Decellularized Kidney Matrix for Perfused Bone Engineering

Abstract

The vascularization of tissue-engineered constructs is yet an unsolved problem. Here, recent work on the decellularization of whole organs has opened new perspectives on tissue engineering. However, existing decellularization protocols last several days and derived biomatrices have only been reseeded with cells from the same tissue origin or stem cells differentiating into these types of tissue. Within the present work, we demonstrate a novel standardized, time-efficient, and reproducible protocol for the decellularization of solid tissues to derive a ready to use biomatrix within only 5 h. Furthermore, we prove that biomatrices are usable as potential scaffolds for tissue engineering of vascularized tissues, even beyond tissue and maybe even species barriers. To prove this, we seeded human primary osteoblasts into a rat kidney bioscaffold. Here, seeded cells spread homogeneously within the matrix and proliferate under dynamic culture conditions. The cells do not only maintain their original phenotype within the matrix, they also show a strong metabolic activity and remodel the biomatrix toward a bone-like extracellular matrix. Thus, the decellularization technique has the ability to become a platform technology for tissue engineering. It potentially offers a universally applicable and easily producible scaffold that addresses the yet unsolved problem of vascularization.

Optimization and Critical Evaluation of Decellularization Strategies to Develop Renal Extracellular Matrix Scaffolds as Biological Templates for Organ Engineering and Transplantation

“…Since the first report describing perfusion of detergents through the coronary arteries of rodent hearts by Ott and Taylor in 2008 (11), several approaches to remove cells from the heart (34), liver (13, 23, 35) and lung (12, 36) have been described. Likewise, a variety of perfusion decellularization protocols for kidney have been tested for rodents, pigs, and humans (reviewed in 10 and described in 14–16, 18, 19, 37, 38), but few studies report data specifically comparing the influence of different chemical-based decellularization procedures on resulting ECM architecture and sufficiency for supporting recellularization (14, 39). …”

Controlling stem cell behavior with decellularized extracellular matrix scaffolds

“…Decellularizing cadaveric organs maintains the macro and micro architectures of the organ along with the natural ECM composition, thereby providing a platform for recellularization and regeneration of the organ [1]. Perfusion decellularization has been used on lungs, livers, kidneys, and hearts from rats, mice, pigs, and humans to create acellular organ scaffolds [10–19]. The process of perfusion decellularization requires flowing the decellularizing reagents through native arteries at physiological pressures in order to enter the innate blood vessel system and reach every cell [20].…”

“…Burgkart et al had a unique experimental set up where they showed that a decellularized kidney could be used for bone engineering because the intact vasculature provided nutrients to the osteoblasts. In this study they perfused a whole decellularized rat kidney with primary human osteoblasts using osteogenic media and saw that the osteoblasts kept their phenotype and reconstructed the matrix [19]. These results highlight that there is still little understood about the kidney ECM niche and how it can affect differentiation.…”

Epithelial Cell Repopulation and Preparation of Rodent Extracellular Matrix Scaffolds for Renal Tissue Development

“…The goal of the described decellularization protocol is to produce an acellular renal ECM that can be used as a 3D scaffolding system for regeneration of kidney structures, including nephron tubules that are repopulated in the present example with human renal cortical tubular epithelial (RCTE) cells. We previously described our rigorous evaluation of an optimal, detergent-based rat kidney decellularization protocol 7 , which is more rapid (approximately one day) than other methods previously reported (Ross et ), and exposes the organ to a considerably lower concentration (0.1%) of the denaturant sodium dodecyl sulfate (SDS) during decellularization than prior reports [12][13][14][15] .…”

“…A limited number of studies have described the use of rodent kidneys for decellularization and subsequent use as a 3D scaffold for cellular repopulation (reviewed elsewhere 1 ) [12][13][14][15][16] . In this protocol, we provide a detailed description of our previously established, optimal decellularization strategy for producing acellular kidney scaffolds from Sprague Dawley rat kidneys 7 .…”