Development of an in vitro coculture of primary sensitive pig neurons and keratinocytes for the study of cutaneous neurogenic inflammation.
Cutaneous neurogenic inflammation (CNI) is often associated with skin disorders. Activated sensory neurons secrete neuropeptides, such as substance P (SP), which initiate or aggravate inflammation in the skin. The discovery of new molecules acting on these neurons is hampered by the difficulty of reproducing the interactions between nerve endings and skin in vitro. We developed an in vitro model based on the coculture of porcine primary keratinocytes and sensory neurons, which mimics skin innervation. To test the relevance of this model, we compared the effects of different substances on CNI by measuring SP secretion in vitro using a sensitive enzyme immunoassay. Collectively, our results indicate that the use of porcine cells could be very useful to perform an in vitro model of CNI. By adding capsaicin, which induces the secretion of SP by neurons, to the culture, we show that our model mimics CNI in vitro, allowing us to screen for molecules that inhibit this inflammatory response. Such a model can be used to test the effects of different substances on CNI and may be useful for dermatological or cosmetic applications. Based on our screen, we found that extracts of Laminaria digitata and Vernonia sublutea inhibit CNI.
Pereira
Ulysse
U
Laboratory of Nervous Factors and Tissue Structure EA 4326, University of Western Brittany, Brest, France.
Boulais
Nicholas
N
Lebonvallet
Nicholas
N
Lefeuvre
Luc
L
Gougerot
Agnès
A
Misery
Laurent
L
D000818:Animals; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods*; D015603:Keratinocytes / Q000166:cytology*; D007795:Laminaria; D008297:Male; D020078:Neurogenic Inflammation / Q000188:drug therapy / Q000276:immunology / Q000473:pathology*; D010936:Plant Extracts / Q000494:pharmacology; D011984:Sensory Receptor Cells / Q000166:cytology* / Q000378:metabolism; D012867:Skin / Q000166:cytology / Q000276:immunology / Q000294:innervation; D013373:Substance P / Q000378:metabolism; D034421:Sus scrofa; D035741:Vernonia
D010936:Plant Extracts; D013373:Substance P