Justet Cristian, Hernández Julio A, Chifflet Silvia
Departamento de Bioquímica, Facultad de Medicina, Universidad de la República, Gral. Flores 2125, 11800, Montevideo, Uruguay.
Sección Biofísica y Biología de Sistemas, Facultad de Ciencias, Universidad de la República, Iguá s/n esq. Mataojo, 11400, Montevideo, Uruguay.
Mol Cell Biochem. 2023 Jan;478(1):89-102. doi: 10.1007/s11010-022-04495-0. Epub 2022 Jun 21.
A mechanical injury in bovine corneal endothelial (BCE) cells in culture induces: (1) a fast calcium wave (FCW); (2) slow increases in cytosolic sodium and calcium, critical for the healing process, and (3) a rise in the apoptotic rate with respect to quiescent cells. In order to investigate the nature of the stimuli that determine the ionic changes and apoptotic response, we performed here studies on a non-injury model of tissue restitution in BCE monolayers. For this, we employed cell cultures grown to confluence in the presence of a Parafilm strip. We observed that, previously to strip removal, most of the border cells had already developed the slow ionic modifications, while in the scratch wounds these changes gradually occur after several hours of healing. This finding suggests that, in BCE cells, the presence of a free edge is sufficient to trigger ionic modifications necessary for wound healing and to elicit an augmented apoptotic response. The apoptotic index of the migrating cells in the Parafilm model (PF) was determined to be approximately two-fold the one of scratch wounds, a result that, in agreement with our previous observations, we attributed to the absence of the FCW in the PF experiments. The findings of this work further contribute to the understanding of epithelial wound healing, a crucial adaptive, and homeostatic response.
培养的牛角膜内皮(BCE)细胞受到机械损伤会引发:(1)快速钙波(FCW);(2)胞质钠和钙缓慢增加,这对愈合过程至关重要;(3)相对于静止细胞,凋亡率上升。为了探究决定离子变化和凋亡反应的刺激因素的本质,我们在此对BCE单层组织修复的非损伤模型进行了研究。为此,我们使用在Parafilm条存在下生长至汇合的细胞培养物。我们观察到,在去除条带之前,大多数边缘细胞已经发生了缓慢的离子变化,而在划痕伤口中,这些变化在愈合数小时后逐渐出现。这一发现表明,在BCE细胞中,自由边缘的存在足以触发伤口愈合所需的离子变化,并引发增强的凋亡反应。Parafilm模型(PF)中迁移细胞的凋亡指数被确定为划痕伤口的约两倍,这一结果与我们之前的观察结果一致,我们将其归因于PF实验中不存在FCW。这项工作的发现进一步有助于理解上皮伤口愈合,这是一种关键的适应性和稳态反应。