Vlado Antonic, Bernd Hartmann, Sandra Münch, Claudia Belfekroun, Gerald Niedobitek, Beate Petschke, Mark Smith D, Richard Thiele, Wolfgang Schaden and Christian Ottomann
The objective of presented study was to evaluate in vitro effects of extracorporeal shockwave therapy (ESWT) on the keratinocyte morphology, cytoskeleton and mitotic activity. To determine in vitro effects of ESWT on keratinocytes, we applied 100 pulses with an energy flux density 0.1 mJ/mm2 and a frequency of 1 Hz at a distance of 5 cm between therapy head and culture flask. The treatment parameters were determined in a pilot study. Haematoxylin and Eosin staining, as well as immunohistochemistry for Ki-67, CK5, CK14 and CK10 was performed on the cultured keratinocytes and results were compared between ESW treated group and Controls. No pathomorphological abnormalities or alterations in the cells monolayer could be observed. When assessing histological images of all HE-stained sections of ESW-treated and untreated keratinocytes (control group), no morphological differences could be seen between or within the groups. The proliferation marker Ki-67 was found at a frequency of 14% for ESW-treated and 10% for untreated keratinocytes. There were also figures of 95% (CK5 staining) and 90% (CK14 staining) for ESW-treated, as well as 85% (CK5 staining) and 85% (CK14 staining) for untreated keratinocytes. CK10-positive cells showed a frequency of 24% in the ESWtreated and 33% in the untreated cell cultures. Immunohistochemical observations indicated an increased proliferation behaviour following treatment with extracorporeal shockwaves in vitro and maturation of the exposed keratinocytes. Our results suggest an increase in proliferation activity of keratinocytes after in vitro treatment with ESWT. Further investigations of the effects of ESWT on the keratinocyte expression of the chemokines and cytokines are underway.
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