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用于骨组织工程的3D打印负载β-磷酸三钙的甲基丙烯酰化明胶/海藻酸盐互穿聚合物网络生物材料墨水

3D Printing β-TCP-laden GelMA/Alginate Interpenetrating-Polymer-Network Biomaterial Inks for Bone Tissue Engineering.

作者信息

de Souza Joyce R, Rahimnejad Maedeh, Soares Igor P Mendes, Anselmi Caroline, de Oliveira Pedro H C, Dos Reis-Prado Alexandre H, Maglaras Victoria, Dal-Fabbro Renan, Trichês Eliandra S, Bottino Marco C

机构信息

Department of Cariology, Restorative Sciences, and Endodontics, University of Michigan School of Dentistry, Ann Arbor, Michigan, USA.

Department of Dental Materials and Prosthodontics, Institute of Science and Technology of São José dos Campos, São Paulo State University (UNESP), São José dos Campos, São Paulo, Brazil.

出版信息

Bioprinting. 2025 Sep;49. doi: 10.1016/j.bprint.2025.e00413. Epub 2025 Apr 14.

Abstract

Bone's capacity for self-repair is limited when large defects arise from trauma or infection. Traditional grafting methods like autografts and allografts often face challenges like immune rejection and limited availability. Traditional scaffold manufacturing techniques for bone tissue engineering frequently lack precise control over the constructs' material composition and pore architecture. Recently, 3D printing technology, particularly with interpenetrating polymer networks (IPNs), has successfully addressed these limitations, improving biocompatibility, strength, and degradation. Our study investigated gelatin methacryloyl (GelMA)/Alginate IPNs laden with beta tri-calcium phosphate (β-TCP) particles in a 3D-printed format to optimize cell proliferation and tissue regeneration conditions. Rheology studies showed shear-thinning viscosity and fast recovery (~90%) to primary viscosity after stress removal, confirming the inks' suitability for extrusion-based printing. Both inks demonstrated high resolution and acceptable printability (0.9-1). Incorporating β-TCP increased the compressive modulus (0.09±0.01 MPa for the control group vs. 0.15±0.01 MPa for 15% (w/v) β-TCP, ***p < 0.001) and swelling ratio, decreasing biodegradation over 35 days. Cell assays showed enhanced cell proliferation over 7 days, with no significant differences between groups. Compared to basal and osteogenic media controls, higher mineralization and osteogenic gene expression were observed in 15% β-TCP-laden 3D-printed constructs on days 14 and 21. Histological analysis showed no signs of inflammation after three weeks, suggesting favorable tissue compatibility. Furthermore, calcium carbonate deposits were identified, evidencing the successful differentiation of mesenchymal stem cells into cells capable of producing a mineralized matrix. This study demonstrated that the (GelMA)/Alginate IPN containing β-TCP could be a successful biomaterial ink with promising bioactive properties for bone tissue engineering.

摘要

当因创伤或感染导致出现大的骨缺损时,骨骼的自我修复能力是有限的。像自体骨移植和异体骨移植这样的传统移植方法常常面临免疫排斥和供应有限等挑战。用于骨组织工程的传统支架制造技术往往对构建体的材料组成和孔隙结构缺乏精确控制。最近,3D打印技术,特别是互穿聚合物网络(IPN),成功地解决了这些局限性,提高了生物相容性、强度和降解性能。我们的研究调查了以3D打印形式负载β-磷酸三钙(β-TCP)颗粒的甲基丙烯酰化明胶(GelMA)/海藻酸盐IPN,以优化细胞增殖和组织再生条件。流变学研究表明,油墨具有剪切变稀粘度,去除应力后能快速恢复至初始粘度的约90%,证实了这些油墨适用于基于挤出的打印。两种油墨均显示出高分辨率和可接受的可打印性(0.9 - 1)。加入β-TCP提高了压缩模量(对照组为0.09±0.01 MPa,15%(w/v)β-TCP组为0.15±0.01 MPa,***p < 0.001)和溶胀率,在35天内降低了生物降解率。细胞试验表明,在7天内细胞增殖增强,各实验组之间无显著差异。与基础培养基和成骨培养基对照组相比,在第14天和第21天,负载15%β-TCP的3D打印构建体中观察到更高的矿化和成骨基因表达。组织学分析显示,三周后没有炎症迹象,表明具有良好的组织相容性。此外,还鉴定出碳酸钙沉积物,证明间充质干细胞成功分化为能够产生矿化基质的细胞。这项研究表明,含有β-TCP的(GelMA)/海藻酸盐IPN可能是一种成功的生物材料油墨,在骨组织工程中具有有前景的生物活性特性。

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