Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran.
Biomed Mater. 2019 Mar 27;14(3):035008. doi: 10.1088/1748-605X/ab0679.
Today advancements in nanotechnology have made extensive progress in tissue engineering. Application of cerium oxide nanoparticles (CeO) has improved regenerative medicine due to their antioxidant properties. In this study, nanoparticles were used to increase the efficacy of skin substitutes. Human skin samples were decellularized using four methods and studied via histological stainings and DNA content analyses. Then CeO dispersing and its stability were investigated. The prepared acellular dermal matrices (ADMs) were immersed in CeO suspension and their effects were evaluated on growth of cultured human adipose derived-mesenchymal stem cells (hAd-MSCs) using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and histological methods. Moreover, their antioxidant properties were assessed based on DPPH degradation. Changes in the collagen contents of the scaffolds containing cells and CeO were also determined by electron microscopy and their tensile strength was compared to ADM. Our results indicated that use of trypsin/NaOH protocol resulted in most efficient cell removal while maintaining extracellular matrix (ECM) architecture. Among different dispersal methods, the approach using Dulbecco's modified Eagle's medium (DMEM), wetting with fetal bovine serum (FBS) and ultrasonic bath resulted in the best stability. Furthermore, it was shown that CeO not only had no toxicity on the cells, but also increased the growth and survival of hAd-MSCs by about 27%, improved free radical scavenging, as well as the amount of collagen and tensile strength of the scaffolds containing nanoparticles compared to the ADM. It can be concluded that the combination of ADM/CeO/hAd-MSCs could be a step forward in skin tissue engineering.
如今,纳米技术的进步在组织工程领域取得了广泛的进展。由于具有抗氧化特性,氧化铈纳米粒子(CeO)的应用改善了再生医学。在这项研究中,使用纳米粒子来提高皮肤替代物的功效。使用四种方法对人体皮肤样本进行脱细胞处理,并通过组织学染色和 DNA 含量分析进行研究。然后研究了 CeO 的分散及其稳定性。将制备的去细胞真皮基质(ADM)浸入 CeO 悬浮液中,并通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴化物(MTT)测定法和组织学方法评估其对培养的人脂肪间充质干细胞(hAd-MSCs)生长的影响。此外,根据 DPPH 降解评估其抗氧化性能。还通过电子显微镜确定了含有细胞和 CeO 的支架中胶原蛋白含量的变化,并将其拉伸强度与 ADM 进行了比较。我们的结果表明,使用胰蛋白酶/NaOH 方案可以有效地去除细胞,同时保持细胞外基质(ECM)结构。在不同的分散方法中,使用 Dulbecco 改良 Eagle 培养基(DMEM)、胎牛血清(FBS)润湿和超声浴的方法可获得最佳的稳定性。此外,结果表明 CeO 不仅对细胞无毒,而且还可使 hAd-MSCs 的生长和存活增加约 27%,提高自由基清除能力以及含有纳米粒子的支架中的胶原蛋白含量和拉伸强度,与 ADM 相比。可以得出结论,ADM/CeO/hAd-MSCs 的组合可能是皮肤组织工程的一个进步。