羊模型中胎儿脊柱裂修复术皮肤闭合用同种异体移植物

Allografts for Skin Closure during In Utero Spina Bifida Repair in a Sheep Model.

作者信息

Mann Lovepreet K, Won Jong Hak, Patel Rajan, Bergh Eric P, Garnett Jeannine, Bhattacharjee Meenakshi B, Narayana Ponnada A, Jain Ranu, Fletcher Stephen A, Lai Dejian, Papanna Ramesha

机构信息

Division of Maternal-Fetal Medicine, The Fetal Center at Children's Memorial Hermann Hospital, Department of Obstetrics, Gynecology and Reproductive Medicine, McGovern Medical School, The University of Texas Health Science Center at Houston (UTHealth), Houston, TX 77030, USA.

Department of Diagnostic and Interventional Imaging, McGovern Medical School, The University of Texas Health Science Center at Houston (UTHealth), Houston, TX 77030, USA.

出版信息

J Clin Med. 2021 Oct 25;10(21):4928. doi: 10.3390/jcm10214928.

DOI:10.3390/jcm10214928

PMID:34768448

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8584988/

Abstract

OBJECTIVES

Use of off-label tissue graft materials, such as acellular dermal matrix (ADM), for in utero repair of severe spina bifida (SB), where primary skin layer closure is not possible, is associated with poor neurological outcomes. The cryopreserved human umbilical cord (HUC) patch has regenerative, anti-inflammatory, and anti-scarring properties, and provides watertight SB repair. We tested the hypothesis that the HUC is a superior skin patch to ADM for reducing inflammation at the repair site and preserving spinal cord function.

METHODS

In timed-pregnant ewes with twins, on gestational day (GD) 75, spina bifida was created without a myelotomy (functional model). On GD 95, repair was performed using HUC vs. ADM patches (randomly assigned) by suturing them to the skin edges. Additionally, full thickness skin closure as a primary skin closure (PSC) served as a positive control. Delivery was performed on GD 140, followed by blinded to treatment neurological assessments of the lambs using the Texas Spinal Cord Injury Scale (TSCIS) for gait, proprioception, and nociception. Lambs without spina bifida were used as controls (CTL). Ex vivo magnetic resonance imaging of spines at the repair site were performed, followed by quantitative pathological assessments. Histological assessments (blinded) included Masson's trichrome, and immunofluorescence for myeloperoxidase (MPO; neutrophils) and for reactive astrocytes (inflammation) by co-staining vimentin and GFAP.

RESULTS

The combined hind limbs' TSCIS was significantly higher in the HUC group than in ADM and PSC groups, = 0.007. Both ADM and PSC groups exhibited loss of proprioception and mild to moderate ataxia compared to controls. MRI showed increased pathological findings in the PSC group when compared to the HUC group, = 0.045. Histologically, the meningeal layer was thickened (inflammation) by 2-3 fold in ADM and PSC groups when compared to HUC and CTL groups, = 0.01. There was lower MPO positive cells in the HUC group than in the ADM group, = 0.018. Posterior column astrocyte activation was increased in ADM and PSC lambs compared to HUC lambs, = 0.03.

CONCLUSION

The HUC as a skin patch for in utero spina bifida repair preserves spinal cord function by reducing underlying inflammation when compared to ADM.

摘要

目的

对于严重脊柱裂（SB）的宫内修复，使用标签外组织移植材料，如脱细胞真皮基质（ADM），在无法进行原皮肤层闭合的情况下，与不良神经学结局相关。冷冻保存的人脐带（HUC）补片具有再生、抗炎和抗瘢痕形成特性，并能实现脊柱裂的防水修复。我们检验了这样一个假设：对于减少修复部位的炎症和保留脊髓功能，HUC是比ADM更优的皮肤补片。

方法

在怀有双胞胎的定时妊娠母羊中，于妊娠第75天（GD75），在不进行脊髓切开术的情况下制造脊柱裂（功能模型）。在GD95时，通过将HUC补片与ADM补片（随机分配）缝合到皮肤边缘来进行修复。此外，全层皮肤闭合作为原皮肤闭合（PSC）作为阳性对照。在GD140时进行分娩，随后使用德克萨斯脊髓损伤量表（TSCIS）对羔羊进行步态、本体感觉和痛觉的盲法治疗后神经学评估。无脊柱裂的羔羊用作对照（CTL）。对修复部位的脊柱进行离体磁共振成像，随后进行定量病理学评估。组织学评估（盲法）包括Masson三色染色，以及通过波形蛋白和胶质纤维酸性蛋白（GFAP）共染色对髓过氧化物酶（MPO；中性粒细胞）和反应性星形胶质细胞（炎症）进行免疫荧光检测。

结果

HUC组后肢TSCIS总分显著高于ADM组和PSC组，P = 0.007。与对照组相比，ADM组和PSC组均出现本体感觉丧失和轻度至中度共济失调。与HUC组相比，MRI显示PSC组的病理结果增加，P = 0.045。组织学上，与HUC组和CTL组相比，ADM组和PSC组的脑膜层增厚（炎症）2至3倍，P = 0.01。HUC组的MPO阳性细胞低于ADM组，P = 0.018。与HUC羔羊相比，ADM和PSC羔羊的后柱星形胶质细胞活化增加，P = 0.03。