Decellularization of porcine small-diameter vascular grafts: evaluation of a latrunculin B-based method and novel perfusion approach.

Abstract

Arteriovenous grafts are widely used for hemodialysis access, yet high failure rates remain a major clinical challenge. Decellularized blood vessels offer promising alternatives, but effective protocols must remove cellular and antigenic components while preserving extracellular matrix (ECM) integrity. This is particularly challenging in elastic arteries, where smooth muscle cells are anchored by dense actin filaments that are difficult to remove without ECM damage from harsh treatments. Here, we present a decellularization strategy based on actin-disrupting agents, specifically Latrunculin B, to facilitate removal of these dense cytoskeletal structures. The protocol integrates osmotic shock, high-ionic-strength salts, and Triton X-100 within a shortened processing time. Full-length porcine carotid arteries were decellularized using a dual-flow perfusion system, and five protocols were evaluated for removal of nuclear and immunogenic material, ECM preservation, mechanical properties, and cytocompatibility with human endothelial cells (HUVECs). All protocols substantially reduced nuclear material while preserving elastin, collagen, and mechanical integrity. Complete removal of immunogenic proteins was achieved with a final alkaline Triton X-100 wash (pH 8-14), highlighting the effectiveness of alkaline treatment in solubilizing membrane-bound antigens. All scaffolds supported HUVEC adhesion and formed a confluent endothelial monolayer within 8 d. Overall, integrating actin disruption via Latrunculin B with dual-salt-Triton processing, alkaline washing, and perfusion enhances decellularization efficiency, preserves ECM structure, reduces processing time and cost, and yields cytocompatible small-diameter vascular scaffolds with strong translational potential.