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氢氧化钠暴露对体外绵羊模型食管上皮细胞的影响:对食管组织工程的启示。

Effects of sodium hydroxide exposure on esophageal epithelial cells in an in vitro ovine model: implications for esophagus tissue engineering.

机构信息

Experimental Fetal Surgery and Tissue Engineering Unit, Department of Pediatric and Adolescent Surgery, Medical University of Graz, Auenbruggerplatz-34, A-8036 Graz, Austria.

出版信息

J Pediatr Surg. 2012 May;47(5):874-80. doi: 10.1016/j.jpedsurg.2012.01.033.

Abstract

BACKGROUND

Esophagus tissue engineering holds promises for esophageal replacement after severe caustic injuries. The aim of this study was to determine whether viable esophageal epithelial cells could be isolated from an esophagus exposed to varying concentrations of alkali with regard to number, viability, and morphology during in vitro culture.

METHODS

Ovine esophagi were exposed to phosphate-buffered saline 2.5%, 15%, or 25% sodium hydroxide (NaOH). The effect of NaOH concentrations on epithelial damage was assessed histologically. Esophageal epithelial cells were then isolated, and cell count and viability were investigated. Finally, cell number, viability, and morphology of esophageal epithelial cells were determined for 24 days of in vitro culture.

RESULTS

Histologic analysis showed a progressive destruction of the epithelium proportional to increasing NaOH concentrations. Esophagi treated with phosphate-buffered saline and 2.5% NaOH showed significantly higher viable cell counts after isolation and culture in comparison with those treated with 15% to 5% NaOH.

CONCLUSION

The evidence presented in this study indicates that epithelial biopsies from an esophagus exposed to low concentrations (2.5%) of NaOH will still yield large numbers of viable cells suitable for tissue engineering applications. In cases of exposure to higher concentrations (15%-25%), alternative cell sources for epithelial regeneration, such as stem cells, will be necessary for tissue engineering applications.

摘要

背景

食管组织工程有望在严重腐蚀性损伤后替代食管。本研究旨在确定暴露于不同浓度碱的食管中是否可以分离出具有活力的食管上皮细胞,以及在体外培养期间细胞数量、活力和形态的变化。

方法

将绵羊食管暴露于磷酸缓冲盐水 2.5%、15%或 25%氢氧化钠(NaOH)中。通过组织学评估 NaOH 浓度对上皮损伤的影响。然后分离食管上皮细胞,并研究细胞计数和活力。最后,在体外培养 24 天,确定食管上皮细胞的数量、活力和形态。

结果

组织学分析表明,上皮的破坏程度与 NaOH 浓度的增加成正比。与用 15%至 5%NaOH 处理的相比,用磷酸缓冲盐水和 2.5%NaOH 处理的食管在分离和培养后显示出明显更高的活细胞计数。

结论

本研究提供的证据表明,暴露于低浓度(2.5%)NaOH 的食管上皮活检仍可获得大量适合组织工程应用的有活力的细胞。在暴露于较高浓度(15%-25%)的情况下,需要其他细胞来源(如干细胞)用于上皮再生,以满足组织工程应用的需要。

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