Matrix Composition and Mechanics of Decellularized Lung Scaffolds

Abstract

The utility of decellularized native tissues for tissue engineering has been widely demonstrated. Here, we examine the production of decellularized lung scaffolds from native rodent lung using two different techniques, principally defined by use of either the detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) or sodium dodecyl sulfate (SDS). All viable cellular material is removed, including at least 99% of DNA. Histochemical staining and mechanical testing indicate that collagen and elastin are retained in the decellularized matrices with CHAPS-based decellularization, while SDS-based decellularization leads to loss of collagen and decline in mechanical strength. Quantitative assays confirm that most collagen is retained with CHAPS treatment but that about 80% of collagen is lost with SDS treatment. In contrast, for both detergent methods, at least 60% of elastin content is lost along with about 95% of native proteoglycan content. Mechanical testing of the decellularized scaffolds indicates that they are mechanically similar to native lung using CHAPS decellularization, including retained tensile strength and elastic behavior, demonstrating the importance of collagen and elastin in lung mechanics. With SDS decellularization, the mechanical integrity of scaffolds is significantly diminished with some loss of elastic function as well. Finally, a simple theoretical model of peripheral lung matrix mechanics is consonant with our experimental findings. This work demonstrates the feasibility of producing a decellularized lung scaffold that can be used to study lung matrix biology and mechanics, independent of the effects of cellular components.

Quantification of Extracellular Matrix Proteins from a Rat Lung Scaffold to Provide a Molecular Readout for Tissue Engineering

“…Lamin A/C appears to be efficiently depleted during the decellularization process, despite known challenges of completely removing nuclear matrix. In contrast, h H2A, appears enriched postdecellularization, despite reports of efficient DNA depletion (5,44,45). Similarly, cellular proteins such as cytoskeletal actins, tubulins, and mitochondrial ATP synthase subunit alpha-1 appear to be inefficiently removed by the decellularization process according to the peptide spectral match comparisons in Table I.…”

Sodium hydroxide based non-detergent decellularizing solution for rat lung

“…9 Petersen and colleagues reported that CHAPS-based decellularization preserved collagen and elastin compared to SDS-based decellularization and how the mechanical integrity of scaffolds significantly diminishes and that some loss of elasticity occurs using SDS decellularization. 8 However, when cells were intratracheally inoculated into the various decellularized lungs, the results of recellularization were comparable for initial binding and short-term (2 wk) proliferation of two different cell types, a stromal progenitor cell and a mouse lung epithelial cell line. 9 We have previously explained the influence of pH of CHAPS solution on ECM preservation.…”

“…6 , 7 Transplantation studies of recellularized lung reveal that the alveolar structure of the bioengineered lung is fragile and the histological integrity of the recellularized lung seems to be determined by damage to the ECM at the thin barrier of the alveoli. 8 Thus, the decellularization process has been considered to be crucial for ECM preservation.…”

“…The most commonly used detergents are sodium dodecyl sulfate (SDS), 3-[(3-cholamidopropyl) dimethylammonio]-1- propanesulfonate (CHAPS) or Triton-X100/sodium deoxycholate (Triton/SDC). 8 , 9 There are few direct comparisons of the various protocols utilizing different detergents, but Wallis and colleagues reported Triton/SDC as been less disruptive to native ECM when compared with CHAPS and SDS based approaches. 9 Petersen and colleagues reported that CHAPS-based decellularization preserved collagen and elastin compared to SDS-based decellularization and how the mechanical integrity of scaffolds significantly diminishes and that some loss of elasticity occurs using SDS decellularization.…”

Stem Cells and Cell Therapies in Lung Biology and Diseases: Conference Report

“…Niklason shared her progress in the development of functional lung constructs. Dr. Niklason described the use of decellularization methods based primarily on the use of vascular and airway perfusion with CHAPS detergent that has been shown to preserve the structure of the lung ECM (48,51). The cell source used by Dr. Niklason was a mixed population of neonatal rat lung cells.…”