猪模型中去细胞化并固定的异种/同种异体基质的血管生物相容性增强及重塑

Enhanced Vascular Biocompatibility and Remodeling of Decellularized and Secured Xenogeneic/Allogeneic Matrices in a Porcine Model.

作者信息

van Steenberghe Mathieu, Schubert Thomas, Bouzin Caroline, Caravaggio Carlo, Guiot Yves, Xhema Daela, Gianello Pierre

机构信息

Pôle de Chirurgie Expérimentale et Transplantation (CHEX), Institut de Recherche Expérimentale et Clinique, Secteur des Sciences de la Santé, Université Catholique de Louvain, Brussels, Belgium.

Banque de Tissus, Unité de Thérapie Cellulaire et Tissulaire de L'Appareil Locomoteur, Cliniques Universitaires Saint-Luc, Brussels, Belgium.

出版信息

Eur Surg Res. 2018;59(1-2):58-71. doi: 10.1159/000487591. Epub 2018 Apr 5.

DOI:10.1159/000487591

PMID:29621750

Abstract

BACKGROUND/PURPOSE: Calcifications and absence of growth potential are the major drawbacks of glutaraldehyde-treated prosthesis. Decellularized and secured xeno-/allogeneic matrices were assessed in a preclinical porcine model for biocompatibility and vascular remodeling in comparison to glutaraldehyde-fixed bovine pericardium (GBP; control).

METHODS

Native human (fascia lata, pericardium) and porcine tissues (peritoneum) were used and treated. In vitro, biopsies were performed before and after treatment to assess decellularization (hematoxylin and eosin/DAPI). In vivo, each decellularized and control tissue sample was implanted subcutaneously in 4 mini-pigs. In addition, 9 mini-pigs received a patch or a tubularized prosthesis interposition on the carotid artery or abdominal aorta of decellularized (D) human fascia lata (DHFL; n = 4), human pericardium (DHP; n = 9), porcine peritoneum (DPPt; n = 7), and control tissue (GBP: n = 3). Arteries were harvested after 1 month and subcutaneous samples after 15-30 days. Tissues were processed for hematoxylin and eosin/von Kossa staining and immunohistochemistry for CD31, alpha-smooth muscle actin, CD3, and CD68. Histomorphometry was achieved by point counting.

RESULTS

A 95% decellularization was confirmed for DHP and DPPt, and to a lower degree for DHFL. In the subcutaneous protocol, CD3 infiltration was significantly higher at day 30 in GBP and DHFL, and CD68 infiltration was significantly higher for GBP (p < 0.05). In intravascular study, no deaths, aneurysms, or pseudoaneurysms were observed. Inflammatory reaction was significantly higher for DHFL and GBP (p < 0.05), while it was lower and comparable for DHP/DPPt. DHP and DPPt showed deeper recellularization, and a new arterial wall was characterized.

CONCLUSIONS

In a preclinical model, DPPt and DHP offered better results than conventional commercialized GBP for biocompatibility and vascular remodeling.

摘要

背景/目的：钙化和缺乏生长潜力是戊二醛处理过的假体的主要缺点。与戊二醛固定的牛心包（GBP；对照）相比，在临床前猪模型中评估了脱细胞并固定的异种/同种异体基质的生物相容性和血管重塑情况。

方法

使用并处理了天然人类组织（阔筋膜、心包）和猪组织（腹膜）。在体外，在处理前后进行活检以评估脱细胞情况（苏木精和伊红染色/DAPI）。在体内，将每个脱细胞组织样本和对照组织样本皮下植入4只小型猪体内。此外，9只小型猪接受了脱细胞的人阔筋膜（DHFL；n = 4）、人心包（DHP；n = 9）、猪腹膜（DPPt；n = 7）和对照组织（GBP：n = 3）制成的补片或管状假体在颈动脉或腹主动脉上的置入。1个月后采集动脉样本，15 - 30天后采集皮下样本。对组织进行苏木精和伊红染色/冯·科萨染色以及针对CD31、α - 平滑肌肌动蛋白、CD3和CD68的免疫组织化学检测。通过点计数进行组织形态计量分析。

结果

证实DHP和DPPt的脱细胞率为95%，DHFL的脱细胞率较低。在皮下实验方案中，GBP和DHFL在第30天时CD3浸润显著更高，GBP的CD68浸润显著更高（p < 0.05）。在血管内研究中，未观察到死亡、动脉瘤或假性动脉瘤。DHFL和GBP的炎症反应显著更高（p < 0.05），而DHP/DPPt的炎症反应更低且相当。DHP和DPPt显示出更深的再细胞化，并形成了新的动脉壁特征。