Biomedical Research Centre, School of Medicine, Universidad de Valparaíso, Valparaíso, Chile.
J Tissue Eng Regen Med. 2013 Jan;7(1):10-9. doi: 10.1002/term.490. Epub 2011 Nov 3.
Several studies have developed efficient oral mucosa constructs using different types of scaffold. However, the changes in the morphology and gene and protein expression profile that could occur in these artificial constructs remain unknown. This study compared the histology and expression of several extracellular matrix molecules in human artificial oral mucosa developed using two different types of scaffolds: fibrin and fibrin-agarose. To that end, bioengineered oral mucosa stromas were constructed from biopsy samples of human oral mucosa and the substitute generated was analyzed at different periods of time in culture. Histological analysis was carried out by light and transmission electron microscopy and the expression of collagen types I, III, and VI, the proteoglycans decorin and biglycan, and the different chains of laminin, were assessed by immunoperoxidase technique. This study found that fibrin scaffolds accelerated fibroblast growth and remodeling of the scaffold, thus enhancing collagen fibrillogenesis. In the fibrin-agarose scaffold, the morphology and organization of the fibroblasts did not change during the culture period. All extracellular matrix proteins analyzed were expressed in both scaffolds. However, in fibrin scaffolds, these proteins were widely distributed and replaced the scaffold during the follow-up period. These results show that the substitutes generated showed histological and molecular similarities with native human oral mucosa stroma. In addition, it was observed that the nature of the biomaterial influenced the behaviour of the oral stromal fibroblasts, thereby modulating their growth, protein synthesis, and collagen fibrillogenesis.
已有多项研究利用不同类型的支架成功构建了高效的口腔黏膜结构，但这些人工构建体的形态和基因及蛋白表达谱可能发生的变化仍不清楚。本研究比较了两种不同类型支架（纤维蛋白和纤维蛋白-琼脂糖）构建的人人工口腔黏膜的组织学和几种细胞外基质分子的表达。为此，从人口腔黏膜活检样本中构建了生物工程口腔黏膜基质，并在培养的不同时间对生成的替代物进行了分析。通过光镜和透射电镜进行组织学分析，并通过免疫过氧化物酶技术评估了 I 型、III 型和 VI 型胶原、蛋白聚糖decorin 和 biglycan 以及层粘连蛋白的不同链的表达。本研究发现纤维蛋白支架加速了成纤维细胞的生长和支架的重塑，从而增强了胶原原纤维的形成。在纤维蛋白-琼脂糖支架中，培养期间成纤维细胞的形态和结构没有改变。两种支架中均表达了所有分析的细胞外基质蛋白。然而，在纤维蛋白支架中，这些蛋白广泛分布并在随访期间取代了支架。这些结果表明，生成的替代物在组织学和分子水平上与人天然口腔黏膜基质具有相似性。此外，还观察到生物材料的性质影响了口腔基质成纤维细胞的行为，从而调节了它们的生长、蛋白合成和胶原原纤维的形成。
