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来自海洋海绵的生物二氧化硅和海绵硬蛋白的3D打印支架:用于骨组织修复的遗传毒性和细胞毒性分析

3D printed scaffolds of biosilica and spongin from marine sponges: analysis of genotoxicity and cytotoxicity for bone tissue repair.

作者信息

Dos Santos Jorge Sousa Karolyne, de Souza Amanda, de Almeida Cruz Matheus, de Lima Lindiane Eloisa, do Espirito Santo Giovanna, Amaral Gustavo Oliva, Granito Renata Neves, Renno Ana Claudia

机构信息

Department of Biosciences, Federal University of São Paulo (UNIFESP), Lab 342, 136 Silva Jardim Street, Santos, SP, 11015020, Brazil.

出版信息

Bioprocess Biosyst Eng. 2024 Sep;47(9):1483-1498. doi: 10.1007/s00449-024-03042-z. Epub 2024 Jun 13.

Abstract

Biosilica (BS) and spongin (SPG) from marine sponges are highlighted for their potential to promote bone regeneration. Moreover, 3D printing is introduced as a technology for producing bone grafts with optimized porous structures, allowing for better cell attachment, proliferation, and differentiation. Thus, this study aimed to characterize the BS and BS/SPG 3D printed scaffolds and to evaluate the biological effects in vitro. The scaffolds were printed using an ink containing 4 wt.% of sodium alginate. The physicochemical characteristics of BS and BS/SPG 3D printed scaffolds were analyzed by SEM, EDS, FTIR, porosity, evaluation of mass loss, and pH measurement. For in vitro analysis, the cellular viability of the MC3T3-E1 cell lineage was assessed using the AlamarBlue assay and confocal microscopy, while genotoxicity and mineralization potential were evaluated through the micronucleus assay and Alizarin Red S, respectively. SEM analysis revealed spicules in BS, the fibrillar structure of SPG, and material degradation over the immersion period. FTIR indicated peaks corresponding to silicon oxide in BS samples and carbon oxide and amine in SPG samples. BS-SPG scaffolds exhibited higher porosity, while BS scaffolds displayed greater mass loss. pH measurements indicated a significant decrease induced by BS, which was mitigated by SPG over the experimental periods. In vitro studies demonstrated the biocompatibility and non-cytotoxicity of scaffold extracts. .Also, the scaffolds promoted cellular differentiation. The micronucleus test further confirmed the absence of genotoxicity. These findings suggest that 3D printed BS and BS/SPG scaffolds may possess desirable morphological and physicochemical properties, indicating in vitro biocompatibility.

摘要

来自海洋海绵的生物二氧化硅(BS)和海绵硬蛋白(SPG)因其促进骨再生的潜力而受到关注。此外,3D打印作为一种生产具有优化多孔结构骨移植体的技术被引入,这种结构有利于更好的细胞附着、增殖和分化。因此,本研究旨在表征BS和BS/SPG 3D打印支架,并评估其体外生物学效应。支架使用含有4 wt.%海藻酸钠的墨水进行打印。通过扫描电子显微镜(SEM)、能谱仪(EDS)、傅里叶变换红外光谱仪(FTIR)、孔隙率、质量损失评估和pH测量来分析BS和BS/SPG 3D打印支架的物理化学特性。对于体外分析,使用alamarBlue检测法和共聚焦显微镜评估MC3T3-E1细胞系的细胞活力,同时分别通过微核试验和茜素红S评估遗传毒性和矿化潜力。SEM分析显示BS中有针状体、SPG有纤维状结构,且在浸泡期间材料有降解。FTIR表明BS样品中对应氧化硅的峰以及SPG样品中对应二氧化碳和胺的峰。BS-SPG支架表现出更高的孔隙率,而BS支架的质量损失更大。pH测量表明BS会导致显著下降,而在实验期间SPG可缓解这种下降。体外研究证明了支架提取物的生物相容性和无细胞毒性。此外,支架促进了细胞分化。微核试验进一步证实了无遗传毒性。这些发现表明3D打印的BS和BS/SPG支架可能具有理想的形态和物理化学性质,显示出体外生物相容性。

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