TY - JOUR A1 - Löpfe, Moira A1 - Duss, Anja A1 - Zafeiropoulou, Katerina-Alexandra A1 - Bjoergvinsdottir, Oddny A1 - Eglin, David A1 - Fortunato, Giuseppino A1 - Klasen, Jürgen A1 - Ferguson, Stephen J. A1 - Würtz-Kozak, Karin A1 - Krupkova, Olga T1 - Electrospray-Based Microencapsulation of Epigallocatechin 3-Gallate for Local Delivery into the Intervertebral Disc JF - Pharmaceutics N2 - Locally delivered anti-inflammatory compounds can restore the homeostasis of the degenerated intervertebral disc (IVD). With beneficial effects on IVD cells, epigallocatechin 3-gallate (EGCG) is a promising therapeutic candidate. However, EGCG is prone to rapid degradation and/or depletion. Therefore, the purpose of this study was to develop a method for controlled EGCG delivery in the degenerated IVD. Primary IVD cells were isolated from human donors undergoing IVD surgeries. EGCG was encapsulated into microparticles by electrospraying of glutaraldehyde-crosslinked gelatin. The resulting particles were characterized in terms of cytocompatibility and anti-inflammatory activity, and combined with a thermoresponsive carrier to produce an injectable EGCG delivery system. Subsequently, electrospraying was scaled up using the industrial NANOSPIDER (TM) technology. The produced EGCG microparticles reduced the expression of inflammatory (IL-6, IL-8, COX-2) and catabolic (MMP1, MMP3, MMP13) mediators in pro-inflammatory 3D cell cultures. Combining the EGCG microparticles with the carrier showed a trend towards modulating EGCG activity/release. Electrospray upscaling was achieved, leading to particles with homogenous spherical morphologies. In conclusion, electrospray-based encapsulation of EGCG resulted in cytocompatible microparticles that preserved the activity of EGCG and showed the potential to control EGCG release, thus favoring IVD health by downregulating local inflammation. Future studies will focus on further exploring the biological activity of the developed delivery system for potential clinical use. KW - degenerative disc disease KW - inflammation KW - drug delivery KW - EGCG KW - microparticles KW - injectable biomaterial KW - electrospraying Y1 - 2019 U6 - https://doi.org/10.3390/pharmaceutics11090435 SN - 1999-4923 VL - 11 IS - 9 PB - MDPI CY - Basel ER - TY - JOUR A1 - Franco-Obregon, Alfredo A1 - Cambria, Elena A1 - Greutert, Helen A1 - Wernas, Timon A1 - Hitzl, Wolfgang A1 - Egli, Marcel A1 - Sekiguchi, Miho A1 - Boos, Norbert A1 - Hausmann, Oliver A1 - Ferguson, Stephen J. A1 - Kobayashi, Hiroshi A1 - Würtz-Kozak, Karin T1 - TRPC6 in simulated microgravity of intervertebral disc cells JF - European Spine Journal N2 - Purpose Prolonged bed rest and microgravity in space cause intervertebral disc (IVD) degeneration. However, the underlying molecular mechanisms are not completely understood. Transient receptor potential canonical (TRPC) channels are implicated in mechanosensing of several tissues, but are poorly explored in IVDs. Methods Primary human IVD cells from surgical biopsies composed of both annulus fibrosus and nucleus pulposus (passage 1-2) were exposed to simulated microgravity and to the TRPC channel inhibitor SKF-96365 (SKF) for up to 5days. Proliferative capacity, cell cycle distribution, senescence and TRPC channel expression were analyzed. Results Both simulated microgravity and TRPC channel antagonism reduced the proliferative capacity of IVD cells and induced senescence. While significant changes in cell cycle distributions (reduction in G1 and accumulation in G2/M) were observed upon SKF treatment, the effect was small upon 3days of simulated microgravity. Finally, downregulation of TRPC6 was shown under simulated microgravity. Conclusions Simulated microgravity and TRPC channel inhibition both led to reduced proliferation and increased senescence. Furthermore, simulated microgravity reduced TRPC6 expression. IVD cell senescence and mechanotransduction may hence potentially be regulated by TRPC6 expression. This study thus reveals promising targets for future studies. KW - Intervertebral disc KW - Simulated microgravity KW - Senescence KW - TRP channels KW - Mechanotransduction KW - Gene expression Y1 - 2018 U6 - https://doi.org/10.1007/s00586-018-5688-8 SN - 0940-6719 SN - 1432-0932 VL - 27 IS - 10 SP - 2621 EP - 2630 PB - Springer CY - New York ER - TY - JOUR A1 - Wu, Yabin A1 - Stoddart, Martin J. A1 - Wuertz-Kozak, Karin A1 - Grad, Sibylle A1 - Alini, Mauro A1 - Ferguson, Stephen J. T1 - Hyaluronan supplementation as a mechanical regulator of cartilage tissue development under joint-kinematic-mimicking loading JF - Interface : journal of the Royal Society KW - synovial fluid KW - cartilage tissue engineering KW - mechanical loading KW - non-Newtonian fluid KW - hyaluronic acid supplementation Y1 - 2017 U6 - https://doi.org/10.1098/rsif.2017.0255 SN - 1742-5689 SN - 1742-5662 VL - 14 PB - Royal Society CY - London ER - TY - JOUR A1 - Sadowska, Aleksandra A1 - Touli, Ermioni A1 - Hitzl, Wolfgang A1 - Greutert, Helen A1 - Ferguson, Stephen J. A1 - Würtz-Kozak, Karin A1 - Hausmann, Oliver N. T1 - Inflammaging in cervical and lumbar degenerated intervertebral discs BT - analysis of proinflammatory cytokine and TRP channel expression JF - European Spine Journal N2 - To investigate and compare the occurrence of inflammatory processes in the sites of disc degeneration in the lumbar and cervical spine by a gene array and subsequent qPCR and to investigate the mechanistic involvement of transient receptor potential channels TRPC6 and TRPV4. The gene expression of inflammatory cytokines and TRP channels was measured in human disc samples obtained from patients undergoing discectomy at the cervical (n = 24) or lumbar (n = 27) spine for degenerative disc disease (DDD) and disc herniation (DH) and analyzed for differences with regard to spinal level, IVD degeneration grade, Modic grade, age, sex, disc region and surgical extent. Aside from genes with known implication in DDD and DH, four previously unreported genes from the interferon and TRP families (IFNA1, IFNA8, IFNB1, TRPC6) could be detected. A correlation between gene expression and age (IL-15) and IVD degeneration grade (IFNA1, IL-6, IL-15, TRPC6), but not Modic grade, was identified. Significant differences were detected between cervical and lumbar discs (IL-15), nucleus and annulus (IL-6, TNF-alpha, TRPC6), single-level and multi-level surgery (IL-6, IL-8) as well as DDD and DH (IL-8), while sex had no effect. Multiple gene-gene pair correlations, either between different cytokines or between cytokines and TRP channels, exist in the disc. This study supports the relevance of IL-6 and IL-8 in disc diseases, but furthermore points toward a possible pathological role of IL-15 and type I interferons, as well as a mechanistic role of TRPC6. With limited differences in the inflammatory profile of cervical and lumbar discs, novel anti-inflammatory or TRP-modulatory strategies for the treatment of disc pathologies may be applicable independent of the spinal region. KW - Cervical and lumbar discs KW - Degenerative disc disease (DDD) and disc herniation (DH) KW - Inflammaging KW - Inflammation KW - Intervertebral disc KW - Transient receptor potential (TRP) channel Y1 - 2017 U6 - https://doi.org/10.1007/s00586-017-5360-8 SN - 0940-6719 SN - 1432-0932 VL - 27 IS - 3 SP - 564 EP - 577 PB - Springer CY - New York ER -