Microfabricated Gaps Reveal the Effect of Geometrical Control in Wound Healing

The geometry (size and shape) of gaps is a key determinant in controlling gap closure during wound healing. However, conventional methods for creating gaps result in un-defined geometries and poorly characterized conditions (cell death factors and cell debris), which can influence the gap closure process. To overcome these limitations, a novel method to create well-defined geometrical gaps is developed. First, smooth muscle cells (SMCs) are seeded in variously shaped micro-containers made out of hyaluronic acid hydrogels. Cell proliferation and cell tension induce fibrous collagen production by SMCs predominantly around the edges of the micro-containers. Upon removal of SMCs, the selectively deposited collagen results in micro-containers with cell-adhesive regions along the edges and walls. Fibroblasts are seeded in these micro-containers, and upon attaching and spreading, they naturally form gaps with different geometries. The rapid proliferation of fibroblasts from the edge results in filling and closure of the gaps. It is demonstrated that gap closure rate as well as closure mechanism is strongly influenced by geometrical features, which points to an important role for cellular tension and cell proliferation in gap closure.