PARP inhibition promotes endothelial-like traits in melanoma cells and modulates pericyte coverage dynamics during vasculogenic mimicry.

Vasculogenic mimicry (VM) describes the ability of highly aggressive tumor cells to develop pseudovascular structures without the participation of endothelial cells. PARP1 is implicated in the activation of hypoxia-inducible factors, which are crucial in tumor neovascularization. We have explored the role of hypoxia and PARP inhibition in VM. In uveal melanoma xenografts, the PARP inhibitor olaparib improved in vivo pericyte coverage specifically of VM channels. This was concomitant with reduced metastasis in olaparib-treated VM+ tumors. PARP inhibition and hypoxia modulated melanoma tube formation in vitro, inducing a more sparse and regular tubular architecture. Whole-transcriptome profiling revealed that olaparib treatment under hypoxic conditions modulated the expression of genes implicated in vasculogenesis during tube formation, enhancing the endothelial-like phenotype of VM+ uveal melanoma cells. PARP inhibition, especially during hypoxia, upregulated PDGFß, which is essential for pericyte recruitment. Our study indicates that PARP inhibitors may enhance the endothelial characteristics of VM+ cells, modulate pericyte coverage, and reduce metastatic spread in VM+ melanoma. © 2022 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.

Periostin in the relation between periodontal disease and atherosclerotic coronary artery disease: A pilot randomized clinical study.

OBJECTIVE: The aim of this study was to analyze the effects of periodontal treatment on markers of atherosclerotic coronary artery disease and circulating levels of periostin. BACKGROUND: Periostin is necessary for periodontal stability, but it is highly present in atherosclerotic plaques. Treatment of periodontal disease, with low levels of local periostin, is thought to reduce systemic levels of periostin. Thus, this may contribute to cardiovascular health. METHODS: A pilot randomized controlled clinical trial was designed to include patients with severe periodontal disease and history of atherosclerotic coronary artery disease. Samples of gingival crevicular fluid (GCF) and serum were collected before and after periodontal treatment by periodontal surgery or non-surgical therapy. The levels of several markers of inflammation and cardiovascular damage were evaluated including CRP, IFN-γ, IL-1ß, IL-10, MIP-1α, periostin, and TNF-α in GCF and CRP, Fibrinogen, IFN-γ, IL-1ß, IL-6, IL-10, L-Selectin, MIP-1α, Periostin, TNF-α, and vWF in serum. RESULTS: A total of 22 patients with an average of 56 years old were recruited for participating in this study. Twenty of them were male. Most of them (82%) had suffered an acute myocardial event and underwent surgery for placing 1, 2, or 3 stents in the coronary arteries more than 6 months ago but less than 1 year. The treatment of periodontal disease resulted in an overall improvement of all periodontal parameters. Regarding the evaluation of GCF and serum, a significant increase of periostin in the GCF was observed after periodontal surgery. In contrast, although other markers in GCF and serum improved, no significant correlations were found. CONCLUSION: Treatment of periodontal disease through periodontal surgery induces a local and transient increase in the levels of periostin in the gingival crevicular fluid. The effects on systemic markers of inflammation and cardiovascular function have not been confirmed.

Correlation between Inflammasomes and Microbiota in Peri-Implantitis.

The activation of inflammasomes is thought to induce the inflammatory process around dental implants. No information is available on the correlation between microbiota and inflammasomes in clinical samples from patients suffering peri-implantitis. For this cross-sectional study, 30 biofilm samples were obtained from 19 patients undergoing surgical treatment for peri-implantitis because of the presence of bleeding on probing, probing depth higher than 6 mm, and radiographic bone loss higher than 3 mm. Then, soft tissue samples from around the implant were also collected. The relative abundance of bacteria and alpha-diversity indexes were calculated after analyzing the 16S rRNA gene using next-generation sequencing. The soft-tissue samples were processed for evaluation of the inflammasomes NLRP3 and AIM2 as well as caspase-1 and IL-1ß. The relative abundance (mean (SD)) of specific species indicated that the most abundant species were Porphyromonas gingivalis (10.95 (14.17)%), Fusobacterium vincentii (10.93 (13.18)%), Porphyromonas endodontalis (5.89 (7.23)%), Prevotella oris (3.88 (4.94)%), Treponema denticola (2.91 (3.19)%), and Tannerella forsythia (2.84 (4.15)%). Several correlations were found between the species and the immunohistochemical detection of the inflammasomes NLRP3 and AIM2 as well as caspase-1 and IL-1ß, both in the epithelium and the lamina propria. A network analysis found an important cluster of variables formed by NLRP3 in the lamina propria and AIM2, caspase-1, and IL-1ß in the lamina propria and the epithelium with Prevotella dentalis, Prevotella tannerae, Tannerella forsythia, or Selenomonas timonae. Thus, it could be concluded that inflammasomes NLRP3 and AIM2 and their downstream effectors caspase-1 and interleukin-1ß can be significantly associated with specific bacteria.

Density mapping of nerve endings in the skin of the palm and flexor retinaculum of the hand. Application to open carpal tunnel release.

In order to re-evaluate the safest area to incise skin and the flexor retinaculum (FR) when performing a carpal tunnel release (CTR), we carried out a mapping study of the nerve endings in the skin and FR on cadaver specimens, which, unlike previous studies for the first time, includes histomorphometry and image digital analysis. After dividing the skin and FR into 20 and 12 sections, respectively, we carried out a histomorphological analysis of nerve endings. The analysis was performed by two neutral observers on 4-µm histological sections stained with hematoxylin-eosin (H-E), and Klüver-Barrera with picrosirius red (KB + PR) methods. A semi-automatic image digital analysis was also used to estimate the percentage of area occupied per nerve. We observed a lower quantity of nerve endings in the skin of the palm of the hand in line with the ulnar aspect of the 4th finger. The ulnar aspect of the FR was the most densely innervated. However, there are no statistically significant differences between sections in the percentage of area occupied per nerve both in the skin and in the FR. We concluded that there is not a safe area to incise when performing carpal tunnel surgery, but taking into account the quantity of nerve endings present in skin and FR, we recommend an incision on the axis of the ulnar aspect of 4th finger when incising skin and on the middle third of the FR for CTR.

Inflammasomes NLRP3 and AIM2 in peri-implantitis: A cross-sectional study.

BACKGROUND: Inflammasome components NLRP3 and AIM2 contribute to inflammation development by the activation of caspase-1 and IL-1ß. They have not been yet evaluated in samples from patients with active peri-implantitis. Thus, the aim of the present study is to analyze the expression of inflammasomes NLRP3 and AIM2 and subsequent caspase 1 and IL-1ß assessing the microenvironment of leukocyte subsets in samples from patients with active peri-implantitis. METHODS: Biopsies were collected from 33 implants in 21 patients being treated for peri-implantitis. Biopsies from gingival tissues from 15 patients with healthy periodontium were also collected for control. These tissues were evaluated through conventional histological stainings. Then, immunohistochemical detection was performed to analyze NLRP3, AIM2, caspase-1, and IL-1ß and markers of different leukocyte subsets. PCR for inflammasomes and related genes was also done. RESULTS: This manuscript reveals a high immunohistochemical and mRNA expression of NLRP3 and AIM2 inflammasomes, caspase-1, and IL-1ß in biopsies collected from human peri-implantitis. The expression of the tested markers was significantly correlated with the increase in inflammatory infiltrate, probing depth, presence of biofilm, and bleeding on probing. In these peri-implantitis lesions, the area of biopsy tissue occupied by inflammatory infiltrate was intense while the area occupied by collagen was significantly lower. In comparison with periodontal healthy tissues, the inflammatory infiltrate was statistically significantly higher in the peri-implantitis biopsies and was mainly composed of plasma cells, followed by T and B lymphocytes. CONCLUSION: In human peri-implantitis, chronic inflammation can be explained in part by the action of IL-1ß/caspase 1 induced through NLRP3 and AIM2 inflammasome activation.

Neuropeptide Cortistatin Regulates Dermal and Pulmonary Fibrosis in an Experimental Model of Systemic Sclerosis.

INTRODUCTION: Scleroderma, or systemic sclerosis, is a complex connective tissue disorder characterized by autoimmunity, vasculopathy, and progressive fibrosis of the skin and internal organs. Because its aetiology is unknown, the identification of genes/factors involved in disease severity, differential clinical forms, and associated complications is critical for understanding its pathogenesis and designing novel treatments. Neuroendocrine mediators in the skin emerge as potential candidates. We investigated the role played by the neuropeptide cortistatin in a preclinical model of scleroderma. METHODS: Dermal fibrosis was induced by repetitive intradermal injections of bleomycin in wild-type and cortistatin-deficient mice. The histopathological signs and expression of fibrotic markers were evaluated in the skin and lungs. RESULTS: An inverse correlation between cortistatin levels and fibrogenic activation exists in the damaged skin and dermal fibroblasts. Bleomycin-challenged skin lesions of mice that are partially and totally deficient in cortistatin showed exacerbated histopathological signs of scleroderma, characterized by thicker and more fibrotic dermal layer, enlarged epidermis, and increased inflammatory infiltration in comparison to those of wild-type mice. Cortistatin deficiency enhanced dermal collagen deposits, connective tissue growth factor expression, loss of microvessels, and predisposition to suffer severe complications that co-occur with dermal exposition to bleomycin, including pulmonary fibrotic disease and increased mortality. Treatment with cortistatin mitigated these pathological processes. DISCUSSION/CONCLUSION: We identify cortistatin as an endogenous break of skin inflammation and fibrosis. Deficiency in cortistatin could be a marker of poor prognosis of scleroderma and associated complications. Cortistatin-based therapies emerge as attractive candidates to treat severe forms of systemic sclerosis and to manage fibrosis-related side effects of bleomycin chemotherapy in oncologic patients.

Limited marginal bone loss in implant-supported fixed full-arch rehabilitations after 5 years of follow-up.

PURPOSE: The aim of the present study was to evaluate the 5-year results in terms of marginal bone level (MBL) around implants supporting fixed full-arch metal-ceramic restorations in a series of cases of patients who had lost their teeth in that dental arch because of severe periodontal disease. MATERIAL AND METHODS: A retrospective cohort study was designed to evaluate the 5-year MBL results of OsseoSpeed™ Astra Tech TX implants with internal tapered conical connection. Age, gender, bone substratum, smoking habits, history of periodontitis, and prosthetic features were recorded. Mixed linear model was used to determine the influence of the different variables on MBL. RESULTS: In this series, a total of 160 implants placed in 19 patients were evaluated. No implant failure was reported during the 5 years of follow-up. Only 14 (8.75%) implants had more than 2 mm of MBL. Abutment height, F(3,142) = 6.917, p < .001, and implant diameter, F(1,141) = 15.059, p < .001, were determined to be statistically associated with MBL. No other effect was significant. Pairwise comparisons showed that MBL was larger for abutment height = 1 (MBL = -0.987, SE = 0.186) compared with the remaining heights [-0.335 (0.171), -0.169 (0.192) and -0.247 (0.267), 2, 4 and 6 mm, respectively]. MBL was larger for narrow (-0.510, SE = 0.169) than for wide implants (-0.364, SE = 0.190). CONCLUSION: The current study demonstrates that the vast majority of internal conical connection implants supporting fixed full-arch metal-ceramic restorations do not suffer from relevant MBL after 5 years in function. Particularly, those implants with transmucosal abutments longer than 2 mm show less than 0.5 mm from the implant shoulder to the marginal bone.

Maxillary sinus floor augmentation comparing bovine versus porcine bone xenografts mixed with autogenous bone graft. A split-mouth randomized controlled trial.

AIM: To compare the effectiveness of two xenografts for maxillary sinus floor augmentation in terms of clinical, radiographical, histologic, and molecular outcomes. MATERIALS AND METHODS: A split-mouth randomized clinical trial was conducted at the University of Granada. Ten consecutive patients in need of bilateral two-staged maxillary sinus floor augmentation were included. Each patient received both biomaterials (porcine bone mineral and anorganic bovine bone), which were randomly assigned for bilateral sinus augmentation. The maxillary autogenous bone scraped from the sinus access window was mixed with each xenograft at a 20:80 ratio. After a healing period of 6 months, bone biopsies were collected with a trephine during the implant placement in the regenerated area. Histologic, histomorphometrical, immunohistochemical, and molecular outcomes were analyzed. Clinical and radiographical data throughout the treatment phases were also evaluated. RESULTS: The resulting anatomic features were similar between both groups. After six months of graft consolidation, the graft resorption rates were similar between both biomaterials. The histologic, histomorphometrical, and immunohistochemical results showed no statistical differences between groups. CONCLUSION: Anorganic bovine bone and porcine bone mineral combined with maxillary autogenous cortical bone show similar biologic and radiologic features in terms of biomaterial resorption, osteoconduction, and osteogenesis when used for maxillary sinus floor augmentation.

The impact of tooth loss on cognitive function.

OBJECTIVE: To investigate if there is epidemiological evidence of an association between edentulism and cognitive decline beside that currently available from limited sample-sized case series and cross-sectional studies considering limited co-variables. MATERIALS AND METHODS: Data from two USA national health surveys [NHIS 2014-2017 and NHANES 2005-2018] were analyzed using multinomial logistic regression to study the impact of type of edentulism and number of remaining teeth on memory and concentration problems. Age, gender, socioeconomic status, education level, cardiovascular health index, body mass index, exercise, alcohol, smoking habits, and anxiety and depression were used as covariates. RESULTS: The combined population sample was 102,291 individuals. Age, socioeconomic status, educational level, anxiety and depression levels, and edentulism showed the highest odds ratios for cognitive decline. Number of teeth present in the mouth was found to be a predictor of cognitive status. This association showed a gradient effect, so that the lower the number of teeth, the greater the risk of exhibiting cognitive decline. CONCLUSIONS: Edentulism was found among the higher ORs for cognitive impairment. CLINICAL RELEVANCE: Maintenance of functional teeth through the promotion of oral health may contribute to the preservation of memory/concentration and other essential cognitive functions. Thus, increasing and efficiently coordinating efforts aimed at preventing of tooth loss in the adult population could substantially contribute to reduce the incidence of cognitive impairment.

The Long-Term Study of Urinary Biomarkers of Renal Injury in Spontaneously Hypertensive Rats.

BACKGROUND: The age-related increase in blood pressure in spontaneously hypertensive rats (SHRs) is associated to cardiac hypertrophy, heart failure, and renal injury. Here, we investigated for the first time the urinary enzymatic activities of glutamil aminopeptidase (GluAp), alanyl aminopeptidase (AlaAp), dipeptidyl peptidase-4 (DPP4), and Klotho urinary levels, proteins that are strongly expressed in the kidney, as early biomarkers of renal injury in SHRs. METHODS: Male SHR and Wistar Kyoto (WKY) rats were studied from 2 to 8 months old. Systolic blood pressure (SBP), the heart rate (HR), metabolic variables, and urinary markers were measured monthly. At the end of the study, a histopathological evaluation of the kidney was performed. RESULTS: Kidneys of SHR did not develop signs of relevant histopathological changes, but showed increased glomerular area and cellularity. Plasma creatinine was decreased, and creatinine clearance was augmented in SHR at the end of the study. Urinary excretion of Klotho was higher in SHR at 5 and 8 months old, whereas plasma Klotho levels were similar to WKY. GluAp, AlaAp, and DPP4 urinary activities were increased in SHR throughout the time-course study. A positive correlation between glomerular area and cellularity with creatinine clearance was observed. Urinary GluAp, AlaAp, DPP4, and Klotho showed positive correlations with SBP. CONCLUSIONS: GluAp, AlaAp, DPP4, and Klotho in the urine are useful tools for the evaluation of renal damage at early stages, before the whole histopathological and biochemical manifestations of renal disease are established. Moreover, these observations may represent a novel and noninvasive diagnostic approach to assess the evolution of kidney function in hypertension and other chronic diseases.

Expression of Musashi-1 Increases in Bone Healing.

Musashi-1 (MSI1) is an RNA-binding protein that regulates progenitor cells in adult and developing organisms to maintain self-renewal capacities. The role of musashi-1 in the bone healing environment and its relation with other osteogenic factors is unknown. In the current study, we analyze the expression of MSI1 in an experimental model of rat femoral bone fractures. We also analyze the relation between MSI1 expression and the expression of two osteogenic markers: periostin (POSTN) and runt-related transcription factor 2 (RUNX2). We use histological, immunohistochemical, and qPCR techniques to evaluate bone healing and the expression of MSI1, POSTN, and RUNX2 over time (4, 7, and 14 days). We compare our findings with non-fractured controls. We find that in bone calluses, the number of cells expressing MSI1 and RUNX2 increase over time and the intensity of POSTN expression decreases over time. Within bone calluses, we find the presence of MSI1 expression in mesenchymal stromal cells, osteoblasts, and osteocytes but not in hypertrophic chondrocytes. After 14 days, the expression of MSI1, POSTN, and RUNX2 was significantly correlated. Thus, we conclude that musashi-1 potentially serves in the osteogenic differentiation of mesenchymal stromal cells and bone healing. Therefore, further studies are needed to determine the possibility of musashi-1's role as a clinical biomarker of bone healing and therapeutic agent for bone regeneration.

Oral health and healthy chewing for healthy cognitive ageing: A comprehensive narrative review.

INTRODUCTION: Ageing leads to physiological cognitive decline that it is worsened in people with neurodegenerative diseases such as Alzheimer's disease. Despite the ongoing search for a solution to this cognitive decline, no effective remedies have been established. It has been determined that modifiable external factors, such as oral health and occlusal function, prevent cognitive decline. OBJECTIVE: To analyse the primary interactions between occlusal function and cognitive functions. MAIN FINDINGS: Masticatory function is related to cognitive functions. In particular, current evidence, from both animal and human studies, suggests that the activation of masticatory muscles and proper mastication, with natural teeth or dental prosthesis, induces the release of several mediators and the activation of specific brain areas. Together, they result in higher neuronal activity, neurotrophic support, blood flow and the prevention of amyloid-beta plaque formation. Thus, all the components of the masticatory system must work together in order to preserve cognitive function. CONCLUSIONS: Available evidence suggests that oral and cognitive health are more interconnected than previously thought. Therefore, maintenance and adequate restoration of the whole masticatory system are important for the prevention of cognitive decline. In summary, oral and chewing health lead to healthy cognitive ageing.

Lactobacillus fermentum CECT5716: a novel alternative for the prevention of vascular disorders in a mouse model of systemic lupus erythematosus.

The aim of the present study was to examine whether the immune-modulatory bacteria Lactobacillus fermentum CECT5716 (LC40) ameliorates disease activity and cardiovascular complications in a female mouse model of lupus. Eighteen-week-old NZBWF1 [systemic lupus erythematosus (SLE)] and NZW/LacJ (control) mice were treated with vehicle or LC40 (5 × 108 colony-forming units/d) for 15 wk. LC40 treatment reduced lupus disease activity, blood pressure, cardiac and renal hypertrophy, and splenomegaly in SLE mice. LC40 reduced the elevated T, B, regulatory T cells (Treg), and T helper (Th)-1 cells in mesenteric lymph nodes of lupus mice. LC40 lowered the higher plasma concentration of proinflammatory cytokines observed in lupus mice. Aortas from SLE mice showed reduced endothelium-dependent vasodilator responses to acetylcholine. Endothelial dysfunction found in SLE is related to an increase of both NADPH oxidase-driven superoxide production and eNOS phosphorylation at the inhibitory Thr495. These activities returned to normal values after a treatment with LC40. Probiotic administration to SLE mice reduced plasma LPS levels, which might be related to an improvement of the gut barrier integrity. LC40 treatment increases the Bifidobacterium count in gut microbiota of SLE mice. In conclusion, our findings identify the gut microbiota manipulation with LC40 as an alternative approach to the prevention of SLE and its associated vascular damage.-Toral, M., Robles-Vera, I., Romero, M., de la Visitación, N., Sánchez, M., O'Valle, F., Rodriguez-Nogales, A., Gálvez, J., Duarte, J., Jiménez, R. Lactobacillus fermentum CECT5716: a novel alternative for the prevention of vascular disorders in a mouse model of systemic lupus erythematosus.

Vasoactive Intestinal Peptide Ameliorates Acute Myocarditis and Atherosclerosis by Regulating Inflammatory and Autoimmune Responses.

Vasoactive intestinal peptide (VIP) is a neuropeptide that exerts various vascular and cardioprotective functions and regulates immune function and inflammatory response at multiple levels. However, its role in inflammatory cardiovascular disorders is largely unknown. Myocarditis and atherosclerosis are two inflammatory and autoimmune cardiovascular diseases that cause important adverse circulatory events. In this study, we investigate the therapeutic effects of VIP in various well-established preclinical models of experimental autoimmune myocarditis and atherosclerosis. Intraperitoneal injection of VIP during the effector phase of experimental autoimmune myocarditis in susceptible BALB/c mice significantly reduced its prevalence, ameliorated signs of heart hypertrophy and injury, attenuated myocardial inflammatory infiltration, and avoided subsequent profibrotic cardiac remodeling. This effect was accompanied by a reduction of Th17-driven cardiomyogenic responses in peripheral lymphoid organs and in the levels of myocardial autoantibodies. In contrast, acute and chronic atherosclerosis was induced in apolipoprotein E-deficient mice fed a hyperlipidemic diet and subjected to partial carotid ligation. Systemic VIP treatment reduced the number and size of atherosclerotic plaques in carotid, aorta, and sinus in hypercholesterolemic mice. VIP reduced Th1-driven inflammatory responses and increased regulatory T cells in atherosclerotic arteries and their draining lymph nodes. VIP also regulated cholesterol efflux in macrophages and reduced the formation of foam cells and their presence in atherosclerotic plaques. Finally, VIP inhibited proliferation and migration of smooth muscle cells and neointima formation in a mouse model of complete carotid ligation. These findings encourage further studies aimed to assess whether VIP can be used as a pharmaceutical agent to treat heart inflammation and atherosclerosis.

Algae-derived hydroxyapatite behavior as bone biomaterial in comparison with anorganic bovine bone: A split-mouth clinical, radiological, and histologic randomized study in humans.

OBJECTIVES: To analyze a modified biphasic phycogenic biomaterial in comparison with anorganic bovine bone in maxillary sinus floor elevation in humans. MATERIAL AND METHODS: Eight male patients in need of bilateral two-stage sinus floor elevation were consecutively recruited for this randomized split-mouth study. A combination of autogenous cortical bone (ACB, 20%) and anorganic bovine bone (ABB, 80%) (ACB + ABB group) or ACB (20%) and modified biphasic phycogenic material (BP, 80%) (ACB + BP group) were randomly assigned to graft each sinus. Patients were followed up for 6 months post-surgery when bone samples were collected for analysis. RESULTS: Radiographically, bone height gain was statistically higher in the ACB + ABB versus the ACB + BP group. While the analysis of the biological compartments showed differences in non-mineralized tissue (39.15 ± 20.97% vs. 65.87 ± 28.59%, ACB + ABB vs. ACB + BP respectively; p = .018) and remnant biomaterial particles (22.62 ± 17.01% vs. 7.96 ± 8.57%, respectively; p = .028), the percentage of mineralized tissue (38.23 ± 17.55% vs. 24.14 ± 24.66%, respectively; p = .398) showed no statistically significant difference. In contrast, ACB + ABB biopsies showed higher Musashi-1-positive cells per mm2 compared to ACB + BP biopsies (811.49 ± 875.30 vs. 236.90 ± 280.81; p < .018), where the fusiform cells corresponded mainly with fibroblasts, as demonstrated by ultrastructural analysis. CONCLUSION: Both combinations of materials exhibited bone formation after 6 months of healing in the maxillary sinus cavity. However, the combination with biphasic phycogenic biomaterial induced a higher radiographical vertical resorption and graft collapse in comparison with the combination with anorganic bovine bone, possibly due to a higher remodeling of the graft.

Correction to: Relationship between periodontal parameters and plasma cytokine profiles in pregnant woman with preterm birth or low birth weight.

The authors are correcting this article because they accidentally included some wrong data in the analysis.

A Rejection Gene Expression Score in Indication and Surveillance Biopsies Is Associated with Graft Outcome.

Rejection-associated gene expression has been characterized in renal allograft biopsies for cause. The aim is to evaluate rejection gene expression in subclinical rejection and in biopsies with borderline changes or interstitial fibrosis and tubular atrophy (IFTA). We included 96 biopsies. Most differentially expressed genes between normal surveillance biopsies (n = 17) and clinical rejection (n = 12) were obtained. A rejection-associated gene (RAG) score was defined as its geometric mean. The following groups were considered: (a) subclinical rejection (REJ-S, n = 6); (b) borderline changes in biopsies for cause (BL-C, n = 13); (c) borderline changes in surveillance biopsies (BL-S, n = 12); (d) IFTA in biopsies for cause (IFTA-C, n = 20); and (e) IFTA in surveillance biopsies (IFTA-S, n = 16). The outcome variable was death-censored graft loss or glomerular filtration rate decline ≥ 30 % at 2 years. A RAG score containing 109 genes derived from normal and clinical rejection (area under the curve, AUC = 1) was employed to classify the study groups. A positive RAG score was observed in 83% REJ-S, 38% BL-C, 17% BL-S, 25% IFTA-C, and 5% IFTA-S. A positive RAG score was an independent predictor of graft outcome from histological diagnosis (hazard ratio: 3.5 and 95% confidence interval: 1.1-10.9; p = 0.031). A positive RAG score predicts graft outcome in surveillance and for cause biopsies with a less severe phenotype than clinical rejection.

Characterization and comparison of neutrophil extracellular traps in gingival samples of periodontitis and gingivitis: A pilot study.

BACKGROUND AND OBJECTIVE: Neutrophil extracellular traps (NETs) are a recently discovered antimicrobial mechanism used by neutrophils that have been proposed as an intervention in the pathogenesis of periodontitis. The objective of our study was to characterize the expression of NETs in gingival tissues with periodontitis and controls and to compare the expression of these traps in gingival tissue samples of patients with gingivitis and periodontitis. MATERIAL AND METHODS: An observational cross-sectional study was conducted on patients with periodontitis, gingivitis, and controls that needed tooth extraction. Gingival tissue biopsies were gathered after clinical examination and tooth extraction. Electron microscopy and immunofluorescence were performed to characterize NETs, comparing periodontitis and control patients. Immunohistochemical analysis was performed to quantify neutrophil extracellular trap expression through extracellular citrullinated histone H3 and myeloperoxidase in biopsies from patients with gingivitis and periodontitis. RESULTS: Thirteen biopsies were gathered from 13 patients: five gingivitis, six periodontitis, and two controls. Electron microscopy and immunofluorescence imaging showed greater expression of neutrophils present in periodontal inflamed tissue compared with controls. Release of nuclear content to the extracellular space was observed, compatible with the formation of NETs. The expression of citrullinated histone H3 was higher in gingivitis samples than periodontitis samples (P = 0.0106). Myeloperoxidase expression was higher in periodontitis than gingivitis, but without achieving statistical significance. CONCLUSION: Neutrophil extracellular traps were found in tissue samples of periodontitis as extracellular components of chromatin, along with neutrophil enzymes, that were not present in healthy controls. The comparison of NETs expression in periodontitis and gingivitis showed higher expression in gingivitis, associating them to acute phases of the periodontal inflammatory process.

Biphasic hydroxyapatite and ß-tricalcium phosphate biomaterial behavior in a case series of maxillary sinus augmentation in humans.

OBJECTIVES: The aim of this study was to evaluate and compare the morphometric components and the histological properties of pristine bone and bone grafted with a biphasic ß-tricalcium phosphate in humans using the maxillary sinus model. Reparative mesenchymal stem cells in the pristine bone and graft were also evaluated. MATERIALS AND METHODS: For this prospective case series, sinus augmentation was performed using a biphasic ß-tricalcium phosphate. After 6 months of healing, a core of remnant native alveolar bone and grafted bone was collected with a trephine. Histological, histomorphometrical, and immunohistochemical techniques were performed. Radiological analysis through cone beam computerized tomography was also conducted. RESULTS: A total of 10 patients were enrolled in this study. Radiologically, patients showed an average increase of crestal bone of 8.03 ± 1.72 mm. Morphologically, the grafted area was composed by 34.93 ± 14.68% of new mineralized tissue, 9.82 ± 11.42% of remnant biomaterial particles, and 55.23 ± 11.03% non-mineralized tissue. Histologically, we found no differences in the number of osteocytes per mm2 (p = 0.674), osteoblasts (p = 0.893), and blood vessels (p = 0.894) in the grafted area compared to the pristine bone. Differences were found on the number of osteoclasts (15.57 ± 27.50 vs. 5.37 ± 16.12, p = 0.027). The number of Musashi-1 positive mesenchymal cells (239.61 ± 177.4 vs. 42.11 ± 52.82, p = 0.027) was also significantly higher in the grafted area than in the pristine bone. CONCLUSION: Biphasic ß-tricalcium phosphate is a suitable biomaterial to be used in the formation of new bone in sinus floor elevation procedures in humans, not only from the histomorphometrical point of view, but also regarding the cellular and vascular quality of the regenerated bone.

Expression of Musashi-1 During Osteogenic Differentiation of Oral MSC: An In Vitro Study.

BACKGROUND: Musashi-1 (MSI1) is a negative regulator of mesenchymal stromal cell (MSC) differentiation which in turn favors cell proliferation. However, little is known about its expression by MSC from the oral cavity and in the context of osteogenic differentiation. AIM: The aim of this study was to analyze the expression of MSI1 in the context of osteogenic differentiation of MSC derived from the oral cavity. MATERIAL/METHODS: For this in vitro study, MSC were isolated from six different origins of the oral cavity. They were extensively characterized in terms of proliferative and clonogenicity potential, expression of stemness genes (MYC, NANOG, POU5F1, and SOX2), expression of surface markers (CD73, CD90, CD105, CD14, CD31, CD34, and CD45) and adipo-, chondro- and osteogenic differentiation potential. Then, osteogenic differentiation was induced and the expression of MSI1 mRNA and other relevant markers of osteogenic differentiation, including RUNX2 and Periostin, were also evaluated. RESULTS: Cell populations from the alveolar bone (pristine or previously grafted with xenograft), dental follicle, dental germ, dental pulp, and periodontal ligament were obtained. The analysis of proliferative and clonogenicity potential, expression of the stemness genes, expression of surface markers, and differentiation potential showed similar characteristics to those of previously published MSC from the umbilical cord. Under osteogenic differentiation conditions, all MSC populations formed calcium deposits and expressed higher SPARC. Over time, the expression of MSI1 followed different patterns for the different MSC populations. It was not significantly different than the expression of RUNX2. In contrast, the expression of MSI1 and POSTN and RUNX2 were statistically different in most MSC populations. CONCLUSION: In the current study, a similar expression pattern of MSI1 and RUNX2 during in vitro osteogenic differentiation was identified.