用于骨再生的 Zn-EGCG 负载壳聚糖支架的制作与评估:从细胞反应到体内性能。

IF 7.7 1区 化学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY International Journal of Biological Macromolecules Pub Date : 2024-11-15 DOI:10.1016/j.ijbiomac.2024.137695
Yu Yang, Wei Sun, Qiang Fu, Zhongyuan Wang, Hui Zhao, Zaijun Wang, Yuzhong Gao, Jian Wang
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引用次数: 0

摘要

我们通过将锌与绿茶中的黄酮类化合物表没食子儿茶素-3-棓酸盐(EGCG)螯合,合成了一种黄酮类金属复合物(FMC),并将其加入壳聚糖(CS)中,通过冷冻干燥法形成三维结构。扫描电子显微镜观察了支架的表面形态和孔隙尺寸,发现支架中存在相互连接的多孔网络。支架显示出最佳孔径(大于 50 μm),有利于骨组织的生长和新生血管的形成。在 CS 基质中加入 Zn-EGCG 可提高抗压强度(0.53±0.045 兆帕),并通过控制膨胀减少酶降解,从而改善力学性能。此外,在最初的一小时内观察到蛋白质吸附增加,这对细胞附着至关重要。此外,FMC 的加入促进了 CS 支架的外源性生物矿化,最早可在模拟体液中持续 4 天。间接细胞毒性测定表明,含有 Zn-EGCG 的支架对小鼠间充质干细胞(mMSCs)没有毒性作用。在成骨环境下,支架通过上调 ALP 活性和增加 Runx2、ColI、OC 和 OPN 等成骨细胞分化标志物的表达,促进了 mMSCs 的钙沉积。我们发现,支架的成骨潜力背后有 miR-15b/smurf-1 信号通路的参与。使用小鸡胚胎 CAM 试验进行的体内评估显示,血管生成得到了增强,并证实了该支架的生物相容性和无毒性。此外,在斑马鱼规模再生模型中,该支架增强了钙沉积和成骨细胞标记表达,与体外研究结果一致。总之,研究结果表明,该支架的成骨潜力如下 壳聚糖
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Fabrication and evaluation of Zn-EGCG-loaded chitosan scaffolds for bone regeneration: From cellular responses to in vivo performance.

We have synthesized a flavonoid metal complex (FMC) by chelating zinc to epigallocatechin-3-gallate (EGCG), a flavonoid present in green tea and incorporated into chitosan (CS) to form 3D constructs by freeze drying method. Scanning electron microscopy characterized The scaffolds for surface morphology and pore dimensions and depicted the presence of interconnected porous network. The scaffolds exhibited optimal pore size (>50 μm), facilitating bone tissue ingrowth and neovascularization. Inclusion of Zn-EGCG into CS matrix improved the mechanical property by increasing compressive strength (0.53±0.045 MPa) and reducing enzymatic degradation with controlled swelling. In addition, increased protein adsorption was observed during the initial hour, which is crucial for cell attachment. Furthermore, the FMC inclusion promoted exogenous biomineralization of CS scaffolds as early as 4d in simulated body fluid. Indirect cytotoxicity measurements indicated the scaffolds with Zn-EGCG had no toxic effects on mouse mesenchymal stem cells (mMSCs). Under osteogenic environment, the scaffold promoted calcium deposition of mMSCs by upregulation of ALP activity and increased expression of osteoblast differentiation markers such as Runx2, ColI, OC and OPN. We found that the involvement of miR-15b/smurf-1 signalling pathway behind the osteogenic potential of the scaffold. In vivo assessments using the chick embryo CAM assay showed enhanced angiogenesis and confirmed the scaffold's biocompatibility with no toxicity. Additionally, in a zebrafish scale regeneration model, the scaffold enhanced calcium deposition and osteoblast marker expression, aligning with the in vitro findings. Overall, form the study it is clear that the osteogenic potential of the scaffold is as follows chitosan < EGCG-Chitosan < Zn-EGCG-Chitosan.

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来源期刊
International Journal of Biological Macromolecules
International Journal of Biological Macromolecules 生物-生化与分子生物学
CiteScore
13.70
自引率
9.80%
发文量
2728
审稿时长
64 days
期刊介绍: The International Journal of Biological Macromolecules is a well-established international journal dedicated to research on the chemical and biological aspects of natural macromolecules. Focusing on proteins, macromolecular carbohydrates, glycoproteins, proteoglycans, lignins, biological poly-acids, and nucleic acids, the journal presents the latest findings in molecular structure, properties, biological activities, interactions, modifications, and functional properties. Papers must offer new and novel insights, encompassing related model systems, structural conformational studies, theoretical developments, and analytical techniques. Each paper is required to primarily focus on at least one named biological macromolecule, reflected in the title, abstract, and text.
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