Hyaluronan (HA), a major component of the cutaneous extracellular-matrix, is involved in tissue repair. Human skin is exposed to and damaged by UVB-irradiation. Here, we investigate the regulation of HA metabolism in human skin during acute UVB-induced inflammation. Expression of HA synthesizing (HAS) and degrading enzymes hyaluronidase (HYAL) as evaluated by quantitative reverse transcribed PCR in response to UVB differed when fibroblasts and HaCaT-keratinocytes, representative cell types in dermis and epidermis, respectively, were compared. Both demonstrated temporally different expression patterns of these genes 3- and 24-hours post-irradiation. This resulted 24-hours post-irradiation in an increase in HAS gene expression in both fibroblasts and HaCaT-keratinocytes, and an increase in HYAL expression only in fibroblasts. HA-production as analyzed by the HA content of conditioned medium was reduced in HaCaT and fibroblast cultures 3-hours post-irradiation, whereas HA increased in HaCaT-cultures 24-hours post-irradiation but remained suppressed in fibroblasts-cultures. Consistently, immunohistochemical staining for HA in human skin 24-hours post-irradiation demonstrated an increased epidermal HA, but a decrease in the dermal compartment. Moreover, analysis of the HA content of dermal microdialysis-fluid revealed increased accumulation of HA degradation products 24-hours post-irradiation. These data demonstrate that there is a complex temporal and spatial regulation of HA-metabolism in skin in response to UVB irradiation.

In skin, an evolving inflammatory or immune response is triggered by early release of a cytokine cascade into the extracellular space. Investigation of extracellular cytokine secretion in situ has been limited by low cut-off filtering membranes and sample volume size and the inability to monitor changes in cytokine protein levels in real-time in situ. Here, we combine for the first time the methods of intradermal microdialysis and antibody protein arraying to profile the early cascade of multiple cytokines in a complex inflammatory response exemplified by ultraviolet B (UVB)-induced inflammation. We observed significant differences of the cytokine and growth factor responses after tissue injury by catheter placement and UVB-induced inflammation. UVB irradiation initiates a rapid proinflammatory response followed by a mixed TH1/TH2 response in which ultimately TH2 cytokines IL-4 and IL10 predominated after 24 h. This most likely indicates the termination and self limitation of the inflammatory response. We conclude that the combination of dermal microdialysis and protein microarray offers a powerful tool to analyze in real-time the complex and rapidly changing interstitial protein milieu during cutaneous inflammatory responses.