Perfusion decellularization of a human limb: A novel platform for composite tissue engineering and reconstructive surgery

Abstract

Muscle and fasciocutaneous flaps taken from autologous donor sites are currently the most utilized approach for trauma repair, accounting annually for 4.5 million procedures in the US alone. However, the donor tissue size is limited and the complications related to these surgical techniques lead to morbidities, often involving the donor sites. Alternatively, recent reports indicated that extracellular matrix (ECM) scaffolds boost the regenerative potential of the injured site, as shown in a small cohort of volumetric muscle loss patients. Perfusion decellularization is a bioengineering technology that allows the generation of clinical-scale ECM scaffolds with preserved complex architecture and with an intact vascular template, from a variety of donor organs and tissues. We recently reported that this technology is amenable to generate full composite tissue scaffolds from rat and non-human primate limbs. Translating this platform to human extremities could substantially benefit soft tissue and volumetric muscle loss patients providing tissue- and species-specific grafts. In this proof-of-concept study, we show the successful generation a large-scale, acellular composite tissue scaffold from a full cadaveric human upper extremity. This construct retained its morphological architecture and perfusable vascular conduits. Histological and biochemical validation confirmed the successful removal of nuclear and cellular components, and highlighted the preservation of the native extracellular matrix components. Our results indicate that perfusion decellularization can be applied to produce human composite tissue acellular scaffolds. With its preserved structure and vascular template, these biocompatible constructs, could have significant advantages over the currently implanted matrices by means of nutrient distribution, size-scalability and immunological response.

Engineering Functional Cardiac Tissues for Regenerative Medicine Applications

“…Some example methods include agitation in solution, thermal shock, convective flow, and manual disruption [119]. One of the more consistent methods is perfusion decellularization [120], which can be applied to cadaver hearts or any other target organ [121]. Importantly, this technique uses the organ's existing vasculature for perfusion, while also preserving the vessels' ECM structure at both macro-and micro-scale.…”

“…Importantly, this technique uses the organ's existing vasculature for perfusion, while also preserving the vessels' ECM structure at both macro-and micro-scale. Native vasculature perfusion-based techniques are well-suited for translating decellularization processes for whole human organs, as they would provide a more even distribution of decellularizing agents, hence avoiding overexposure and potential toxic effects [118,121].…”

Whole Organ Tissue Vascularization: Engineering the Tree to Develop the Fruits

“…The process is usually achieved by means of detergents and enzymes, coupled with physical stress. Nearly every tissue of the human body has been decellularized, and very recently, whole human limbs have been used to produce acellular scaffolds (Gerli et al, 2018 ). This technique has the unique advantage of generating a scaffold that closely resembles the native environment from both a biochemical and anatomical point of view (Crapo et al, 2011 ).…”

New insight into functional limb regeneration: A to Z approaches

“…This group, for the first time, decellularized forelimbs to provide a natural ECM scaffold for limb regeneration (Jank et al, ). More recently, they succeeded in decellularization of an entire cadaveric human limb and preserved its structure, ECM components, and vascular network (Gerli, Guyette, Evangelista‐Leite, Ghoshhajra, & Ott, ). Although they overcame the scaffold issue and preserved ECM positional identity and vascular network, the challenge of one appropriate cell source, cell‐loading into deeper layers of the scaffold remained.…”