Non-enzymatic replication of brown-rotted wood: physicochemical and mechanical characterization for laboratory experiments

Abstract Replication of brown-rotted wood is essential for researchers across disciplines who require suitable and reliable substrates for laboratory experiments. While enzymatic protocols are commonly used, their inherent heterogeneity limits reproducibility and compromises replicate consistency. As a result, researchers are turning to non-enzymatic protocols, particularly the Chelator-Mediated Fenton (CMF) reaction. Although these protocols are successful, they are typically evaluated based solely on the chemical properties of decayed wood, while other key properties remain largely unexamined. This study therefore aimed to examine the physical and mechanical properties of CMF-treated wood, alongside its chemistry, to evaluate its suitability as a reliable substrate for laboratory studies. Scots pine ( Pinus sylvestris L.) samples were CMF-treated and analyzed chemically by ATR-FTIR spectroscopy, mechanically through Shore D hardness and micro three-point static bending tests, and physically by measuring equilibrium moisture content, apparent density, mass loss, and color. For comparison, samples decayed by the brown rot fungus Coniophora puteana were also examined. Results showed that CMF-treated samples closely mimic fungal decay, exhibiting selective polysaccharide depolymerization, pronounced mechanical strength loss, and distinctive physical and morphological changes. These findings demonstrate that non-enzymatic decay protocols can reliably replicate the physicochemical and mechanical properties of brown-rotted wood, offering consistent substrates for experimental research.