Topical formulation containing exosomes derived from mesenchymal cells for treatment of skin wounds

• Grant number: 21/12681-9
• Support Opportunities: Research Grants - Innovative Research in Small Business - PIPE
• Start date: May 01, 2022
• End date: April 30, 2024
• Field of knowledge: Health Sciences - Medicine
• Principal Investigator: Adriana Oliveira Manfiolli
• Grantee: Adriana Oliveira Manfiolli
• Company: In Situ Terapia Celular Ltda
• CNAE: Pesquisa e desenvolvimento experimental em ciências físicas e naturais
• City: Ribeirão Preto
• Associated researchers: Carolina Caliári Oliveira
• Associated research grant: 19/00782-5 - The use of exosomes derived from mesenchymal cells in the regenerative medicine: utilization of by-products of the cell therapy for the formulation of topical use compound for wounds healing, AP.PIPE
• Associated scholarship(s): 22/04778-5 - Topical formulation containing exosomes derived from mesenchymal cells for the treatment of skin wounds, BP.PIPE

Abstract

Skin wounds, both chronic and acute, represent a growing problem not only for patients but also for health systems in general, and there is a range of patients who are refractory to conventional therapies. Therefore, new technologies using the principles of regenerative medicine and tissue engineering are constantly being incorporated into innovative therapies for the treatment of skin wounds. In this context, regenerative medicine through cell therapy, and more recently, exosome therapy, has emerged as an alternative treatment for these patients. Among the cells used therapeutically, mesenchymal cells (CMs) stand out mainly for their immunomodulatory and regenerative properties. Much of the therapeutic potential of these cells has been attributed to the secretion of bioactive molecules often contained in their extracellular vesicles (EVs), or more specifically in exosomes. In a first approach, an In Situ Cell Therapy developed a 3D biodressing containing CMs for the treatment of patients with chronic wounds and severe burns. However, knowing the therapeutic potential of EVs that are secreted by cells in the culture medium, we started the development of our second product, which is a topical formulation for the treatment of skin wounds, containing exosomes that are recycled from the cell culture medium during the production process of the biodressings. In the first phase of this project, we showed that exosomes purified on a laboratory scale using the ultrafiltration/ultracentrifugation protocol had satisfactory characteristics and the ability to stimulate fibroblast proliferation in vitro, and that the topical formulation containing these exosomes was non-phototoxic and non-irritating in in vitro safety assays. The integrity of exosomes is essential for its biological activity and is therefore one of the main requirements for the development of large-scale isolation methods for purification of these nanovesicles. Therefore, the goal of this project is to optimize a platform for obtaining exosomes secreted by CMs through the implementation of large-scale isolation protocol that ensures the maintenance of the regenerative properties of these EVs, as well as incorporating these exosomes in a topical formulation for the treatment of skin wounds. For this purpose, CMs derived from human umbilical cord will be isolated and cultured following GMP (Good Manufacturing Practice) conditions. The exosomes secreted by CMs in the culture medium will be purified through a tangential flow filtration system (TFF) and characterized. The characterization of exosomes will be done by size, exosomal markers and microRNA expression analysis. After characterization, the exosomes will be incorporated into topical formulations that will be tested in the experimental model of skin wound. The acceleration of the wound healing process will indicate the therapeutic effectiveness of the formulations containing the exosomes. The mechanisms involved in the tissue regeneration process will be investigated using histopathological and immunological analyses. Thus, based on the results obtained in this project, and continuing with clinical trials at a further stage, we believe that we will bring to the market a safe and effective regenerative medicine product based on CM exosomes for the treatment of skin wounds. Our purpose is to develop an innovative product that will benefit patients in need of effective wound care solutions. (AU)