Department of Obstetrics and Gynecology, Tufts Medical Center, Boston, MA 02111, USA.
Tissue Eng Part A. 2012 Mar;18(5-6):499-507. doi: 10.1089/ten.TEA.2011.0309. Epub 2011 Nov 1.
Cervical dysfunction contributes to a significant number of preterm births and is a common cause of morbidity and mortality in newborn infants. Cervical dysfunction is related to weakened load bearing properties of the collagen-rich cervical stroma. However, the mechanisms responsible for cervical collagen changes during pregnancy are not well defined. It is known that blood flow and oxygen tension significantly increase in reproductive tissues during pregnancy. To examine the effect of oxygen tension, a key mediator of tissue homeostasis, on the formation of cervical-like tissue in vitro, we grew primary human cervical cells in both two-dimensional (2D) and three-dimensional (3D) culture systems at 5% and 20% oxygen. Immunofluorescence studies revealed a stable fibroblast phenotype across six passages in all subjects studied (n=5). In 2D culture for 2 weeks, 20% oxygen was associated with significantly increased collagen gene expression (p<0.01), increased tissue wet weight (p<0.01), and increased collagen concentration (p=0.046). 3D cultures could be followed for significantly longer time frames than 2D cultures (12 weeks vs. 2 weeks). In contrast to 2D cultures, 20% oxygen in 3D cultures was associated with decreased collagen concentration (p<0.01) and unchanged collagen gene expression, which is similar to cervical collagen changes seen during pregnancy. We infer that 3D culture is more relevant for studying cervical collagen changes in vitro. The data suggest that increased oxygen tension may be related to significant cervical collagen changes seen in pregnancy.
宫颈功能障碍导致大量早产，是新生儿发病率和死亡率的常见原因。宫颈功能障碍与富含胶原蛋白的宫颈基质的承载能力减弱有关。然而，导致怀孕期间宫颈胶原变化的机制尚不清楚。众所周知，怀孕期间生殖组织的血流量和氧张力显著增加。为了研究氧张力这一组织内环境的关键调节因子对体外宫颈样组织形成的影响，我们在 5%和 20%氧浓度下，将原代人宫颈细胞分别在二维（2D）和三维（3D）培养系统中进行培养。免疫荧光研究显示，所有研究对象（n=5）在六个传代中均保持稳定的成纤维细胞表型。在 2D 培养 2 周时，20%的氧浓度与胶原基因表达显著增加（p<0.01）、组织湿重增加（p<0.01）和胶原浓度增加（p=0.046）相关。3D 培养可以比 2D 培养跟踪更长的时间框架（12 周 vs. 2 周）。与 2D 培养相反，3D 培养中的 20%氧浓度与胶原浓度降低（p<0.01）和胶原基因表达不变相关，这与怀孕期间宫颈胶原的变化相似。我们推断 3D 培养更适合体外研究宫颈胶原的变化。数据表明，氧张力增加可能与怀孕期间宫颈胶原的显著变化有关。
