Abstract The effect of bioresorbable materials on aging in cultured mouse NIH 3T3 fibroblasts treated with elevated glucose concentration was investigated. The cells were grown on films produced from the silkworm fibroin and rS1/9, a recombinant analog of Nephila clavipes spidroin 1. Exposure to 50 mM glucose of the cells grown on uncoated glass support resulted in the cell growth retardation. The average areas of the cells and nuclei and the percentage of apoptotic cells increased, whereas the amount of soluble collagen decreased. In contrast, on the fibroin and spidroin films, the cell density and the percentage of 5-bromo-2′-deoxyuridine (BrdU)-positive cells were higher vs. the cells grown on the glass support. The films protected NIH 3T3 fibroblasts from the glucose-induced death. The most prominent effects on the cell density, BrdU incorporation, and apoptosis prevention were observed in the cells cultured on spidroin films. Unlike the cells grown on glass support (decrease in the soluble collagen production) or fibroin (no effect), production of soluble collagen by the cells grown on spidroin films increased after cell exposure to 50 mM glucose. Molecular analysis demonstrated that 50 mM glucose upregulated phosphorylation of the NFκB heterodimer p65 subunit in the cells grown on the glass support. The treatment of cells grown on fibroin films with 5.5 mM or 50 mM glucose had no effect on p65 phosphorylation. The same treatment decreased p65 phosphorylation in the cells on the spidroin films. These results demonstrate the anti-aging efficacy of biomaterials derived from the silk proteins and suggest that spidroin is more advantageous for tissue engineering and therapy than fibroin.

中文翻译：

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重组蜘蛛丝膜减弱了 NIH 3T3 成纤维细胞中葡萄糖诱导衰老的个体标志物

摘要 研究了生物可吸收材料对用升高的葡萄糖浓度处理的培养的小鼠 NIH 3T3 成纤维细胞衰老的影响。细胞在由蚕丝蛋白和 rS1/9（一种 Nephila clavipes spidroin 1 的重组类似物）制成的薄膜上生长。在未包被的玻璃支持物上生长的细胞暴露于 50 mM 葡萄糖导致细胞生长迟缓。细胞和细胞核的平均面积以及凋亡细胞的百分比增加，而可溶性胶原蛋白的量减少。相比之下，在丝心蛋白和蜘蛛蛋白膜上，细胞密度和 5-溴-2'-脱氧尿苷 (BrdU) 阳性细胞的百分比高于玻璃支持物上生长的细胞。这些薄膜保护 NIH 3T3 成纤维细胞免受葡萄糖诱导的死亡。对细胞密度最显着的影响，在蜘蛛蛋白膜上培养的细胞中观察到 BrdU 掺入和细胞凋亡预防。与在玻璃支持物上生长的细胞（可溶性胶原蛋白的产生减少）或丝心蛋白（无影响）不同，在细胞暴露于 50 mM 葡萄糖后，在蛛丝蛋白膜上生长的细胞产生的可溶性胶原蛋白增加。分子分析表明，在玻璃支持物上生长的细胞中，50 mM 葡萄糖上调了 NFκB 异二聚体 p65 亚基的磷酸化。用 5.5 mM 或 50 mM 葡萄糖处理在丝心蛋白膜上生长的细胞对 p65 磷酸化没有影响。相同的处理降低了蜘蛛蛋白膜上细胞中的 p65 磷酸化。