接种于人类胶原蛋白基质上的自体脂肪基质细胞用于慢性伤口的真皮再生：概念验证性临床研究

Autologous Adipose Stromal Cells Seeded onto a Human Collagen Matrix for Dermal Regeneration in Chronic Wounds: Clinical Proof of Concept.

作者信息

Lafosse Aurore, Desmet Céline, Aouassar Najima, André Wivine, Hanet Marie-Sophie, Beauloye Christophe, Vanwijck Romain, Poirel Hélène A, Gallez Bernard, Dufrane Denis

机构信息

Brussels, Belgium From the Endocrine Cell Therapy Unit, Center of Tissue/Cell Therapy, Institut de Recherche Expérimentale et Clinique, Plastic and Reconstructive Surgery Unit, Louvain Drug Research Institute, Biomedical Magnetic Resonance Research Unit, Center for Human Genetics and Human Molecular Genetics, de Duve Institute, Cliniques Universitaires Saint-Luc, Cardiovascular Research Unit, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain.

出版信息

Plast Reconstr Surg. 2015 Aug;136(2):279-295. doi: 10.1097/PRS.0000000000001437.

DOI:10.1097/PRS.0000000000001437

PMID:25946602

Abstract

BACKGROUND

Nonhealing wounds are unable to integrate skin autografts by avascular and fibrotic dermal tissue. Adipose-derived stromal cells can improve the local environment of the wound bed by angiogenesis and immunomodulation. This work aimed to develop a biological dressing made of adipose-derived stromal cells onto a human acellular collagen matrix.

METHODS

Adipose-derived stromal cells were isolated from human adipose tissue (n = 8). In vitro, the genetic stability during early and late passages (1, 4, 10, and 16) and vascular endothelial growth factor (VEGF) secretion were assessed. Adipose-derived stromal cell adhesion and spreading on collagen matrix were preliminarily studied. In vivo tumorigenicity, angiogenesis, and tissue oxygenation were assessed after implantation of the construct in nude rats (n = 10). The biological dressing was manufactured and implanted in three patients with chronic wounds.

RESULTS

In vitro, aneuploidies, but no clonal transformation, were detected up to late cellular passages. VEGF was secreted more during hypoxia (0.1% oxygen) than during normoxia (21% oxygen). Adipose-derived stromal cells can adhere and spread on the scaffold within 18 to 20 days. No tumor development occurred 3 months after implantation in immunocompromised rats. Vessel counts and tissue oxygenation were higher after adipose-derived stromal cell implantation. In patients, granulation tissue was found (276 percent of vessel density), followed by epithelialization or split-thickness skin engraftment up to 22 months after implantation.

CONCLUSIONS

Implantation of adipose-derived stromal cells seeded onto human acellular collagen matrix (biological dressing) represents a promising therapy for nonhealing wounds, offering improvement in dermal angiogenesis and remodeling. This therapy using autologous stromal cells is safe, without significant genetic alterations after in vitro expansion.

摘要

背景

难愈性伤口无法通过无血管和纤维化的真皮组织整合自体皮肤移植。脂肪来源的基质细胞可通过血管生成和免疫调节改善伤口床的局部环境。本研究旨在开发一种由脂肪来源的基质细胞制成的生物敷料，并将其应用于人体脱细胞胶原基质上。

方法

从人脂肪组织中分离脂肪来源的基质细胞（n = 8）。在体外，评估早期和晚期传代（第1、4、10和16代）期间的遗传稳定性以及血管内皮生长因子（VEGF）的分泌情况。初步研究脂肪来源的基质细胞在胶原基质上的黏附和铺展情况。将构建体植入裸鼠（n = 10）后，评估其体内致瘤性、血管生成和组织氧合情况。制备生物敷料并将其植入3例慢性伤口患者体内。

结果

在体外，直至细胞传代后期均检测到非整倍体，但未检测到克隆转化。低氧（0.1%氧气）条件下VEGF的分泌量高于常氧（21%氧气）条件。脂肪来源的基质细胞可在18至20天内在支架上黏附和铺展。免疫缺陷大鼠植入后3个月未发生肿瘤。脂肪来源的基质细胞植入后血管计数和组织氧合情况更佳。在患者中，植入后发现有肉芽组织（血管密度增加276%），随后在长达22个月的时间内出现上皮化或中厚皮片移植。