USE OF SILK FIBROIN FOR ENCAPSULATION AND DIFFERENTIATION OF STEM CELLS AS A STARTING POINT FOR THE DEVELOPMENT OF ADVANCED CARTILAGE TREATMENT

In the field of regenerative medicine and tissue engineering, intensive research is being conducted on the regeneration of cartilage that is unable to regenerate itself when injured. One of the most important cartilage diseases is osteoarthritis, which is a problem for humans and animals worldwide and a great burden to society in personal, social and economic terms. It is one of the most common diseases in dogs. Conventional therapy with NSAIDs is often unsatisfactory and has many undesirable side effects. The development of cartilage therapy is focused on the use of mesenchymal stem cells/medical signaling cells (MSCs) to improve existing or develop new cell therapies. An important area of research related to cartilage regeneration is the search for ways to exploit the differentiation potential of MSC for use in surgical approaches to cartilage treatment. These are indicated in cases of severe cartilage damage or inadequate analgesic and anti-inflammatory drug therapy. In addition, options are being sought for the development of systems that would allow cells to be immobilised at the target site without surgical intervention, by a minimally invasive injection method. Administration of cells by injection and simultaneous immobilization of cells at the target site using a cellular carrier would make it possible to avoid surgically induced tissue trauma, reduce damage to adjacent tissue, shorten the duration of surgery, and accelerate patient recovery. In addition, the new therapeutic options also aim to develop pre-prepared, readily available ("off-the-shelf") cell products for treatment, as the therapeutic use of MSC is associated with a time-consuming cell preparation process and a short lifespan. Readily available, prepared products would greatly facilitate clinical decision-making at the point-of-care, thereby facilitating access to MSC for treatment and making them available to a broad spectrum of patients. The project proposal addresses all three of the above research areas for cartilage regeneration. This project involves the use of a silk fibroin biomaterial as a cell carrier in the form of an injectable hydrogel for the encapsulation and cartilage differentiation of MSCs from canine adipose tissue. We will produce a hydrogel of silk fibroin by an innovative process that we expect will contribute to chondrogenic differentiation of MSCs. We will also optimise the freezing and thawing of the cell-hydrogel construct, which has not been described before and will contribute significantly to the production of a pre-prepared, readily available product for potential therapeutic use. The proposed study represents continuation of the research we have conducted in the past. The results of the study could make an important contribution to the understanding of the mechanisms of differentiation of MSCs, thus enabeling progress in overcoming the obstacles to exploiting the differentiation potential of MSC in the clinic. The results of the project, together with the developing therapeutic use of donor MSCs, may bring the routine therapeutic use of MSCs closer to reality. The development of hydrogels with the ability to encapsulate other cells or therapeutic agents such as growth factors or extracellular vesicles would also enable their use for the potential treatment of many other diseases. At the same time, the project will also be relevant for human medicine, as the course and mechanisms of disease development are similar in many diseases in animals nad humans. Also, research into new veterinary treatments is an important basis for translational studies in human medicine, as they provide important data on the efficacy and safety of the use of cell products.