寸，设计与之匹配的钙化软骨层支架以及软骨支架三维模型，并转化为.stl格式文件，经分层软件处理，将数据导入光固化快速成型机；Step 2. According to the size of the ceramic bone scaffold, design the matching calcified cartilage layer scaffold and the three-dimensional model of the cartilage scaffold, and convert it into a .stl format file. After processing by layering software, import the data into the light-curing rapid prototyping machine;

步骤三，采用光敏材料聚乙二醇双丙烯酸酯，配制含陶瓷粉末的水凝胶溶液，按照体积比将陶瓷粉末∶聚乙二醇双丙烯酸酯∶光引发剂I-1173∶去离子水＝90∶(30-50)∶(0.3-0.5)∶(50-70)的比例均匀混合，采用球磨机球磨2-3小时，置于真空注型机中除去气泡；Step 3, adopt photosensitive material polyethylene glycol diacrylate, prepare the hydrogel solution containing ceramic powder, ceramic powder according to volume ratio: polyethylene glycol diacrylate: photoinitiator I-1173: deionized water= The ratio of 90:(30-50):(0.3-0.5):(50-70) is uniformly mixed, ball milled for 2-3 hours with a ball mill, and placed in a vacuum injection molding machine to remove air bubbles;

步骤四，采用光敏材料聚乙二醇双丙烯酸酯，配制水凝胶溶液，按照体积比将聚乙二醇双丙烯酸酯∶光引发剂I-1173∶去离子水＝(30-50)∶(0.3-0.5)∶(50-70)的比例均匀混合，采用磁力搅拌机搅拌匀速搅拌30-60min，置于真空注型机中除去气泡；Step 4, using the photosensitive material polyethylene glycol diacrylate to prepare a hydrogel solution, polyethylene glycol diacrylate: photoinitiator I-1173: deionized water=(30-50):( The ratio of 0.3-0.5): (50-70) is uniformly mixed, stirred with a magnetic stirrer at a constant speed for 30-60min, and placed in a vacuum injection molding machine to remove air bubbles;

步骤五，将陶瓷骨支架置于快速成型机中具有调节校准装置的托板上，置于步骤三所制备的含陶瓷粉末的水凝胶溶液中，调节陶瓷骨支架与钙化软骨层支架三维数据轮廓对准，调整液面零位，进行光斑扫描，直接成型三维结构；Step five, place the ceramic bone scaffold on the supporting plate with the adjustment and calibration device in the rapid prototyping machine, place it in the hydrogel solution containing ceramic powder prepared in step three, adjust the three-dimensional data of the ceramic bone scaffold and the calcified cartilage layer scaffold Contour alignment, adjust the zero position of the liquid level, perform spot scanning, and directly form a three-dimensional structure;

步骤六，将托板升起，将含陶瓷粉末的水凝胶溶液替换为步骤四所制备的水凝胶溶液，将数据更换为水凝胶软骨支架三维数据，调节陶瓷骨支架与钙化软骨层支架三维数据轮廓对准，调整液面零位，进行光斑扫描，直接成型三维结构，制得多层复合支架，置于去离子水中反复溶胀、消毒处理。Step 6, lift the supporting plate, replace the hydrogel solution containing ceramic powder with the hydrogel solution prepared in step 4, replace the data with the 3D data of the hydrogel cartilage scaffold, and adjust the ceramic bone scaffold and calcified cartilage layer Align the three-dimensional data profile of the bracket, adjust the zero position of the liquid surface, perform spot scanning, and directly form a three-dimensional structure to make a multi-layer composite bracket, which is repeatedly swollen and sterilized in deionized water.

所述的光引发剂I-1173为2-羟基-2-甲基-1-苯基-1-丙基酮。The photoinitiator I-1173 is 2-hydroxy-2-methyl-1-phenyl-1-propyl ketone.

所制备的组织工程骨软骨多层复合支架，包括多孔陶瓷骨支架1，在多孔陶瓷骨支架1上固化有含陶瓷成分的水凝胶支架2，在含陶瓷成分的水凝胶支架2上固化有纯水凝胶支架3。The prepared tissue engineered osteocartilage multilayer composite scaffold includes a porousceramic bone scaffold 1, ahydrogel scaffold 2 containing ceramic components is solidified on the porousceramic bone scaffold 1, and ahydrogel scaffold 2 containing ceramic components is solidified on the porousceramic bone scaffold 1. There are pure hydrogel holders3.

多孔陶瓷骨支架1表面具有预先设计的类似自然骨表面的孔隙结构，有利于类钙化软骨层支架在制备过程中的渗入，以及复合支架植入后血液从陶瓷骨支架向水凝胶软骨支架的有限渗透。The surface of the porousceramic bone scaffold 1 has a pre-designed pore structure similar to the surface of natural bone, which is conducive to the infiltration of the calcified cartilage-like layer scaffold during the preparation process, and the flow of blood from the ceramic bone scaffold to the hydrogel cartilage scaffold after the composite scaffold is implanted. Limited penetration.

含陶瓷成分的水凝胶支架2为生物陶瓷与水凝胶的混合体，能够促使复合支架植入后钙化软骨层组织的形成，起到具有类似自然骨与软骨之间的钙化软骨层的作用，阻挡血管从陶瓷骨支架向水凝胶软骨支架的侵入，促使复合支架植入后软骨的形成。Thehydrogel scaffold 2 containing ceramic components is a mixture of bioceramics and hydrogel, which can promote the formation of calcified cartilage layer tissue after the composite scaffold is implanted, and play a role similar to the calcified cartilage layer between natural bone and cartilage , to block the invasion of blood vessels from the ceramic bone scaffold to the hydrogel cartilage scaffold, and promote the formation of cartilage after the composite scaffold is implanted.

水凝胶软骨支架3为具有生物相容性的聚乙二醇双丙烯酸酯固化形成，具有与植入部位相匹配的曲面结构，便于复合支架植入后与自然关节表面的匹配，为细胞的贴附及组织的生长提供一个良好的环境。Thehydrogel cartilage scaffold 3 is formed by solidification of biocompatible polyethylene glycol diacrylate, and has a curved surface structure that matches the implantation site, which facilitates the matching of the composite scaffold with the natural joint surface after implantation, and is a support for cells. Attachment and tissue growth provide a favorable environment.

本发明在快速成型制备陶瓷骨支架的基础上，提出以陶瓷支架表面为基底，利用光固化快速成型技术制备类钙化软骨层支架以及水凝胶软骨支架，实现多层多材料复合支架的制备，可以快速制备出定制化、结合强度高的骨软骨多层复合支架，为骨软骨组织工程复合支架的制备提供了定制化、工艺简单、精度高的工艺方法。On the basis of rapid prototyping to prepare ceramic bone scaffolds, the present invention proposes to use the surface of ceramic scaffolds as a base, and use photocuring rapid prototyping technology to prepare calcified cartilage-like layer scaffolds and hydrogel cartilage scaffolds to realize the preparation of multi-layer multi-material composite scaffolds. The multi-layered osteochondral composite scaffold can be quickly prepared with customization and high bonding strength, which provides a customized, simple and high-precision process method for the preparation of composite scaffolds for osteochondral tissue engineering.

附图说明Description of drawings

图1是本发明的工艺流程图。Fig. 1 is a process flow diagram of the present invention.

图2是陶瓷支架定位机构图。Fig. 2 is a diagram of the positioning mechanism of the ceramic bracket.

图3是本发明的结构示意图，其中：图3(a)是多层复合支架正视图；图3(b)是复合支架等轴测图。Fig. 3 is a schematic structural view of the present invention, wherein: Fig. 3(a) is a front view of a multi-layer composite bracket; Fig. 3(b) is an isometric view of a composite bracket.

图4是骨软骨多层复合支架的植入部位示意图。Fig. 4 is a schematic diagram of the implantation site of the osteochondral multilayer composite scaffold.

具体实施方式Detailed ways

下面结合附图对本发明作进一步的详细描述。The present invention will be described in further detail below in conjunction with the accompanying drawings.

参照图1，按照本发明提出的利用光固化快速成型技术，以基于快速成型技术制备的陶瓷支架为基底，依托其多孔表面，直接成型类钙化软骨层支架及水凝胶软骨支架，其具体步骤包括：Referring to Fig. 1, according to the light-curing rapid prototyping technology proposed by the present invention, the ceramic support prepared based on the rapid prototyping technology is used as the base, relying on its porous surface, directly forming the calcified cartilage-like layer support and the hydrogel cartilage support, and the specific steps include:

一种组织工程骨软骨复合支架的一体化光固化成型方法，包括以下步骤：An integrated light-curing molding method for a tissue-engineered osteochondral composite scaffold, comprising the following steps:

步骤一、采用凝胶注模或光固化成型技术或其他方法制备多孔陶瓷骨支架1，多孔陶瓷骨支架1表面均匀分布孔径不小于200μm的孔隙；Step 1. The porousceramic bone scaffold 1 is prepared by gel injection molding or photocuring molding technology or other methods, and pores with a diameter of not less than 200 μm are evenly distributed on the surface of the porousceramic bone scaffold 1;

多孔陶瓷骨支架1根据支架植入部位的需求，利用三维反求技术，提取关节轮廓，经Geomagics及Pro/E软件处理，设计多孔陶瓷支架，其上表面轮廓源自关节曲面，并且具有均匀分布的至少200μm孔隙，孔隙占40-50％面积，制备原型或负型；类钙化软骨层支架厚度为300μm，水凝胶软骨支架厚度为1000μm，三维模型存为.stl格式文件，采用快速成型机制备陶瓷骨支架负型，采用凝胶注模技术，制备陶瓷素坯；或者采用快速成型技术，直接固化陶瓷浆料，成型陶瓷素坯。经干燥、脱树脂脱胶等工艺，高温烧结，制备陶瓷骨支架，或采用其他方法，如三维打印成型法，熔融沉积法等制备陶瓷骨支架。Porousceramic bone scaffold 1 According to the requirements of the implanted site, the three-dimensional inversion technology is used to extract the joint contour, and after Geomagics and Pro/E software processing, the porous ceramic scaffold is designed. The upper surface contour is derived from the articular surface and has a uniform distribution. The pores of at least 200 μm, the pores account for 40-50% of the area, and the prototype or negative type is prepared; the thickness of the calcified cartilage layer scaffold is 300 μm, and the thickness of the hydrogel cartilage scaffold is 1000 μm. The three-dimensional model is saved as a .stl format file, using a rapid prototyping mechanism Prepare the negative shape of the ceramic bone bracket, and use the gel injection molding technology to prepare the ceramic green body; or use the rapid prototyping technology to directly solidify the ceramic slurry to form the ceramic green body. After drying, resin degumming and other processes, high-temperature sintering, the ceramic bone scaffold is prepared, or other methods, such as three-dimensional printing method, fused deposition method, etc., are used to prepare the ceramic bone scaffold.

步骤二，根据陶瓷骨支架尺寸，设计与之匹配的钙化软骨层支架以及软骨支架三维模型，并转化为.stl格式文件，经分层软件处理，将数据导入光固化快速成型机；Step 2. According to the size of the ceramic bone scaffold, design the matching calcified cartilage layer scaffold and the three-dimensional model of the cartilage scaffold, and convert it into a .stl format file. After processing by layering software, import the data into the light-curing rapid prototyping machine;

步骤三，采用光敏材料聚乙二醇双丙烯酸酯，配制含陶瓷粉末的水凝胶溶液，按照体积比将陶瓷粉末∶聚乙二醇双丙烯酸酯∶光引发剂I-1173∶去离子水＝90∶40∶0.5∶60的比例均匀混合，采用球磨机球磨3小时，置于真空注型机中除去气泡；Step 3, adopt photosensitive material polyethylene glycol diacrylate, prepare the hydrogel solution containing ceramic powder, ceramic powder according to volume ratio: polyethylene glycol diacrylate: photoinitiator I-1173: deionized water= Mix evenly at a ratio of 90:40:0.5:60, mill with a ball mill for 3 hours, and place in a vacuum injection molding machine to remove air bubbles;

步骤四，采用光敏材料聚乙二醇双丙烯酸酯，配制水凝胶溶液，按照体积比将聚乙二醇双丙烯酸酯∶光引发剂I-1173∶去离子水＝40∶0.5∶60的比例均匀混合，采用磁力搅拌机搅拌匀速搅拌50min，置于真空注型机中除去气泡；Step 4, using the photosensitive material polyethylene glycol diacrylate to prepare a hydrogel solution, according to the volume ratio polyethylene glycol diacrylate: photoinitiator I-1173: deionized water=40:0.5:60 ratio Mix evenly, use a magnetic stirrer to stir at a constant speed for 50 minutes, and place in a vacuum injection molding machine to remove air bubbles;

步骤五，将陶瓷骨支架置于快速成型机中具有调节校准装置的托板上，置于步骤三所制备的含陶瓷粉末的水凝胶溶液中，调节陶瓷骨支架与钙化软骨层支架三维数据轮廓对准，调整液面零位，进行光斑扫描，直接成型三维结构；Step five, place the ceramic bone scaffold on the supporting plate with the adjustment and calibration device in the rapid prototyping machine, place it in the hydrogel solution containing ceramic powder prepared in step three, adjust the three-dimensional data of the ceramic bone scaffold and the calcified cartilage layer scaffold Contour alignment, adjust the zero position of the liquid level, perform spot scanning, and directly form a three-dimensional structure;

本步骤的具体实施方式为：参照图2，采用二维定位机构，将陶瓷骨支架根据模型在计算机软件magics中的位置而精确定位于快速成型机托板之上，调整零位，置于步骤三所制备的含陶瓷粉末的水凝胶溶液中，，进行类钙化软骨层数据的扫描，扫描工艺参数为：固体激光器波长355nm，光斑直径0.20mm，模型分层厚度0.10mm，填充扫描速度2500.00mm/s，填充向量间距0.10mm，支撑扫描速度500mm/s，制备时跳过支撑扫描此步骤，跨跳速度12000.00mm/s，轮廓扫描速度3000mm/s，补偿直径0.1mm，工作台升降速度2.0mm/s，点支撑扫描时间1.66ms，绞结构扫描时间1.37ms；The specific implementation of this step is as follows: referring to Figure 2, using a two-dimensional positioning mechanism, the ceramic bone bracket is accurately positioned on the rapid prototyping machine pallet according to the position of the model in the computer software magics, and the zero position is adjusted, and placed in the step In the hydrogel solution containing ceramic powder prepared by San, scan the data of the calcified cartilage-like layer. The scanning process parameters are: solid-state laser wavelength 355nm, spot diameter 0.20mm, model layer thickness 0.10mm, filling scanning speed 2500.00 mm/s, filling vector spacing 0.10mm, support scanning speed 500mm/s, skip the support scanning step during preparation, jump speed 12000.00mm/s, contour scanning speed 3000mm/s, compensation diameter 0.1mm, table lifting speed 2.0mm/s, point support scanning time 1.66ms, twisted structure scanning time 1.37ms;

步骤六，将托板升起，将含陶瓷粉末的水凝胶溶液替换为步骤四所制备的水凝胶溶液，将数据更换为水凝胶软骨支架三维数据，调节陶瓷骨支架与钙化软骨层支架三维数据轮廓对准，调整液面零位，进行光斑扫描，直接成型三维结构，制得多层复合支架，置于去离子水中反复溶胀、消毒处理；Step 6, lift the supporting plate, replace the hydrogel solution containing ceramic powder with the hydrogel solution prepared in step 4, replace the data with the 3D data of the hydrogel cartilage scaffold, and adjust the ceramic bone scaffold and calcified cartilage layer Align the three-dimensional data profile of the bracket, adjust the zero position of the liquid surface, perform spot scanning, and directly form a three-dimensional structure to make a multi-layer composite bracket, which is repeatedly swollen and sterilized in deionized water;

本步骤的具体实施方式为：上升托板，将溶液替换为步骤四所制备的水凝胶溶液，将数据更换为水凝胶软骨支架三维数据，调节陶瓷骨支架与钙化软骨层支架三维数据轮廓对准，调整液面零位，进行水凝胶软骨支架的扫描固化，扫描工艺参数为：固体激光器波长355nm，光斑直径0.20mm，模型分层厚度0.10mm，填充扫描速度5500.00mm/s，填充向量间距0.10mm，支撑扫描速度1000mm/s，制备时跳过支撑扫描此步骤，跨跳速度12000.00mm/s，轮廓扫描速度6000mm/s，补偿直径0.1mm，工作台升降速度2.0mm/s，点支撑扫描时间0.50ms，绞结构扫描时间0.50ms；复合支架制备完成后，取出，置于去离子水中清洗，对其进行后续消毒等处理，进行体外植入。此时可不必调整二维定位机构位置，置换陶瓷支架即可重复制备复合支架。The specific implementation of this step is: raise the supporting plate, replace the solution with the hydrogel solution prepared in step 4, replace the data with the three-dimensional data of the hydrogel cartilage support, and adjust the three-dimensional data profile of the ceramic bone support and the calcified cartilage layer support Align, adjust the zero position of the liquid level, and scan and solidify the hydrogel cartilage scaffold. The scanning process parameters are: solid-state laser wavelength 355nm, spot diameter 0.20mm, model layer thickness 0.10mm, filling scanning speed 5500.00mm/s, filling The vector spacing is 0.10mm, the support scanning speed is 1000mm/s, the support scanning step is skipped during preparation, the jump speed is 12000.00mm/s, the contour scanning speed is 6000mm/s, the compensation diameter is 0.1mm, and the table lifting speed is 2.0mm/s. The scan time for point supports is 0.50 ms, and the scan time for twisted structures is 0.50 ms; after the composite scaffold is prepared, it is taken out, cleaned in deionized water, subjected to subsequent disinfection and other treatments, and implanted in vitro. At this time, it is not necessary to adjust the position of the two-dimensional positioning mechanism, and the composite stent can be repeatedly prepared by replacing the ceramic stent.

上述步骤中所述的光引发剂I-1173为2-羟基-2-甲基-1-苯基-1-丙基酮。The photoinitiator I-1173 described in the above steps is 2-hydroxy-2-methyl-1-phenyl-1-propyl ketone.

参照图3，所制备的组织工程骨软骨多层复合支架，包括多孔陶瓷骨支架1，在多孔陶瓷骨支架1上固化有含陶瓷成分的水凝胶支架2，在含陶瓷成分的水凝胶支架2上固化有纯水凝胶支架3。Referring to Fig. 3, the prepared tissue engineered osteochondral multilayer composite scaffold includes a porousceramic bone scaffold 1, ahydrogel scaffold 2 containing ceramic components is solidified on the porousceramic bone scaffold 1, and ahydrogel scaffold 2 containing ceramic components is solidified on the porousceramic bone scaffold 1. Apure hydrogel support 3 is solidified on thesupport 2 .

图4是本发明植入新西兰兔的股骨远端部位示意图，图中显示支架表面与关节表面曲面匹配。Fig. 4 is a schematic diagram of the distal end of the femur implanted in New Zealand rabbits according to the present invention, which shows that the surface of the bracket matches the curved surface of the articular surface.

Claims (4)

Translated fromChinese

1.一种组织工程骨软骨复合支架的一体化光固化成型方法，其特征在于，包括以下步骤：1. an integrated photocuring molding method of tissue engineering osteochondral composite support, is characterized in that, comprises the following steps:步骤一、采用凝胶注模或光固化成型技术或其他方法制备多孔陶瓷骨支架1，多孔陶瓷骨支架(1)表面均匀分布孔径不小于200μm的孔隙；Step 1. The porous ceramic bone scaffold 1 is prepared by gel injection molding or photocuring molding technology or other methods, and pores with a diameter of not less than 200 μm are uniformly distributed on the surface of the porous ceramic bone scaffold (1);步骤二，根据陶瓷骨支架尺寸，设计与之匹配的钙化软骨层支架以及软骨支架三维模型，并转化为.stl格式文件，经分层软件处理，将数据导入光固化快速成型机；Step 2. According to the size of the ceramic bone scaffold, design the matching calcified cartilage layer scaffold and the three-dimensional model of the cartilage scaffold, and convert it into a .stl format file. After processing by layering software, import the data into the light-curing rapid prototyping machine;步骤三，采用光敏材料聚乙二醇双丙烯酸酯，配制含陶瓷粉末的水凝胶溶液，按照体积比将陶瓷粉末∶聚乙二醇双丙烯酸酯∶光引发剂I-1173∶去离子水＝90∶(30-50)∶(0.3-0.5)∶(50-70)的比例均匀混合，采用球磨机球磨2-3小时，置于真空注型机中除去气泡；Step 3, adopt photosensitive material polyethylene glycol diacrylate, prepare the hydrogel solution containing ceramic powder, ceramic powder according to volume ratio: polyethylene glycol diacrylate: photoinitiator I-1173: deionized water= The ratio of 90:(30-50):(0.3-0.5):(50-70) is uniformly mixed, ball milled for 2-3 hours with a ball mill, and placed in a vacuum injection molding machine to remove air bubbles;步骤四，采用光敏材料聚乙二醇双丙烯酸酯，配制水凝胶溶液，按照体积比将聚乙二醇双丙烯酸酯∶光引发剂I-1173∶去离子水＝(30-50)∶(0.3-0.5)∶(50-70)的比例均匀混合，采用磁力搅拌机搅拌匀速搅拌30-60min，置于真空注型机中除去气泡；Step 4, using the photosensitive material polyethylene glycol diacrylate to prepare a hydrogel solution, polyethylene glycol diacrylate: photoinitiator I-1173: deionized water=(30-50):( The ratio of 0.3-0.5): (50-70) is uniformly mixed, stirred with a magnetic stirrer at a constant speed for 30-60min, and placed in a vacuum injection molding machine to remove air bubbles;步骤五，将陶瓷骨支架(1)置于快速成型机中具有调节校准装置的托板上，置于步骤三所制备的含陶瓷粉末的水凝胶溶液中，调节陶瓷骨支架与钙化软骨层支架三维数据轮廓对准，调整液面零位，进行光斑扫描，直接成型三维结构；Step 5, place the ceramic bone scaffold (1) on the supporting plate with the adjustment and calibration device in the rapid prototyping machine, place it in the hydrogel solution containing ceramic powder prepared in step 3, adjust the ceramic bone scaffold and the calcified cartilage layer Align the three-dimensional data profile of the bracket, adjust the zero position of the liquid level, perform spot scanning, and directly form the three-dimensional structure;步骤六，将托板升起，将含陶瓷粉末的水凝胶溶液替换为步骤四所制备的水凝胶溶液，将数据更换为水凝胶软骨支架三维数据，调节陶瓷骨支架与钙化软骨层支架三维数据轮廓对准，调整液面零位，进行光斑扫描，直接成型三维结构，制得多层复合支架，置于去离子水中反复溶胀、消毒处理。Step 6, lift the supporting plate, replace the hydrogel solution containing ceramic powder with the hydrogel solution prepared in step 4, replace the data with the 3D data of the hydrogel cartilage scaffold, and adjust the ceramic bone scaffold and calcified cartilage layer Align the three-dimensional data profile of the bracket, adjust the zero position of the liquid surface, perform spot scanning, and directly form a three-dimensional structure to make a multi-layer composite bracket, which is repeatedly swollen and sterilized in deionized water.2.根据权利要求1所述的一种组织工程骨软骨复合支架的一体化光固化成型方法，其特征在于，包括以下步骤：2. The integrated photocuring molding method of a kind of tissue engineering osteochondral composite scaffold according to claim 1, it is characterized in that, comprising the following steps:步骤一、采用凝胶注模或光固化成型技术或其他方法制备多孔陶瓷骨支架1，多孔陶瓷骨支架1表面均匀分布孔径不小于200μm的孔隙；Step 1. The porous ceramic bone scaffold 1 is prepared by gel injection molding or photocuring molding technology or other methods, and pores with a diameter of not less than 200 μm are evenly distributed on the surface of the porous ceramic bone scaffold 1;多孔陶瓷骨支架(1)根据支架植入部位的需求，利用三维反求技术，提取关节轮廓，经Geomagics及Pro/E软件处理，设计多孔陶瓷支架，其上表面轮廓源自关节曲面，并且具有均匀分布的至少200μm孔隙，孔隙占40-50％面积，制备原型或负型；类钙化软骨层支架厚度为300μm，水凝胶软骨支架厚度为1000μm，三维模型存为.stl格式文件，采用快速成型机制备陶瓷骨支架负型，采用凝胶注模技术，制备陶瓷素坯；或者采用快速成型技术，直接固化陶瓷浆料，成型陶瓷素坯。经干燥、脱树脂脱胶等工艺，高温烧结，制备陶瓷骨支架，或采用其他方法，如三维打印成型法，熔融沉积法等制备陶瓷骨支架。Porous ceramic bone scaffold (1) According to the requirements of the scaffold implantation site, the three-dimensional inverse technology is used to extract the joint contour, and after Geomagics and Pro/E software processing, the porous ceramic scaffold is designed. The upper surface contour is derived from the articular surface, and has The pores of at least 200 μm are evenly distributed, and the pores account for 40-50% of the area, and the prototype or negative type is prepared; the thickness of the calcified cartilage layer scaffold is 300 μm, and the thickness of the hydrogel cartilage scaffold is 1000 μm. The three-dimensional model is saved as a .stl format file, and the rapid The forming machine prepares the negative type of the ceramic bone bracket, and adopts the gel injection molding technology to prepare the ceramic green body; or adopts the rapid prototyping technology to directly solidify the ceramic slurry to form the ceramic green body. After drying, resin degumming and other processes, high-temperature sintering, the ceramic bone scaffold is prepared, or other methods, such as three-dimensional printing method, fused deposition method, etc., are used to prepare the ceramic bone scaffold.步骤二，根据陶瓷骨支架尺寸，设计与之匹配的钙化软骨层支架以及软骨支架三维模型，并转化为.stl格式文件，经分层软件处理，将数据导入光固化快速成型机；Step 2. According to the size of the ceramic bone scaffold, design the matching calcified cartilage layer scaffold and the three-dimensional model of the cartilage scaffold, and convert it into a .stl format file. After processing by layering software, import the data into the light-curing rapid prototyping machine;步骤三，采用光敏材料聚乙二醇双丙烯酸酯，配制含陶瓷粉末的水凝胶溶液，按照体积比将陶瓷粉末∶聚乙二醇双丙烯酸酯∶光引发剂I-1173∶去离子水＝90∶40∶0.5∶60的比例均匀混合，采用球磨机球磨3小时，置于真空注型机中除去气泡；Step 3, adopt photosensitive material polyethylene glycol diacrylate, prepare the hydrogel solution containing ceramic powder, ceramic powder according to volume ratio: polyethylene glycol diacrylate: photoinitiator I-1173: deionized water= Mix evenly at a ratio of 90:40:0.5:60, mill with a ball mill for 3 hours, and place in a vacuum injection molding machine to remove air bubbles;步骤四，采用光敏材料聚乙二醇双丙烯酸酯，配制水凝胶溶液，按照体积比将聚乙二醇双丙烯酸酯∶光引发剂I-1173∶去离子水＝40∶0.5∶60的比例均匀混合，采用磁力搅拌机搅拌匀速搅拌50min，置于真空注型机中除去气泡；Step 4, using the photosensitive material polyethylene glycol diacrylate to prepare a hydrogel solution, according to the volume ratio polyethylene glycol diacrylate: photoinitiator I-1173: deionized water=40:0.5:60 ratio Mix evenly, use a magnetic stirrer to stir at a constant speed for 50 minutes, and place in a vacuum injection molding machine to remove air bubbles;步骤五，将陶瓷骨支架置于快速成型机中具有调节校准装置的托板上，置于步骤三所制备的含陶瓷粉末的水凝胶溶液中，调节陶瓷骨支架与钙化软骨层支架三维数据轮廓对准，调整液面零位，进行光斑扫描，直接成型三维结构；Step five, place the ceramic bone scaffold on the supporting plate with the adjustment and calibration device in the rapid prototyping machine, place it in the hydrogel solution containing ceramic powder prepared in step three, adjust the three-dimensional data of the ceramic bone scaffold and the calcified cartilage layer scaffold Contour alignment, adjust the zero position of the liquid level, perform spot scanning, and directly form a three-dimensional structure;本步骤的具体实施方式为：采用二维定位机构，将陶瓷骨支架根据模型在计算机软件magics中的位置而精确定位于快速成型机托板之上，调整零位，置于步骤三所制备的含陶瓷粉末的水凝胶溶液中，，进行类钙化软骨层数据的扫描，扫描工艺参数为：固体激光器波长355nm，光斑直径0.20mm，模型分层厚度0.10mm，填充扫描速度2500.00mm/s，填充向量间距0.10mm，支撑扫描速度500mm/s，制备时跳过支撑扫描此步骤，跨跳速度12000.00mm/s，轮廓扫描速度3000mm/s，补偿直径0.1mm，工作台升降速度2.0mm/s，点支撑扫描时间1.66ms，绞结构扫描时间1.37ms。The specific implementation of this step is: use a two-dimensional positioning mechanism to accurately position the ceramic bone bracket on the rapid prototyping machine pallet according to the position of the model in the computer software magics, adjust the zero position, and place it on the prepared in step 3 In the hydrogel solution containing ceramic powder, scan the data of the calcified cartilage-like layer. The scanning process parameters are: solid-state laser wavelength 355nm, spot diameter 0.20mm, model layer thickness 0.10mm, filling scanning speed 2500.00mm/s, The filling vector spacing is 0.10mm, the support scanning speed is 500mm/s, the support scanning step is skipped during preparation, the jump speed is 12000.00mm/s, the contour scanning speed is 3000mm/s, the compensation diameter is 0.1mm, and the table lifting speed is 2.0mm/s , the scanning time of the point support is 1.66ms, and the scanning time of the twisted structure is 1.37ms.步骤六，将托板升起，将含陶瓷粉末的水凝胶溶液替换为步骤四所制备的水凝胶溶液，将数据更换为水凝胶软骨支架三维数据，调节陶瓷骨支架与钙化软骨层支架三维数据轮廓对准，调整液面零位，进行光斑扫描，直接成型三维结构，制得多层复合支架，置于去离子水中反复溶胀、消毒处理。Step 6, lift the supporting plate, replace the hydrogel solution containing ceramic powder with the hydrogel solution prepared in step 4, replace the data with the 3D data of the hydrogel cartilage scaffold, and adjust the ceramic bone scaffold and calcified cartilage layer Align the three-dimensional data profile of the bracket, adjust the zero position of the liquid surface, perform spot scanning, and directly form a three-dimensional structure to make a multi-layer composite bracket, which is repeatedly swollen and sterilized in deionized water.本步骤的具体实施方式为：上升托板，将溶液替换为步骤四所制备的水凝胶溶液，将数据更换为水凝胶软骨支架三维数据，调节陶瓷骨支架与钙化软骨层支架三维数据轮廓对准，调整液面零位，进行水凝胶软骨支架的扫描固化，扫描工艺参数为：固体激光器波长355nm，光斑直径0.20mm，模型分层厚度0.10mm，填充扫描速度5500.00mm/s，填充向量间距0.10mm，支撑扫描速度1000mm/s，制备时跳过支撑扫描此步骤，跨跳速度12000.00mm/s，轮廓扫描速度6000mm/s，补偿直径0.1mm，工作台升降速度2.0mm/s，点支撑扫描时间0.50ms，绞结构扫描时间0.50ms；复合支架制备完成后，取出，置于去离子水中清洗，对其进行后续消毒等处理，进行体外植入。此时可不必调整二维定位机构位置，置换陶瓷支架即可重复制备复合支架。The specific implementation of this step is: raise the supporting plate, replace the solution with the hydrogel solution prepared in step 4, replace the data with the three-dimensional data of the hydrogel cartilage support, and adjust the three-dimensional data profile of the ceramic bone support and the calcified cartilage layer support Align, adjust the zero position of the liquid level, and scan and solidify the hydrogel cartilage scaffold. The scanning process parameters are: solid-state laser wavelength 355nm, spot diameter 0.20mm, model layer thickness 0.10mm, filling scanning speed 5500.00mm/s, filling The vector spacing is 0.10mm, the support scanning speed is 1000mm/s, the support scanning step is skipped during preparation, the jump speed is 12000.00mm/s, the contour scanning speed is 6000mm/s, the compensation diameter is 0.1mm, and the table lifting speed is 2.0mm/s. The scan time for point support is 0.50 ms, and the scan time for twisted structure is 0.50 ms; after the composite scaffold is prepared, it is taken out, cleaned in deionized water, subjected to subsequent disinfection and other treatments, and implanted in vitro. At this time, it is not necessary to adjust the position of the two-dimensional positioning mechanism, and the composite stent can be repeatedly prepared by replacing the ceramic stent.3.根据权利要求1或2所述的一种组织工程骨软骨复合支架的一体化光固化成型方法，其特征在于，步骤中所述的光引发剂I-1173为2-羟基-2-甲基-1-苯基-1-丙基酮。3. The integrated photocuring molding method of a tissue engineered osteochondral composite scaffold according to claim 1 or 2, wherein the photoinitiator I-1173 described in the step is 2-hydroxyl-2-formazan Base-1-phenyl-1-propyl ketone.4.一种组织工程骨软骨复合支架，包括多孔陶瓷骨支架(1)，其特征在于，在多孔陶瓷骨支架(1)上固化有含陶瓷成分的水凝胶支架(2)，在含陶瓷成分的水凝胶支架(2)上固化有纯水凝胶支架(3)。4. A tissue engineering osteochondral composite support, comprising a porous ceramic bone support (1), is characterized in that, on the porous ceramic bone support (1), a hydrogel support (2) containing ceramic components is solidified, and The pure hydrogel support (3) is cured on the hydrogel support (2) of the components.

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