A Multi-targeted Approach for the Impaired Healing of Diabetic Foot Ulcers Through Combined Electric Field Stimulation and Nitric Oxide Therapy - Project Summary Diabetic Foot Ulcer (DFU) is a major complication of diabetes, often leading to lower extremity amputations. DFUs are characterized by a complex pathology resulting from peripheral vascular disease, neuropathy, and infections, all exacerbated by prolonged hyperglycemia, which collectively impede normal wound healing. Standard DFU treatments, such as surgical debridement, wound dressing, and infection management, primarily address individual aspects of the pathology, leading to only about 25% of cases achieving complete healing within 12 weeks. This highlights the limitations of current treatments and the need for innovative solutions that comprehensively address multiple aspects of DFU. Diabetes impairs mitochondrial activity, reducing cellular energy and hindering wound healing and closure. Our previous research showed that electric field stimulation significantly enhanced mitochondrial activity, ATP production, and the proliferation of human dermal fibroblasts. Additionally, nitric oxide (NO) has been shown to promote angiogenesis, prevents ischemia, and reduces inflammation and infection, as also demonstrated in our study with a NO releasing prohealing peptide amphiphile (PA) nanomatrix. However, in diabetic conditions, decreased endothelial NO synthase (eNOS) activity lowers NO levels. Thus, as a multi-targeted strategy, combining electric field stimulation with NO therapy may enhance DFU healing by stimulating cellular energy for wound closure and tissue regeneration, while also reducing complications, such as vascular disease, inflammation, and infection in hyperglycemic conditions. In this study, we will evaluate the therapeutic effects of electric field stimulation and NO therapy, investigating the healing mechanisms of diabetic wounds. To conduct this evaluation, we will develop a bio-engineered cell culture platform that mimics a 3D human skin extracellular matrix (ECM) environment, integrating electric stimulation and NO delivery. This platform will be designed using a peptide amphiphile (PA)-based NO-releasing nanomatrix coating on a highly porous 3D electrospun nanofiber scaffold with two-electrode arrangement (semi- capacitive coupling) for electric stimulation, as proposed in Specific Aim 1. In Specific Aim 2, we will evaluate the effects of combined treatments, particularly electric field stimulation, on behaviors related to wound healing, such as cell proliferation and ECM production, in diabetic skin cells derived from DFU patients under hyperglycemia using the platform. This evaluation will focus on the ATP binding purinergic (P2) receptor-mediated pathway. We will also test the combined treatment, particularly NO, to enhance diabetic endothelial activities, reduce inflammation, and inhibit DFU related bacterial growth under hyperglycemia using the platform in Specific Aim 3. Successful completion of this study will offer a novel DFU treatment strategy by delivering electric field stimulation and NO, validated with an innovative wound healing cell culture platform, enhancing current treatment methods. Additionally, it will enhance Rowan University's biomedical research capabilities and provide valuable hands-on research opportunities for undergraduate students, fostering their development as future scientists.
