Multi-Omics Comparison of Proline-Rich Peptide-Enhanced Hyaluronic Acid Gels versus Conventional Regenerative Materials: An Early Wound-Healing Model

Aims: To compare the early wound-healing responses to crosslinked hyaluronic acid enriched with two proline-rich peptides (P2, P6) against unmodified hyaluronic acid and the enamel-matrix derivative (EMD) in a porcine gingival-detachment model. Methods: In six pigs, defects around premolars were treated with HA, HA + P2, HA + P6 or EMD. After 6 days, the sites were harvested and evaluated using histology, immunohistochemistry, multiplex cytokine assay and untargeted proteomics of the gels, which were examined, informing an integrated multiomics approach analysis. Results: Both peptide formulations improved the composite histological score compared to HA alone, and HA + P6 matched EMD in suppressing oedema, TNF-α staining and CD80+ macrophages while increasing mannose-receptor labelling. Proteomics revealed that P2 upregulated actin (Q6QAQ1) and histone H2A (F2Z5L5), consistent with active remodelling, whereas EMD reduced trypsin and trypsinogen, indicating a more stabilised wound milieu; HA + P6 showed an intermediate yet favourable profile. IL-6 levels were lower in both peptide groups and EMD than in HA, and all gels proved noncytotoxic, with HA + P6 enhancing cell viability. Conclusion: HA + P6 suppressed TNF-α and trypsin signalling while boosting M2-linked markers and proresolution proteins to the same extent as EMD, whereas plain HA did not. Thus, adding a short proline-rich peptide confers EMD-like early immunomodulation on a fully synthetic HA carrier. These observations, although limited to 6-day soft tissue healing, warrant longer studies in true periodontal regeneration models.