Autologous Exosomes From Platelet-rich Plasma as a Modern Tool of Regenerative Dentistry

The research focuses on the laboratory assessment of the content and the regenerative and anti-inflammatory potential of autologous exosomes from platelet-rich plasma, as well as their comparison with other autologous products used in intra-articular injections (PRP, PRF).

The work will include:

1. Production of autologous materials (PRP, PRF, EXO); appropriate centrifuges and the AutologIX system (Biological Innovations) will be used for isolation.
2. Characterization of physico-chemical properties of EXO (concentration, size, morphology), using analytical methods such as nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM).
3. Analysis of content using flow cytometry and spectrophotometry.
4. Assessment of uptake by cells (using confocal microscopy) and promotion of proliferation and migration of human chondrocytes and fibroblasts.
5. Comparative analysis of the results obtained (1) for the study and control groups and (2) for PRP, PRF, and EXO.

The implementation of the project will constitute a significant contribution to the development of knowledge about the molecular mechanisms of the regenerative action of plasma-derived exosomes. The obtained results will allow a better understanding of the action of the evaluated systems, and, consequently, better effects of intra-articular injections through the optimal selection of autologous injection material. It should be emphasized that autologous injection therapies are safe for patients and are associated with the lowest risk of complications among all injection therapies used in the treatment of TMJ disorders, which is why their precise investigation is very important.

• Laboratory analysis of the content and biological activity of PRP, PRF, and exosomes obtained from blood samples (Diagnostic test): Laboratory analysis include in vitro tests aimed to: (1) evaluation of the size and concentration of exosomes, (2) the measurement of the level of cytokines and growth factors in the samples of PRP, PRF and EXO, (3) comparison of the impact of exosomes, PRF and PRP (from the study and control groups) on the proliferation and migration of human chondrocytes and fibroblasts.

Primary outcomes

• Comparison of the number and size of exosomes obtained from the study and control groups [04.2026 - 12.2027]
• Comparison of the level of cytokines and growth factors in exosomes, PRF and PRP samples obtained from the study and control groups [04.2026 - 12.2027]
• Comparison of the impact of exosomes, PRF and PRP (from the study and control groups) on the proliferation and migration of human chondrocytes and fibroblasts [04.2026 - 12.2027]

Sex: All, ages: 18 Years to any, accepts healthy volunteers

Inclusion Criteria:

* chronic pain of the temporomandibular joints and/or masticatory muscles and/or chronic dysfunction of the temporomandibular joints (lasting more than 3 months, diagnosed based on the standardized, international Diagnostic Criteria for Temporomandibular Disorders (DC/TMD) protocol) - criterion for the study group
* consent to participate in the study - criterion for both groups
* age over 18 years - criterion for both groups

Exclusion Criteria:

* cancer,
* pregnancy
* uncontrolled diabetes
* cardiovascular diseases (platelet dysfunction syndrome, critical thrombocytopenia, hemodynamic instability, sepsis, hemoglobin < 10 g/dl, platelet count < 105/µl)
* severe autoimmune diseases
* taking anticoagulants
* active skin lesions at the planned puncture site
* infections, patient undergoing antibiotic therapy, or within 3 weeks of its completion.