Augmented dried versus cryopreserved amniotic membrane as an ocular surface dressing.
Augmented dried versus cryopreserved amniotic membrane as an ocular surface dressing.
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DOI：
10.1371/journal.pone.0078441
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Hopkinson A
中科院分区：
文献类型：
--
作者：
Allen CL;Clare G;Stewart EA;Branch MJ;McIntosh OD;Dadhwal M;Dua HS;Hopkinson A
干羊膜（AM）可作为眼科手术中的一种有用的治疗辅助物，并且与冷冻保存的AM相比具有后勤优势。保存技术的差异可以显着影响AM的生化组成和物理性质，可能影响临床疗效。本研究旨在研究在不存在和存在糖冻干保护剂的情况下干燥AM的生物化学和结构效应及其与冷冻保存材料相比的生物相容性。将AM冷冻保存或干燥，用海藻糖或棉子糖和抗氧化剂表没食子儿茶素（EGCG）进行预处理或不进行预处理。使用电子显微镜评估结构和视觉比较。使用免疫测定和免疫荧光法确定AM生物因子的定位、表达和释放。采用细胞增殖、细胞毒性、细胞凋亡和细胞迁移试验评价AM与角膜上皮细胞（CEC）或角膜基质细胞单层共培养的生物相容性。干燥灭活AM上皮，但低于冷冻保存和细胞损伤减少干燥AM预处理海藻糖或棉子糖。单独干燥的AM，与海藻糖或棉子糖相比，冷冻保存的AM表现出更大的因子保留效率和生物利用度，并表现出更持久的生化因子体外时间释放。细胞健康分析表明，与冷冻保存的AM相比，具有海藻糖或棉子糖的干燥AM是相容的和上级的底物，用于初级CEC扩增，具有增加的增殖和降低的LDH和半胱天冬酶-3水平。与冷冻保存和新鲜AM相比，与干燥和海藻糖或棉子糖膜共培养的永生化人CEC系（hiCEC）中的伤口愈合改善支持了这一概念。我们改进的保存工艺和我们得到的优化的干燥AM具有增强的结构特性和生化稳定性，并且是常规冷冻保存AM的上级基质。此外，该产品稳定且易于运输，使其能够在全球范围内用于临床和军事领域。
Dried amniotic membrane (AM) can be a useful therapeutic adjunct in ophthalmic surgery and possesses logistical advantages over cryopreserved AM. Differences in preservation techniques can significantly influence the biochemical composition and physical properties of AM, potentially affecting clinical efficacy. This study was established to investigate the biochemical and structural effects of drying AM in the absence and presence of saccharide lyoprotectants and its biocompatibility compared to cryopreserved material. AM was cryopreserved or dried with and without pre-treatment with trehalose or raffinose and the antioxidant epigallocatechin (EGCG). Structural and visual comparisons were assessed using electron microscopy. Localisation, expression and release of AM biological factors were determined using immunoassays and immunofluorescence. The biocompatibility of the AM preparations co-cultured with corneal epithelial cell (CEC) or keratocyte monolayers were assessed using cell proliferation, cytotoxicity, apoptosis and migration assays. Drying devitalised AM epithelium, but less than cryopreservation and cellular damage was reduced in dried AM pre-treated with trehalose or raffinose. Dried AM alone, and with trehalose or raffinose showed greater factor retention efficiencies and bioavailability compared to cryopreserved AM and demonstrated a more sustained biochemical factor time release in vitro. Cellular health assays showed that dried AM with trehalose or raffinose are compatible and superior substrates compared to cryopreserved AM for primary CEC expansion, with increased proliferation and reduced LDH and caspase-3 levels. This concept was supported by improved wound healing in an immortalised human CEC line (hiCEC) co-cultured with dried and trehalose or raffinose membranes, compared to cryopreserved and fresh AM. Our modified preservation process and our resultant optimised dried AM has enhanced structural properties and biochemical stability and is a superior substrate to conventional cryopreserved AM. In addition this product is stable and easily transportable allowing it to be globally wide reaching for use in clinical and military sectors.
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影响因子：
2.4
作者：
Kitagawa, Kiyotaka;Okabe, Motonori;Hayashi, Atsushi
通讯作者：
Hayashi, Atsushi
影响因子：
2
作者：
Koizumi, N;Inatomi, T;Kinoshita, S
通讯作者：
Kinoshita, S
DOI：
10.1016/s0301-2115(00)00345-6
发表时间：
2001-02-01
期刊：
EUROPEAN JOURNAL OF OBSTETRICS GYNECOLOGY AND REPRODUCTIVE BIOLOGY
影响因子：
--
作者：
Kjaergaard, N;Hein, M;Madsen, H
通讯作者：
Madsen, H
影响因子：
4.4
作者：
Hopkinson, Andrew;McIntosh, Richard S.;Dua, Harminder S.
通讯作者：
Dua, Harminder S.
影响因子：
4.4
作者：
Hopkinson, Andrew;McIntosh, Richard S.;Dua, Harminder S.
通讯作者：
Dua, Harminder S.