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[三维生物打印支架拓扑结构介导的免疫反应对小鼠毛囊周期的影响]

[Effects of immune responses mediated by topological structures of three-dimensional bioprinted scaffolds on hair follicle cycle in mice].

作者信息

Liu Q H, Li Z, Gala Enhejiri, Zhang C, Song W, Wang Y Z, Liang L T, Zhang M D, Huang Y Y, Li X H, Huang S

机构信息

School of Basic Medical Sciences, Inner Mongolia Medical University, Hohhot 010110, China.

Research Center for Wound Repair and Tissue Regeneration, Medical Innovation Research Department, the PLA General Hospital, Beijing 100048, China.

出版信息

Zhonghua Shao Shang Yu Chuang Mian Xiu Fu Za Zhi. 2024 Jan 20;40(1):43-49. doi: 10.3760/cma.j.cn501225-20231020-00125.

Abstract

To explore the effects of the immune responses mediated by topological structures of three-dimensional bioprinted scaffolds on hair follicle cycle in mice. The study was an experimental research. The alginate-gelatin composite hydrogels were printed into scaffolds using a three-dimensional bioprinter and named T45 scaffolds, T60 scaffolds, and T90 scaffolds according to the 3 topological structures of the scaffolds (the rotation angles of the printhead during printing were 45°, 60°, and 90°, respectively), and the morphology of the three scaffolds was observed after cross-linking by naked eyes. Nine 8-week-old female C57BL/6J mice were divided into T45 group, T60 group, and T90 group, according to the random number table, with three mice in each group, and the T45, T60, and T90 scaffolds were subcutaneously implanted on the back of mice, respectively. On post implantation day (PID) 7, the hair growth in the dorsal depilated area of mice was observed, the thickness of the fiber capsule around the scaffolds was observed by hematoxylin-eosin staining, and the expression levels of CD68, bone morphogenetic protein-2 (BMP-2), and tumor necrosis factor (TNF) protein in the tissue surrounding the scaffolds were observed by immunofluorescence staining. The samples of the above experiments were all 3. The topological structures of the three scaffolds were all clear with high fidelity after cross-linking. On PID 7, the hair growth was obvious in the dorsal depilated area of mice in T45 group and T90 group, while hair growth was slow in the scaffold implantation area of mice in T60 group, which was significantly different from that of the unimplanted area. On PID 7, compared with (18±4) μm in T90 group, the thickness of both the fiber capsule around the scaffolds ((39±4) and (55±8) μm) of mice in T45 group and T60 group was significantly increased (<0.05); the thickness of the fiber capsule around the scaffolds of mice in T60 group was also significantly increased compared with that in T45 group (<0.05). On PID 7, the expression level of CD68 protein in the tissue surrounding the scaffolds of mice in T60 group was significantly higher than the levels in T45 group and T90 group (with both values <0.05). The expression level of BMP-2 protein in the tissue surrounding the scaffolds of mice in T60 group was significantly higher than the levels in T45 group and T90 group (with both values <0.05), and the expression level of BMP-2 protein in the tissue surrounding the scaffolds of mice in T45 group was significantly higher than that in T90 group (<0.05). The expression level of TNF protein in the tissue surrounding the scaffolds of mice in T60 group was significantly lower than the levels in T45 group and T90 group (with both values <0.05). Three-dimensional bioprinted scaffolds with different topological structures mediate different degrees of immune responses after being implanted in mice. A moderate immune response promotes hair growth in depilated area of mice, while an excessive immune response results inhibits the hair follicle entering into the anagen phase.

摘要

探讨三维生物打印支架的拓扑结构介导的免疫反应对小鼠毛囊周期的影响。本研究为实验性研究。使用三维生物打印机将海藻酸钠 - 明胶复合水凝胶打印成支架,并根据支架的3种拓扑结构(打印过程中喷头的旋转角度分别为45°、60°和90°)将其命名为T45支架、T60支架和T90支架,交联后肉眼观察三种支架的形态。将9只8周龄雌性C57BL/6J小鼠按随机数字表分为T45组、T60组和T90组,每组3只,分别将T45、T60和T90支架皮下植入小鼠背部。在植入后第7天(PID 7),观察小鼠背部脱毛区的毛发生长情况,苏木精 - 伊红染色观察支架周围纤维囊的厚度,免疫荧光染色观察支架周围组织中CD68、骨形态发生蛋白 - 2(BMP - 2)和肿瘤坏死因子(TNF)蛋白的表达水平。上述实验样本均为3个。交联后三种支架的拓扑结构均清晰且保真度高。在PID 7时,T45组和T90组小鼠背部脱毛区毛发生长明显,而T60组小鼠支架植入区毛发生长缓慢,与未植入区有显著差异。在PID 7时,T45组和T60组小鼠支架周围纤维囊的厚度(分别为(39±4)μm和(55±8)μm)均显著高于T90组的(18±4)μm(P<0.05);T60组小鼠支架周围纤维囊的厚度也显著高于T45组(P<0.05)。在PID 7时,T60组小鼠支架周围组织中CD68蛋白的表达水平显著高于T45组和T90组(P值均<0.05)。T60组小鼠支架周围组织中BMP - 2蛋白的表达水平显著高于T45组和T90组(P值均<0.05),T45组小鼠支架周围组织中BMP - 2蛋白的表达水平显著高于T90组(P<0.05)。T60组小鼠支架周围组织中TNF蛋白的表达水平显著低于T45组和T90组(P值均<0.05)。不同拓扑结构的三维生物打印支架植入小鼠后介导不同程度的免疫反应。适度的免疫反应促进小鼠脱毛区毛发生长,而过度的免疫反应则抑制毛囊进入生长期。

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