Background

New materials are currently designed for efficient treatment of oral tissue lesions by guided tissue regeneration. The aim of this study was to develop a multifunctional 3D hybrid biomaterial consisting of extracellular matrix components, collagen, chondroitin 4-sulfate and fibronectin, functionalised with silver nanoparticles, intended to improve periodontitis treatment protocols.

Methods

Structural observations were performed by autometallography, scanning and transmission electron microscopy. In vitro tests of 3D constructs of embedded gingival fibroblasts within hybrid biomaterial were performed by MTS and Live/Dead assays. Genotoxicity was assessed by comet assay. In vivo experiments using chick embryo chorioallantoic membrane (CAM) assay analysed the degradation and nanoparticles release, but also angiogenesis, new tissue formation in 3D constructs and the regenerative potential of the hybrid material. Biological activity was investigated in experimental models of inflamed THP-1 macrophages and oral specific bacterial cultures.

Results

Light micrographs showed distribution of silver nanoparticles on collagen fibrils. Scanning electron micrographs revealed a microstructure with interconnected pores, which favoured cell adhesion and infiltration. Cell viability and proliferation were significantly higher within the 3D hybrid biomaterial than in 2D culture conditions, while absence of the hybrid material's genotoxic effect was found. In vivo experiments showed that the hybrid material was colonised by cells and blood vessels, initiating synthesis of new extracellular matrix. Besides the known effect of chondroitin sulfate, incorporated silver nanoparticles increased the anti-inflammatory activity of the hybrid biomaterial. The silver nanoparticles maintained their antibacterial activity even after embedding in the polymeric scaffold and inhibited the growth of F. nucleatum and P. gingivalis.

Conclusion

The novel biomimetic scaffold functionalised with silver nanoparticles presented regenerative, anti-inflammatory and antimicrobial potential for oral cavity lesions repair.

中文翻译：

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植入银纳米颗粒的仿生支架改善口腔损伤的体外和体内评价

背景

当前设计了新材料，用于通过引导的组织再生来有效治疗口腔组织损伤。这项研究的目的是开发一种多功能3D杂化生物材料，该材料由细胞外基质成分，胶原蛋白，4-硫酸软骨素和纤连蛋白组成，并用银纳米颗粒进行功能化，旨在改善牙周炎的治疗方案。

方法

通过自动金相，扫描和透射电子显微镜进行结构观察。通过MTS和活/死分析对混合生物材料中嵌入的牙龈成纤维细胞的3D构建体进行了体外测试。基因毒性通过彗星试验评估。使用鸡胚绒膜尿囊膜（CAM）分析的体内实验分析了降解和纳米颗粒释放，还分析了血管生成，3D构造中新组织的形成以及杂化材料的再生潜力。在发炎的THP-1巨噬细胞和口腔特定细菌培养物的实验模型中研究了生物活性。

结果

光学显微照片显示了银纳米颗粒在胶原纤维上的分布。扫描电子显微照片显示具有相互连接的孔的微结构，这有利于细胞粘附和浸润。3D杂交生物材料中的细胞活力和增殖明显高于2D培养条件，而没有发现杂交材料的遗传毒性作用。体内实验表明，杂化材料被细胞和血管定殖，开始合成新的细胞外基质。除了硫酸软骨素的已知作用外，掺入的银纳米颗粒还增强了杂化生物材料的抗炎活性。银纳米粒子甚至嵌入聚合物支架后仍保持其抗菌活性，并抑制了F. nucleatum和gingivalis的生长。

结论

用银纳米颗粒功能化的新型仿生支架具有再生，消炎和抗菌潜力，可修复口腔病变。