PROJECT SUMMARY
Autologous bone grafting, the current gold standard for treating critical sized bone defects and chronic
nonunions, has limited success, leaving a need for adjunct treatments to contribute to overall bone healing.
Extracellular vesicles, of which there are many types, are a promising new cell-free, membrane-bound
therapeutic in the field of regenerative medicine. Specific to the field of bone regeneration and repair are matrix
vesicles; small extracellular vesicles released by mineralization-competent cells that become anchored in the
extracellular matrix and are a necessary component of mineralized bone formation via endochondral ossification.
They are involved in cellular signaling via small, non-coding microRNA in the growth plate, suggesting that they
may play a similar role in bone. Previous research indicates that chondrocyte-derived matrix vesicles are
enriched with microRNA and other cell signaling molecules that contribute to their ability to influence proliferation
and differentiation of target cells, however the role and mechanism of action of osteoblast-derived matrix
vesicles, particularly in bone as opposed to the growth plate, remains to be elucidated. With this information, the
biological mediation of bone development and regeneration that matrix vesicles provide may translate into novel
therapeutic potential for orthopedic pathologies, including critical size bone defects and fracture nonunions.
Therefore, we hypothesize that matrix vesicles produced by osteoblast-lineage cells, as a specific subset of
extracellular vesicle, use their microRNA cargo to enhance osteogenic differentiation and proliferation, drive
osteogenesis, and aid in bone defect healing. First, we aim to determine the relationship between osteoblast-
derived matrix vesicles and another class of extracellular vesicle, the exosome. Next, we aim to determine the
pathways targeted by matrix vesicle microRNA cargo. Finally, we aim to determine the extent to which
osteoblast-derived matrix vesicles modulate osteogenesis and bone defect healing. To test these aims, we will
use osteoblast-lineage cells to derive matrix vesicles and characterize protein expression, size, and morphology.
We will also use in vitro experimental models to assess pathway involvement and evaluate co-culture response
with osteoblast-like cells. To test the therapeutic potential, we will use a translatable in vivo model of a mouse
long bone defect and a biorthogonal injectable hydrogel to deliver matrix vesicles. We expect to find that
osteoblast-derived matrix vesicles are a specialized subclass of extracellular vesicle with microRNA cargo that
targets the canonical Wnt pathway to activate cellular signaling and leads to increased osteoblastic differentiation
of target cells when co-cultured in vitro. Our in vivo model is expected to demonstrate improved healing upon
treatment with hydrogel-delivered matrix vesicles. All of which together demonstrates the role matrix vesicles
play in the coordinated effort to form new bone and their viability as a therapeutic option to improve bone healing.
