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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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Immunofluorescent detection of DPT protein in normal skin sections.(a) Sections treated with FITC conjugated secondary antibody showing strong signals in the dermis (indicated by arrows). ii) Magnified image of the same section revealing the absence of DPT protein in the epidermis. Scale bar – 100 μm.
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f8: Immunofluorescent detection of DPT protein in normal skin sections.(a) Sections treated with FITC conjugated secondary antibody showing strong signals in the dermis (indicated by arrows). ii) Magnified image of the same section revealing the absence of DPT protein in the epidermis. Scale bar – 100 μm.

Mentions: DPT protein expression was analyzed using western blotting and immunohistological studies. The results of the western blotting shown in the fig. 6b clearly depicted that DPT is not expressed in epidermis and HaCaT cells, in contrast to the conspicuous expression in the dermis and normal fibroblasts. In histological analysis, the sections treated with anti – human DPT and subsequent HRP or FITC conjugated secondary antibody revealed that DPT protein levels are markedly low in epidermis than in dermis (fig. 7 and fig. 8). The brown color developed due to HRP-DAB reaction was present throughout the dermis localizing on the collagen bundles. The epidermis showed a very feeble or no color development indicating the absence of antigen (DPT protein). The bright signals in the immunofluorescence studies also corroborated that DPT is absent and abundant in the epidermis and dermis respectively.


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

Krishnaswamy VR, Korrapati PS - Sci Rep (2014)

Immunofluorescent detection of DPT protein in normal skin sections.(a) Sections treated with FITC conjugated secondary antibody showing strong signals in the dermis (indicated by arrows). ii) Magnified image of the same section revealing the absence of DPT protein in the epidermis. Scale bar – 100 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f8: Immunofluorescent detection of DPT protein in normal skin sections.(a) Sections treated with FITC conjugated secondary antibody showing strong signals in the dermis (indicated by arrows). ii) Magnified image of the same section revealing the absence of DPT protein in the epidermis. Scale bar – 100 μm.
Mentions: DPT protein expression was analyzed using western blotting and immunohistological studies. The results of the western blotting shown in the fig. 6b clearly depicted that DPT is not expressed in epidermis and HaCaT cells, in contrast to the conspicuous expression in the dermis and normal fibroblasts. In histological analysis, the sections treated with anti – human DPT and subsequent HRP or FITC conjugated secondary antibody revealed that DPT protein levels are markedly low in epidermis than in dermis (fig. 7 and fig. 8). The brown color developed due to HRP-DAB reaction was present throughout the dermis localizing on the collagen bundles. The epidermis showed a very feeble or no color development indicating the absence of antigen (DPT protein). The bright signals in the immunofluorescence studies also corroborated that DPT is absent and abundant in the epidermis and dermis respectively.

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