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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

Expression of DPT in epidermis and dermis.(a) Agarose gel images showing DPT PCR products, 168 bp. (i–v) and RPL32 PCR products, 147 bp. (vii–xi). Lanes: i & xi) No template control for DPT and RPL32 respectively, ii & x) NSF, iii & ix) HaCaT cells, iv & viii) Epidermis, v & vii) Dermis, vi) 100 bp DNA ladder. (b) Western blotting analysis of protein homogenates from i) HaCaT cells and ii) epidermis revealing the absence of DPT protein in contrary to the prominent expression in v) dermal and vi) NSF protein extracts. The bands were compared with iv) rDPT and iii) protein molecular weight marker to determine the band size.
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f6: Expression of DPT in epidermis and dermis.(a) Agarose gel images showing DPT PCR products, 168 bp. (i–v) and RPL32 PCR products, 147 bp. (vii–xi). Lanes: i & xi) No template control for DPT and RPL32 respectively, ii & x) NSF, iii & ix) HaCaT cells, iv & viii) Epidermis, v & vii) Dermis, vi) 100 bp DNA ladder. (b) Western blotting analysis of protein homogenates from i) HaCaT cells and ii) epidermis revealing the absence of DPT protein in contrary to the prominent expression in v) dermal and vi) NSF protein extracts. The bands were compared with iv) rDPT and iii) protein molecular weight marker to determine the band size.

Mentions: Semi quantitative PCR analysis revealed that DPT gene is not expressed in epidermis and HaCaT cells (fig. 6a). On the other hand, normal fibroblasts and dermis showed a prominent expression of DPT. The expression of internal control gene, RPL32, was observed in all the samples imparting that the mRNA expression of DPT is restricted to dermis. The absence of non-specific amplicons clearly indicated the specificity of the primers used.


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

Krishnaswamy VR, Korrapati PS - Sci Rep (2014)

Expression of DPT in epidermis and dermis.(a) Agarose gel images showing DPT PCR products, 168 bp. (i–v) and RPL32 PCR products, 147 bp. (vii–xi). Lanes: i & xi) No template control for DPT and RPL32 respectively, ii & x) NSF, iii & ix) HaCaT cells, iv & viii) Epidermis, v & vii) Dermis, vi) 100 bp DNA ladder. (b) Western blotting analysis of protein homogenates from i) HaCaT cells and ii) epidermis revealing the absence of DPT protein in contrary to the prominent expression in v) dermal and vi) NSF protein extracts. The bands were compared with iv) rDPT and iii) protein molecular weight marker to determine the band size.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f6: Expression of DPT in epidermis and dermis.(a) Agarose gel images showing DPT PCR products, 168 bp. (i–v) and RPL32 PCR products, 147 bp. (vii–xi). Lanes: i & xi) No template control for DPT and RPL32 respectively, ii & x) NSF, iii & ix) HaCaT cells, iv & viii) Epidermis, v & vii) Dermis, vi) 100 bp DNA ladder. (b) Western blotting analysis of protein homogenates from i) HaCaT cells and ii) epidermis revealing the absence of DPT protein in contrary to the prominent expression in v) dermal and vi) NSF protein extracts. The bands were compared with iv) rDPT and iii) protein molecular weight marker to determine the band size.
Mentions: Semi quantitative PCR analysis revealed that DPT gene is not expressed in epidermis and HaCaT cells (fig. 6a). On the other hand, normal fibroblasts and dermis showed a prominent expression of DPT. The expression of internal control gene, RPL32, was observed in all the samples imparting that the mRNA expression of DPT is restricted to dermis. The absence of non-specific amplicons clearly indicated the specificity of the primers used.

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