PURIFICATION OF SAPONIN-PDRN COMPLEX DERIVED FROM PLATYCODON GRANDIFLORUS BASED ON A DUAL-FUNCTIONAL MAGNETIC BEAD-COLUMN SYSTEM AND VALIDATION OF ITS IN VITRO / IN VIVO SKIN REGENERATION, BARRIER ENHANCEMENT, AND WSSV ANTIVIRAL EFFICACY

Dong Myong Kim*, Jun Ku Lee, Won Kyu Lee, Ji Yeon Kim, Sae Rom Park, Young Ho Choi

Despite the tissue regeneration effects of fish-derived polydeoxyribonucleotide (PDRN), the paradigm is shifting towards plant-derived PDRN (Phyto DNA) and complexes of plant-specific active ingredients due to the instability of marine resource supply and the risk of zoonotic infectious diseases. This study aims to purify on a large scale the Platycodon grandiflorus Saponin-PDRN complex, in which pharmacologically active oleanane-type triterpenoid saponins (such as Platycodin D) chemically and physically interact with low-molecular-weight DNA from the crude extract of Platycodon grandiflorus. To achieve this, a dual-functional magnetic bead-column-magnet integrated platform was established by introducing a magnetic core (Fe3O4), a cationic capture layer (PEI/DEAE), and a hydrophilic PEG spacer. Through the electrostatic primary capture and structure-specific secondary capture mechanisms of this purification platform, a high-purity complex (A260/A280 = 1.88 ± 0.02) with a molecular weight ranging from 100 to 1200 bp was recovered at a yield of 92.4%. In vitro evaluations revealed that the purified Saponin-PDRN complex activated the adenosine A2A receptor pathway, promoting human dermal fibroblast (HDF) proliferation by 148.5% and cell migration by 88.3%, while significantly inhibiting the expression of inflammatory cytokines (TNF-α, IL-6) in macrophages. Simultaneously, regarding skin barrier enhancement, a plant DNA-specific mechanism was confirmed, which upregulated the expression of antioxidant enzymes (SOD1) and barrier proteins (FLG, LOR) while inhibiting collagen-degrading enzymes (MMP-1). Furthermore, in an infection model of White Spot Syndrome Virus (WSSV), a fatal pathogen in aquaculture, the complex blocked the expression of early replication genes (ie1, vp28) and recorded an in vivo survival rate of 85.0%, driven by the synergistic effect of the saponin's viral envelope membrane fusion inhibition and PDRN's host innate immunity enhancement. In a mouse wound healing model, the complex demonstrated its high utility as a skin regeneration cosmeceutical and natural antiviral biomaterial by inducing 96.8% complete re-epithelialization and CD31-positive neovascularization by day 14 of treatment.