WP06: Biochemical, structural and molecular-biological quality control of skin substitutes

Objectives

The main objective of work package 06 is the molecular evaluation of the performance of the skin substitutes. This will be assessed starting with biochemical analysis focusing on matrix molecules involved in skin functioning like elasticity and scar formation. Subsequently, gene expression analysis using high density gene expression arrays will be performed. In addition the skin substitutes will be analysed on its morphology before and after implantation. Together, the obtained results will lead to an objective evaluation of the clinical applicability of the skin substitutes.

Workpackage Description

The scaffold will be constructed by Partner 01a and Partner 05. The three skin substitutes, denovoDerm, denovoSkin and NovoMaix will be analysed before and after implantation. Punch biopsies will be taken at two time points after transplantation. Punch biopsies from a healthy control will be used as a control. Partner 04 will perform the biochemical and gene-expression analyses. The morphological characterization will be performed by Partner 01a, Partner 02 and Partner 04.

 

Task 1:

Biochemical analysis of skin substitutes before and after implantation focusing on matrix molecules, involved in normal skin functioning like elasticity and scar formation. The following molecules, making up the larger part of the neodermis, will be quantitatively assayed: collagens, elastins and glycosaminoglycans.

 

Task 2:

Molecular-biological analysis of skin substitutes after implantation. Gene expression of the skin scaffolds after implantation will be compared to control skin, giving an objective analysis of the status of the skin formation. Genes involved in epidermal skin formation, like keratins, involucrin and loricrin will be analysed. The data obtained from the gene expression arrays will be validated with quantitative polymerase chain reaction. These results indicate to what extent the skin substitutes have induced normal skin formation. Furthermore, it will give an overview of the cellular processes taking place in the skin substitute. Combing these results, this may set new standards to evaluate the performance of the skin transplants in patients.

 

Task 3:

Histological analyses of the three skin substitutes before and after implantation. Pore size, collagen fibril integrity and cell distribution before implantation will be studied using scanning and transmission electron microscopy. The degradation of the skin substitute and formation of the new skin will be analysed with routine stainings, immuno histrochemistry and electron microscopy. Cooperation on the histological level takes place with Partner 01a and Partner 02. The results will indicate the degree of formation of new skin, and the degradation of the skin substitute applied.

 

Task 4:

Combining results obtained in task 1-3 and other work packages to objectively evaluate the clinical performance of the skin substitutes.