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3D打印中空管道双交联水凝胶组织工程支架

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背景:在培养对结构、氧需求度较高的细胞时,需要构建一个具有中空管道结构的立体生物支架来保证细胞得到足够的营养与氧气.近年来,具有中空管道结构的水凝胶组织工程支架受到广泛关注.目的:以海藻酸钠为基础的生物支架材料结合同轴打印技术制备具有中空管道结构的组织工程支架,通过灌注方式接种细胞验证其生物相容性.方法:制备海藻酸钠-丙烯酰胺混合打印溶液,控制同轴打印过程中的内外层打印速度、海藻酸钠浓度、接收皿中氯化钙浓度等参数实现打印具有中空管道组织工程支架——海藻酸钠-聚丙烯酰胺双交联水凝胶,表征支架的微观形貌与弹性模量;将小鼠成纤维细胞灌注至组织工程支架中空管道内,利用活死细胞染色观察支架的细胞相容性.结果与结论:①通过探究打印过程中的打印参数,当内层打印速度不变时,中空管道的外径随着外层打印溶液的流速提升而提升,内径有略微提升;当外层打印溶液流速不变、内层溶液流速提升时,中空管道支架的外径基本不变,内径有明显提升;②实验结果显示海藻酸钠的最佳浓度为2.5%,过高的浓度不利于多层结构层与层之间的融合,过低的浓度制备出的水凝胶机械性能不足;③海藻酸钠-聚丙烯酰胺双交联水凝胶的弹性模量较高,普遍高于200 kPa,并且随着氯化钙浓度的提升而增大,在内层氯化钙浓度为2%、接收皿氯化钙浓度为0.3%时双交联水凝胶的弹性模量达到最大值(375 kPa);④组织工程支架灌注细胞体外培养后的活死细胞染色显示,细胞沿着中空管道轴向分布且具有较高的存活率,但细胞浓度小于灌注时的细胞浓度;⑤结果表明,海藻酸钠-聚丙烯酰胺双交联水凝胶具有较强的机械性能与良好的生物相容性,可应用在构建具有中空管道的组织工程支架中,并且"先制备支架、后接种细胞"的方式也避免了传统"细胞与打印溶液混合再进行制备"方式对支架材料及加工方式的限制.
3D printed hollow pipe double-crosslinked hydrogel tissue engineering scaffold
BACKGROUND:When cultivating cells with high demand for structure and oxygen,it is necessary to construct a three-dimensional biological scaffold with hollow pipe structure to make sure the cells get enough nutrients and oxygen.In recent years,hydrogel tissue engineering scaffolds with hollow pipe structure have been paid more and more attention.OBJECTIVE:The biological scaffold material based on sodium alginate was combined with coaxial printing technology to prepare a tissue engineering scaffold with a hollow pipe structure,and the cells were inoculated by perfusion to verify its biological properties.METHODS:The sodium alginate-acrylamide mixed printing solution was prepared,and the parameters such as the printing speed of the inner and outer layers in the coaxial printing process,sodium alginate concentration,and calcium chloride concentration in the receiving dish were controlled to realize the printing of the tissue engineering scaffold with a hollow pipe—sodium alginate-polyacrylamide double-crosslinked hydrogel.The microstructure and elastic modulus of the scaffold were characterized.Mouse fibroblasts were injected into hollow pipes of tissue engineering scaffolds.Cell compatibility was observed by living/dead cell staining.RESULTS AND CONCLUSION:(1)By exploring the printing parameters in the printing process,when the inner printing speed was constant,the outer diameter of the hollow pipe increased with the increase of the flow rate of the outer printing solution,and the inner diameter increased slightly.When the flow rate of the outer layer printing solution was constant,and the flow rate of the inner layer solution was increased,the outer diameter of the hollow pipe was basically unchanged,and the inner diameter was significantly improved.(2)Experimental results showed that the concentration of sodium alginate was 2.5%.Excessive concentration was not conducive to the fusion of multi-layer structure layers,and the mechanical properties of hydrogels prepared at too low concentration were insufficient.(3)The elastic modulus of the double-crosslinked hydrogel was higher,generally higher than 200 kPa,and increased with the increase of the concentration of calcium chloride,and reached the maximum value of 375 kPa when the concentration of calcium chloride in the inner layer was 2%and the concentration of calcium chloride in the receiving dish was 0.3%.(4)The staining of live and dead cells after the tissue engineering scaffold perfusion cells in vitro showed that the cells were distributed along the axis of the hollow pipe and had a higher survival rate,but the cell concentration was lower than that during perfusion.(5)The results show that the sodium alginate-polyacrylamide double-crosslinked hydrogel has strong mechanical properties while retaining good biocompatibility,and can be used in the construction of tissue engineering scaffolds with hollow pipes,and the method of"first preparing the scaffold,then inoculating the cells"also avoids the traditional"cells and printing solution are mixed and then prepared"method to limit the scaffold material and processing method.

coaxial printingtissue engineeringdouble-crosslinked hydrogelsodium alginateacrylamideion crosslinkingscaffold

王仁智、陈远汾、李金玮

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广西大学机械工程学院,广西壮族自治区 南宁市 530004

同轴打印 组织工程 双交联水凝胶 海藻酸钠 丙烯酰胺 离子交联 支架

2025

中国组织工程研究
中国康复医学会,《中国组织工程研究与临床康复》杂志社

中国组织工程研究

北大核心
影响因子:1.387
ISSN:2095-4344
年,卷(期):2025.29(16)