Effects of JWH015 in cytokine secretion in primary human keratinocytes and fibroblasts and its suitability for topical/transdermal delivery

Abstract

BackgroundJWH015 is a cannabinoid (CB) receptor type 2 agonist that produces immunomodulatory effects. Since skin cells play a key role in inflammatory conditions and tissue repair, we investigated the ability of JWH015 to promote an anti-inflammatory and pro-wound healing phenotype in human primary skin cells.MethodsHuman primary keratinocytes and fibroblasts were stimulated with lipopolysaccharide. The mRNA expression of cannabinoid receptors was determined using RT-PCR. The effects of JWH015 (0.05, 0.1, 0.5, and 1 µM) in pro- and anti-inflammatory factors were tested in lipopolysaccharide-stimulated cells. A scratch assay, using a co-culture of keratinocytes and fibroblasts, was used to test the effects of JWH015 in wound healing. In addition, the topical and transdermal penetration of JWH015 was studied in Franz diffusion cells using porcine skin and LC-MS.ResultsThe expression of CB1 and CB2 receptors (mRNA) and the production of pro- and anti-inflammatory factors enhanced in keratinocytes and fibroblasts following lipopolysaccharide stimulation. JWH015 reduced the concentration of major pro-inflammatory factors (IL-6 and MCP-1) and increased the concentration of a major anti-inflammatory factor (TGF-β) in lipopolysaccharide-stimulated cells. JWH015 induced a faster scratch gap closure. These JWH015’seffects were mainly modulated through both CB1 and CB2 receptors. Topically administered JWH015 was mostly retained in the skin and displayed a sustained and low level of transdermal permeation.ConclusionsOur findings suggest that targeting keratinocytes and fibroblasts with cannabinoid drugs could represent a therapeutic strategy to resolve peripheral inflammation and promote tissue repair.

Fas/FasL pathway and cytokines in keratinocytes in atopic dermatitis – Manipulation by the electromagnetic field

“…In this study, the response of keratinocytes was evaluated. Time points of 4h and 24h were selected because literature data indicate that after stimulation, secretion of preformed cytokines occurs between 2 nd and 4 th hour, and the secretion of de novo formed cytokines occurs between 20 th and 24 th hour after exposure [ 20 , 21 ]. What is more, a pilot kinetic study of cytokine secretion was performed allowing for the selection of time points.…”

Targeting Peripherally Restricted Cannabinoid Receptor 1, Cannabinoid Receptor 2, and Endocannabinoid-Degrading Enzymes for the Treatment of Neuropathic Pain Including Neuropathic Orofacial Pain

“…In a sciatic nerve CCI model, CB2 receptor expression was observed in the spinal cord associated with injury-induced reactive microglia [105]. CB2 receptor expression in peripheral tissues was also reported to be increased under inflammatory conditions [118,119]. Under a local CFA injection-induced inflammatory pain condition, CB2 receptor expression at the mRNA and protein levels was increased in the inflamed skin tissue, and the receptors were mainly distributed in keratinocytes, macrophages and T-lymphocytes in the epidermis and dermis of the inflamed skin tissues [118].…”

“…Under a local CFA injection-induced inflammatory pain condition, CB2 receptor expression at the mRNA and protein levels was increased in the inflamed skin tissue, and the receptors were mainly distributed in keratinocytes, macrophages and T-lymphocytes in the epidermis and dermis of the inflamed skin tissues [118]. CB2 receptors expressed in keratinocytes were increased under inflammatory or infectious conditions [118,119]. The presence of CB2 receptors in glial and inflammatory cells makes them attractive targets to reduce pain under inflammatory and neuropathic pain conditions [111,120].…”

Characterization of Endocannabinoid System and Interleukin Profiles in Ovine AEC: Cannabinoid Receptors Type-1 and Type-2 as Key Effectors of Pro-Inflammatory Response

“…CB modulation was carried out using a CB1 agonist and antagonists ACEA (1 µM) and AM281 (1 µM) [45]. CB2 activation and inhibition [46,47] were induced by adding the selective agonist JWH-015 (1 µM) and the antagonist SR1445282 (1 µM).…”