Karim Aos S, Yan Amy, Ocotl Edgar, Bennett Daniel D, Wang Ziyue, Kendziorski Christina, Gibson Angela L F
Department of Surgery, University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin.
Department of Dermatology, University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin.
Wound Repair Regen. 2019 Mar;27(2):150-161. doi: 10.1111/wrr.12692. Epub 2018 Dec 26.
The regenerative capacity of burn wounds, and the need for surgical intervention, depends on wound depth. Clinical visual assessment is considered the gold standard for burn depth assessment but it remains a subjective and inaccurate method for tissue evaluation. The purpose of this study was to compare visual assessment with microscopic and molecular techniques for human burn depth determination, and illustrate differences in the evaluation of tissue for potential regenerative capacity. Using intraoperative visual assessment, patients were identified as having deep partial thickness or full thickness burn wounds. Tangential excisions of burn tissue were processed with hematoxylin and eosin to visualize tissue morphology, lactate dehydrogenase assay to ascertain cellular viability, and Keratin-15 and Ki67 to identify epidermal progenitor cells and proliferative capacity, respectively. RNA from deep partial and full thickness burn tissue as well as normal tissue controls were submitted for RNA sequencing. Lactate dehydrogenase, Keratin-15, and Ki67 were found throughout the excised burn wound tissue in both deep partial thickness burn tissues and in the second tangential excision of full thickness burn tissues. RNA sequencing demonstrated regenerative capacity in both deep partial and full thickness burn tissue, however a greater capacity for regeneration was present in deep partial thickness compared with full thickness burn tissues. In this study, we highlight the discordance that exists between the intraoperative clinical identification of burn injury depth, and microscopic and molecular determination of viability and regenerative capacity. Current methods utilizing visual assessment for depth of injury are imprecise, and can lead to removal of viable tissue. Additionally, hematoxylin and eosin microscopic analysis should not be used as the sole method in research or clinical determination of depth, as there are no differences in staining between viable and nonviable tissue.
烧伤创面的再生能力以及手术干预的必要性取决于创面深度。临床视觉评估被认为是烧伤深度评估的金标准，但它仍然是一种主观且不准确的组织评估方法。本研究的目的是比较视觉评估与微观和分子技术在确定人类烧伤深度方面的差异，并阐明在评估具有潜在再生能力的组织时存在的差异。通过术中视觉评估，确定患者患有深二度或三度烧伤创面。对烧伤组织进行削痂处理，用苏木精和伊红染色以观察组织形态，进行乳酸脱氢酶测定以确定细胞活力，分别用角蛋白-15和Ki67来鉴定表皮祖细胞和增殖能力。将深二度和三度烧伤组织以及正常组织对照的RNA送去进行RNA测序。在深二度烧伤组织和三度烧伤组织的第二次削痂组织中，均在切除的烧伤创面组织中发现了乳酸脱氢酶、角蛋白-15和Ki67。RNA测序表明深二度和三度烧伤组织均具有再生能力，然而与三度烧伤组织相比，深二度烧伤组织的再生能力更强。在本研究中，我们强调了术中临床对烧伤损伤深度的识别与微观和分子对活力及再生能力的测定之间存在的不一致。目前利用视觉评估损伤深度的方法并不精确，可能导致切除有活力的组织。此外，苏木精和伊红微观分析不应作为研究或临床确定深度的唯一方法，因为有活力和无活力组织之间的染色没有差异。
