Department of Dental Materials and Biomaterials Research, RWTH Aachen University Hospital, Pauwelsstrasse 30, 52074, Aachen, Germany.
Macromol Biosci. 2024 Feb;24(2):e2300162. doi: 10.1002/mabi.202300162. Epub 2023 Sep 27.
Collagen-type I gels are widely used for the fabrication of 3D in vitro gingival models. Unfortunately, their long-term stability is low, which limits the variety of in vitro applications. To overcome this problem and achieve better hydrolytic stability of 3D gingival models, fibrin-based hydrogel blends with increased long-term stability in vitro are investigated. Two different fibrin-based hydrogels are tested: fibrin 2.5% (w/v) and fibrin 1% (w/v)/gelatin 5% (w/v). Appropriate numbers of primary human gingival fibroblasts (HGFs) and OKG4/bmi1/TERT (OKG) keratinocytes are optimized to achieve a homogeneous distribution of cells under the assumed 3D conditions. Both hydrogels support the viability of HGFs and the stability of the hydrogel over 28 days. In vitro cultivation at the air-liquid interface triggers keratinization of the epithelium and increases its thickness, allowing the formation of multiple tissue-like layers. The presence of HGFs in the hydrogel further enhances epithelial differentiation. In conclusion, a fibrin-based 3D gingival model mimics the histology of native gingiva in vitro and ensures its long-term stability in comparison with the previously reported collagen paralogs. These results open new perspectives for extending the period within which specific biological or pathological conditions of artificial gingival tissue can be evaluated.
I 型胶原凝胶广泛用于制造 3D 体外牙龈模型。不幸的是，其长期稳定性较低，限制了体外应用的多样性。为了解决这个问题，并实现更好的 3D 牙龈模型的水解稳定性，研究了具有更高体外长期稳定性的纤维蛋白基水凝胶混合物。测试了两种不同的纤维蛋白基水凝胶：2.5%（w/v）纤维蛋白和 1%（w/v）纤维蛋白/5%（w/v）明胶。优化了适当数量的原代人牙龈成纤维细胞（HGFs）和 OKG4/bmi1/TERT（OKG）角质形成细胞，以在假设的 3D 条件下实现细胞的均匀分布。两种水凝胶都支持 HGFs 的活力和水凝胶在 28 天内的稳定性。在气液界面的体外培养可引发上皮的角化作用，并增加其厚度，从而形成多个组织样层。水凝胶中存在 HGFs 可进一步增强上皮分化。总之，基于纤维蛋白的 3D 牙龈模型在体外模拟了天然牙龈的组织学，并与以前报道的胶原类似物相比，确保了其长期稳定性。这些结果为延长人工牙龈组织特定生物或病理条件的评估期开辟了新的前景。
