ارزيابي قابليت توليد پوست مصنوعي مبتني بر تركيب سه عنصر: داربست (پرده‌ي آمنيوتيك)، سلول (سلول‌هاي بنيادي و فيبروبلاست) و فاكتورهاي محلول (عصاره پوست)

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

زيست شناسي گياهي و جانوري

Effective healing of skin wounds remains one of the major challenges in regenerative medicine an‎d tissue engineering. The use of natural scaffolds such as the human amniotic membrane, due to its high biocompatibility an‎d favorable matrix properties, can offer a novel an‎d efficient approach for skin tissue regeneration. In this study, with the aim of investigating the effects of human skin tissue extract on the growth, organization, an‎d matrix synthesis of skin cells, a decellularized human amniotic membrane was used as a biological scaffold. First, the amniotic membrane was isolated from fresh human placentas an‎d decellularized using physical an‎d chemical methods. The success of the decellularization process was confirmed through H&E, DAPI, an‎d immunofluorescence staining. Subsequently, human skin tissue extract was prepared, an‎d its optimal concentration was determined using the MTT assay. In the next stage, epidermal stem cells an‎d fibroblasts were cultured on the epithelial an‎d stromal surfaces of the membrane, respectively, an‎d treatment with the tissue extract an‎d conditioned medium was applied in different groups. Histological analyses using H&E, DAPI, an‎d immunofluorescence staining revealed that treatment with the skin tissue extract led to increased cell density, improved regeneration of the epithelial layer, an‎d greater organization in the arrangement of matrix fibers. Moreover, Masson’s trichrome staining an‎d software-based analysis of collagen content showed that collagen synthesis in the treated groups was significantly higher than in the control group. These findings indicate the positive effect of tissue extract treatment on enhancing fibroblast activity an‎d extracellular matrix regeneration. Overall, the results of this study suggest that the use of human skin extract alongside the natural scaffold of the amniotic membrane can be an effective strategy for improving the conditions of three-dimensional culture of skin cells an‎d for the development of bioactive dressings an‎d tissue-engineered systems for skin wound healing.

4