Preconditioning Methods to Improve Mesenchymal Stromal Cell-Derived Extracellular Vesicles in Bone Regeneration—A Systematic Review

SIMPLE SUMMARY: The evidence of the therapeutic effects of mesenchymal stromal cells (MSCs), so-called stem cells, in several diseases relies mostly on the substances they secrete, including their extracellular vesicles (EVs). EVs are an important component of cell communication and they carry a cargo that is similar to their parent cell. Cells respond differently based on their microenvironment, and so it is expected that the therapeutic potential of these vesicles can be modulated by the enrichment of their parent cell microenvironment. With this in mind, we conducted a systematic search for papers that preconditioned MSCs and collected their EVs to assess their potential to favor bone formation. The results showed different methods for MSC preconditioning, including chemical induction, culture conditions, and genetic modifications. All methods were able to improve the therapeutic effects of the derived EVs for bone formation. However, the heterogeneity among studies—regarding the type of cell, EV concentration, and scaffolds—made it difficult to compare fairly the types of preconditioning methods. In summary, the microenvironment greatly influences MSCs, and using preconditioning methods can potentially improve the therapeutic effects of their derived EVs in bone regeneration and other bone diseases. ABSTRACT: Mesenchymal stromal cells (MSCs) have long been used in research for bone regeneration, with evidence of their beneficial properties. In the segmental area of MSC-based therapies, MSC-derived extracellular vesicles (EVs) have also shown great therapeutic effects in several diseases, including bone healing. This study aimed to assess whether the conditioning of MSCs improves the therapeutic effects of their derived extracellular vesicles for bone regeneration. Electronic research was performed until February 2021 to recover the studies in the following databases: PubMed, Scopus, and Web of Science. The studies were screened based on the inclusion criteria. Relevant information was extracted, including in vitro and in vivo experiments, and the animal studies were evaluated for risk of bias by the SYRCLE tool. A total of 463 studies were retrieved, and 18 studies met the inclusion criteria (10 studies for their in vitro analysis, and 8 studies for their in vitro and in vivo analysis). The conditioning methods reported included: osteogenic medium; dimethyloxalylglycine; dexamethasone; strontium-substituted calcium silicate; hypoxia; 3D mechanical microenvironment; and the overexpression of miR-375, bone morphogenetic protein-2, and mutant hypoxia-inducible factor-1α. The conditioning methods of MSCs in the reported studies generate exosomes able to significantly promote bone regeneration. However, heterogeneity regarding cell source, conditioning method, EV isolation and concentration, and defect model was observed among the studies. The different conditioning methods reported in this review do improve the therapeutic effects of MSC-derived EVs for bone regeneration, but they still need to be addressed in larger animal models for further clinical application.