Keywords
Silver
Venous Ulcer
Wound Healing
Metalloproteinases
How to Cite
Abstract
Introduction. Chronic venous leg ulcers disproportionately affect older adults and are characterized by delayed healing, high recurrence rates, and substantial impacts on quality of life and healthcare costs. Persistent inflammation and an imbalance between matrix metalloproteinases and their tissue inhibitors contribute to healing failure, maintaining the wound in an inflammatory state. Oxidized regenerated cellulose/collagen dressings, with and without silver, accelerate healing in venous ulcers, but the cellular mechanisms linking these outcomes to changes in the wound microenvironment are not fully understood. This study evaluated how oxidized regenerated cellulose/collagen and oxidized regenerated cellulose/collagen/silver modulate venous ulcer fibroblasts, focusing on proteases, their endogenous inhibitors and key growth factors, to connect in vitro findings to potential clinical applications.
Methods. Fibroblasts were isolated from chronic venous leg ulcers and cultured for 72 hours in control medium, oxidized regenerated cellulose/collagen (2.8 mg/mL) or oxidized regenerated cellulose/collagen/silver (4.2 mg/mL). Cell metabolic activity was assessed by MTT assay. MMP 1, MMP 2, MMP 3, MMP 10, MMP 13, TIMP 1, TIMP 2, TIMP 3 and TGF β1, TGF β2, TGF β3 were quantified in conditioned media using bead based multiplex immunoassays. Vimentin, collagen type I and collagen type III expression were evaluated by immunofluorescence. Statistical analysis used the paired Wilcoxon signed rank test (p< 0.05).
Results: Both dressings maintained venous leg ulcer fibroblast metabolic activity at levels comparable to, or slightly higher than, control medium, without evidence of overt cytotoxicity. Oxidized regenerated cellulose/collagen alone did not significantly alter MMPs, TIMPs or TGF β isoforms. Oxidized regenerated cellulose/collagen/silver significantly increased MMP 1 (p = 0.0002) and reduced MMP 13 (p = 0.0155) versus control, while decreasing TIMP 2 and TIMP 3 and preserving TIMP 1. TGF β1, TGF β2, TGF β3 and collagen type I and III expression remained unchanged across groups, and vimentin expression was preserved.
Discussion. In this in vitro model, oxidized regenerated cellulose/collagen and oxidized regenerated cellulose/collagen/silver preserved fibroblast viability and did not overtly interfere with TGF β signaling or collagen type I and III expression. The silver free matrix did not modify the analyzed protease inhibitor levels, supporting a predominantly extracellular protease sequestering mechanism. The silver containing matrix induced a shift in the fibroblast derived protease inhibitor profile, characterized by increased MMP 1, decreased MMP 13 and reduced TIMP 2 and TIMP 3 with preserved TIMP 1, a pattern that may rebalance protease activity toward matrix remodeling under non cytotoxic conditions. These findings offer a mechanistic, hypothesis generating framework with translational implications for the clinical use of silver-containing oxidized regenerated cellulose/collagen dressings in venous leg ulcers.
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Copyright (c) 2026 Julia Teixeira Nicolosi, Silvana Cereijido Altran, Liliana Scorzoni, André Oliveira Paggiaro, Viviane Fernandes de Carvalho , Debora Levy, Rolf Gemperli, Cesar Isaac