Richter Gresham T, Mehta Deepak, Albert David, Elluru Ravindhra G
Department of Pediatric Otolaryngology, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio 45208, USA.
Arch Otolaryngol Head Neck Surg. 2009 Jan;135(1):45-52. doi: 10.1001/archoto.2008.516.
To develop a novel mouse model of acquired subglottic stenosis (SGS) using heterotopic transplanted laryngotracheal complexes (LTCs).
Pilot randomized controlled animal study.
Forty-eight C57BL/6 mice.
Twenty-four donor C57BL/6 mice underwent LTC harvesting. The LTCs were then implanted deep to a cutaneous dorsal flap in 24 allogenic recipients. Sixteen LTCs underwent direct subglottic injury before transplantation, while 8 control LTCs were transplanted without injury. Transplanted LTCs were harvested 1, 2, 3, and 4 weeks after surgery. Tissues were fixed and cut transversely in 6-microm sections from the larynx to the second tracheal ring. Movat pentachrome staining was performed for connective tissue and morphometric analysis. Digital images of the subglottis were captured at x 20 magnification. The thicknesses of the lamina propria and the epithelium were measured at 5 random and equally spaced locations within the subglottic lumen. Vascular endothelial growth factor 164 (VEGF 164) and transforming growth factor beta1 (TGF-beta1) immunohistochemistry was performed on representative sections.
Lamina propria thickness was significantly greater in transplanted LTCs 3 and 4 weeks after injury compared with controls (P = .03, P = .01, respectively). Combined results (groups harvested at 1-4 weeks) revealed a significant difference between all 8 control animals and all 16 experimental animals (P < .001). Epithelial thickness was also greater in the transplanted LTCs 2, 3, and 4 weeks after injury to the subglottis compared with controls (P = .04 for weeks 2 through 4). Movat pentachrome staining showed random distributions and high concentrations of connective tissue within the lamina propria of the subglottis. The VEGF 164 and TGF-beta1 staining patterns were consistent with previous in vivo models of SGS.
Heterotopic transplanted LTCs in mice can provide an inexpensive and flexible model for experimental investigation of acquired SGS.
利用异位移植喉气管复合体（LTCs）建立一种新型获得性声门下狭窄（SGS）小鼠模型。
初步随机对照动物研究。
48只C57BL/6小鼠。
24只供体C57BL/6小鼠接受LTC采集。然后将LTC植入24只同种异体受体的皮肤背侧皮瓣深部。16个LTC在移植前接受直接声门下损伤，而8个对照LTC未受伤直接移植。术后1、2、3和4周采集移植的LTC。组织固定后，从喉至第二气管环横向切成6微米切片。进行Movat五色染色以进行结缔组织和形态计量分析。在20倍放大倍数下拍摄声门下的数字图像。在声门下腔内5个随机且等距的位置测量固有层和上皮的厚度。对代表性切片进行血管内皮生长因子164（VEGF 164）和转化生长因子β1（TGF-β1）免疫组织化学检测。
与对照组相比，损伤后3周和4周移植的LTC中固有层厚度显著增加（分别为P = 0.03，P = 0.01）。综合结果（1 - 4周采集的组）显示，所有8只对照动物与所有16只实验动物之间存在显著差异（P < 0.001）。与对照组相比，声门下损伤后2、3和4周移植的LTC中上皮厚度也更大（第2至4周P = 0.04）。Movat五色染色显示声门下固有层内结缔组织分布随机且浓度高。VEGF 164和TGF-β1染色模式与先前的SGS体内模型一致。
小鼠异位移植LTC可为获得性SGS的实验研究提供一种廉价且灵活的模型。
