Authors:

Kyle Jeffery Heaton

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About Paper:

Volumetric muscle loss (VML) injuries are highly prevalent and often result in fibrotic scarring that reduces muscle function and compromises quality of life. Although many biomaterial therapies have been developed to treat VML, an understanding of how biomaterial features affect endogenous fibroblast activation and matrix deposition remains unexplored. One challenge lies in identifying fibroblasts in preserved tissue sections. It has been reported that alpha-smooth muscle actin (?-SMA), a well-known marker of activated fibroblasts in other tissues, does not accurately stain muscle-resident fibroblasts. To address this, we qualitatively and quantitatively compared the presence of fibroblasts stained with either alpha smooth muscle actin (?-SMA) or platelet-derived growth factor receptor alpha (PDGFRA) in bulk or porous hydrogels implanted into a mouse tibialis anterior VML defect. PDGFRA is a well-known marker for muscle-resident fibro/adipogenic progenitor cells (FAPs), a cell type believed to play a role in muscle fibrosis after injury. Muscle cryosections were processed using an optimized immunohistochemistry protocol. Additionally, the samples were visualized using fluorescence microscopy and the image processing tool ImageJ. Our data revealed fibroblast integration into the granular hydrogel injury models; as well as, a difference in staining abundance between ?-SMA and PDGFRA revealing a difference in identified staining elements. Our findings demonstrate the importance of antibody selection to stain for fibroblasts during muscle repair. Future work will correlate fibroblast presence with collagen staining to determine patterns of fibrosis around or within implanted biomaterials. Keywords: Volumetric Muscle Loss; Biomaterials; Immunohistochemistry; Fibrosis; Antibodies

Source:

Purdue University / 2024

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Co-authors:

Kyle Jeffery Heaton