Morphological evaluation of the amniotic membrane decellularization

O. Ignatov, Adrian Cociug, O. Pascal, V. Nacu
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Abstract

Background: Biological materials derived from decellularized tissues could be a good basis for progress in regenerative medicine while maintaining the main components of the extracellular matrix. A promising scaffold for tissue-engineered is the human amniotic membrane. It is one of the oldest biomaterials used for scaffolds. Material and methods: 3 placentas were obtained through Human Tissue Bank. Under sterile condition human amniotic membrane was collected. The human amniotic membrane was treated with 0.5% of sodium dodecyl sulphate (SDS), 1% Triton for 24 and 5 hours. Amniotic membrane decellularization was also carried out in combination with ultrasound bath for 20 minutes 3 times. For morphological and structure evaluation of human amniotic membrane the scanning electron microscopy of native amniotic membrane and histology of decellularized and native amniotic membrane were performed. Results: The human amniotic membrane decellularization process with 0.5% SDS solution and 1% Triton solution showed that decellularization for 24 hours is too aggressive for human amniotic membrane structure. The decellularization for 5h with 1% Triton solution was incomplete. Conclusions: The method of decellularization with 0.5% SDS solution is more suitable for amniotic membrane decellularization and can be performed in only 5 hours. The use of ultrasound bath did not have a significant effect on the obtained results.
羊膜脱细胞的形态学评价
背景:从脱细胞组织中提取的生物材料可以为再生医学的发展奠定良好的基础,同时保持细胞外基质的主要成分。人羊膜是一种很有前途的组织工程支架。它是最古老的用于支架的生物材料之一。材料和方法:通过人体组织库获得3块胎盘。在无菌条件下采集人羊膜。人羊膜分别用0.5%十二烷基硫酸钠(SDS)和1% Triton处理24和5小时。同时联合超声浴进行羊膜脱细胞术,每次20分钟,共3次。对天然羊膜的扫描电镜、脱细胞羊膜和天然羊膜的组织学进行了形态学和结构评价。结果:0.5% SDS溶液和1% Triton溶液的人羊膜脱细胞过程表明,24小时的脱细胞对人羊膜结构的破坏太大。1% Triton溶液5h的脱细胞不完全。结论:0.5% SDS溶液脱细胞法更适合羊膜脱细胞,仅需5小时即可完成。超声浴的使用对所获得的结果没有显著影响。
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