Synthesis of a retro-GFOGER Adamantane-Based Collagen Mimetic Peptide Imbibed in a Hyaluronic Acid Hydrogel for Enhanced Wound Healing

Abstract

This study reported the synthesis and formulation of an adamantane-based collagen mimetic peptide (CMP) hydrogel containing the integrin-binding motif retro-GFOGER, designed to enable the controlled delivery of CMPs with the ability of direct wound healing for the potential treatment of acute wounds. Initially, two adamantane-functionalized CMPs (peptides NL008 and NL010) were synthesized, characterized, and comparatively screened for their in vitro biocompatibility and bioactivity. In vitro evaluations of scratch closure and biocompatibility were assessed on human-derived keratinocytes. Release and permeation of the peptides were evaluated in vitro and ex vivo. Wound closure rates and histological evaluations were performed on male Sprague-Dawley rats over 3, 7, and 14 days for the NL010-HAgel formulation. Peptide NL010 was found to be the most suitable candidate among the adamantane CMPs. For a comparative study, peptide NL010 and its palmitic acid analogue, NL009, were loaded into a hyaluronic acid (HA) hydrogel and lyophilized. The CMP hydrogels exhibited porosity (<30 μm) and were viscoelastic solids. The physicomechanical properties of the formulations showed optimal characteristics for application as wound dressings in terms of textural profile. Peptide NL008 exhibited lower bioactivity and cell viability compared to NL009 and NL010 across various concentrations and cell lines. Peptide release from NL009-HAgel and NL010-HA gel was 74% and 83%, respectively. Across an ex vivo porcine skin membrane, the CMP-HAgel showed good permeation and was retained in the epidermis and superficial dermis. CMP-HAgel at 0.1% (w/v) showed better HaCaT cell viabilities. In vitro assays demonstrated that the NL010-HA gel achieved scratch closure (99.9%) within 24 h, while the NL009-HAgel showed scratch closure (93.7%) within the same time frame. In vivo, NL010-HAgel improved healing by enhancing epithelialization and granulation tissue deposition (via fibroblast and collagen responses). The findings of this study suggested that the CMP cell-instructive hydrogel is a promising platform with the potential to accelerate wound healing.