Assessment of skin viability: is it necessary to use different methodologies?
Assessment of skin viability: is it necessary to use different methodologies?
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DOI：
10.1034/j.1600-0846.2003.00039.x
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发表时间：
2003-11-01
影响因子：
2.2
通讯作者：
Maillard, JY
中科院分区：
文献类型：
--
作者：
Messager, S;Hann, AC;Maillard, JY
背景/目的：皮肤是复杂的，可能会显示不同的结构和代谢变化'离体'。本研究的目的是遵循皮肤活力的措施，并评估其有用性作为violence.Materials和方法的标记：我们评估了新鲜或冷冻后，随后解冻的皮肤样本的活力。评估包括组织病理学外观，乳酸脱氢酶（LDH）的活性，耗氧量和皮肤pH.Results：新鲜和冷冻的皮肤样本，使用光学和电子显微镜的形态学调查显示，相对明确的表皮和真皮样品。冷冻样本显示出角质层碎裂的一些迹象，尽管这并不明显。在4 ℃保存的新鲜样品中测得的LDH活性较低，但可稳定长达7天。在32 ℃下保存的新鲜样品与在冰箱中保存长达4天的样品具有相当的LDH活性。冷冻样品，解冻，然后保持在4 ℃孵育24小时后显示稳定的LDH活性。然而，在32 ℃下孵育的冷冻样品显示出结果的高度可变性，孵育5天后LDH活性高达800 U/L。新鲜切除以及新鲜解冻的样品显示出最高的呼吸速率。新鲜和解冻的样品储存了很长一段时间有一个显着较低的（有时不存在）的耗氧率。我们的研究结果还表明，新鲜样品在32 ℃孵育24 h后，耗氧速率增加。所有样品的耗氧速率在15分钟测量期内达到平台，即使新鲜样品也没有耗尽培养基中的所有氧气。离体皮肤样品的pH值明显高于体内人体皮肤，在32 ℃下孵育46 h后，新鲜样品的pH值明显低于冷冻样品。所有的协议是可重复的，新鲜切除和新鲜解冻的皮肤样品表现出最高的速率violence.Conclusion：离体皮肤显示随着时间的推移的几个参数的变化。建议使用两种或三种技术来评估皮肤活力，至少包括氧测量和酶测定。
Background/aim: Skin is complex and may display variable structural and metabolic change 'ex vivo'. The present study aimed to follow measures of skin viability and evaluate their usefulness as markers of viability.Materials and methods: We evaluated the viability of skin samples fresh or after being frozen and subsequently thawed. Assessments included histopathological appearance, lactate dehydrogenase (LDH) activity, oxygen consumption and skin pH.Results: Morphological investigations of fresh and frozen skin samples using light and electron microscopy showed samples with relatively well-defined epidermis and dermis. Frozen samples showed some sign of stratum corneum fragmentation, although this was not obvious. LDH activity measured in fresh samples kept at 4degreesC was low, but it was stable up to 7 days. Fresh samples kept at 32degreesC had a comparable LDH activity to the ones kept in the fridge up to 4 days. Frozen samples, thawed and then kept at 4degreesC showed a stable LDH activity after 24 h of incubation. However, frozen samples incubated at 32degreesC demonstrated a high variability in results, with up to 800 U/L of LDH activity after 5 days of incubation. Freshly excised as well as freshly thawed samples showed the highest respiration rates. Fresh and thawed samples stored for a long period of time had a significantly lower (sometimes non-existent) oxygen consumption rate. Our results also showed an increase in the oxygen consumption rate of fresh samples being incubated at 32degreesC for 24 h. The oxygen consumption rate for all samples reached a plateau within the 15-min measurement period and even the fresh samples did not deplete all the oxygen from the medium. Skin samples ex vivo showed a significantly higher pH than human skin in vivo, and when incubated for 46 h at 32degreesC, fresh samples had a significantly lower pH than frozen samples. All protocols were reproducible and freshly excised and freshly thawed skin samples showed the highest rates of viability.Conclusion: ex vivo skin shows variation of several parameters over time. It is recommended to use two or three techniques for evaluation of skin viability including at least oxygen measurement and an enzyme assay.