This subproject is one of many research subprojects utilizing theresources provided by a Center grant funded by NIH/NCRR. The subproject andinvestigator (PI) may have received primary funding from another NIH source,and thus could be represented in other CRISP entries. The institution listed isfor the Center, which is not necessarily the institution for the investigator.Wound healing is the process by which the body repairs damaged tissue. Through the work of specialized cells called fibroblasts, connective tissue is regenerated at the site of injury, as fibroblasts produce collagen, fibronectin, elastin, and peptidoglycan to stabilize and heal the wound. Recent studies have shown that in vitro skin-equivalent, collagen-based tissue constructs called RAFTs can be used to model damaged tissue.We developed a model raft culture model based on a collagen/fibronectin gel mixture. Specifically, we constructed a normal collagen-based raft with a circular fibronectin ?plug? in the center to simulate the introduction of a wound into the collagen gel. Fibronectin, unlike collagen, does not generate second-harmonic generation (SHG) signals under multiphoton microscopy (MPM). Thus, the newly synthesized collagen in the fibronectin gel region can be distinguished from the old collagen already present in the outer regions of the construct. This model facilitates the analysis of the overall collagen production and 3-D structure.
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