Addition of PRG4 to Optimize the Anti-Adhesive Properties of Tissue Barrier Seprafilm^®

Postsurgical adhesion remains a significant clinical challenge. Seprafilm^® is widely used as a barrier to reduce adhesion formation, however its efficacy may be inconsistent. Recombinant human proteoglycan-4 (rhPRG4), a surface-active, anti-inflammatory glycoprotein found in synovial fluid inhibits cell and protein adhesion. This study investigated whether coating Seprafilm with rhPRG4 could enhance its anti-adhesive potential.

Macrophages (J774), human fibroblast-like synoviocytes (HFLS), and melanoma (A375) cells were cultured on tissue culture wells treated with rhPRG4 or bovine submaxillary mucin (BSM) to determine optimal seeding density. Seprafilm sheets were coated with rhPRG4 or BSM (0.78-200 µg/mL). Cell adhesion was quantified using CellTiter-Glo^® while non-adherent macrophages were assessed for viability and re-adhesion. Migration was tested with the Oris^™ Universal Cell Migration assay and oxidative stress with MitoSOX^™ Red staining. ANOVA with multiple comparisons was used for statistical analysis.

Seprafilm alone promoted HFLS and macrophage adhesion, whereas application of rhPRG4 significantly reduced adherence compared to both control and BSM coated surfaces. The combination of rhPRG4 with Seprafilm showed the greatest reduction in adhesion without compromising viability or re-adherence. Seprafilm alone increased macrophage migration and ROS production, while rhPRG4 coated surfaces alone or in combination with Seprafilm suppressed both.

Coating Seprafilm with rhPRG4 enhances its anti-adhesive properties by reducing adhesion, migration, and oxidative stress in vitro. These findings suggest rhPRG4 may improve the biological performance of adhesion barriers by suppressing early cellular infiltration and inflammatory activation. Anti-adhesive bioactive surface modifications may advance postsurgical adhesion prevention and wound healing.