Sustained delivery of small interfering RNA (siRNA) is a challenge in gene silencing for managing gene‐related disorders. Although nanoparticle‐mediated electrospun fibers enable sustainable gene silencing, low efficiency, loss of biological activity, toxicity issues, and complex electrospinning techniques are all bottlenecks of these systems. Preventing peritendinous adhesion is crucial for their successful use, which involves blocking cellular signaling via physical barriers. Here, a multifunctional, yet structurally simple, cationic 2,6‐pyridinedicarboxaldehyde‐polyethylenimine (PDA)‐mediated extracellular signal‐regulated kinase (ERK)2‐siRNA polymeric delivery system is reported, in the form of peritendinous antiadhesion electrospun poly‐l‐lactic acid/hyaluronan membranes (P/H), with the ability to perform sustained release of bioactive siRNA for long‐term prevention of adhesions and ERK2 silencing. After 4 days of culture, the cell area and proliferation rate of chicken embryonic fibroblasts on siRNA+PDA+P/H membrane are significantly less than those on P/H and siRNA+P/H membranes. The in vivo results of average optical density of collagen type III (Col III) and gene expression of ERK2 and its downstream SMAD3 in the siRNA+PDA+P/H group are less than those of P/H and siRNA+P/H groups. Consequently, siRNA+PDA+P/H electrospun membrane can protect the bioactivity of ERK2‐siRNA and release it in a sustained manner. Moreover, adhesion formation is inhibited by reducing fibroblast proliferation and Col III deposition, and downregulating ERK2 and its downstream SMAD3.

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