Sher Ada C, Liu Lumei, Izem Melwan, Calyeca Jazmin, Meeker Molly O, Chiang Tendy
The Ohio State University College of Medicine, Columbus, Ohio, USA.
Center of Regenerative Medicine, Abigail Wexner Research Institute, Nationwide Children's Hospital, Columbus, Ohio, USA.
Laryngoscope. 2025 Feb;135(2):614-621. doi: 10.1002/lary.31801. Epub 2024 Oct 5.
A critical barrier to successful tracheal transplantation is poor vascularization. Despite its importance, little is known about microvascular regeneration in tissue-engineered grafts. We have demonstrated that partially decellularized tracheal grafts (PDTG) support neotissue formation including new submucosal microvasculature (CD31+). However, the perfusion of this neovasculature is unknown. In this study, we used a mouse model of tracheal replacement to measure the microvascular regeneration and perfusion of PDTG.
PDTG and syngeneic tracheal grafts (STG, surgical control) (n = 5 for each group) were orthotopically transplanted into C5BL/6 J mice. We quantified vascularity of STG and PDTG samples at 1 and 3 months with conventional histology (N = 3 ~ 10/group). At 1, 3, and 6 months, animals were injected with fluorescein isothiocyanate (FITC) tomato lectin into the left ventricle. After perfusion, tracheas were fixed, harvested, mounted, stained for CD31 expression, and imaged with resonant scanning confocal microscopy. Percent CD31+, FITC area was compared between groups and endpoints compared with native trachea. Microvascular intersections were quantified using Sholl analysis.
Functional microvasculature was seen in both groups. Although percent vascularization (CD31) in PDTG was restored by 3 months, microvascular pattern in PDTG displayed a unique morphology compared with control. Surgery alone appeared to globally change microvascular pattern and perfusion. PDTG demonstrated equivalent perfusion to surgical control by 6 months. Sholl analysis revealed a reduction of microvessel intersectionality that persisted in PDTG and was not seen in surgical or native controls.
PDTG exhibited microvascular regeneration. Perfusion was present in PDTG, improved, and persisted over long-term time points.
NA Laryngoscope, 135:614-621, 2025.
气管移植成功的一个关键障碍是血管化不良。尽管其很重要，但关于组织工程移植物中的微血管再生知之甚少。我们已经证明，部分脱细胞气管移植物（PDTG）可支持新组织形成，包括新的黏膜下微血管（CD31+）。然而，这种新生血管的灌注情况尚不清楚。在本研究中，我们使用气管置换小鼠模型来测量PDTG的微血管再生和灌注情况。
将PDTG和同基因气管移植物（STG，手术对照）（每组n = 5）原位移植到C57BL/6J小鼠体内。我们在1个月和3个月时用传统组织学方法（每组N = 3~10）对STG和PDTG样本的血管化情况进行定量分析。在1个月、3个月和6个月时，将异硫氰酸荧光素（FITC）番茄凝集素注入动物左心室。灌注后，固定气管，取出，包埋，进行CD31表达染色，并用共振扫描共聚焦显微镜成像。比较各组之间CD31+、FITC面积百分比，并将各时间点与天然气管进行比较。使用Sholl分析对微血管交叉点进行定量分析。
两组均可见功能性微血管。虽然PDTG中的血管化百分比（CD31）在3个月时恢复，但与对照组相比，PDTG中的微血管模式呈现出独特的形态。单纯手术似乎会全面改变微血管模式和灌注情况。到6个月时，PDTG的灌注情况与手术对照组相当。Sholl分析显示，微血管交叉性降低，这种情况在PDTG中持续存在，而在手术或天然对照组中未观察到。
PDTG表现出微血管再生。PDTG中存在灌注，灌注情况得到改善，并在长期时间点持续存在。
无《喉镜》，2025年，第135卷，第614 - 621页
