Fibroblasts treated with macrophage conditioned medium results in phenotypic shifts and changes in collagen organization

Published on Jan 29, 2021in Materials Science and Engineering: C5.88
· DOI :10.1016/J.MSEC.2021.111915
Zhuqing Li2
Estimated H-index: 2
(Iowa State University),
Kaitlin M. Bratlie22
Estimated H-index: 22
(Iowa State University)
Abstract In tissue regeneration, the goal is to regenerate tissue similar to what was damaged or missing while preventing fibrotic scarring, which may lead to decreased mechanical strength and dissimilar tissue characteristics compared to native tissue. We believe collagen orientation plays a critical role in wound contraction and scarring and that it is modulated by myofibroblasts. We used macrophage conditioned medium to simulate complex events that can influence the fibroblast phenotype during the wound healing process. In addition to examining the effect of macrophage phenotype on fibroblasts, we inhibited focal adhesion kinase (FAK), Rho-associated protein kinase (ROCK), and myosin II for fibroblasts cultured on both tissue culture plastic and methacrylated gellan gum to understand how different pathways and materials influence fibroblast responses. Collagen orientation, α-SMA expression, focal adhesion area, and cell migration were altered by inhibition of FAK, ROCK, or myosin II and macrophage phenotype, along with the substrate. An increase in either focal adhesion area or α-smooth muscle actin (α-SMA) expression correlated with an aligned collagen orientation. Gellan gum hydrogels upregulated α-SMA expression in ROCK inhibited conditioned media and downregulated the FAK area in FAK and ROCK inhibited conditioned media. Myosin II had no impact on the α-SMA expression on the substrate compared to coverslip except for M2 conditioned medium. Gellan gum hydrogel significantly increased cell migration under FAK and Myosin II mediated conditioned media and unconditioned media. Collectively, our study examined how macrophage phenotype influences fibroblast response, which would be beneficial in controlling scar tissue formation.
#1Martin JakobssonH-Index: 60
#2Larry A. MayerH-Index: 59
Last. June PadmanH-Index: 3
view all 31 authors...
The processes controlling advance and retreat of outlet glaciers in fjords draining the Greenland Ice Sheet remain poorly known, undermining assessments of their dynamics and associated sea-level rise in a warming climate. Mass loss of the Greenland Ice Sheet has increased six-fold over the last four decades, with discharge and melt from outlet glaciers comprising key components of this loss. Here we acquired oceanographic data and multibeam bathymetry in the previously uncharted Sherard Osborn ...
#1Diah Gemala Ibrahim (Hasanuddin University)H-Index: 2
#2Ji-Ae Ko (Hiroshima University)H-Index: 16
Last. Yoshiaki Kiuchi (Hiroshima University)H-Index: 24
view all 5 authors...
: Scar formation is the most common cause for failure of glaucoma filtration surgery because of increased fibroblast proliferation and activation. We have now examined the effect of Y-27632, a Rho-associated protein kinase (ROCK) inhibitor, on postsurgical scarring formation in human Tenon fibroblasts (HTFs). Collagen gel contraction assay was used to compare contractility activity of Y-27632 with several antiglaucoma drugs. Immunofluorescence and western blotting were used to examine expression...
6 CitationsSource
#1Boris Hinz (U of T: University of Toronto)H-Index: 69
#2Christopher A. McCulloch (U of T: University of Toronto)H-Index: 86
Last. Nuno M. Coelho (U of T: University of Toronto)H-Index: 10
view all 3 authors...
Abstract Activated fibroblasts promote physiological wound repair following tissue injury. However, dysregulation of fibroblast activation contributes to the development of fibrosis by enhanced production and contraction of collagen-rich extracellular matrix. At the peak of their activities, fibroblasts undergo phenotypic conversion into highly contractile myofibroblasts by developing muscle-like features, including formation of contractile actin-myosin bundles. The phenotype and function of fib...
35 CitationsSource
#1Claire E. Witherel (Drexel University)H-Index: 7
#2Daniel Abebayehu (UVA: University of Virginia)H-Index: 9
Last. Kara L. Spiller (Drexel University)H-Index: 25
view all 4 authors...
Biomaterial-mediated inflammation and fibrosis remain a prominent challenge in designing materials to support tissue repair and regeneration. Despite the many biomaterial technologies that have been designed to evade or suppress inflammation (i.e. delivery of anti-inflammatory drugs, hydrophobic coatings, etc.) many materials are still subject to a foreign body response, resulting encapsulation of dense, scar-like extracellular matrix. The primary cells involved in biomaterial-mediated fibrosis ...
40 CitationsSource
#1Zhuqing Li (Iowa State University)H-Index: 2
#2Kaitlin M. Bratlie (Iowa State University)H-Index: 22
In tissue engineering scaffolds, macrophages play a critical role in determining the host response to implanted biomaterials. Macrophage phenotype is dynamic throughout the host response, and a balance of phenotypes is essential for timely progression from injury to proper wound healing. Therefore, it is important to predict how materials will modulate the response of macrophages. In this study, we investigated the effect of methacrylated gellan gum (GG) hydrogels on macrophage phenotype and pro...
1 CitationsSource
#1Monika Lodyga (U of T: University of Toronto)H-Index: 19
#2Elizabeth Cambridge (U of T: University of Toronto)H-Index: 3
Last. Boris Hinz (U of T: University of Toronto)H-Index: 69
view all 12 authors...
Macrophages contribute to the activation of fibroblastic cells into myofibroblasts, which secrete collagen and contract the collagen matrix to acutely repair injured tissue. Persistent myofibroblast activation leads to the accumulation of fibrotic scar tissue that impairs organ function. We investigated the key processes that turn acute beneficial repair into destructive progressive fibrosis. We showed that homotypic cadherin-11 interactions promoted the specific binding of macrophages to and pe...
53 CitationsSource
#2Ritva HeljasvaaraH-Index: 25
Last. Taina PihlajaniemiH-Index: 82
view all 5 authors...
#1Wenwen Li (TJU: Tianjin University)H-Index: 2
#2Changyue Sun (TJU: Tianjin University)H-Index: 5
Last. Dachao Li (TJU: Tianjin University)H-Index: 13
view all 8 authors...
This paper proposes a novel fiber attenuated total reflection (ATR) sensor with silver nanoparticles (AgNPs) on the flattened structure based on mid-infrared spectroscopy for detecting low concentration of glucose with high precision. The flattened structure was designed to add the effective optical path length to improve the sensitivity. AgNPs were then deposited on the surface of the flattened area of the fiber via chemical silver mirror reaction for further improving the sensitivity by enhanc...
8 CitationsSource
#1Anuraag Boddupalli (Iowa State University)H-Index: 4
#2Kaitlin M. Bratlie (Iowa State University)H-Index: 22
: The pH of dermal wounds shifts from neutral during the inflammatory phase to slightly basic in the tissue remodeling phase. Stage specific wound treatment can be developed using environmentally responsive alginate hydrogels. The chemistry of these networks dictates swelling behavior. Here, we fabricated alginate hydrogels using chain growth, step growth, and combined mixed mode gelation methods to crosslink methacrylated alginate (ALGMA) and gain control over swelling responses. Methacrylation...
5 CitationsSource
#1Fei Chang (SDU: Shandong University)H-Index: 1
#2Yunpeng Zhang (SDU: Shandong University)H-Index: 1
Last. Xunwei Wu (Harvard University)H-Index: 28
view all 9 authors...
: Rho-associated protein kinase (ROCK) plays crucial roles in the proliferation and migration of different types of cells. ROCK inhibitor Y-27632 was previously reported to inhibit melanoma cell growth, and ROCK signaling was suggested to be a therapeutic target for treating melanoma. However, the negative effect of Y-27632 on melanoma cells was mainly seen in studies on murine B16 melanoma cells. Here, we reported that ROCK inhibitor actually promoted human melanoma cell growth and migration in...
18 CitationsSource
Cited By0