Comparative assessment of residual DNA quantification methods in human acellular dermal grafts: a systematic review

Human acellular dermal grafts (HADGs) are widely used in regenerative medicine and tissue banking because of their biocompatibility and structural support. Residual DNA is an important quality attribute for assessing cellular material removal, and its interpretation should be integrated with evaluation of extracellular matrix (ECM) integrity and scaffold performance. A systematic review (search date: October 15, 2025) of PubMed, Scopus, Web of Science, Embase, and grey literature was conducted. English-language in vitro or experimental studies (2010-2025) reporting quantitative residual DNA levels and at least one structural, functional, or biological assessment were included. Data on decellularization protocols, DNA quantification methods, analytical sensitivity, validation, and scaffold performance were extracted. Study quality was assessed using the Joanna Briggs Institute checklist. Ten studies met the inclusion criteria from 1,250 screened records. Residual DNA varied with decellularization method and quantification technique. Enzymatic and hybrid protocols achieved near-complete DNA removal (> 98%) while preserving extracellular matrix (ECM) structure, basement membrane integrity, and cytocompatibility. Fluorometric and quantitative polymerase chain reaction (qPCR) assays offered higher sensitivity and reproducibility than spectrophotometry, which often overestimated DNA content. Detergent- or alkaline-only methods showed greater variability and sometimes compromised ECM integrity or cell attachment. HADG quality and translational suitability are strongly influenced by decellularization and the DNA quantification approaches. Evidence supports enzymatic or hybrid methods integrating nuclease treatment with controlled detergent exposure, combined with sensitive DNA assays, as the most effective framework for standardized quality assessment, regulatory compliance, and reproducible clinical application.