OMFS IMPATH Research Group, Faculty of Medicine, Department of Imaging and Pathology, KU Leuven and Oral and Maxillofacial Surgery, University Hospitals Leuven, Kapucijnenvoer 33, 3000, Leuven, Belgium.
Department of Oral Health Sciences, KU Leuven and Paediatric Dentistry and Special Dental Care, University Hospitals Leuven, Kapucijnenvoer 33, 3000, Leuven, Belgium.
Sci Rep. 2021 Mar 11;11(1):5687. doi: 10.1038/s41598-021-84405-4.
Tailored hydrogels mimicking the native extracellular environment could help overcome the high variability in outcomes within regenerative endodontics. This study aimed to evaluate the effect of the chemokine-binding and antimicrobial polymer, chlorite-oxidized oxyamylose (COAM), on the microstructural properties of fibrin and self-assembling peptide (SAP) hydrogels. A further goal was to assess the influence of the microstructural differences between the hydrogels on the in vitro behavior of human dental pulp stem cells (hDPSCs). Structural and mechanical characterization of the hydrogels with and without COAM was performed by atomic force microscopy and scanning electron microscopy to characterize their microstructure (roughness and fiber length, diameter, straightness, and alignment) and by nanoindentation to measure their stiffness (elastic modulus). Then, hDPSCs were encapsulated in hydrogels with and without COAM. Cell viability and circularity were determined using confocal microscopy, and proliferation was determined using DNA quantification. Inclusion of COAM did not alter the microstructure of the fibrin hydrogels at the fiber level while affecting the SAP hydrogel microstructure (homogeneity), leading to fiber aggregation. The stiffness of the SAP hydrogels was sevenfold higher than the fibrin hydrogels. The viability and attachment of hDPSCs were significantly higher in fibrin hydrogels than in SAP hydrogels. The DNA content was significantly affected by the hydrogel type and the presence of COAM. The microstructural stability after COAM inclusion and the favorable hDPSCs' response observed in fibrin hydrogels suggest this system as a promising carrier for COAM and application in endodontic regeneration.
模仿天然细胞外环境的定制水凝胶可以帮助克服再生牙髓学中结果的高度变异性。本研究旨在评估趋化因子结合和抗菌聚合物亚氯酸盐氧化羟胺（COAM）对纤维蛋白和自组装肽（SAP）水凝胶微观结构特性的影响。进一步的目标是评估水凝胶之间微观结构差异对人牙髓干细胞（hDPSCs）体外行为的影响。通过原子力显微镜和扫描电子显微镜对含和不含 COAM 的水凝胶进行结构和机械特性表征，以表征其微观结构（粗糙度和纤维长度、直径、直线度和取向），并通过纳米压痕测量其刚度（弹性模量）。然后，将 hDPSCs 包埋在含和不含 COAM 的水凝胶中。使用共聚焦显微镜测定细胞活力和圆度，并使用 DNA 定量法测定增殖。COAM 的包含并未改变纤维蛋白水凝胶的纤维水平的微观结构，而影响 SAP 水凝胶的微观结构（均匀性），导致纤维聚集。SAP 水凝胶的刚度比纤维蛋白水凝胶高七倍。hDPSCs 的活力和附着在纤维蛋白水凝胶中明显高于 SAP 水凝胶。DNA 含量受水凝胶类型和 COAM 的存在显著影响。在包含 COAM 后微观结构的稳定性以及在纤维蛋白水凝胶中观察到的 hDPSCs 的良好反应表明该系统作为 COAM 的有前途的载体，并可应用于牙髓再生。
