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The CXC chemokine cCAF stimulates precocious deposition of ECM molecules by wound fibroblasts, accelerating development of granulation tissue.

Feugate JE, Wong L, Li QJ, Martins-Green M - BMC Cell Biol. (2002)

Bottom Line: In contrast, stimulation of fibronectin and collagen I both require the entire molecule and do not involve changes in gene expression.Fibronectin accumulation appears to be linked to tenascin production, and collagen I to decreased MMP-1 levels.In addition, cCAF is chemotactic for fibroblasts and accelerates their migration.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Cell Biology and Neuroscience, University of California, Riverside, California, USA. feugate@citrus.ucr.edu

ABSTRACT

Background: During wound repair, fibroblasts orchestrate replacement of the provisional matrix formed during clotting with tenascin, cellular fibronectin and collagen III. These, in turn, are critical for migration of endothelial cells, keratinocytes and additional fibroblasts into the wound site. Fibroblasts are also important in the deposition of collagen I during scar formation. The CXC chemokine chicken Chemotactic and Angiogenic Factor (cCAF), is highly expressed by fibroblasts after wounding and during development of the granulation tissue, especially in areas where extracellular matrix (ECM) is abundant. We hypothesized that cCAF stimulates fibroblasts to produce these matrix molecules.

Results: Here we show that this chemokine can stimulate precocious deposition of tenascin, fibronectin and collagen I, but not collagen III. Studies in culture and in vivo show that tenascin stimulation can also be achieved by the N-terminal 15 aas of the protein and occurs at the level of gene expression. In contrast, stimulation of fibronectin and collagen I both require the entire molecule and do not involve changes in gene expression. Fibronectin accumulation appears to be linked to tenascin production, and collagen I to decreased MMP-1 levels. In addition, cCAF is chemotactic for fibroblasts and accelerates their migration.

Conclusions: These previously unknown functions for chemokines suggest that cCAF, the chicken orthologue of human IL-8, enhances healing by rapidly chemoattracting fibroblasts into the wound site and stimulating them to produce ECM molecules, leading to precocious development of granulation tissue. This acceleration of the repair process may have important application to healing of impaired wounds.

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Effect of cCAF on collagen levels.(A&B) Immunoblot analysis for Colls I and III, respectively. All lanes contain equal amounts of total protein, as measured by the DC protein assay (BioRad). Treatment of fibroblasts with cCAF, but not the C- or N-peptides, stimulates a small but reproducible increase in protein levels of Coll I when compared with untreated cells (A). Neither cCAF nor peptide treatments changed Coll III levels (B).(C) Levels of Colls I and III in wounds treated with cCAF. Immunoblot analysis of protein extracts from wounded wings treated with 1 μg of cCAF were performed as described in Materials and Methods section. Coll I levels are slightly higher than those of the control after 3 days of treatment with cCAF. However, they are similar to those of the control at 7 days. Coll III levels do not differ between control and cCAF-treated wings.
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Figure 6: Effect of cCAF on collagen levels.(A&B) Immunoblot analysis for Colls I and III, respectively. All lanes contain equal amounts of total protein, as measured by the DC protein assay (BioRad). Treatment of fibroblasts with cCAF, but not the C- or N-peptides, stimulates a small but reproducible increase in protein levels of Coll I when compared with untreated cells (A). Neither cCAF nor peptide treatments changed Coll III levels (B).(C) Levels of Colls I and III in wounds treated with cCAF. Immunoblot analysis of protein extracts from wounded wings treated with 1 μg of cCAF were performed as described in Materials and Methods section. Coll I levels are slightly higher than those of the control after 3 days of treatment with cCAF. However, they are similar to those of the control at 7 days. Coll III levels do not differ between control and cCAF-treated wings.

Mentions: To determine whether cCAF stimulates fibroblasts to produce increased levels of Colls I and III, we treated cultures as for TN. Immunoblot analysis for Coll I showed that cCAF, but not the N- or C-peptides, caused a small increase in Coll I protein levels (Fig. 6A). Probing the same blots for Coll III revealed no cCAF-induced difference in the level of production of this protein (Fig. 6B). In vivo, Coll I was also slightly increased by treatment with the chemokine at 3 days after wounding but did not increase to higher levels than the control at 7 days, whereas the same blots probed for Coll III showed similar levels in treated and control at both 3 and 7 days (Figure 6C).


The CXC chemokine cCAF stimulates precocious deposition of ECM molecules by wound fibroblasts, accelerating development of granulation tissue.

Feugate JE, Wong L, Li QJ, Martins-Green M - BMC Cell Biol. (2002)

Effect of cCAF on collagen levels.(A&B) Immunoblot analysis for Colls I and III, respectively. All lanes contain equal amounts of total protein, as measured by the DC protein assay (BioRad). Treatment of fibroblasts with cCAF, but not the C- or N-peptides, stimulates a small but reproducible increase in protein levels of Coll I when compared with untreated cells (A). Neither cCAF nor peptide treatments changed Coll III levels (B).(C) Levels of Colls I and III in wounds treated with cCAF. Immunoblot analysis of protein extracts from wounded wings treated with 1 μg of cCAF were performed as described in Materials and Methods section. Coll I levels are slightly higher than those of the control after 3 days of treatment with cCAF. However, they are similar to those of the control at 7 days. Coll III levels do not differ between control and cCAF-treated wings.
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Related In: Results  -  Collection

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Figure 6: Effect of cCAF on collagen levels.(A&B) Immunoblot analysis for Colls I and III, respectively. All lanes contain equal amounts of total protein, as measured by the DC protein assay (BioRad). Treatment of fibroblasts with cCAF, but not the C- or N-peptides, stimulates a small but reproducible increase in protein levels of Coll I when compared with untreated cells (A). Neither cCAF nor peptide treatments changed Coll III levels (B).(C) Levels of Colls I and III in wounds treated with cCAF. Immunoblot analysis of protein extracts from wounded wings treated with 1 μg of cCAF were performed as described in Materials and Methods section. Coll I levels are slightly higher than those of the control after 3 days of treatment with cCAF. However, they are similar to those of the control at 7 days. Coll III levels do not differ between control and cCAF-treated wings.
Mentions: To determine whether cCAF stimulates fibroblasts to produce increased levels of Colls I and III, we treated cultures as for TN. Immunoblot analysis for Coll I showed that cCAF, but not the N- or C-peptides, caused a small increase in Coll I protein levels (Fig. 6A). Probing the same blots for Coll III revealed no cCAF-induced difference in the level of production of this protein (Fig. 6B). In vivo, Coll I was also slightly increased by treatment with the chemokine at 3 days after wounding but did not increase to higher levels than the control at 7 days, whereas the same blots probed for Coll III showed similar levels in treated and control at both 3 and 7 days (Figure 6C).

Bottom Line: In contrast, stimulation of fibronectin and collagen I both require the entire molecule and do not involve changes in gene expression.Fibronectin accumulation appears to be linked to tenascin production, and collagen I to decreased MMP-1 levels.In addition, cCAF is chemotactic for fibroblasts and accelerates their migration.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Cell Biology and Neuroscience, University of California, Riverside, California, USA. feugate@citrus.ucr.edu

ABSTRACT

Background: During wound repair, fibroblasts orchestrate replacement of the provisional matrix formed during clotting with tenascin, cellular fibronectin and collagen III. These, in turn, are critical for migration of endothelial cells, keratinocytes and additional fibroblasts into the wound site. Fibroblasts are also important in the deposition of collagen I during scar formation. The CXC chemokine chicken Chemotactic and Angiogenic Factor (cCAF), is highly expressed by fibroblasts after wounding and during development of the granulation tissue, especially in areas where extracellular matrix (ECM) is abundant. We hypothesized that cCAF stimulates fibroblasts to produce these matrix molecules.

Results: Here we show that this chemokine can stimulate precocious deposition of tenascin, fibronectin and collagen I, but not collagen III. Studies in culture and in vivo show that tenascin stimulation can also be achieved by the N-terminal 15 aas of the protein and occurs at the level of gene expression. In contrast, stimulation of fibronectin and collagen I both require the entire molecule and do not involve changes in gene expression. Fibronectin accumulation appears to be linked to tenascin production, and collagen I to decreased MMP-1 levels. In addition, cCAF is chemotactic for fibroblasts and accelerates their migration.

Conclusions: These previously unknown functions for chemokines suggest that cCAF, the chicken orthologue of human IL-8, enhances healing by rapidly chemoattracting fibroblasts into the wound site and stimulating them to produce ECM molecules, leading to precocious development of granulation tissue. This acceleration of the repair process may have important application to healing of impaired wounds.

Show MeSH
Related in: MedlinePlus