Fibroblasts Close a Void in Free Space by a Purse-String Mechanism.

The mechanism by which stromal cells fill voids in injured tissue remains a fundamental question in regenerative medicine. While it is well-established that fibroblasts fill voids by depositing extracellular matrix (ECM) proteins as they migrate toward the wound site, little is known about their ability to adopt an epithelial-like purse-string behavior. To investigate fibroblast behavior during gap closure, we created an artificial wound with a large void space. We discovered that fibroblasts could form a free-standing bridge over deep microvoids, closing the void via purse-string contraction, a mechanism previously thought to be unique to epithelial wound closure. The findings also revealed that myosin II mediated contractility and intercellular adherent junctions were required for the closure of the fibroblast gap in our fabricated three-dimensional artificial wound. To fulfill their repair function under the specific microenvironmental conditions of wounds, fibroblasts appeared to acquire the structural features of epithelial cells, namely, contractile actin bundles that span over multiple cells along the boundary. These findings shed light on a novel mechanism by which stromal cells bridge the 3D gap during physiological processes such as morphogenesis and wound healing.

An injectable, self-assembled multicellular microsphere with the incorporation of fibroblast-derived extracellular matrix for therapeutic angiogenesis.

Decellularized human lung fibroblast-derived matrix (hFDM) has demonstrated its excellent proangiogenic capability. In this study, we propose a self-assembled, injectable multicellular microspheres containing human umbilical vein endothelial cells (HUVECs) and mesenchymal stem cell (MSCs), collagen hydrogel (Col), and hFDM toward therapeutic angiogenesis. Those multicellular microspheres are spontaneously formed by the mixtures of cell and hydrogel after being dropped on the parafilm for several hours. The size of microspheres can be manipulated via adjusting the initial volume of droplets and the culture period. The cells in the microspheres are highly viable. Multicellular microspheres show good capability of cell migration on 2D culture plate and also exhibit active cell sprouting in 3D environment (Col) forming capillary-like structures. We also find that multiple angiogenic-related factors are significantly upregulated with the multicellular microspheres prepared via Col and hFDM (Col/hFDM) than those prepared using Col alone or single cells (harvested from cocultured HUVECs/MSCs in monolayer). For therapeutic efficacy evaluation, three different groups of single cells, Col and Col/hFDM microspheres are injected to a hindlimb ischemic model, respectively, along with PBS injection as a control group. It is notable that Col/hFDM microspheres significantly improve the blood reperfusion and greatly attenuate the fibrosis level of the ischemic regions. In addition, Col/hFDM microspheres show higher cell engraftment level than that of the other groups. The incorporation of transplanted cells with host vasculature is detectable only with the treatment of Col/hFDM. Current results suggest that hFDM plays an important role in the multicellular microspheres for angiogenic cellular functions in vitro as well as in vivo. Taken together, our injectable multicellular microspheres (Col/hFDM) offer a very promising platform for cell delivery and tissue regenerative applications.