Diseases of the salivary glands in infants and adolescents

Background

Current approaches in bone regeneration combine osteoconductive scaffolds with bioactive cytokines like BMP or VEGF. The idea of our in-vitro trial was to apply VEGF₁₆₅ in gradient concentrations to an equine collagen carrier and to study pharmacological and morphological characteristics of the complex in a circulation model.

Methods

Release kinetics of VEGF₁₆₅ complexed in different quantities in a collagen matrix were determined in a circulation model by quantifying protein concentration with ELISA over a period of 5 days. The structural changes of the collagen matrix were assessed with light microscopy, native scanning electron microscopy (SEM) as well as with immuno-gold-labelling technique in scanning and transmission electron microscopy (TEM).

Results

We established a biological half-life for VEGF₁₆₅ of 90 minutes. In a half-logarithmic presentation the VEGF₁₆₅ release showed a linear declining gradient; the release kinetics were not depending on VEGF₁₆₅ concentrations. After 12 hours VEGF release reached a plateau, after 48 hours VEGF₁₆₅ was no longer detectable in the complexes charged with lower doses, but still measurable in the 80 μg sample. At the beginning of the study a smear layer was visible on the surface of the complex. After the wash out of the protein in the first days the natural structure of the collagen appeared and did not change over the test period.

Conclusions

By defining the pharmacological and morphological profile of a cytokine collagen complex in a circulation model our data paves the way for further in-vivo studies where additional biological side effects will have to be considered. VEGF₁₆₅ linked to collagen fibrils shows its improved stability in direct electron microscopic imaging as well as in prolonged release from the matrix. Our in-vitro trial substantiates the position of cytokine collagen complexes as innovative and effective treatment tools in regenerative medicine and and may initiate further clinical research.