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β2-adrenoceptor activation modulates skin wound healing processes to reduce scarring.

Le Provost GS, Pullar CE - J. Invest. Dermatol. (2014)

Bottom Line: Here we identify a β2AR-mediated mechanism for scar reduction. β2ARag significantly reduced HDF differentiation, via multiple cAMP and/or fibroblast growth factor 2 or basic FGF (FGF2)-dependent mechanisms, in the presence of transforming growth factor betaβ1, reduced contractile function, and inhibited mRNA expression of a number of profibrotic markers. β2ARag also reduced inflammation and angiogenesis in zebrafish and CAMs in vivo, respectively.In Red Duroc pig full-thickness wounds, β2ARag reduced both scar area and hyperpigmentation by almost 50% and significantly improved scar quality.Both macrophage infiltration and angiogenesis were initially decreased, whereas DF function was impaired in the β2ARag-treated porcine wound bed.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Physiology and Pharmacology, University of Leicester, Leicester, UK.

ABSTRACT
During wound healing, excessive inflammation, angiogenesis, and differentiated human dermal fibroblast (HDF ) function contribute to scarring, whereas hyperpigmentation negatively affects scar quality. Over 100 million patients heal with a scar every year. To investigate the role of the beta 2 adrenergic receptor (β2AR) in wound scarring, the ability of beta 2 adrenergic receptor agonist (β2ARag) to alter HDF differentiation and function, wound inflammation, angiogenesis, and wound scarring was explored in HDFs, zebrafish, chick chorioallantoic membrane assay (CAM), and a porcine skin wound model, respectively. Here we identify a β2AR-mediated mechanism for scar reduction. β2ARag significantly reduced HDF differentiation, via multiple cAMP and/or fibroblast growth factor 2 or basic FGF (FGF2)-dependent mechanisms, in the presence of transforming growth factor betaβ1, reduced contractile function, and inhibited mRNA expression of a number of profibrotic markers. β2ARag also reduced inflammation and angiogenesis in zebrafish and CAMs in vivo, respectively. In Red Duroc pig full-thickness wounds, β2ARag reduced both scar area and hyperpigmentation by almost 50% and significantly improved scar quality. Indeed, mechanisms delineated in vitro and in other in vivo models were evident in the β2ARag-treated porcine scars in vivo. Both macrophage infiltration and angiogenesis were initially decreased, whereas DF function was impaired in the β2ARag-treated porcine wound bed. These data collectively reveal the potential of β2ARag to improve skin scarring.

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Immunohistochemistry (IHC) reveals a reduction in profibrotic mechanisms underpinning the β2AR agonist (β2ARag)-mediated scar reduction. α-Naphtyl acetate esterase (α-NAE) staining (all biopsies) (a) and CD163 IHC (day 14 biopsies) (b) determined the percentage of wound bed macrophage-infiltrated area. Angiogenesis was assessed by von Willebrand factor (vWF) IHC (all biopsies) (c) and day 56 excised scars (d). vWF-positive discrete blood vessel density was determined within defined wound bed areas. Masson's trichrome staining, smooth muscle α actin (SMA), fibronectin (FN) EDA, and fibroblast growth factor 2 or basic FGF (FGF2) IHC were performed on day 56 excised scars. Wound bed area (defined from H&E staining (not shown)) without collagen fibers (red) was measured. (e) SMA and FN EDA positive-stained wound bed area was measured (f, g). FGF2 expression was evaluated using an intensity score (0–2) (h). Representative figures and graphical quantification are shown (2 biopsies/N=5 (biopsies); 10 scars/N=5 (excised scars)). Lines demarcate the wound bed. Scale bar=1 mm. Data are presented as mean±SEM (*P<0.05, **P<0.01, ***P<0.001). NS, not significant.
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fig5: Immunohistochemistry (IHC) reveals a reduction in profibrotic mechanisms underpinning the β2AR agonist (β2ARag)-mediated scar reduction. α-Naphtyl acetate esterase (α-NAE) staining (all biopsies) (a) and CD163 IHC (day 14 biopsies) (b) determined the percentage of wound bed macrophage-infiltrated area. Angiogenesis was assessed by von Willebrand factor (vWF) IHC (all biopsies) (c) and day 56 excised scars (d). vWF-positive discrete blood vessel density was determined within defined wound bed areas. Masson's trichrome staining, smooth muscle α actin (SMA), fibronectin (FN) EDA, and fibroblast growth factor 2 or basic FGF (FGF2) IHC were performed on day 56 excised scars. Wound bed area (defined from H&E staining (not shown)) without collagen fibers (red) was measured. (e) SMA and FN EDA positive-stained wound bed area was measured (f, g). FGF2 expression was evaluated using an intensity score (0–2) (h). Representative figures and graphical quantification are shown (2 biopsies/N=5 (biopsies); 10 scars/N=5 (excised scars)). Lines demarcate the wound bed. Scale bar=1 mm. Data are presented as mean±SEM (*P<0.05, **P<0.01, ***P<0.001). NS, not significant.

Mentions: Wound inflammation was investigated using both the macrophage α-naphtyl acetate esterase (α-NAE) stain and an anti-macrophage antibody, CD163, specific for the M2 alternatively activated/healer macrophages (Weisser et al., 2013). α-NAE staining revealed an 18% decrease in biopsy macrophage-infiltrated area, after 7 days, in β2ARag-treated wounds (Figure 5a). In contrast, after 14 days, β2ARag treatment increased the macrophage-infiltrated area by 28% (Figure 5a). From 21 to 56 days post wounding, there was no difference in macrophage-infiltrated area between groups (Figure 5a). At 14 days post wounding, CD163-specific staining appeared similar to the α-NAE staining (Figure 5b).


β2-adrenoceptor activation modulates skin wound healing processes to reduce scarring.

Le Provost GS, Pullar CE - J. Invest. Dermatol. (2014)

Immunohistochemistry (IHC) reveals a reduction in profibrotic mechanisms underpinning the β2AR agonist (β2ARag)-mediated scar reduction. α-Naphtyl acetate esterase (α-NAE) staining (all biopsies) (a) and CD163 IHC (day 14 biopsies) (b) determined the percentage of wound bed macrophage-infiltrated area. Angiogenesis was assessed by von Willebrand factor (vWF) IHC (all biopsies) (c) and day 56 excised scars (d). vWF-positive discrete blood vessel density was determined within defined wound bed areas. Masson's trichrome staining, smooth muscle α actin (SMA), fibronectin (FN) EDA, and fibroblast growth factor 2 or basic FGF (FGF2) IHC were performed on day 56 excised scars. Wound bed area (defined from H&E staining (not shown)) without collagen fibers (red) was measured. (e) SMA and FN EDA positive-stained wound bed area was measured (f, g). FGF2 expression was evaluated using an intensity score (0–2) (h). Representative figures and graphical quantification are shown (2 biopsies/N=5 (biopsies); 10 scars/N=5 (excised scars)). Lines demarcate the wound bed. Scale bar=1 mm. Data are presented as mean±SEM (*P<0.05, **P<0.01, ***P<0.001). NS, not significant.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4263603&req=5

fig5: Immunohistochemistry (IHC) reveals a reduction in profibrotic mechanisms underpinning the β2AR agonist (β2ARag)-mediated scar reduction. α-Naphtyl acetate esterase (α-NAE) staining (all biopsies) (a) and CD163 IHC (day 14 biopsies) (b) determined the percentage of wound bed macrophage-infiltrated area. Angiogenesis was assessed by von Willebrand factor (vWF) IHC (all biopsies) (c) and day 56 excised scars (d). vWF-positive discrete blood vessel density was determined within defined wound bed areas. Masson's trichrome staining, smooth muscle α actin (SMA), fibronectin (FN) EDA, and fibroblast growth factor 2 or basic FGF (FGF2) IHC were performed on day 56 excised scars. Wound bed area (defined from H&E staining (not shown)) without collagen fibers (red) was measured. (e) SMA and FN EDA positive-stained wound bed area was measured (f, g). FGF2 expression was evaluated using an intensity score (0–2) (h). Representative figures and graphical quantification are shown (2 biopsies/N=5 (biopsies); 10 scars/N=5 (excised scars)). Lines demarcate the wound bed. Scale bar=1 mm. Data are presented as mean±SEM (*P<0.05, **P<0.01, ***P<0.001). NS, not significant.
Mentions: Wound inflammation was investigated using both the macrophage α-naphtyl acetate esterase (α-NAE) stain and an anti-macrophage antibody, CD163, specific for the M2 alternatively activated/healer macrophages (Weisser et al., 2013). α-NAE staining revealed an 18% decrease in biopsy macrophage-infiltrated area, after 7 days, in β2ARag-treated wounds (Figure 5a). In contrast, after 14 days, β2ARag treatment increased the macrophage-infiltrated area by 28% (Figure 5a). From 21 to 56 days post wounding, there was no difference in macrophage-infiltrated area between groups (Figure 5a). At 14 days post wounding, CD163-specific staining appeared similar to the α-NAE staining (Figure 5b).

Bottom Line: Here we identify a β2AR-mediated mechanism for scar reduction. β2ARag significantly reduced HDF differentiation, via multiple cAMP and/or fibroblast growth factor 2 or basic FGF (FGF2)-dependent mechanisms, in the presence of transforming growth factor betaβ1, reduced contractile function, and inhibited mRNA expression of a number of profibrotic markers. β2ARag also reduced inflammation and angiogenesis in zebrafish and CAMs in vivo, respectively.In Red Duroc pig full-thickness wounds, β2ARag reduced both scar area and hyperpigmentation by almost 50% and significantly improved scar quality.Both macrophage infiltration and angiogenesis were initially decreased, whereas DF function was impaired in the β2ARag-treated porcine wound bed.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Physiology and Pharmacology, University of Leicester, Leicester, UK.

ABSTRACT
During wound healing, excessive inflammation, angiogenesis, and differentiated human dermal fibroblast (HDF ) function contribute to scarring, whereas hyperpigmentation negatively affects scar quality. Over 100 million patients heal with a scar every year. To investigate the role of the beta 2 adrenergic receptor (β2AR) in wound scarring, the ability of beta 2 adrenergic receptor agonist (β2ARag) to alter HDF differentiation and function, wound inflammation, angiogenesis, and wound scarring was explored in HDFs, zebrafish, chick chorioallantoic membrane assay (CAM), and a porcine skin wound model, respectively. Here we identify a β2AR-mediated mechanism for scar reduction. β2ARag significantly reduced HDF differentiation, via multiple cAMP and/or fibroblast growth factor 2 or basic FGF (FGF2)-dependent mechanisms, in the presence of transforming growth factor betaβ1, reduced contractile function, and inhibited mRNA expression of a number of profibrotic markers. β2ARag also reduced inflammation and angiogenesis in zebrafish and CAMs in vivo, respectively. In Red Duroc pig full-thickness wounds, β2ARag reduced both scar area and hyperpigmentation by almost 50% and significantly improved scar quality. Indeed, mechanisms delineated in vitro and in other in vivo models were evident in the β2ARag-treated porcine scars in vivo. Both macrophage infiltration and angiogenesis were initially decreased, whereas DF function was impaired in the β2ARag-treated porcine wound bed. These data collectively reveal the potential of β2ARag to improve skin scarring.

Show MeSH
Related in: MedlinePlus