Heterologous production and evaluation of the wound healing potential of Pangasius hypophthalmus Annexin A4 Academic Article in Scopus uri icon

abstract

  • Chronic wounds are injuries that significantly affect 2 % of the global population, with a 25$ billion USD annual quota for the US healthcare system, making cost-effective solutions necessary. Chronic wounds become stalled due to chronic inflammation. Annexins have been shown to modulate inflammation and migration in macrophages and fibroblasts, respectively, important steps during wound healing. Also, Pangasius hypophthalmus ANXA4 (PH-ANXA4) wound healing potential was previously prospected due to its structural similarity to its human counterpart. In this study, the potential of recombinant PH-ANXA4 (rPH-ANXA4) as a wound healing promoter is explored. First, its recombinant production using Pichia pastoris was assessed. Furthermore, its biochemical activity was studied through isothermal titration calorimetry (ITC) and differential scanning calorimetry (DSC). In vitro wound healing potential was evaluated by fibroblast (HDFa) cell migration after scratch, and by measuring nitric oxide production in murine macrophages (RAW 267.4). Regarding production, the established process yielded 6 mg with 95 % purity. ITC revealed a Ca2+-dependent binding of rPH-ANXA4 to negatively charged multilamellar vesicles (MLVs). DSC showed that Ca2+-dependent interaction protects MLVs from Ca2+ destabilization. Moreover, in vitro analysis showed that rPH-ANXA4 reduced nitric oxide production by up to 70 % in macrophages, while enhancing fibroblast migration compared to the positive control (Centella asiatica 10 ¿g/mL) at low doses (0.78 and 1.56 ¿g/mL). The obtained findings shed light on the preliminary potential of rPH-ANXA4 to be characterized as a bioactive in wound-healing formulations. © 2025 Elsevier B.V.

publication date

  • January 1, 2026