募集抗原特异性T细胞的3D打印支架促进血管化

李翠笛 , 马振江 , 李文韬 , 颉天阳 , 钟丽萍 , 陈宏方 , 王文豪 , 王金武 , 崔文国 , 赵永祥

工程(英文) ›› 2022, Vol. 17 ›› Issue (10) : 183 -195.

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工程(英文) ›› 2022, Vol. 17 ›› Issue (10) : 183 -195. DOI: 10.1016/j.eng.2021.05.018
研究论文

募集抗原特异性T细胞的3D打印支架促进血管化

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3D-Printed Scaffolds Promote Angiogenesis by Recruiting Antigen-Specific T Cells

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摘要

3D打印支架植入后激起的免疫反应是组织修复效果的主要决定性因素。因此,巧妙调控免疫反应的支架材料具有高效修复组织的潜能。本研究受抗原特异性免疫反应对血管新生促进作用的启发,通过动态共价键将介孔二氧化硅微棒(MSR)/聚乙烯亚胺(PEI)/卵清蛋白(OVA)自组装疫苗与3D打印自固化磷酸钙骨水泥(CPC)基支架结合,构建了一种具有调控局部体液免疫反应功能的组织修复支架。通过局部释放OVA激活体液免疫反应,支架可有效募集抗原特异性CD4+ 2 型辅助性T细胞(Th2 细胞),从而促进支架植入早期的血管新生;在提供充足和丰富的血供环境的同时,MSR微棒中释放的硅离子还能有效加速骨再生。在室温下稳定成型的自固化磷酸钙基打印浆料能够促进功能性氧化透明质酸在材料中的均匀分布和结构保持,并构建具有均匀连通孔道的支架。将MSR/PEI 作为抗原载体共价结合于富含醛基的支架,可实现OVA的稳定释放。在体外实验中,该疫苗负载支架有效募集并激活了树突细胞使树突细胞呈递抗原,还促进了骨髓间充质干细胞(BMSC)成骨分化。在皮下包埋实验中,疫苗负载支架增加了Th2 细胞在脾脏中的比例,局部募集了抗原特异性T细胞并促进了支架内的血管新生。此外,原位颅骨缺损修复模型结果表明,负载疫苗的支架可促进血管化骨组织再生。总之,该方法为具有促进血管新生功能个性化植入体的设计提供了一种新思路。

Abstract

The immune response after implantation is a primary determinant of the tissue-repair effects of three-dimensional (3D)-printed scaffolds. Thus, scaffolds that can subtly regulate immune responses may display extraordinary functions. Inspired by the angiogenesis promotion effect of humoral immune response, we covalently combined mesoporous silica microrod (MSR)/polyethyleneimine (PEI)/ovalbumin (OVA) self-assembled vaccines with 3D-printed calcium phosphate cement (CPC) scaffolds for local antigen-specific immune response activation. With the response activated, antigen-specific CD4+ T helper 2 (Th2) cells can be recruited to promote early angiogenesis. The silicon (Si) ions from MSRs can accelerate osteogenesis, with an adequate blood supply being provided. At room temperature, scaffolds with uniformly interconnected macropores were printed using a self-setting CPC-based printing paste, which promoted the uniform dispersion and structural preservation of functional polysaccharides oxidized hyaluronic acid (OHA) inside. Sustained release of OVA was achieved with MSR/PEI covalently attached to scaffolds rich in aldehyde groups as the vaccine carrier. The vaccine-loaded scaffolds effectively recruited and activated dendritic cells (DCs) for antigen presentation and promoted the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in vitro. When embedded subcutaneously in vivo, the vaccine-loaded scaffolds increased the proportion of Th2 cells in the spleen and locally recruited antigen-specific T cells to promote angiogenesis in and around the scaffold. Furthermore, the result in a rat skull defect-repair model indicated that the antigen-specific vaccine-loaded scaffolds promoted the regeneration of vascularized bone. This method may provide a novel concept for patient-specific implant design for angiogenesis promotion.

关键词

3D打印 / 免疫微环境调控 / 血管新生 / 骨再生

Key words

3D printing / Immune microenvironment regulating / Angiogenesis / Bone regeneration

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李翠笛,马振江,李文韬,颉天阳,钟丽萍,陈宏方,王文豪,王金武,崔文国,赵永祥. 募集抗原特异性T细胞的3D打印支架促进血管化[J]. 工程(英文), 2022, 17(10): 183-195 DOI:10.1016/j.eng.2021.05.018

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