Abstract:
Reconstructive techniques to repair severe tissue defects include the use of autologous fasciocutaneous flaps, which may be limited due to donor site availability or lead to complications such as donor site morbidity. A number of synthetic or natural dermal substitutes are in use clinically, but none have the architectural complexity needed to reconstruct deep tissue defects. The perfusion decellularization of fasciocutaneous flaps is an emerging technique that yields a scaffold with the necessary composition and vascular microarchitecture and serves as an alternative to autologous flaps. In this study, we show the perfusion decellularization of porcine fasciocutaneous flaps using sodium dodecyl sulfate (SDS) at three different concentrations, and identify that 0.2% SDS results in a decellularized flap that is efficiently cleared of its cellular material at 86%, has maintained its collagen and glycosaminoglycan content, and preserved its microvasculature architecture. We further demonstrate that the decellularized graft has the porous structure and growth factors that would facilitate repopulation with cells. Finally, we show the biocompatibility of the decellularized flap using human dermal fibroblasts, with cells migrating as deep as 150 µm into the tissue over a 7-day culture period. Overall, our results demonstrate the promise of decellularized porcine flaps as an interesting alternative for reconstructing complex soft tissue defects, circumventing the limitations of autologous skin flaps.
摘要:
修复严重组织缺损的重建技术包括使用自体筋膜皮瓣,这可能是有限的,由于供体部位的可用性或导致并发症,如供体部位的发病率。许多合成或天然真皮替代品在临床上使用,但是没有一个具有重建深层组织缺陷所需的结构复杂性。筋膜皮瓣的灌注脱细胞化是一种新兴的技术,可产生具有必要组成和血管微结构的支架,并可替代自体皮瓣。在这项研究中,我们显示了使用三种不同浓度的十二烷基硫酸钠(SDS)对猪筋膜皮瓣的灌注去细胞化,并确定0.2%SDS导致去细胞化的皮瓣,其细胞材料在86%被有效清除,保持了它的胶原蛋白和糖胺聚糖含量,并保留了它的微血管结构。我们进一步证明,脱细胞移植物具有多孔结构和生长因子,可促进细胞再增殖。最后,我们显示了使用人真皮成纤维细胞的脱细胞皮瓣的生物相容性,在7天的培养期内,细胞迁移到组织中的深度为150µm。总的来说,我们的结果证明了去细胞猪皮瓣作为重建复杂的软组织缺损的一个有趣的替代方案的前景,规避自体皮瓣的局限性。