Development of age-less tissue-engineered oral mucosa substitute

WST-8活性降低和GLUT-1表达增加提示角质形成细胞在低氧培养中代谢由氧化代谢向无氧糖酵解转变。细胞周期图显示低氧条件下细胞G0/G1期停滞。此外，我们结合细胞周期分析评估了p21的表达，因为处于G1期的“周期细胞”不表达p21。低氧培养中p21的表达较低，提示低氧培养中存在较多的G1细胞。因此，细胞在低氧条件下表现出显著较高的集落形成效率(CFE)是由于循环细胞的比例较高。最后，利用组织工程口腔黏膜(TEOM)的等效物，我们研究了低氧对常规和长期培养的口腔黏膜上皮结构的影响。组织学检查显示，在低氧条件下，TEOM上皮细胞的动态平衡得到较好的维持。

Decreasing of WST-8 activity and increasing of GLUT-1 expression suggested that keratinocytes metabolism were shift oxidative metabolism to anaerobic glycolysis in hypoxic culture. Cell cycle profile shows G0/G1 arrest in hypoxic culture. In addition, we assessed p21 expression concomitant with cell cycle analysis because "cycling cells" in G1 phase do not express p21. p21 expression were lower in hypoxic culture suggested that greater number of G1 cells existed in hypoxic culture. Thus, significant higher colony forming efficiency (CFE) shown in cells under hypoxic conditions was due to a higher proportion of cycling cells. Finally, using a tissue-engineered oral mucosa (TEOM) equivalent, we investigated the effect of hypoxia on the epithelial structure for regular and long-term cultivation. The histological examinations showed that the epithelial homeostasis of the TEOM was better sustained in hypoxic condition.