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用于骨组织工程的BMP-2模拟肽共价锚定到聚醚醚酮表面的策略

Strategies for the Covalent Anchoring of a BMP-2-Mimetic Peptide to PEEK Surface for Bone Tissue Engineering.

作者信息

Cassari Leonardo, Zamuner Annj, Messina Grazia Maria Lucia, Marsotto Martina, Chang Hao-Chen, Coward Trevor, Battocchio Chiara, Iucci Giovanna, Marletta Giovanni, Di Silvio Lucy, Dettin Monica

机构信息

Department of Industrial Engineering, University of Padova, Via Marzolo 9, 35131 Padova, Italy.

Department of Civil, Environmental, and Architectural Engineering, University of Padova, Via Marzolo 9, 35131 Padova, Italy.

出版信息

Materials (Basel). 2023 May 21;16(10):3869. doi: 10.3390/ma16103869.

Abstract

Researchers in the field of tissue engineering are always searching for new scaffolds for bone repair. Polyetheretherketone (PEEK) is a chemically inert polymer that is insoluble in conventional solvents. PEEK's great potential in tissue engineering applications arises from its ability to not induce adverse reactions when in contact with biological tissues and its mechanical properties, which are similar to those of human bone. These exceptional features are limited by the bio-inertness of PEEK, which causes poor osteogenesis on the implant surface. Here, we demonstrated that the covalent grafting of the sequence (48-69) mapped on the BMP-2 growth factor (GBMP1α) significantly enhances the mineralization and gene expression of human osteoblasts. Different chemical methods were employed for covalently grafting the peptide onto 3D-printed PEEK disks: (a) the reaction between PEEK carbonyls and amino-oxy groups inserted in the peptides' N-terminal sites (oxime chemistry) and (b) the photoactivation of azido groups present in the peptides' N-terminal sites, which produces nitrene radicals able to react with PEEK surface. The peptide-induced PEEK surface modification was assessed using X-ray photoelectron measurements, while the superficial properties of the functionalized material were analyzed by means of atomic force microscopy and force spectroscopy. Live and dead assays and SEM measurements showed greater cell cover on functionalized samples than the control, without any cytotoxicity induction. Moreover, functionalization improved the rate of cell proliferation and the amount of calcium deposits, as demonstrated by the AlamarBlue™ and alizarin red results, respectively. The effects of GBMP1α on h-osteoblast gene expression were assayed using quantitative real-time polymerase chain reaction.

摘要

组织工程领域的研究人员一直在寻找用于骨修复的新型支架。聚醚醚酮(PEEK)是一种化学惰性聚合物,不溶于传统溶剂。PEEK在组织工程应用中的巨大潜力源于其与生物组织接触时不会引发不良反应的能力以及其机械性能,这些性能与人类骨骼相似。然而,这些卓越特性受到PEEK生物惰性的限制,这导致植入物表面的成骨作用较差。在此,我们证明了映射在BMP-2生长因子(GBMP1α)上的序列(48-69)的共价接枝显著增强了人成骨细胞的矿化和基因表达。采用了不同的化学方法将该肽共价接枝到3D打印的PEEK盘上:(a)PEEK羰基与插入肽N端位点的氨氧基之间的反应(肟化学),以及(b)肽N端位点存在的叠氮基团的光活化,其产生能够与PEEK表面反应的氮烯自由基。使用X射线光电子能谱测量评估肽诱导的PEEK表面改性,同时通过原子力显微镜和力谱分析功能化材料的表面性质。活细胞和死细胞检测以及扫描电子显微镜测量表明,功能化样品上的细胞覆盖度比对照更高,且未诱导任何细胞毒性。此外,如AlamarBlue™和茜素红结果分别所示,功能化提高了细胞增殖速率和钙沉积量。使用定量实时聚合酶链反应测定GBMP1α对人成骨细胞基因表达的影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8f2b/10222618/6a18d163ffbc/materials-16-03869-g001.jpg

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