Response regarding “Assessing the effects of bladder decellularization protocols on extracellular matrix (ECM) structure, mechanics, and biology”

As both of our astute commenters noted, there is currently not a standard protocol for bladder decellularization. This variability presents a challenge for surgeons and researchers who use bladder acellular matrix (BAM) products, as well as those who seek to interpret translational studies that have applied BAM in tissue engineering or surgical reconstruction. Highlighting the myriad structural, mechanical, and biological factors that may differ among resultant products is the aim of our manuscript [1]. Two methods were selected based on previous literature and represent the vast number reported [2]. However, this manuscript is not intended as a reference for optimal decellularization methods. Specific methods should be selected based on the intended application. Additionally, studies applying decellularized matrices should report a relevant description of the methods applied and a comprehensive characterization of the BAM product to aid in study interpretation [3,4].

Specific to the commenter’s concerns regarding the use of an enzymatic and detergent combination in one cohort and a detergent only in the second cohort, this selection was based on the efficacy considerations of each agent, as demonstrated in prior literature. Specifically, the detergent Triton X-100 was combined with a 0.25% trypsin step, as previously reported, as Triton X-100 alone was inadequate for complete decellularization when applied in isolation [4]. While prolonged exposure to trypsin can disrupt collagen and elastin, this time-dependent process can likewise be harnessed to increase the efficiency of subsequent decellularization agents [2]. Thus, the agents selected for this study were designed to ensure efficient and effective (>90%) target decellularization to minimize the risk of antigenic components [5], while applying the minimum hypothesized required exposure to achieve this aim.

Given the varied decellularization methods, we propose that this manuscript is viewed as a step toward creating a standardized assessment strategy to assist in selecting a decellularization method that yields a product aligned with each researcher’s unique needs. Regardless of the method selected, we recommend a comprehensive analysis and reporting of BAM mechanical, structural, and biological parameters to maximize translatability and scientific interpretation.