ارزيابي قابليت توليد پوست مصنو‌عي مبتني بر تركيب سه عنصر: داربست (پرده آمنيوتيك )، سلول (سلول‌هاي بنيادي و فيبروبلاست) و فاكتورهاي محلول (محيط كشت شرطي)

eva‎luation of the ability to produce artificial skin based on the combination of three elements: scaffold (amniotic membrane), cells (stem cells an‎d fibroblast) an‎d soluble factors (conditioned medium)

زيست شناسي گياهي و جانوري

In the pursuit of developing innovative strategies for replacing damaged skin tissues, the use of biologically derived scaffolds from human sources particularly the human amniotic membrane (HAM) has attracted considerable attention as a promising option in skin tissue engineering. The aim of this study was to design an‎d eva‎luate a bilayer artificial skin using decellularized human amniotic membrane, human fibroblasts, an‎d epidermal stem cells, in the presence of bioactive factors derived from conditioned culture medium. Following a controlled decellularization process, the amniotic membrane was prepared in a manner that preserved the three-dimensional extracellular matrix structure, including type I, III, an‎d IV collagens, as well as its basal layer. The resulting scaffold was subsequently used as a substrate for cell culture, with fibroblasts seeded on the chorionic surface an‎d epidermal stem cells seeded on the amniotic surface. To assess biological responses, multiple assays were performed: the MTT assay to eva‎luate cell viability an‎d proliferation; hematoxylin an‎d eosin (H&E) an‎d Masson’s trichrome staining for histological examination; an‎d immunofluorescence analysis to determine proliferative indices. The findings indicated that the HAM scaffold retained its structural an‎d biological properties after decellularization an‎d was capable of supporting cell growth, although cell proliferation in certain groups was limited. Treatment with conditioned medium containing secreted factors had a positive effect on cell viability an‎d adhesion. These results are consistent with previous studies on the clinical application of HAM in skin repair, highlighting its potential as an effective scaffold for skin tissue regeneration. Overall, the design of this preliminary HAM-based artificial skin model represents a significant step forward in the development of tissue engineered products. Nevertheless, to comprehensively eva‎luate the efficacy of this system under in vivo conditions an‎d biomechanical dynamic models, further animal studies an‎d improvements toward creating multicellular an‎d vascularized artificial skin structures are recommended.

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