Brink Hallie E, Stalling Simone S, Nicoll Steven B
Department of Bioengineering, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
In Vitro Cell Dev Biol Anim. 2005 Sep-Oct;41(8-9):252-7. doi: 10.1290/0503023R.1.
The wound healing response to injury can be affected by many factors such as cell migration and extracellular matrix elaboration. The objective of this study was to examine the serum- and age-dependent effects on cell migration, adhesion, and collagen expression by skin fibroblasts. Dermal fibroblasts were isolated and plated with and without serum for up to 7 d. Cell migration was determined by quantitative image analysis, adhesion was quantified using a centrifugation assay, and collagen expression was assessed by PCR and immunohistochemical staining. Both adult and fetal fibroblasts migrated significantly faster in serum-containing medium compared to serum-free medium. There was no significant difference in migration between the two cell types in either serum-containing or serum-free medium. There was no significant difference in adhesion in the presence of serum, although there was a greater fraction of adherent fetal skin fibroblasts than adult fibroblasts in serum-free medium. Moreover, the adherent fraction of fetal fibroblasts in serum-free medium was not significantly different from that in serum-containing medium, suggesting that fetal skin fibroblasts possess serum-independent adhesion properties. Collagen mRNA expression was significantly up-regulated in serum-free compared to serum-containing medium for both cell types. With respect to collagen immunohistochemistry, both dermal fibroblast populations exhibited greater type I collagen compared to type III collagen staining. Quantitative assessment of collagen staining indicated significantly enhanced type I collagen secretion in the presence of serum by fetal skin fibroblasts. These findings suggest that intrinsic cellular characteristics may govern the observed differences in adult and fetal wound healing.
伤口对损伤的愈合反应会受到许多因素的影响,如细胞迁移和细胞外基质的形成。本研究的目的是检测血清和年龄对皮肤成纤维细胞的细胞迁移、黏附及胶原蛋白表达的影响。分离真皮成纤维细胞,在有血清和无血清条件下培养长达7天。通过定量图像分析确定细胞迁移,使用离心试验对黏附进行定量,并通过PCR和免疫组织化学染色评估胶原蛋白表达。与无血清培养基相比,成年和胎儿成纤维细胞在含血清培养基中的迁移速度均显著更快。在含血清或无血清培养基中,两种细胞类型之间的迁移没有显著差异。在有血清存在的情况下,黏附没有显著差异,尽管在无血清培养基中,贴壁的胎儿皮肤成纤维细胞比例高于成年成纤维细胞。此外,胎儿成纤维细胞在无血清培养基中的贴壁比例与在含血清培养基中的贴壁比例没有显著差异,这表明胎儿皮肤成纤维细胞具有不依赖血清的黏附特性。与含血清培养基相比,两种细胞类型在无血清培养基中胶原蛋白mRNA表达均显著上调。关于胶原蛋白免疫组织化学,与III型胶原蛋白染色相比,两种真皮成纤维细胞群体均表现出更强的I型胶原蛋白染色。胶原蛋白染色的定量评估表明,胎儿皮肤成纤维细胞在有血清存在的情况下I型胶原蛋白分泌显著增加。这些发现表明,内在细胞特性可能决定了观察到的成年和胎儿伤口愈合的差异。