Active Nanofibrous Membrane Effects on Gingival Cell Inflammatory Response

Abstract

Alpha-melanocyte stimulating hormone (α-MSH) is involved in normal skin wound healing and also has anti-inflammatory properties. The association of α-MSH to polyelectrolyte layers with various supports has been shown to improve these anti-inflammatory properties. This study aimed to evaluate the effects of nanofibrous membrane functionalized with α-MSH linked to polyelectrolyte layers on gingival cell inflammatory response. Human oral epithelial cells (EC) and fibroblasts (FB) were cultured on plastic or electrospun Poly-ε-caprolactone (PCL) membranes with α-MSH covalently coupled to Poly-L-glutamic acid (PGA-α-MSH), for 6 to 24 h. Cells were incubated with or without Porphyromonas gingivalis lipopolysaccharide (Pg-LPS). Cell proliferation and migration were determined using AlamarBlue test and scratch assay. Expression of interleukin-6 (IL-6), tumor necrosis factor (TNF-α), and transforming growth factor-beta (TGF-β) was evaluated using RT-qPCR method. Cell cultures on plastic showed that PGA-α-MSH reduced EC and FB migration and decreased IL-6 and TGF-β expression in Pg-LPS stimulated EC. PGA-α-MSH functionalized PCL membranes reduced proliferation of Pg-LPS stimulated EC and FB. A significant decrease of IL-6, TNF-α, and TGF-β expression was also observed in Pg-LPS stimulated EC and FB. This study showed that the functionalization of nanofibrous PCL membranes efficiently amplified the anti-inflammatory effect of PGA-α-MSH on gingival cells.

Synthesis of a Novel Electrospun Polycaprolactone Scaffold Functionalized with Ibuprofen for Periodontal Regeneration: An In Vitro andIn Vivo Study

“…Herein, this dose was able to reduce the expression of inflammatory markers induced by Pg -LPS stimulation in both cell types significantly. Pg -LPS is a strong inducer of pro-inflammatory responses in gingival EC and FB [ 28 , 47 ]. In this inflammatory model, Pg -LPS increased COX-2 and IL-8 expression as previously observed in a cell-dependent manner [ 48 , 49 , 50 ].…”

“…The scratch was captured immediately and after 24 h with an optical microscope (Nikon inverted microscope, Eclipse TS100, Nikon, Champigny-sur-Marne, France), and the area of the scratch was calculated with Photoshop CS4. The closure percentage of the scratch was calculated as ((surface of the scratch at time 0 h and surface of the scratch at time 24 h)/(surface of the scratch at time 0 h × 100)), as described in [ 28 ]. Only ECs, being the first cell type to migrate to the periodontal wound and hindering tissue regeneration owing to their high proliferation rate, were selected for performing the scratch test.…”

“…The functionalization of scaffolds with drugs could be obtained with several methods such as impregnation, incorporation, encapsulation, coating and grafting, hence imparting different advantageous characteristics to the membrane [ 27 ]. In this context, a poly-ε-caprolactone (PCL) membrane was evaluated and demonstrated for its pro-regenerative ability in periodontal applications [ 28 ]. The PCL membrane has been reported to be biodegradable and biocompatible with enhanced mechanical properties to stabilize the initial clot [ 29 ].…”

“…The PCL membrane has been reported to be biodegradable and biocompatible with enhanced mechanical properties to stabilize the initial clot [ 29 ]. Interestingly, functionalization with anti-inflammatory compounds such as alpha-melanocyte-stimulating hormone (α-MSH) has already displayed amplified anti-inflammatory effects associated with anti-soft tissue invasion and anti-fibrotic characteristics [ 28 ]. Fascinated by this concept, NSAID-loaded electrospun membranes (ketoprofen/PCL and piroxicam/chitosan) have also been successfully tested for periodontal regeneration in vitro with promising results [ 30 , 31 , 32 ].…”

Cytokines during periodontal wound healing: potential application for new therapeutic approach

“…Visualization of an electrospun polycaprolactone nanofibrous membrane. Macroscopic observation ( a ) and scanning electron microscopy ( b , c , d ) (Morand et al , )…”