INTRODUCTION
The prospect of using decellularized organs that have been recellularized by patient-specific progenitor cells for organ and tissue replacement opens the possibility for future clinical applications wherein an essentially autologous transplant occurs.1-4 Retention of extracellular matrix (ECM) components during the decellularized process is crucial for influencing the behavior of cells that are subsequently placed on the decellularized scaffold.5 ECM components play a major role in the proper migration, protein expression, and active signaling pathways of the donor cells.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 overall decellularization process involves cell destruction by detergents and extensive rinsing of residual cell components and DNA debris. The ability of detergents have been compared because it is one of the important factors for ECM retention. 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.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. In the study, DNA depletion was correlated with high pH levels of CHAPS solution.10 Although there are many decellularization methods available at the time of writing including physical, chemical, enzymatic, and proteinase approaches, however high pH alone may be able to achieve sufficient decellularization.11
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Based on this possibility, we evaluated the decellularization ability of high pH sodium hydroxide (NaOH)-PBS solution without using detergents. We showed that high pH NaOH-PBS solution alone has sufficient ability for decellularization and provides comparable ECM preservation when compared to other detergent based decellularization solutions. We also confirmed comparable cell attachment and gas exchange function after the recellularization process in the NaOH group when compared to the SDS group. We collaterally achieved cost reduction in the decellularization process with NaOH-PBS, which will be a significant issue as decellularization based lung regeneration studies are scaled-up for application on larger models such as porcine or human organs and for future clinical usage.
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