异种移植心脏瓣膜的再细胞化可降低体内大鼠模型中的异种反应性免疫应答。

Recellularization of xenograft heart valves reduces the xenoreactive immune response in an in vivo rat model.

机构信息

Division of Cardiac Surgery, Department of Surgery, University of Alberta, Edmonton, AB, Canada.

Faculty of Medicine and Dentistry, University of Alberta, Edmonton, AB, Canada.

出版信息

Eur J Cardiothorac Surg. 2022 Jan 24;61(2):427-436. doi: 10.1093/ejcts/ezab439.

DOI:10.1093/ejcts/ezab439

PMID:34633028

Abstract

OBJECTIVES

Our aim was to address the role of autologous mesenchymal stem cell recellularization of xenogenic valves on the activation of the xenoreactive immune response in an in vivo rat model.

METHODS

Explanted aortic valve constructs from female Hartley guinea pigs were procured and decellularized, followed by recellularization with autologous Sprague-Dawley rat mesenchymal stem cells. Aortic valve xenografts were then implanted into the infrarenal aorta of recipient rats. Grafts were implanted as either autologous grafts, non-decellularized (NGP), decellularized and recellularized xenografts (RGP). Rats were euthanized after 7 and 21 days and exsanguinated and the grafts were explanted.

RESULTS

The NGP grafts demonstrated significant burden of granulocytes (14.3 cells/HPF) and CD3+ T cells (3.9 cells/HPF) compared to the autologous grafts (2.1 granulocytes/HPF and 0.72 CD3+ T cells/HPF) after 7 days. A lower absolute number of infiltrating granulocytes (NGP vs autologous, 6.4 vs 2.4 cells/HPF) and CD3+ T cells (NGP vs autologous, 2.8 vs 0.8 cells/HPF) was seen after 21 days. Equivalent granulocyte cell infiltration in the RGP grafts (2.4 cells/HPF) compared to the autologous grafts (2.1 cells/HPF) after 7 and 21 days (2.8 vs 2.4 cells/HPF) was observed. Equivalent CD3+ T-cell infiltration in the RGP grafts (0.63 cells/HPF) compared to the autologous grafts (0.72 cells/HPF) after 7 and 21 days (0.7 vs 0.8 cells/HPF) was observed. Immunoglobulin production was significantly greater in the NGP grafts compared to the autologous grafts at 7 (123.3 vs 52.7 mg/mL) and 21 days (93.3 vs 71.6 mg/mL), with a similar decreasing trend in absolute production. Equivalent immunoglobulin production was observed in the RGP grafts compared to the autologous grafts at 7 (40.8 vs 52.7 mg/mL) and 21 days (29.5 vs 71.6 mg/mL).

CONCLUSIONS

Autologous mesenchymal stem cell recellularization of xenogenic valves reduces the xenoreactive immune response in an in vivo rat model and may be an effective approach to decrease the progression of xenograft valve dysfunction.

摘要

目的

本研究旨在探讨同种异体骨髓间充质干细胞再细胞化异种瓣膜在体内大鼠模型中对异种反应性免疫应答的激活作用。

方法

从雌性 Hartley 豚鼠中取出已被取出的主动脉瓣组织，并进行去细胞化处理，随后用同种异体 Sprague-Dawley 大鼠骨髓间充质干细胞进行再细胞化。然后将异种移植物植入受体大鼠的肾下主动脉。移植物作为自体移植物、非去细胞化移植物（NGP）、去细胞化和再细胞化异种移植物（RGP）进行植入。在第 7 天和第 21 天，大鼠被安乐死并放血，然后取出移植物。

结果

与自体移植物（2.1 个粒细胞/高倍视野和 0.72 个 CD3+T 细胞/高倍视野）相比，在第 7 天，NGP 移植物的粒细胞（14.3 个/高倍视野）和 CD3+T 细胞（3.9 个/高倍视野）的负担明显更重。在第 21 天，观察到浸润性粒细胞（NGP 与自体，6.4 与 2.4 个/高倍视野）和 CD3+T 细胞（NGP 与自体，2.8 与 0.8 个/高倍视野）的绝对数量减少。在第 7 天和第 21 天，RGP 移植物的粒细胞浸润与自体移植物相似（2.4 个/高倍视野）（2.1 个/高倍视野）（2.8 个/高倍视野），CD3+T 细胞浸润也相似（0.63 个/高倍视野）（0.72 个/高倍视野）（0.7 个/高倍视野）。与自体移植物相比，在第 7 天（123.3 与 52.7mg/ml）和第 21 天（93.3 与 71.6mg/ml），NGP 移植物的免疫球蛋白产生明显更高，绝对产量呈下降趋势。在第 7 天（40.8 与 52.7mg/ml）和第 21 天（29.5 与 71.6mg/ml），RGP 移植物的免疫球蛋白产生与自体移植物相似。