Litwiniuk M, Radowicka M, Krejner A, Grzela T
PhD student, Department of Histology and Embryology, Medical University of Warsaw.
PhD student, First Department of Gynaecology and Obstetrics, Medical University of Warsaw.
J Wound Care. 2017 Aug 2;26(8):498-503. doi: 10.12968/jowc.2017.26.8.498.
The amniotic membrane (AM) is a rich source of biologically active factors, important for wound healing and is widely used in various clinical applications, including tissue engineering, reconstructive surgery and wound management. The aim of the present proof-of-concept study was to assess the influence of amniotic membrane extracts on in vitro proliferation of main cells involved in tissue regeneration. The assessment was done in regards to the content of selected biologically active factors in amniotic membrane extracts.
The quantitative analysis of EGF, TGF-β and TIMP-1 in tested samples was assayed by enzyme-linked immunosorbent assay (ELISA) tests. The influence of amniotic membrane extracts on proliferation of keratinocytes (HaCaT), fibroblasts (Wi-38) and endothelial cell lines (HECa-10) was assessed using a colorimetric tetrazolium salt reduction assay.
In all of the amnion samples high amounts of EGF, TGF-β and TIMP-1 were detected. However, the content of these factors varied between placental and cervical portions of the same membrane. Moreover, various concentrations of biologically active factors between physiological at-term delivery and caesarean section-derived membranes were also observed. All of the assessed amnion extracts stimulated proliferation of HaCaT and Wi-38 cells, although samples prepared from caesarean section-derived cervical portion of amniotic membrane stimulated more proliferation of keratinocytes than of fibroblasts. In contrast to HaCaT and Wi-38 cells, proliferation of HECa-10 cell line was inhibited by all tested extracts.
The results of our proof-of-concept study confirm that biological dressings prepared from amniotic membrane, especially its placental portion, since they stimulated both fibroblasts and keratinocytes, may provide relevant support for wound healing. On the other hand, dressings prepared from caesarean section-derived cervical portion of amniotic membrane, since they stimulate mainly epidermal cells, may be suitable for some specific applications, where more selective action is required, such as in ocular surgery. However, verification of this observation requires further studies.
羊膜是生物活性因子的丰富来源，对伤口愈合很重要，并且广泛应用于各种临床领域，包括组织工程、重建手术和伤口处理。本概念验证研究的目的是评估羊膜提取物对组织再生中主要细胞体外增殖的影响。该评估是针对羊膜提取物中选定生物活性因子的含量进行的。
通过酶联免疫吸附测定（ELISA）试验对测试样品中的表皮生长因子（EGF）、转化生长因子-β（TGF-β）和金属蛋白酶组织抑制因子-1（TIMP-1）进行定量分析。使用比色四唑盐还原试验评估羊膜提取物对角质形成细胞（HaCaT）、成纤维细胞（Wi-38）和内皮细胞系（HECa-10）增殖的影响。
在所有羊膜样本中均检测到大量的EGF、TGF-β和TIMP-1。然而，这些因子的含量在同一膜的胎盘部分和宫颈部分之间有所不同。此外，还观察到足月顺产和剖宫产来源的羊膜之间生物活性因子的浓度各异。所有评估的羊膜提取物均刺激了HaCaT和Wi-38细胞的增殖，尽管从剖宫产来源的羊膜宫颈部分制备的样品对角质形成细胞的刺激增殖作用比对成纤维细胞更强。与HaCaT和Wi-38细胞相反，所有测试提取物均抑制了HECa-10细胞系的增殖。
我们的概念验证研究结果证实，由羊膜制备的生物敷料，尤其是其胎盘部分，由于它们能刺激成纤维细胞和角质形成细胞，可能为伤口愈合提供相关支持。另一方面，由剖宫产来源的羊膜宫颈部分制备的敷料，由于它们主要刺激表皮细胞，可能适用于一些需要更具选择性作用（例如眼科手术）的特定应用。然而，这一观察结果的验证需要进一步研究。
