How to evaluate effectiveness of healing on dermis and epidermis?

Wound healing is a very complex process, specific to every organism and tissue, and addresses several factors and different cellular and tissular groups.

Cutaneous wound healing involves many processes of dermis and epidermis regeneration for which physical forces play a crucial role. Indeed, dermis regeneration concerns fibroblasts ability to engage its traction forces and to produce and organize new tissues.

Epidermal wound healing depends, in part, on newly-formed cell-cell junctions robustness.

About tight protein junctions studies and traction forces involved, AFM is a unique tool to :

  • Carry out experiments in medium culture to make possible cell growth in quasi-physiological conditions ; temperature may be controlled and drugs or actives added in situ during the experiment,
  • Get High Resolution Imaging (at nanoscale) fitting perfectly cells junctions imaging,
  • Follow up mechanical properties modifications in real time.

DermoMeca® provides solutions to study active ingredients or finished products effect in the 2 skin compartiments involved in wound healing.

 

Epidermal wound healing efficacy test

DermoMeca® provides a solution to investigate actives or products effect on cell-cell junction robustness and cell motility.

Models : primary keratinocytes at high confluence, making a notch mimicking a wound.

Methods :

  • AFM imaging of newly-formed cell cell junctions
  • Mechanical measurements of cell junction stiffness and actin cytoskeleton.

Readouts :

  • Images and mechanical properties of newly-formed junctions and cells
  • Quantification of cell-cell junctions number, morphometry (thickness, lenght), stiffness measurements (junctions and cells),
  • Cell motility / cytoskeleton activity study

 

Murins keratinocytes AFM images. Data catched at the cell-cell junction to visualize adhesion structure

 

Dermal wound healing efficacy test

DermoMeca® provides a solution to evaluate an active or product effect on : fibroblasts activity (traction force and motility) and quality/amount of newly-synthetised extracellular matrix.

Models : primary fibroblastes at medium confluence on collagen I matrix, making a notch mimicking a wound.

Methods : mechanical properties measurements (elastic modulus) and AFM imaging.

Readouts :

  • Mechanical mapping of extracellular matrix
  • Quantitative data of extracellular matrix stiffness properties (elastic modulus) and fibroblasts traction forces
  • Mechanical mapping and quantitative data of fibroblasts
  • Extracellular matrix imaging and morphometric parameters (fiber density, fiber thickness, fiber orientation)

Strain stiffening mapping of collagen network near a fibroblast cytoplasmic extension showing traction force

CONCLUSION

This solution is perfect to highlight the actives or products effect for empowering fibroblasts ability to restore damaged dermis.

 

Epidermal and dermal wound healing efficacy test

Complete solution to study newly-formed cell junctions, extracellular matrix characteristics and fibroblasts activity.

Models :

  • Full reconstructed skin, making a notch mimicking a wound.
  • Cryosections from previous skin models at different times of healing

Methods : AFM imaging and Seconde Harmonic Generation (SHG)

Readouts :

  • Mechanical mapping of extracellular matrix
  • Quantitative data of extracellular matrix stiffness properties (elastic modulus) and fibroblasts traction forces
  • Mechanical mapping and quantitative data of fibroblasts
  • Collagen network imaging and morphometric parameters (fiber density, fiber thickness, fiber orientation)
  • Images and mechanical properties of newly-formed junctions and cells
  • Quantification of cell-cell junctions number, morphometry (thickness, lenght), stiffness measurements (junctions and cells),
  • SHG images and quantitative data about extracellular matrix density

Skin explant SHG image. A. Elastin. B. Collagen. C. Epidermis. D. Merge

A. Fluorescence Image of human skin section. B. AFM topographical image taken in the epidermis to visualize the detailed structure of the cell-cell adhesion structure and of the cytoskeleton. C. Mechanical mapping of the same regions showing the detailed mechanical properties of cell-cell adhesion structure and of the cytoskeleton.

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