Effects of bone marrow mesenchymal stem cells (BM-MSCs) on rat pial microvascular remodeling after transient middle cerebral artery occlusion

Previous studies have shown that the pial microcirculation remodeling improves neurological outcome after middle cerebral artery occlusion (MCAO), accompanied by higher expression of vascular endothelial growth factor (VEGF) and endothelial nitric oxide synthase (eNOS), modulating in vivo angiogenesis. This study was aimed to assess the effects of bone marrow mesenchymal stem cells (BM-MSCs) infused after MCAO on rat pial microcirculation. Animals were subjected to 2 h MCAO followed by BM-MSCs infusion into internal carotid artery. Pial microcirculation was observed at different reperfusion times by fluorescence microscopy. Geometric characteristics of arteriolar networks, permeability increase, leukocyte adhesion, perfused capillary density, VEGF, and endothelial nitric oxide synthase (e-NOS) expression were evaluated. Green fluorescent protein (GFP)-BM-MSCs were used to evaluate their distribution and cell phenotype development during reperfusion. BM-MSCs stimulated a geometric rearrangement of pial networks with formation of new anastomotic vessels sprouting from preexistent arterioles in the penumbra at 7-14-28 days of reperfusion. At the same time VEGF and eNOS expression increased. GFP-BM-MSCs appear to be involved in endothelial and smooth muscle cell programming in the infarcted area. In conclusion, transient MCAO induced pial vascular remodeling characterized by arteriolar anastomotic arcades (originated from preexistent arterioles in penumbra area) able to overlap the ischemic core supplying blood to the neuronal tissue. BM-MSCs appear to accelerate angiogenic processes facilitating new vessel formation; this mechanism was promoted by an increase in VEGF and eNOS expression

Occurrence of hashimoto thyroiditis among the first- and second-degree relatives of systemic lupus erythematosus patients with Hashimoto thyroiditis

Occurrence of Hashimoto thyroiditis among the first- and second-degree relatives of systemic lupus erythematosus patients with Hashimoto thyroiditis

Early treatment with hydroxychloroquine prevents the development of endothelial dysfunction in a murine model of systemic lupus erythematosus

INTRODUCTION: Accelerated atherosclerosis is one of the major causes of morbidity in patients with systemic lupus erythematosus (SLE). Endothelial dysfunction (ED) is considered an early marker of atherosclerosis. It is a reversible alteration, thus representing an attractive target for prevention strategies against cardiovascular disease. Studies have shown that ED occurs in patients with SLE even in the absence of severe, active disease. Hydroxychloroquine (HCQ) is widely used in SLE to control disease activity, but its use is also associated with an improvement in long-term prognosis. Beyond the beneficial effect in well-established disease, our hypothesis is that treatment with HCQ might have a beneficial impact on ED prevention in SLE. The aim of this study was to assess the impact of early treatment with HCQ on ED in a murine model of SLE. METHODS: Twelve-week-old NZB/W F1 (NZ) and C57BL/6 J mice (controls) were allocated to receive HCQ or vehicle for 6, 12, or 18 weeks. Proteinuria and anti-double-stranded DNA autoantibodies were determined. ED was assessed in mesenteric arteries (pressurized myography). Nitric oxide (NO) availability and reactive oxygen species (ROS) production were evaluated. Vascular ROS production was measured with dihydroethidium (DHE) fluorescent dye. RESULTS: Starting from 18 weeks of age, NZ mice showed a progressive reduction in NO availability, which was normalized by ascorbic acid and apocynin in the up to 24-week-old group, and partly ameliorated in older animals. HCQ administration normalized the NO availability in the up to 24-week-old group, with a partial amelioration in the 30-week-old group. DHE analysis revealed a progressive increment of vascular ROS generation among NZ groups, which was prevented by apocynin. Similarly, in the NZ HCQ-treated group, vascular ROS production was abrogated. CONCLUSIONS: The ED that characterizes this mouse model of SLE is caused by the nicotinamide adenine dinucleotide phosphate oxidase-driven ROS excess. Very early treatment with HCQ is able to exert vascular protection via an antioxidant effect

Efficacy and safety of infliximab or adalimumab in severe mucocutaneous Behçet’s syndrome refractory to traditional immunosuppressants: a 6-month, multicentre, randomised controlled, prospective, parallel group, single-blind trial

Introduction: Evidence from randomised controlled trials on anti-tumour necrosis factor (TNF) agents in patients with Behçet's syndrome (BS) is low. Method: We conducted a phase 3, multicentre, prospective, randomised, active-controlled, parallel-group study to evaluate the efficacy and safety of either infliximab (IFX) or adalimumab (ADA) in patients with BS. Adults patients with BS presenting with active mucocutaneous manifestations, occurring while on therapy with either azathioprine or cyclosporine for at least 3 months prior to study entry, were eligible. Participants were randomly assigned (1:1) to receive IFX or ADA for 6 months. The primary study outcome was the time to response of manifestations over 6-month anti-TNF alpha agents' treatment. Results: 42 patients underwent screening visits, of whom 40 were randomly assigned to the IFX group (n=22) or to the ADA group (n=18). All patients at the time of randomisation had active mucocutaneous manifestations and a smaller proportion had concomitant vital organ involvement (ie, six and three patients with ocular and neurological involvement, respectively). A total of 14 (64%) responders in the IFX group and 17 (94%) in the ADA group were observed. Retention on treatment was 95% and 94% in the IFX and in the ADA group, respectively. Quality of life resulted to be significantly improved in both groups from baseline, as well as Behçet's Disease Current Activity Form assessment. We registered two adverse events (one serious) in the ADA group and three non-serious adverse events in the IFX group. Discussion: The overall results of this study confirm the effectiveness of both IFX and ADA in achieving remission in patients with BS affected by mucocutaneous involvement

Effetti del trapianto di cellule staminali mesenchimali sulle alterazioni del microcircolo piale del ratto dopo occlusione transitoria dell'arteria cerebrale media

L’occlusione dell’arteria cerebrale media (MCAO) induce la formazione di tre zone che possono essere classificate in base all’apporto ematico: il core ischemico non perfuso, la zona di penombra con ridotto flusso e la zona più periferica normalmente irrorata. Lo studio dell’area di penombra ha sempre destato molto interesse in quanto la riduzione del flusso sanguigno non è così marcata da causare una perdita irreversibile del metabolismo energetico e necrosi cellulare. Pertanto durante la riperfusione, la penombra definita metaforicamente “la bella addormentata” può manifestare una potenziale capacità di ripresa dell’attività neuronale. Le terapie attualmente impiegate nella pratica clinica per la cura dell’ictus limitano i danni da non reflusso nella fase acuta ma a lungo termine non risultano efficaci nel risolvere i deficit neurologici. Pertanto la ricerca di nuove terapie per la risoluzione del danno ischemico è ancora argomento di controversie e di notevole interesse scientifico. Al momento quelle più promettenti sembrano essere le terapie cellulari. Il presente progetto di ricerca ha come obiettivo quello di valutare in vivo quantitativamente e qualitativamente gli effetti delle cellule staminali mesenchimali (MSCs) sulle alterazioni vascolari a breve e lungo termine che si verificano in seguito all’occlusione transitoria dell’arteria cerebrale media. Per questo studio è stato messo a punto un modello sperimentale animale che permette la visualizzazione diretta del microcircolo cerebrale piale sia in fase acuta che a differenti tempi di riperfusione. Tale metodica consente di valutare l’organizzazione del network piale e le alterazioni vascolari indice di danno. Lo studio sperimentale è stato condotto su ratti Wistar maschi del peso di 250-300 g nei quali è stata eseguita sotto anestesia l’occlusione transitoria dell’arteria cerebrale media mediante inserzione di un nanofilamento ricoperto di silicone. È stata preparata una finestra cranica aperta a livello della corteccia parietale per esporre il network microvascolare piale. L’osservazione del microcircolo è stata condotta con una metodica di microscopia in fluorescenza utilizzando come tracciante l’isotiocianato di fluoresceina legato al destrano (70kDa). Le arteriole sono state classificate secondo una metodica centripeta (metodo di Strahler, modificato secondo il diametro). Il danno vascolare è stato quantificato attraverso l’analisi computerizzata di 4 parametri: formazione dell’edema, adesione leucocitaria, perfusione capillare e vasomotilità arteriolare. Il trattamento cellulare è consistito nell’infusione intracarotidea di cellule staminali mesenchimali precedentemente caratterizzate fenotipicamente. Infine gli animali utilizzati sono stati sottoposti a test comportamentali per verificare le funzioni motorie e sensoriali. In condizioni fisiologiche il network microvascolare piale è organizzato in 5 ordini di vasi arteriolari. Ai capillari è stato assegnato ordine 0, alle arteriole da cui originano i capillari è stato assegnato ordine 1 e ordini progressivamente maggiori fino ad arrivare alle arteriole più grandi del preparato (ordine 5). La distribuzione dei diametri, delle lunghezze e delle ramificazioni dei diversi ordini di vasi risponde alla legge di Horton. Questo andamento viene descritto dal rapporto segmenti/elementi che quando assume valore di 1 indica simmetria di biforcazione mentre valori superiori ad 1 indicano asimmetria. I vasi di ordine maggiore risultano più asimmetrici. Tale asimmetria è descritta in dettaglio dalla matrice di connettività. Nessuna alterazione microvascolare è stata rilevata durante l’intero periodo di osservazione (60 min). L’occlusione transitoria dell’arteria cerebrale media e la successiva riperfusione (1h) determina una marcata alterazione dell’organizzazione geometrica del network microvascolare ed importanti danni microvascolari come aumentata permeabilità, numerosi leucociti adesi alle pareti venulari, una ridotta perfusione capillare ed un’alterata funzionalità arteriolare. A vari tempi di riperfusione (7, 14, 28 giorni) le arteriole piali si riorganizzano geometricamente in arteriole anastomotiche fino a raggiungere un modello vascolare simile a quello osservato in condizioni fisiologiche. I danni vascolari si attenuano con il prolungare della riperfusione anche se persiste una pronunciata permeabilità vascolare, un’elevata adesione leucocitaria e una ridotta perfusione capillare. Inoltre l’attività di vasomotilità è risultata leggermente inferiore a quella osservata nei basali così come la risposta all’Acetilcolina e Papaverina. Il trattamento con le cellule staminali mesenchimali dopo l’occlusione transitoria dell’arteria cerebrale media e successiva riperfusione induce una riorganizzazione in tempi più brevi del network microvascolare. Le arteriole ai diversi tempi di riperfusione si riorganizzano in vasi anastomotici che attraversano l’intera regione ischemica mantenendo le caratteristiche geometriche osservate nel basale. I danni microvascolari sono meno rilevanti in tutte le fasi della riperfusione, le arteriole rispondono dilatandosi alla somministrazione di Acetilcolina e Papaverina e dopo 28 giorni riacquistano le stesse frequenze di vasomotilità proprie delle condizioni fisiologiche. I test comportamentali eseguiti ai diversi tempi di riperfusione hanno evidenziato che i ratti trattati con MSCs mostrano una ripresa in tempi più brevi della funzionalità motoria e sensoriale se confrontati con gli animali non trattati. Concludendo la somministrazione di cellule staminali mesenchimali induce una più rapida riorganizzazione del network microvascolare secondo una distribuzione frattale come osservata nel basale; inoltre le arteriole si riorganizzazione in vasi anastomotici e quelle più piccole (ordine 1 e 2) emettono numerosi vasi che penetrano nello strato corticale. E’ possibile ipotizzare che le MSCs siano in grado di indurre angiogenesi tramite l’espressione di fattori di crescita e citochine quali BDNF, NGF, VEGF. Tale attività è finalizzata a ripristinare un adeguato apporto ematico sia nella zona di penombra che in quella ischemica in modo da permettere una normale attività neuronale. La terapia cellulare potrà essere una promettente alternativa e avere un impatto drastico nella terapia acuta dell'ictus cerebrale limitando la disabilità permanente

Leukopenia, lymphopenia, andneutropenia in systemic lupus erythematosus: Prevalence and clinical impact--A systematic literature review

OBJECTIVE: To systematically review the available evidence to evaluate (1) the prevalence and degree of leukopenia, lymphopenia, and neutropenia in patients with systemic lupus erythematosus (SLE), (2) whether these conditions carry a major infection risk for patients, and (3) whether a treatment with colony stimulating factors (CSF) can be an effective and safe option in SLE patients with leukopenia. MATERIAL AND METHODS: MedLine and Embase were searched by including MeSH terms, text words, and subheadings "systemic lupus erythematosus," "leukopenia" (first search), and "colony stimulating factor" (second search). Inclusion and exclusion criteria were a priori defined and two reviewers screened the retrieved articles for selection criteria; data from the included studies were recorded in ad hoc standard forms; the results were synthesized and transported to evidence tables. RESULTS: A total of 17 articles were included in the systematic literature review: nine articles were retrieved for the first research question and 11 for the second while no articles satisfied the inclusion criteria for the third research question. The prevalence of leukopenia is reported in 22-41.8% of cases and lymphopenia is reported cumulatively from 15% to 82% of the patients while neutropenia is described in 20-40% of the patients. There is no evidence of a significant association between overall reduction of white blood cells and infection occurrence while some studies found a strong association between low lymphocytes/neutrophils count and the risk of major infections. Only case reports and case series have been found to investigate the safety of CSF in SLE patients. CONCLUSIONS: The results of this systematic literature review are inconclusive for many aspects related to the original research questions and highlight the need for further studies. Indeed, the strength of the evidence is not sufficiently robust to draw specific recommendations on how to balance between the need to treat the patient with SLE with immunosuppressive drugs and the risk of severe infections

Rat pial microvascular responses to melatonin during bilateral common carotid artery occlusion and reperfusion

The present study assessed the in vivo rat pial microvascular responses induced by melatonin during brain hypoperfusion and reperfusion (RE) injury. Pial microcirculation of male Wistar rats was visualized by fluorescence microscopy through a closed cranial window. Hypoperfusion was induced by bilateral common carotid artery occlusion (BCCAO, 30 min); thereafter, pial microcirculation was observed for 60 min. Arteriolar diameter, permeability increase, leukocyte adhesion to venular walls, perfused capillary length (PCL), and capillary red blood cell velocity (V(RBC) ) were investigated by computerized methods. Melatonin (0.5, 1, 2 mg/kg b.w.) was intravenously administered 10 min before BCCAO and at the beginning of RE. Pial arterioles were classified in five orders according to diameter, length, and branchings. In control group, BCCAO caused decrease in order 2 arteriole diameter (by 17.5 ± 3.0% of baseline) that was reduced by 11.8 ± 1.2% of baseline at the end of RE, accompanied by marked leakage and leukocyte adhesion. PCL and capillary V(RBC) decreased. At the end of BCCAO, melatonin highest dosage caused order 2 arteriole diameter reduction by 4.6 ± 2.0% of baseline. At RE, melatonin at the lower dosages caused different arteriolar responses. The highest dosage caused dilation in order 2 arteriole by 8.0 ± 1.5% of baseline, preventing leakage and leukocyte adhesion, while PCL and V(RBC) increased. Luzindole (4 mg/kg b.w.) prior to melatonin caused order 2 arteriole constriction by 12.0 ± 1.5% of baseline at RE, while leakage, leukocyte adhesion, PCL and V(RBC) were not affected. Prazosin (1 mg/kg b.w.) prior to melatonin did not significantly change melatonin's effects. In conclusion, melatonin caused different responses during hypoperfusion and RE, modulating pial arteriolar tone likely by MT1 and MT2 melatonin receptors while preventing blood-brain barrier changes through its free radical scavenging action

TNF-alpha inhibitors in Systemic Lupus Erythematosus. A case report and a systematic literature review

Joint involvement is a common manifestation of systemic lupus erythematosus (SLE) and is described as a non-erosive mild synovitis. However some SLE patients may present a more severe joint involvement requiring aggressive therapy. We describe the case of a SLE patient with a severe arthritis unresponsive to methotrexate, successfully treated with anti-TNF-alpha drug as induction therapy and we report the results of a systematic literature review on the use of TNF-alpha inhibitors in SLE

Impact of joint involvement on patients reported outcomes in systemic lupus erythematosus

Joint involvement is common among patients with systemic lupus erythematosus (SLE). Aim of this work was to evaluate the correlation between the presence of joint involvement and patient-reported pain, perception of disease activity, general health and quality of life