Proof of concept for the use of dog mesenchymal stem cells secretoma in regenerative therapies

Abstract

Fibrin Heterologous Sealant (FHS), developed by a Brazilian public university, is a low cost-effective, coagulant, adhesive, sealant biopolymer with properties already tested in the Phase I/II clinical trial in chronic venous ulcers. Recently we also discovered its capacity for cellular scaffold and drug delivery system. On the other hand, regenerative stem cell-based therapies have been widely used in veterinary medicine. Currently, this treatment is much sought after, since Brazil has more than 70 million animals including dogs and cats, although this market is not yet regulated by the Ministry of Agriculture, Livestock and Supply (MAPA). This leads to dubious, ineffective and even inadequate treatments. To circumvent this, R4D Biotech, a spinoff derived from research group UNESP Fibrin Sealant and Stem Cell, will use the final conditioned medium resulting from in vitro stem cell growth called secretoma. It is rich in vesicles called exosomes, plus growth factors and signaling molecules. The use of these vesicles for bone repair has the following advantages: they can be easily isolated from in vitro mesenchymal stem cell (MSC) culture; they are safer than living cell-based treatments, avoiding cellular toxicity and immune rejection; do not express MHCI or MHCII proteins on the surface of MSCs, avoiding immunogenicity caused by living cell-based therapy; stimulate bone regeneration in vivo and in vitro; they are stable compared to living cells and are not at risk for aneuploidy, preventing the development of tumors. Thus, we will use these SHF-associated exosomes to produce a new product, which upon standardization and clinical approval will be required to MAPA register. It can be used as an aid in orthopedic surgery of dogs, as well as surgical and skin wounds. The aim of the product will be to accelerate tissue healing and regeneration. To achieve these results, we need to evaluate, characterize, and standardize the MSCs secretoma extracted from the bone marrow, adipose tissue, and umbilical cord matrix of dogs. Mass spectrometry, which uses proteomic analysis tools, is a powerful technology for identifying and quantifying protein components and will therefore be used for characterization to standardize and validate a new product of these vesicles. The optimal concentration of FHS to be used will also be investigated and formulated. This proof-of-concept will develop a prototype to treat acute bone lesions, accelerating bone callus formation, shortening recovery and improving the animal's quality of life. The company, based on established partnerships, will build on a consolidated platform (FHS), will add stem cell secretoma components to be standardized by this research, validating by mass spectrometry - all innovative and at the frontier of knowledge. To make this project viable, an R&D contract was signed with the UNESP Innovation Agency - AUIN 19DIT025, "Development of a veterinary product based on fibrin and stem cells". This agreement allows access to UNESP patent no BR1020140114327. In addition, it allows the supply of snake venom-derived fibrin sealant by the Center for Studies of Venoms and Venomous Animals (CEVAP) at Sao Paulo State University (UNESP), Campus Botucatu. We will create a nonexistent and globally competitive product. This will generate wealth for the country, which has been losing competitiveness in the high technology and innovation market. (AU)