In Vitro Effects of Three Blood Derivatives on Human Corneal Epithelial Cells
In Vitro Effects of Three Blood Derivatives on Human Corneal Epithelial Cells
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DOI：
10.1167/iovs.11-7340
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发表时间：
2012-08-01
影响因子：
4.4
通讯作者：
Morales, Maria-Celia
中科院分区：
文献类型：
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作者：
Freire, Vanesa;Andollo, Noelia;Morales, Maria-Celia
目的.我们比较了三种血液衍生物，自体血清（AS），富血小板血浆（PRP）和富含生长因子血浆衍生血清（PRGF）对人角膜上皮（HCE）细胞系的影响，以评估其作为角膜上皮疾病有效治疗的潜力。通过ELISA定量表皮生长因子（EGF）、成纤维细胞生长因子（FGF）、血管内皮生长因子（VEGF）、肝细胞生长因子（HGF）、血小板衍生生长因子（PDGF）和纤连蛋白的浓度。用MTT比色法检测HCE细胞的增殖和活力。通过相差显微镜评估细胞形态。几个连接蛋白，外皮蛋白，整合素α 6基因的表达模式进行了分析，通过实时RT-PCR。我们发现PRGF中的EGF水平显著高于AS和PRP。然而，AS和PRGF诱导HCE细胞的稳健增殖。此外，PRGF培养的细胞生长为异质集落，表现出分化和未分化的细胞表型，而AS和PRP处理的培养物表现出相当均匀的集落。最后，与AS或PRP相比，PRGF上调了与通讯和细胞分化相关的几个基因的表达。PRGF促进角膜上皮形成所需的生物过程，如增殖和分化。由于PRGF的作用与常规使用的血液衍生物相似，因此目前的研究结果需要进一步研究PRGF作为眼表疾病的新替代治疗方法。（Invest Ophthalmol维斯科学。2012; 53：5571-5578）DOI：10.1167/iovs.11-7340
PURPOSE. We compared the effects of three blood derivatives, autologous serum (AS), platelet-rich plasma (PRP), and serum derived from plasma rich in growth factors (PRGF), on a human corneal epithelial (HCE) cell line to evaluate their potential as an effective treatment for corneal epithelial disorders.METHODS. The concentrations of epidermal growth factor (EGF), fibroblast growth factor (FGF), vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), platelet-derived growth factor (PDGF), and fibronectin were quantified by ELISA. The proliferation and viability of HCE cells were measured by an 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) colorimetric assay. Cell morphology was assessed by phase-contrast microscopy. The patterns of expression of several connexin, involucrin, and integrin alpha 6 genes were analyzed by real-time RT-PCR.RESULTS. We found significantly higher levels of EGF in PRGF compared to AS and PRP. However, AS and PRGF induced robust proliferation of HCE cells. In addition, PRGF cultured cells grew as heterogeneous colonies, exhibiting differentiated and non-differentiated cell phenotypes, whereas AS-and PRP-treated cultures exhibited quite homogeneous colonies. Finally, PRGF upregulated the expression of several genes associated with communication and cell differentiation, in comparison to AS or PRP.CONCLUSIONS. PRGF promotes biological processes required for corneal epithelialization, such as proliferation and differentiation. Since PRGF effects are similar to those associated with routinely used blood derivatives, the present findings warrant further research on PRGF as a novel alternative treatment for ocular surface diseases. (Invest Ophthalmol Vis Sci. 2012; 53:5571-5578) DOI:10.1167/iovs.11-7340