Gibson Angela L F, Shatadal Shalini
Department of Surgery, University of Wisconsin-Madison School of Medicine and Public Health, Madison, Wisconsin.
Department of Surgery, University of Wisconsin-Madison School of Medicine and Public Health, Madison, Wisconsin.
J Surg Res. 2017 Jul;215:83-87. doi: 10.1016/j.jss.2017.03.064. Epub 2017 Apr 7.
Cell viability is paramount to wound healing in burn injury. Current methods to determine depth of burn injury in the research setting are based on the subjective visualization of cell viability using hematoxylin and eosin staining. The purpose of this study was to develop a simplified method of lactate dehydrogenase (LDH) staining to identify viable cells in frozen sections of human burn tissue that can be used in the research setting.
After surgical excision, human burn tissue was processed for histologic evaluation. Tissues were fixed and protected with sucrose incubation before cryopreservation. An LDH staining method was developed and evaluated for prolonged stain stability. To evaluate cellular viability in the tissues as demonstrated by enzymatic activity of LDH, digital images of tissue sections were obtained immediately after and 1 mo after staining.
The cryopreserved sections of deep partial thickness human burn tissue revealed cellular viability throughout the tissue with the exception of the most superficial region of the tissue. Unlike the hematoxylin and eosin-stained sections, clear demarcation of cellular viability was evident in the LDH-stained sections.
Our simplified protocol identifies, without ambiguity, the viability of the cellular elements in deep partial thickness and full thickness burn injured tissue.
细胞活力对于烧伤创面愈合至关重要。目前在研究中确定烧伤深度的方法是基于苏木精和伊红染色主观观察细胞活力。本研究的目的是开发一种简化的乳酸脱氢酶（LDH）染色方法，以识别可用于研究的人烧伤组织冰冻切片中的活细胞。
手术切除后，对人烧伤组织进行组织学评估。在冷冻保存前，组织经固定并用蔗糖孵育进行保护。开发并评估了一种LDH染色方法以延长染色稳定性。为了通过LDH的酶活性评估组织中的细胞活力，在染色后立即和染色后1个月获取组织切片的数字图像。
人深Ⅱ度烧伤组织的冷冻切片显示，除组织最表层区域外，整个组织均有细胞活力。与苏木精和伊红染色切片不同，LDH染色切片中细胞活力的界限清晰可见。
我们的简化方案能够明确鉴定深Ⅱ度和全层烧伤损伤组织中细胞成分的活力。
