Abstract:
The most recent progress in reconstructive therapy for the management of periodontitis and peri-implantitis bone defects has relied on the development of highly porous biodegradable bioaerogels for guided bone regeneration. The objective of this work was to evaluate in vitro the osteoinduction of periodontal-originating cells (human dental follicle mesenchymal cells, DFMSCs) promoted by a nano-hydroxyapatite/chitosan (nHAp/CS) bioaerogel, which was purified and sterilized by a sustainable technique (supercritical CO2). Moreover, the in vivo bone regeneration capacity of the nHAp/CS bioaerogel was preliminarily assessed as a proof-of-concept on a rat calvaria bone defect model. The quantification of DNA content of DFMSCs seeded upon nHAp/CS and CS scaffolds (control material) showed a significant increase from the 14th to the 21st day of culture. These results were corroborated through confocal laser scanning microscopy analysis (CLSM). Furthermore, the alkaline phosphatase (ALP) activity increased significantly on the 21st day, similarly for both materials. Moreover, the presence of nHAp promoted a significantly higher expression of osteogenic genes after 21 days when compared to CS scaffolds and control. CLSM images of 21 days of culture also showed an increased deposition of OPN over the nHAp/CS surface. The in vivo bone formation was assessed by microCT and histological analysis. The in vivo evaluation showed a significant increase in bone volume in the nHAp/CS test group when compared to CS and the empty control, as well as higher new bone formation and calcium deposition within the nHAp/CS structure. Overall, the present study showed that the nHAp/CS bioaerogel could offer a potential solution for periodontal and peri-implant bone regeneration treatments since the in vitro results demonstrated that it provided favorable conditions for DFMSC proliferation and osteogenic differentiation, while the in vivo outcomes confirmed that it promoted higher bone ingrowth.
摘要:
用于牙周炎和种植体周围炎的骨缺损的重建治疗的最新进展依赖于高度多孔的可生物降解的生物气凝胶的开发,用于引导骨再生。这项工作的目的是评估体外诱导牙周起源细胞(人牙囊间充质细胞,DFMSC)由纳米羟基磷灰石/壳聚糖(nHAp/CS)生物气凝胶促进,通过可持续技术(超临界CO2)进行纯化和灭菌。此外,初步评估了nHAp/CS生物气凝胶的体内骨再生能力,作为大鼠颅骨骨缺损模型的概念验证。接种在nHAp/CS和CS支架(对照材料)上的DFMSC的DNA含量的定量显示从培养的第14天到第21天显著增加。通过共聚焦激光扫描显微镜分析(CLSM)证实了这些结果。此外,碱性磷酸酶(ALP)活性在第21天显著增加,这两种材料也是如此。此外,与CS支架和对照相比,nHAp的存在在21天后促进了成骨基因的表达。培养21天的CLSM图像还显示OPN在nHAp/CS表面上的沉积增加。通过显微CT和组织学分析评估体内骨形成。体内评估显示,与CS和空对照相比,nHAp/CS测试组的骨体积显着增加,以及nHAp/CS结构内更高的新骨形成和钙沉积。总的来说,本研究表明,nHAp/CS生物气凝胶可以为牙周和种植体周围骨再生治疗提供潜在的解决方案,因为体外结果表明它为DFMSC增殖和成骨分化提供了有利条件,而体内结果证实,它促进了更高的骨向内生长。