SCARLESS WOUND HEALING: Increased Collagen Deposition Along Wound Edge in Human Fibroblasts vs. hEScells

Roxanne Piol Bartolome, Tam Thanh Nguyen, Eileen Rosalyn Ing

Abstract


Scarless wound healing is the ideal result to all who suffer from minor to major wounds. Scars occur as a result of excess collagen being assembled in a wound. Prolyl-4-hydroxylase (P4H) is the enzyme that modifies single collagen strands so that they can assemble into larger fibrils, and together with overall collagen levels determine the extent of scarring. Documenting the P4H levels in a wound and finding a more refined measurement of scar potential may give insights into wound healing. One process in which P4H was observed in wounds involved in vitro scratch assays on human primary fibroblasts containing TGFβ1.  A wound was created by using a micropipette tip that was dragged across the diameter along the most confluent region of the plate of primary fibroblast cells.   Immunohistochemistry (IHC) was performed on the in vitro scratch assays using antibody 5B5, a reliable marker for P4H. The staining at the edges of the scratch of the in vitro scratch assay was positive for the 5B5 marker, indicating changes in collagen chain assembly.  Human embryonic stem cells (hEScells) were also used in comparison to the human primary fibroblasts cells. The IHC of the hEScells showed fluorescence of the 5B5 marker throughout the entire plate instead of just on the edges of the scratch, indicating that the signaling responsible for turning on P4H is missing in the hEScells tested. These results suggest that collagen deposition and assembly in human primary fibroblast cells are more localized compared to hES cells, and may explain in part why hES cells demonstrate the ability to heal without scarring.  Our results suggest that targeting the border cells of wounds should be pursued to prevent the formation of scars.

Keywords


scarless wound healing, collagen deposition, human embryonic stem cells (hEScells), human primary fibroblasts, prolyl-4-hydroxylase (P4H), 5B5, TGFβ1, Vimentin

Full Text: PDF

Refbacks

  • There are currently no refbacks.