Department of Anatomy, Poznań University of Medical Science, 6 Święcickiego St., 60-781 Poznań, Poland.
Department of Biomaterials and Experimental Dentistry, Poznań University of Medical Sciences, 61-701 Poznań, Poland.
Int J Mol Sci. 2018 Mar 29;19(4):1027. doi: 10.3390/ijms19041027.
The oral mucosal tissue is a compound structure composed of morphologically and physiologically different cell types. The morphological modification involves genetically determined lifespan, which may be recognized as the balance between cell survival and apoptosis. Although the biochemical processes and pathways in oral mucosa, with special regards to drug transport, delivery, and metabolism, are well known, the cellular physiological homeostasis in this tissue requires further investigation. The porcine buccal pouch mucosal cells (BPMCs) collected from 20 pubertal crossbred Landrace gilts, were used in this study. Immediately after recovery, the oral mucosa was separated micro-surgically, and treated enzymatically. The dispersed cells were transferred into primary in vitro culture systems for a long-term cultivation of 30 days. After each step of in vitro culture (IVC), the cells were collected for isolation of total RNA at 24 h, 7, 15, and 30 days of IVC. While the expression was analyzed for days 7, 15, and 30, the 24th hour was used as a reference for outcome calibration. The gene expression profile was determined using Affymetrix microarray assays and necessary procedures. In results, we observed significant up-regulation of , , , , , , , , , , , , , , and genes, belonging to two ontological groups, namely "positive regulation of metabolic process", and "regulation of homeostatic process" at 7 day of IVC as compared to down-regulation at days 15 and 30. These findings suggest that the metabolic processes and homeostatic regulations are much more intense in porcine mucosal cells at day 7 of IVC. Moreover, the increased expression of marker genes, for both of these ontological groups, may suggest the existence of not only "morphological lifespan" during tissue keratinization, but also "physiological checkpoint" dedicated to metabolic processes in oral mucosa. This knowledge may be useful for preclinical experiments with drugs delivery and metabolism in both animals and humans.
口腔黏膜组织是一种由形态和生理上不同的细胞类型组成的复合结构。形态学改变涉及遗传决定的寿命，这可能被认为是细胞存活和凋亡之间的平衡。尽管口腔黏膜的生化过程和途径，特别是药物的转运、传递和代谢，已经被很好地了解，但该组织中的细胞生理稳态需要进一步研究。本研究中使用了从 20 头青春期杂交长白母猪采集的猪颊囊黏膜细胞（BPMCs）。口腔黏膜在恢复后立即进行显微外科分离，并进行酶处理。分散的细胞被转移到原代体外培养系统中进行长达 30 天的长期培养。在体外培养（IVC）的每一步后，在 IVC 的第 24、7、15 和 30 天收集细胞以分离总 RNA。虽然在第 7、15 和 30 天分析了表达情况，但第 24 小时被用作结果校准的参考。使用 Affymetrix 微阵列测定和必要的程序确定基因表达谱。结果，我们观察到在第 7 天的 IVC 中，属于两个本体论组的基因 、 、 、 、 、 、 、 、 、 、 、 和 显著上调，与第 15 和 30 天的下调相比。这些发现表明，在第 7 天的 IVC 中，猪黏膜细胞的代谢过程和体内平衡调节更为剧烈。此外，这两个本体论组的标记基因表达增加，可能不仅表明组织角化过程中存在“形态寿命”，而且还表明口腔黏膜中存在专门用于代谢过程的“生理检查点”。这些知识可能对动物和人类的药物输送和代谢的临床前实验有用。
