Evaluation of TGF β -3-loaded eggshell membranes as potential therapeutic biomaterials for wound healing applications

Abstract Chronic wounds impose a serious healthcare burden with current options limited by efficacy, high cost, and risks of infection or scarring. Transforming growth factor β 3 (TGF- β 3) is attractive for regulating healing, limiting scarring, and promoting extracellular matrix remodeling, yet its translation ability impacted by rapid degradation and need for controlled local delivery. The eggshell membrane (ESM) is a biocompatible, collagen-rich, low-cost natural scaffold suitable for growth factor delivery. Although chicken ESM (CESM) is better studied, comparative assessment of duck (DESM) and ostrich (OESM) remains scarce. Here we systematically evaluate CESM, DESM, and OESM to establish their suitability for localized TGF- β 3 delivery. Using Fourier-transform infrared spectroscopy (FTIR), differential scanning calorimetry, dynamic mechanical analysis, water contact angle measurement, and scanning electron microscopy, we show that all three membranes possess collagen-rich fibrous networks, high porosity, and thermal stability ( T m > 135 °C). Hydrated membranes were hydrophilic, supporting a moist wound milieu. DESM resisted hydration-induced change, whereas OESM showed superior extensibility that may benefit dynamic wound regions. In vitro release revealed sustained TGF- β 3 delivery within 72 h across all membranes, with CESM and DESM providing more stable release than OESM. Cytotoxicity lactate dehydrogenase (LDH), viability (MTS), and live/dead assays indicated safety; dermal fibroblasts maintained ∼ 105% viability at 0.01–300 ng ml −1 . In ovo chorioallantoic membrane (CAM) assays showed that TGF- β 3 increased vessel thickness, with DESM plus TGF- β 3 yielding the greatest effect, while branching responses were variable. This first comparative evaluation of CESM, DESM, and OESM as TGF- β 3 delivery platforms integrates materials characterization with in vitro and in ovo assays and supports ESM-based dressings for chronic wound care.