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Role of dermatopontin in re-epithelialization: implications on keratinocyte migration and proliferation.

Krishnaswamy VR, Korrapati PS - Sci Rep (2014)

Bottom Line: Re-epithelialization is a key event in wound healing and any impairment in that process is associated with various pathological conditions.The results showed that DPT promotes keratinocyte migration in a dose dependant fashion but fail to induce proliferation.To conclude, DPT has a profound role in wound healing specifically during re-epithelialization by promoting keratinocyte migration via paracrine action from the underlying dermis.

View Article: PubMed Central - PubMed

Affiliation: Biomaterials Department, CSIR - Central Leather Research Institute, Adyar, Chennai - 600 020, Tamil Nadu, India.

ABSTRACT
Re-epithelialization is a key event in wound healing and any impairment in that process is associated with various pathological conditions. Epidermal keratinocyte migration and proliferation during re-epithelialization is largely regulated by the cytokines and growth factors from the provisional matrix and dermis. Extracellular matrix consists of numerous growth factors which mediate cell migration via cell membrane receptors. Dermatopontin (DPT), a non-collagenous matrix protein highly expressed in dermis is known for its striking ability to promote cell adhesion. DPT also enhances the biological activity of transforming growth factor beta 1 which plays a central role in the process of wound healing. This study was designed to envisage the role of DPT in keratinocyte migration and proliferation along with its mRNA and protein expression pattern in epidermis. The results showed that DPT promotes keratinocyte migration in a dose dependant fashion but fail to induce proliferation. Further, PCR and immunodetection studies revealed that the mRNA and protein expression of DPT is considerably negligible in the epidermis in contrast to the dermis. To conclude, DPT has a profound role in wound healing specifically during re-epithelialization by promoting keratinocyte migration via paracrine action from the underlying dermis.

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Related in: MedlinePlus

Phalloidin staining of cells treated with and without DPT.The number of thick and extended lamellipodia forming cells was higher in DPT treated cells (a - d) than in control cells (e - h). A typical representation of images are (a & e) Phase contrast image showing the cytoplasmic protrusions formed. (b & f) F-Actin filaments stained with Phalloidin (green) showing the thick actin network formed at the migrating edge, a typical phenotype of the migratory cells. (c & g) Nucleus stained with DAPI (blue). (d & h) Fused image showing the stained nucleus and F-actin. The focal adhesion points formed in the direction of migration are indicated by red arrows in the subset of image b. Scale bar – 50 μm.
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f3: Phalloidin staining of cells treated with and without DPT.The number of thick and extended lamellipodia forming cells was higher in DPT treated cells (a - d) than in control cells (e - h). A typical representation of images are (a & e) Phase contrast image showing the cytoplasmic protrusions formed. (b & f) F-Actin filaments stained with Phalloidin (green) showing the thick actin network formed at the migrating edge, a typical phenotype of the migratory cells. (c & g) Nucleus stained with DAPI (blue). (d & h) Fused image showing the stained nucleus and F-actin. The focal adhesion points formed in the direction of migration are indicated by red arrows in the subset of image b. Scale bar – 50 μm.

Mentions: The extension of lamellipodia, an indicator of cell migration was assessed by staining the actin fibers. Phalloidin staining of F- actin showed the formation of thick fiber assembly and focal adhesion points in the DPT treated cells (fig. 3) confirming the involvement of DPT in keratinocyte migration. The lamellipodia formation captured after scratch assay is shown in the fig. 4. The untreated cells show very few lamellipodia formations when compared to the treated cells which displayed multiple lamellipodial extensions indicating the enhancement in the migratory rate of HaCaT cells by DPT.


Role of dermatopontin in re-epithelialization: implications on keratinocyte migration and proliferation.

Krishnaswamy VR, Korrapati PS - Sci Rep (2014)

Phalloidin staining of cells treated with and without DPT.The number of thick and extended lamellipodia forming cells was higher in DPT treated cells (a - d) than in control cells (e - h). A typical representation of images are (a & e) Phase contrast image showing the cytoplasmic protrusions formed. (b & f) F-Actin filaments stained with Phalloidin (green) showing the thick actin network formed at the migrating edge, a typical phenotype of the migratory cells. (c & g) Nucleus stained with DAPI (blue). (d & h) Fused image showing the stained nucleus and F-actin. The focal adhesion points formed in the direction of migration are indicated by red arrows in the subset of image b. Scale bar – 50 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4260223&req=5

f3: Phalloidin staining of cells treated with and without DPT.The number of thick and extended lamellipodia forming cells was higher in DPT treated cells (a - d) than in control cells (e - h). A typical representation of images are (a & e) Phase contrast image showing the cytoplasmic protrusions formed. (b & f) F-Actin filaments stained with Phalloidin (green) showing the thick actin network formed at the migrating edge, a typical phenotype of the migratory cells. (c & g) Nucleus stained with DAPI (blue). (d & h) Fused image showing the stained nucleus and F-actin. The focal adhesion points formed in the direction of migration are indicated by red arrows in the subset of image b. Scale bar – 50 μm.
Mentions: The extension of lamellipodia, an indicator of cell migration was assessed by staining the actin fibers. Phalloidin staining of F- actin showed the formation of thick fiber assembly and focal adhesion points in the DPT treated cells (fig. 3) confirming the involvement of DPT in keratinocyte migration. The lamellipodia formation captured after scratch assay is shown in the fig. 4. The untreated cells show very few lamellipodia formations when compared to the treated cells which displayed multiple lamellipodial extensions indicating the enhancement in the migratory rate of HaCaT cells by DPT.

Bottom Line: Re-epithelialization is a key event in wound healing and any impairment in that process is associated with various pathological conditions.The results showed that DPT promotes keratinocyte migration in a dose dependant fashion but fail to induce proliferation.To conclude, DPT has a profound role in wound healing specifically during re-epithelialization by promoting keratinocyte migration via paracrine action from the underlying dermis.

View Article: PubMed Central - PubMed

Affiliation: Biomaterials Department, CSIR - Central Leather Research Institute, Adyar, Chennai - 600 020, Tamil Nadu, India.

ABSTRACT
Re-epithelialization is a key event in wound healing and any impairment in that process is associated with various pathological conditions. Epidermal keratinocyte migration and proliferation during re-epithelialization is largely regulated by the cytokines and growth factors from the provisional matrix and dermis. Extracellular matrix consists of numerous growth factors which mediate cell migration via cell membrane receptors. Dermatopontin (DPT), a non-collagenous matrix protein highly expressed in dermis is known for its striking ability to promote cell adhesion. DPT also enhances the biological activity of transforming growth factor beta 1 which plays a central role in the process of wound healing. This study was designed to envisage the role of DPT in keratinocyte migration and proliferation along with its mRNA and protein expression pattern in epidermis. The results showed that DPT promotes keratinocyte migration in a dose dependant fashion but fail to induce proliferation. Further, PCR and immunodetection studies revealed that the mRNA and protein expression of DPT is considerably negligible in the epidermis in contrast to the dermis. To conclude, DPT has a profound role in wound healing specifically during re-epithelialization by promoting keratinocyte migration via paracrine action from the underlying dermis.

Show MeSH
Related in: MedlinePlus