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Proliferation and wound healing studies. (A-F) Cell proliferation of human corneal epithelial cells (HCE cell line) after treatment with recombinant human plasma gelsolin (rhu-pGSN) assessed with fluorescenceactivated cell sorting (FACS). (A) Stimulation with rhu-pGSN and (B) devoid of rhu-pGSN (bovine serum albumin (BSA) protein control). (C,E) Stimulation with lipopolysaccharide (LPS) and tumor necrosis factor α (TNFα) in combination with rhu-pGSN. (D,F) Stimulation with LPS and TNFα in combination with BSA. (G) Normalized impedance after stimulation with rhu-pGSN assessed with Electric Cell-Substrate Impedance Sensing (ECIS ® ). Stimulations were BSA, 30 µg/ml rhu-pGSN, 300 µg/ml rhu-pGSN. Start of stimulation 0 hours. Statistical significance: *p≤0.05, **p≤0.005, ***p≤0.0005. (H) Scratch assay (n = 3) on HCE. Cells were wounded using a pippet tip. Wounded areas (red stripes) after 0 hours and 24 hours of incubation. (I) Restored wound area after scratch (n = 3) and 24 hour incubation with rhu-pGSN or BSA, compared to control values. The wound healing rates were significantly higher under stimulation with 300 µl/ml rhu-pGSN compared with no rhu-pGSN control as well as BSA protein control. Statistical significance: ***p < 0.001. (J) rhu-pGSN promotes re-epithelialization of corneal wounds in combined in vivo/ex vivo model. The measured wound areas are highlighted in red.
Source publication
Woundhealing disorders characterized by impaired or delayed re-epithelialization are a serious medical problem that is painful and difficult to treat. Gelsolin (GSN), a known actin modulator, supports epithelial cell regeneration and apoptosis. The aim of this study was to estimate the potential of recombinant gelsolin (rhu-pGSN) for ocular surface...
Contexts in source publication
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... on cell proliferation. In order to mimic inflammatory conditions similar to bacterial infection or DED, we challenged rhu-pGSN stimulated HCE cells with the bacterial toxin LPS and the inflammatory cytokine TNFα. Flow cytometry analysis based on BrdU incorporation demonstrated increased HCE cell proliferation in cells treated with rhu-pGSN (Fig. 3A,B). Stimulation with 300 µg/ml rhu-pGSN yielded an increased cell proliferation rate (34.6%) compared to treatment with 300 µg/ml BSA (20.7%). LPS and TNFα were used to simulate a bacterial inflammation and an immune response, respectively. An additional treatment with 1 µg/ml lipopolysaccharides (LPS) or 10 ng tumor necrosis factor alpha ...
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... proliferation rate (34.6%) compared to treatment with 300 µg/ml BSA (20.7%). LPS and TNFα were used to simulate a bacterial inflammation and an immune response, respectively. An additional treatment with 1 µg/ml lipopolysaccharides (LPS) or 10 ng tumor necrosis factor alpha (TNFα) had an expected diminishing effect on the proliferation rates ( Fig. 3C-F). However, rates under rhu-pGSN stimulation were still markedly higher than under BSA control treatment (LPS: 26.5% versus 18.2%; TNFα: 20.8% versus 13.4%) ( Table ...
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... of the measured impedance or simplified as cell proliferation under rhu-pGSN stimulation was also analyzed by Electric Cell-Substrate Impedance Sensing (ECIS ® ) ( (Fig. 3H,I). After 24 hours incubation time, the remaining wound area was significantly (p < 0.001) narrowed with a more than 2.5 fold increase in wound area restoration (Supplement Statistic ...
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... effects of rhu-pGSN with regard to wound healing were also visible in a corneal wound healing mouse model (in vivo/ex vivo cornea defect model) (Fig. 3J). Red coloration indicates the wound surface. To compare the corneal wound healing process between placebo-treated and rhu-pGSN groups, we performed Kaplan-Meier analysis using digitized areas of the remaining corneal defect area over time. No significant reduction of the wounded area over time was observed in the untreated group as ...
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... epithelial cells (n = 3); HMGEC SFM = human meibomian gland epithelial cells cultured with serum free medium (n = 3); HMGEC SCM = human meibomian gland epithelial cells cultured with serum containing medium to induce differentiation (n = 3)) using an anti-GSN antibody. Stomach, liver and lung tissue samples served as positive controls. (Fig. 3J). These results were statistically significant (p = 0.0005) compared to the other treatment groups (Supplement Fig. ...
Citations
... This observation was consistent with previous studies in echinoderms [19,54,55]. It has been well documented that mammalian GSN plays an important role in the modulation of actin filaments in growing cells [56,57]. A scratch assay in vitro revealed that the recombinant human gelsolin (rhuGSN) promoted the proliferation and migration of fibroblasts and caused wound closure with the production of cytokine IL-6 in a 3T3-L1 mouse embryo fibroblast cell line [58]. ...
... Gene function data obtained from UniProtKB/Swiss-Prot database [15], unless stated otherwise. Implicated in TGF-β-dependent smooth muscle actin synthesis in myofibroblasts and epithelial-mesenchymal transition [17]. ...
... Fibrillin-1 (FBLN1) is implicated in stabilizing ECM proteins, including periostin, through regulation of the biological availability of latent TGF-β [16]. In turn, signaling through TGF-β results in the expression of CNN1 [54], involved in ECM remodeling and airway hyperreactivity [55], as well as GSN [17] (gelsolin) and TMEFF implicated in epithelial-mesenchymal transition [56]. ATPIF1 promotes transcriptional activation of NFKB, resulting in a proliferative response and tissue remodeling [57]. ...
Asthma heterogeneity complicates the search for targeted treatment against airway inflammation and remodeling. We sought to investigate relations between eosinophilic inflammation, a phenotypic feature frequent in severe asthma, bronchial epithelial transcriptome, and functional and structural measures of airway remodeling. We compared epithelial gene expression, spirometry, airway cross-sectional geometry (computed tomography), reticular basement membrane thickness (histology), and blood and bronchoalveolar lavage (BAL) cytokines of n = 40 moderate to severe eosinophilic (EA) and non-eosinophilic asthma (NEA) patients distinguished by BAL eosinophilia. EA patients showed a similar extent of airway remodeling as NEA but had an increased expression of genes involved in the immune response and inflammation (e.g., KIR3DS1), reactive oxygen species generation (GYS2, ATPIF1), cell activation and proliferation (ANK3), cargo transporting (RAB4B, CPLX2), and tissue remodeling (FBLN1, SOX14, GSN), and a lower expression of genes involved in epithelial integrity (e.g., GJB1) and histone acetylation (SIN3A). Genes co-expressed in EA were involved in antiviral responses (e.g., ATP1B1), cell migration (EPS8L1, STOML3), cell adhesion (RAPH1), epithelial–mesenchymal transition (ASB3), and airway hyperreactivity and remodeling (FBN3, RECK), and several were linked to asthma in genome- (e.g., MRPL14, ASB3) or epigenome-wide association studies (CLC, GPI, SSCRB4, STRN4). Signaling pathways inferred from the co-expression pattern were associated with airway remodeling (e.g., TGF-β/Smad2/3, E2F/Rb, and Wnt/β-catenin).
... Our results indicate that pGSN exerts pleiotropic effects on the BBB and is associated with several different cell signaling pathways. The complex action of pGSN and interaction with molecular mechanisms includes the wound-healing process associated with the ability of pGSN to accelerate cell migration [67,68], as well as anti-inflammatory and antioxidant properties that may limit the exaggerated inflammatory response of endothelial cells forming the blood-brain barrier. Since the ability of the S1 protein to inhibit cell migration was also reversed by pGSN, we suggest that this property of pGSN was at least partly due to its ability to induce VEGFR2 expression. ...
Background
Plasma gelsolin (pGSN) is an important part of the blood actin buffer that prevents negative consequences of possible F-actin deposition in the microcirculation and has various functions during host immune response. Recent reports reveal that severe COVID-19 correlates with reduced levels of pGSN. Therefore, using an in vitro system, we investigated whether pGSN could attenuate increased permeability of the blood–brain barrier (BBB) during its exposure to the portion of the SARS-CoV-2 spike protein containing the receptor binding domain (S1 subunit).
Materials and methods
Two- and three-dimensional models of the human BBB were constructed using the human cerebral microvascular endothelial cell line hCMEC/D3 and exposed to physiologically relevant shear stress to mimic perfusion in the central nervous system (CNS). Trans-endothelial electrical resistance (TEER) as well as immunostaining and Western blotting of tight junction (TJ) proteins assessed barrier integrity in the presence of the SARS-CoV-2 spike protein and pGSN. The IncuCyte Live Imaging system evaluated the motility of the endothelial cells. Magnetic bead-based ELISA was used to determine cytokine secretion. Additionally, quantitative real-time PCR (qRT-PCR) revealed gene expression of proteins from signaling pathways that are associated with the immune response.
Results
pGSN reversed S1-induced BBB permeability in both 2D and 3D BBB models in the presence of shear stress. BBB models exposed to pGSN also exhibited attenuated pro-inflammatory signaling pathways (PI3K, AKT, MAPK, NF-κB), reduced cytokine secretion (IL-6, IL-8, TNF-α), and increased expression of proteins that form intercellular TJ (ZO-1, occludin, claudin-5).
Conclusion
Due to its anti-inflammatory and protective effects on the brain endothelium, pGSN has the potential to be an alternative therapeutic target for patients with severe SARS-CoV-2 infection, especially those suffering neurological complications of COVID-19.
... However, the biological significance of the interaction between pGSN and fibronectin remains unknown treatment can also support epithelial cell regeneration and enhance re-epithelialization, resulting in faster wound healing in vitro, ex vivo, and in vivo. 31 Furthermore, the recombinant pGSN treatment significantly increased the expression of α-SMA, a morphological marker for differentiation of fibroblasts into myofibroblasts, suggesting that pGSN supplementation-induced wound healing effects are partly attributed to fibroblast transformation. 31 Of note, the production of several pro-inflammatory cytokines, tumor necrosis factor α (TNFα), transforming growth factor β (TGFβ), and IL-6, can be increased in human chondrocytes after treatment with recombinant pGSN. ...
... 31 Furthermore, the recombinant pGSN treatment significantly increased the expression of α-SMA, a morphological marker for differentiation of fibroblasts into myofibroblasts, suggesting that pGSN supplementation-induced wound healing effects are partly attributed to fibroblast transformation. 31 Of note, the production of several pro-inflammatory cytokines, tumor necrosis factor α (TNFα), transforming growth factor β (TGFβ), and IL-6, can be increased in human chondrocytes after treatment with recombinant pGSN. 32 Besides, the pGSN treatment also modulates the expression of cartilage ECM-related genes, providing an environment to support wound healing. ...
The protein expression of gelsolin, an actin scavenger controlling cytoskeletal remodeling, cell morphology, differentiation, movement, and apoptosis, has been found to be significantly decreased in several pathological conditions including neurodegenerative diseases, inflammatory disorders, and cancers. Its extracellular isoform, called plasma gelsolin (pGSN), is one of the most abundant plasma proteins in the circulation, and has emerged as a novel diagnostic biomarker for early disease detection. Current evidence reveals that gelsolin can function as either an oncoprotein or a tumor suppressor depending on the carcinoma type. Interestingly, recent studies have shown that pGSN is also involved in immunomodulation, revealing the multifunctional roles of pGSN in tumor progression. In this review, we discuss the current knowledge focusing on the roles of gelsolin in inflammation and wound healing, cancers, and tumor microenvironment. Future prospects of pGSN related studies and clinical application are also addressed.
... It has been proved that GSN is one of the important components of the cytoskeleton (Jun et al. 2004). It plays critical roles in cellular activity, and plays an important role in cell movement, cell-cell communication, death during tissue morphogenesis and inflammatory response (Gao et al. 2017;Wittmann et al. 2018;Yeh et al. 2016). Many studies reported that the GSN has a vital impact on the occurrence and treatment of disease, such as promoting the occurrence and development of pneumonia and pulmonary fibrosis (Oikonomou et al. 2009), alleviate cardiovascular disease (Rouayrenc et al. 1984), mitigation of the occurrence and development of AD (Hirko et al. 2007), and inhibits the occurrence of ALD (alcoholic liver Communicated by Joan Cerda. ...
The aim of this study was to assess the potential of 21 bp mutation in the second intron of the GSN gene as a molecular marker-assisted by exploring the effect of 21 bp mutation on growth traits in four beef cattle breeds. Gelsolin (GSN), a member of the superfamily of gel proteins, is involved in the regulation of a variety of cellular activities in the organism and plays an important function in cell motility, apoptosis, signal transduction and inflammatory responses. Gelatin can not only negatively regulate the pro-apoptotic effect of P53 protein, but also promote apoptosis by blocking the interaction between actin and deoxyribonuclease, so, the GSN gene was selected as a candidate gene in this study. In this study, a 21 bp mutation on the second intron to the GSN gene was verified in 573 individuals of Yunling (YL, n = 220), Jiaxian (JX, n = 140), Xianan (XN, n = 114) and Qinchuan (QC, n = 97) cattle breeds using Once PCR and agarose gel electrophoresis. The association analysis of polymorphisms in the GSN gene with growth traits in four breeds was revealed: in YL cattle, the heart girth and forehead size of heterozygous ID genotype were significantly higher than II genotype (P < 0.05). In JX cattle, the withers height, body length and heart girth of II and ID genotype were significantly highest than DD genotype (P < 0.01); the height at hip cross and height at sacrum of II genotype was significantly highest than DD genotype (P < 0.01), but ID genotype was significantly higher than DD genotype. In XN cattle, the abdominal girth and circumference of the cannon bone of II genotype were significantly higher than ID genotype (P < 0.05). In QC cattle, the hucklebone width of ID genotype was significantly the highest than II genotype (P < 0.01). The results suggest that GSN may be an important candidate gene and that a 21 bp mutation on the second intron to the GSN gene can be used for molecular marker-assisted selection of four beef cattle breeds.
... MLS form in the bone marrow and migrate from embryonic progenitor cells into the synovial membrane. As recently shown, the majority of MLS form a dynamic membrane-like structure around the synovial cavity, forming an internal immunological barrier at the binding protein involved in cell mobility, cell shape, metabolism and wound healing processes [34,35]. Three isoforms are known today, with the plasmatic form (pGSN) being the most common, involved in various immune system processes as well as wound healing [34,36]. ...
Gelsolin (GSN) is an actin-binding protein involved in cell formation, metabolism and wound closure processes. Since this protein is known to play a role in arthritis, here we investigate how the synovial membrane with its specific synoviocytes contributes to the expression of GSN and how the amount of GSN expressed is modulated by different types of arthritis. Synovial membranes from adult healthy subjects and patients with rheumatoid arthritis (RA) and osteoarthritis (OA) are analyzed by immunofluorescence, Western blot and ELISA. Macrophage-like synoviocytes (MLS) and fibroblast-like synoviocytes (FLS) were isolated, cultured and analyzed for their potential to produce and secrete GSN. In addition, the GSN concentrations in the synovial fluid of various forms of arthritis are determined by ELISA. GSN is produced by the healthy and arthritic synovial membranes. Both forms of synoviocytes (MLS and FLS) release GSN. The results show that there is a significant reduction in GSN in the synovial fluid in adult patients with OA. This reduction is also detectable in adult patients with RA but is not as evident. In juvenile arthritis, there is a slight increase in GSN concentration in the synovial fluid. This study shows that primary MLS and FLS express GSN and that these cells, in addition to articular chondrocytes, contribute to GSN levels in synovial fluid. Furthermore, GSN concentrations are modulated in different types of arthritis. Further studies are needed to fully understand how GSN is involved in joint homeostasis.
... Selected in vivo, ex vivo, and in vitro studies investigating actin cytoskeletal protein effects upon wound-healing processes Gelsolin In vivo Generation of GSN-null mice Defects in hemostasis and platelet activation, inflammatory response and leukocyte motility, and defects in fibroblast function Witke et al. 1995 GelsolinIn vitro Addition of recombinant gelsolin (rGSN) to mouse fibroblast GSN siRNA knockdown in human corneal fibroblast Enhanced migration and increased antioxidant effect, decreased SMA synthesisWittmann et al. 2018;Vaid et al. 2020 Gelsolin Ex vivo Addition of rGSN to mice cornea defect model Elevated cell proliferation and increased wound closure rate ...
Wound healing requires a complex cascade of highly controlled and conserved cellular and molecular processes. These involve numerous cell types and extracellular matrix molecules regulated by the actin cytoskeleton. This microscopic network of filaments is present within the cytoplasm of all cells and provides the shape and mechanical support required for cell movement and proliferation. Here, an overview of the processes of wound healing are described from the perspective of the cell in relation to the actin cytoskeleton. Key points of discussion include the role of actin, its binding proteins, signaling pathways, and events that play significant roles in the phases of wound healing. The identification of cytoskeletal targets that can be used to manipulate and improve wound healing is included as an emerging area of focus that may inform future therapeutic approaches to improve healing of complex wounds.
... Hornhautepithel oder der Haut ist GSN an regenerationsfördernden Prozessen im Knorpel beteiligt (Cowin et al., 2003;Wittmann et al., 2018) RhuGSN hat eine direkte Auswirkung auf die Zytokinproduktion phCs in vitro (Abb. 14). ...
... rekombinantes Gelsolin (rhuGSN) hat bereits positive Effekte bei der Reepithalisierung von Korneaverletzungen gezeigt(Wittmann et al., 2018). Daher wird in der vorliegendenArbeit analysiert, ob Gelsolin (GSN) eine vergleichbare positive Wirkung auf Gelenkknorpel als einem weiteren avaskulären Gewebe hat. ...
... Vorarbeiten aus dem Institut für Funktionelle und Klinische Anatomie der FAU Erlangen-Nürnberg haben einen signifikant positiven Effekt von rhuGSN auf die korneale Wundheilung gezeigt(Wittmann et al., 2018). Daher soll in der vorliegenden Arbeit der Frage nachgegangen werden, welche Auswirkung rhuGSN auf die Chondrozyten im avaskulären Gelenkknorpel hat. ...
Mit fortgeschrittenem Alter sind Organsimen nicht mehr in der Lage Wundheilungs- und Regenerationsprozesse regelrecht aufrecht zu erhalten. Besonders avaskuläre Gewebe sind davon betroffen, weshalb vermehrt degenerativer Veränderungen des Gelenkknorpels, wie z.B. Arthrose (Osteoarthrose, OA), aber auch Erkrankungen der Augenoberfläche, wie Trockenes Auge und korneale Defekte, auftreten. In der vorliegenden Arbeit sollen das Protein Gelsolin (GSN) und eine gold-basierte Serumtherapie (Goldic®) auf ihre Wirkung in diesen Geweben sowie ein neuer Ansatz zur Erklärung vom Trockenen Auge untersucht werden. Da für GSN bereits signifikante Ergebnisse bei der kornealen Wundheilung erzielt wurden, wurde dieses Aktin-bindende Protein auf seine Wirkung in primären humanen Chondrozyten (phCs) getestet. Eine Stimulation mit 3 µg rhuGSN/ml fördert signifikant die Wundverschlussrate in phCs. Zusätzlich werden Gene, welche den extrazellulären Matrixaufbau unterstützen, wie ACAN, COL2A1 sowie die TIMPs, stärker exprimiert. Die angeblich auf einer GSN-Anreicherung basierende Goldic®-Therapie erzielte im in vitro Scratch OA Modell auf phCs eine signifikant wundheilende Wirkung. In geringen Konzentrationen zeigt Goldic® in humanen Hornhautepithelzellen (HCE) Zellen ebenfalls eine Wundheilung unterstützende Wirkung. Besonders die Komposition der peripheren Blutzellen sowie die Zytokinkonzentrationen im Serum von Patienten werden positiv durch Goldic® beeinflusst. Die Anreicherung von GSN im Blutserum konnte in dieser Arbeit nicht bestätigt werden. In einem Nebenprojekt wurde SPARC (Secreted Protein Acidic And Cysteine Rich) in der murinen Tränendrüse nachgewiesen. Dabei zeigte sich, dass es aufgrund altersbedingter Methylierungsprozesse zu einer Reduktion von SPARC in der Tränendrüse kommt. Das Vorkommen von SPARC in Myoepithelzellen der Tränendrüse sowie der Verlust im Rahmen des Sjögrens Syndroms, lässt auf den funktionellen Einfluss von SPARC auf die Kontraktion der Myoepithelzellen und konsekutiv die Sekretion von Tränenflüssigkeit vermuten. Zusammenfassend deuten die Ergebnisse der präsentierten Arbeit unter Einbeziehung der aktuellen Literatur darauf hin, dass GSN in avaskulären Geweben konzentrationsabhängig die Wundheilung unterstützt, Goldic® eine vielversprechende Serumtherapie für Gelenkerkrankungen ist sowie dass SPARC eine bislang unbekannte Rolle in der Sekretion der wässrigen Komponente des Tränenfilms und damit auf die Pathogenese des Trockenen Auges einnimmt. Weiterführende Untersuchungen sind daher notwendig, um detaillierte Einblicke in die dargestellten therapeutischen Möglichkeiten zu bekommen.
... It is responsible for cell differentiation, epithelial cell regeneration and apoptosis, and is expressed in all tissues of the ocular system and is secreted in the TF. It is more abundant in tissues belonging to the ocular surface compared to parenchymal organs [26,27]. As mentioned above, since actin can be modified to facilitate the entry of T. gondii into host cells, possibly the increase in gelsolin was associated with this. ...
... Responsible for cell differentiation, epithelial cell regeneration and apoptosis. Expressed in all tissues of the ocular system, secreted by TF [26,27]. ...
Background
Tear film (TF) helps maintain and protect ocular function against damage to the ocular surface. Proteins are one of its main constituents, whose expression pattern can be used as a biomarker of ocular changes and systemic diseases. The aim of this study was to evaluate the expression of proteins in the TF of domestic cats before and after infection with Toxoplasma gondii , in the phases of acute infection and chronicity. Twelve healthy cats received orally homogenized brain matter obtained from mice inoculated with T. gondii oocysts, strain ME49. Cat feces were collected daily from the third day after infection to assess the release of oocysts. TF samples were obtained from cats, by Schirmer’s Tear Test 1, on day 0 (before infection), day 5 after infection (acute phase of infection, with maximum peak release of oocysts in feces) and on day 21 after infection (start of chronic phase, 7 days after total absence of oocyst release in feces). Tear samples were also submitted to proteomic analysis in a Q-Tof-Premier mass spectrometer.
Results
A total of 37 proteins with scores equal to or greater than 100 were identified on D0, followed by 36 on D5 and 42 on D21. Of these, 27 were common to D0 and D5, 33 to D0 and D21, 27 to D5 and D21, and 26 were common to the three groups, totaling 54 proteins. The most abundant proteins were lipocalin allergen Fel d, serum albumin, aldehyde dehydrogenase, lactoperoxidase and lactotransferrin. There was no significant difference in the abundance of proteins found on D0 and D5, but there was a statistical difference between D0 and D21 for ACT1_AEDAE, CERU_HUMAN and GELS_HUMAN. Regarding D5 and D21, there were significant differences for KV1_CANLF, LAC_PIG, TRFL_PIG, ACT1_AEDAE, CERU_HUMAN, GELS_HUMAN and OVOS2_HUMAN.
Conclusions
The main proteins identified in the TF of domestic cats are similar to those found in humans and other animal species. Most are part of the ocular surface defense system against injuries. The most expressed proteins in animals in the chronic phase of T. gondii infection are associated with the immune response to the parasite.
... GSN is a promising biomarker for osteoporosis [16] and is often analysed in age-related studies [5,17]. It was recently shown, that GSN is involved in wound healing processes in the cornea, another non-vascular tissue, where it significantly promotes the re-epithelialisation after wounding [18]. As the supportive effect of GSN on non-vascular tissues was already demonstrated, we investigated the influence of GSN on wound healing and migration processes in human articular cartilage using recombinant human gelsolin (rhuGSN). ...
... In this study, we present a new approach for a therapeutic methodology to support the own repair processes of cartilage by treating phCs with rhuGSN. With our experiments we could show that in phCs, similar to other non-vascular tissue cells, like human corneal cells (HCE) [18] and skin, GSN was found to be involved in wound closure processes [20]. This indicates that GSN might is a major factor in overall wound-healing and migration processes. ...
... We could show that stimulation with 3 μg rhuGSN significantly supports wound closure in vitro (Fig. 4B). In comparison, in mouse fibroblast a concentration up to 12.5 μg of GSN [22] and in HCE cells up to 300 μg of rhuGSN is needed to get positive wound healing effects [18]. This indicates that GSN needs to be evaluated tissue specific to determine which concentrations show the most beneficial effect. ...
Objective
It is known that recombinant human gelsolin (rhuGSN) supports wound closure and migration processes in avascular tissue. Since articular cartilage degradation plays an important role in osteoarthritis (OA), we are investigating how rhuGSN affects regeneration processes in human articular cartilage and represents a promising new therapeutic approach for the treatment of OA.
Methods
Primary human chondrocytes (phCs) from articular knee cartilage were cultured with different concentrations of rhuGSN to analyse its direct effect in vitro. In addition, phCs were stimulated with 10 ng/mL IL-1β or TNF-α to simulate osteoarthritis in vitro and treated with different concentrations of rhuGSN to investigate the beneficial effect in disease treatment. Cytokine secretion and gene expression as well as wound assays were performed.
Results
GSN significantly promotes wound closure in phCs after 60 h compared to untreated cells. After 24 h treatment with 30 μg/mL rhuGSN, TGF-β secretion increases significantly in the in vitro osteoarthritis model. Gene expression of MMP1 as well as SPARC is reduced in chondrocytes due to treatment with GSN in the OA model. At the same time, CXCR4 expression increases significantly after 24 h treatment with 3 μg/mL GSN.
Conclusion
In the in vitro model of osteoarthritis, rhuGSN promotes wound closure of chondrocytes by a supported migration as well as expression of reconstructive and down regulated expression of deconstructive genes concentration dependently. Further experiments are needed to fully understand the beneficial effect of gelsolin on human chondrocytes and to verify this promising approach for a pharmacological treatment of osteoarthritis.
