Tooth loss and depression in Chilean participants of the National Health Survey 2016-2017: Oral and social functions mediation analysis.

BACKGROUND: Previous studies have indicated the association between poor oral health and depression in adults. This study evaluated oral and social functions contribution to the association between tooth loss and depressive symptoms in Chilean individuals. METHODS: We used data from the Chilean National Health Survey. The number of remaining teeth (≤19 versus ≥20 teeth) and anterior tooth losses were the exposure variables. Outcome was depression, measured through a self-report question and with the Composite International Diagnostic Interview - Short Form (CIDI SF). Mediating variables were determined by five questions, including problems regarding "speaking", "pain and suffering", "eating", "daily activities", and "social relationships". We performed logistic regression models adjusted by multiple confounders variables. Finally, we calculated indirect, direct effect, total effect, and the proportion mediated (PM). RESULTS: We included 5383 participants. The self-reported depression and suspected depression prevalence were 22,1 % and 14,0 % respectively. The total effect of fewer remaining teeth (≤19) on self-reported depression was 1.21 (95 % CI 1.02-1.44), and 1.09 (95 % CI 0.90-1.33) for suspected depression. All five variables of oral and social functions significantly mediated the association between tooth loss and depression. Feeling uncomfortable when speaking or eating discomfort were the most significant mediators. LIMITATIONS: The mediation analysis should be interpreted with caution due to the cross-sectional design. CONCLUSIONS: Deterioration of oral and social functions was a significant mediator in the association between tooth loss and depression, in particular feeling uncomfortable when speaking or eating. This mechanism should be considered in interventions to improve mental health.

L-PRF Secretome from Both Smokers/Nonsmokers Stimulates Angiogenesis and Osteoblast Differentiation In Vitro.

Leukocyte and Platelet-Rich Fibrin (L-PRF) is part of the second generation of platelet-concentrates. L-PRF derived from nonsmokers has been used in surgical procedures, with its beneficial effects in wound healing being proven to stimulate biological activities such as cell proliferation, angiogenesis, and differentiation. Cigarette smoking exerts detrimental effects on tissue healing and is associated with post-surgical complications; however, evidence about the biological effects of L-PRF derived from smokers is limited. This study evaluated the impact of L-PRF secretome (LPRFS) derived from smokers and nonsmokers on angiogenesis and osteoblast differentiation. LPRFS was obtained by submerging L-PRF membranes derived from smokers or nonsmokers in culture media and was used to treat endothelial cells (HUVEC) or SaOs-2 cells. Angiogenesis was evaluated by tubule formation assay, while osteoblast differentiation was observed by alkaline phosphatase and osterix protein levels, as well as in vitro mineralization. LPRFS treatments increased angiogenesis, alkaline phosphatase, and osterix levels. Treatment with 50% of LPRFS derived from smokers and nonsmokers in the presence of osteogenic factors stimulates in vitro mineralization significantly. Nevertheless, differences between LPRFS derived from smokers and nonsmokers were not found. Both LPRFS stimulated angiogenesis and osteoblast differentiation in vitro; however, clinical studies are required to determine the beneficial effect of LPRFS in smokers.

FOXO1 regulates wound-healing responses in human gingival fibroblasts.

BACKGROUND AND OBJECTIVE: Forkhead box-O 1 (FOXO1) is a transcription factor actively involved in oral wound healing at the epithelial barrier. However, less is known regarding the role of FOXO1 during the tissue repair response in the connective tissue compartment. This study explored the involvement of FOXO1 in the modulation of fibroblast activity related to wound healing. METHODS: Primary cultures of human gingival fibroblasts were obtained from four healthy young donors. Myofibroblastic differentiation, collagen gel contraction, cell migration, cell spreading, and integrin activation were evaluated in the presence or absence of a FOXO1 inhibitor (AS1842856). Variations in mRNA and proteins of interest were evaluated through qRT-PCR and western blot, respectively. Distribution of actin, α-smooth muscle actin, and ß1 integrin was evaluated using immunofluorescence. FOXO1 and TGF-ß1 expression in gingival wound healing was assessed by immunohistochemistry in gingival wounds performed in C57BL/6 mice. Images were analyzed using ImageJ/Fiji. ANOVA or Kruskal-Wallis test followed by Tukey's or Dunn's post-hoc test was performed. All data are expressed as mean ± SD. p < .05 was considered statistically significant. RESULTS: FOXO1 inhibition caused a decrease in the expression of the myofibroblastic marker α-SMA along with a reduction in fibronectin, type I collagen, TGF-ß1, and ß1 integrin mRNA level. The FOXO1 inhibitor also caused decreases in cell migration, cell spreading, collagen gel contraction, and ß1 integrin activation. FOXO1 and TGF-ß1 were prominently expressed in gingival wounds in fibroblastic cells located at the wound bed. CONCLUSION: The present study indicates that FOXO1 plays an important role in the modulation of several wound-healing functions in gingival fibroblast. Moreover, our findings reveal an important regulatory role for FOXO1 on the differentiation of gingival myofibroblasts, the regulation of cell migration, and collagen contraction, all these functions being critical during tissue repair and fibrosis.

MicroRNAs: The Missing Link between Hypertension and Periodontitis?

Cardiovascular diseases are the leading cause of death worldwide, and arterial hypertension is a recognized cardiovascular risk factor that is responsible for high morbidity and mortality. Arterial hypertension is the result of an inflammatory process that results in the remodeling and thickening of the vascular walls, which is associated with an immunological response. Previous studies have attempted to demonstrate the relationship between oral disease, inflammation, and the development of systemic diseases. Currently, the existence of an association between periodontitis and hypertension is a controversial issue because the underlying pathophysiological processes and inflammatory mechanisms common to both diseases are unknown. This is due to the fact that periodontitis is a chronic inflammatory disease that affects the interface of teeth and surrounding tissues. However, the most likely explanation for understanding this association is related to low-grade chronic inflammation. An initial path in the study of the relationship between the mentioned pathologies is the possibility of an epigenetic influence, mediated by noncoding RNAs as microRNAs. Thus, in the present review we describe the role of microRNAs related to arterial hypertension and/or periodontitis. In addition, we identified 13 common microRNAs between periodontitis and hypertension. According to the predictions of the DIANA-mirPath program, they can regulate genes involved in 52 signaling pathways.

Leukocyte- and Platelet-Rich Fibrin (L-PRF) Obtained from Smokers and Nonsmokers Shows a Similar Uniaxial Tensile Response In Vitro.

We evaluated and compared the biomechanical properties of Leukocyte-and Platelet Rich Fibrin L-PRF clots and membranes derived from smoker and nonsmoker donors. Twenty venous-blood donors (aged 18 to 50 years) were included after signing informed consent forms. L-PRF clots were analyzed and then compressed to obtain L-PRF membranes. L-PRF clot and membrane samples were tested in quasi-static uniaxial tension and the stress-stretch response was registered and characterized. Furthermore, scanning electron microscope representative images were taken to see the fibrin structure from both groups. The analysis of stress-stretch curves allowed us to evaluate the statistical significance in differences between smoker and nonsmoker groups. L-PRF membranes showed a stiffer response and higher tensile strength when compared to L-PRF clots. However, no statistically significant differences were found between samples from smokers and nonsmokers. With the limitations of our in vitro study, we can suggest that the tensile properties of L-PRF clots and membranes from the blood of smokers and nonsmokers are similar. More studies are necessary to fully characterize the effect of smoking on the biomechanical behavior of this platelet concentrate, to further encourage its use as an alternative to promote wound healing in smokers.

Association between number of remaining teeth and incident depression in a rural Chilean cohort.

OBJECTIVES: Previous studies have established an association between tooth loss and depression. However, longitudinal evidence is scarce and needs to be verified in other populations. The aim of this study was to examine the longitudinal association between the number of remaining teeth and incident depression at 2- and 4-years follow-up in individuals enrolled in the Maule cohort (MAUCO) in Chile. METHODS: This prospective study used the information of individuals, aged 38 to 74 years, excluding those with depression at baseline. The number of remaining teeth at baseline was determined in four groups: "20 or more teeth", "10 to 19 teeth", "1 to 9 teeth" and "no natural teeth". Depression was measured through the PHQ-9. Logistic regression was performed to calculate the odds ratios (OR) for incidence depression at both periods of follow-ups, adjusting for age, sex, educational attainment, diabetes mellitus II, and stressful events at follow-up. Also, we performed adjusted multinomial logistic models to analysis the association between the number of remaining teeth and depression severity. RESULTS: In total individuals (n = 3335 at follow 1, n = 2461 at follow 2), all groups have ORs for incident depression above 1 considering 20 or more teeth as reference. In men, those with 10-19 teeth have 2.44 times higher odds of incident depression than those with 20 or more teeth (OR 2.44, CI 95% 1.33-4.50). Edentulous subjects at 4 years follow-up had 2.24 times higher odds of depression than those with more than 20 teeth (OR 2.24 CI 95%1.35-3.72). In women, the ORs (CI 95%) of incident depression were 2.56 (1.50-4.39), 1.56 (1.02-2.40) and 1.27 (0.90-1.81) for "none", "1-9", "10-19" respectively in comparison to the reference group. In edentulous individuals at baseline, the odds for each of the comparisons "mild vs no", "moderate vs no", "moderately severe vs no" and "severe vs no" were above 1, at both follow-ups. CONCLUSION: Individuals with less than 20 teeth in the mouth could had higher odds of incident depression at 2- and 4-years follow-up, with differences between men and women. Also, in our study, edentulism was associated with increased odds of incident depression at 4-years follow-up in women, and with higher levels of severity of depressive symptoms.

Smoking habits do not affect biological responses induced by leucocyte and platelet-rich fibrin in periodontal ligament cells.

BACKGROUND AND OBJECTIVE: Leucocyte- and platelet-rich fibrin has been developed to stimulate wound healing response. However, it is currently unknown whether smoking affects the biological responses elicited by leucocyte- and platelet-rich fibrin on periodontal ligament-derived mesenchymal stromal cells. This study analyzes the kinetics of biomolecule release from leucocyte- and platelet-rich fibrin derived from smokers and nonsmokers and their effect on periodontal ligament cell proliferation and migration as essential biological activities during wound healing. METHODS: Biomolecules present in leucocyte- and platelet-rich fibrin exudates and conditioned media collected from smokers and nonsmokers were analyzed by Luminex arrays. Periodontal ligament-derived mesenchymal stromal cell obtained from one nonsmoker were treated with leucocyte- and platelet-rich fibrin exudates or leucocyte- and platelet-rich fibrin conditioned media derived from both smokers and nonsmokers. The parameters evaluated included cell proliferation, determined by Ki67 immunostaining and migration assessed using transwell assays. Also, cells were treated with nicotine in the presence of fetal bovine serum 10% or leucocyte- and platelet-rich fibrin conditioned media. RESULTS: A similar biomolecular profile was detected in leucocyte- and platelet-rich fibrin exudates and leucocyte- and platelet-rich fibrin conditioned media from smokers and nonsmokers, stimulating (periodontal ligament-derived mesenchymal stromal cell) proliferation, and migration to a comparable degree. Nicotine reduced cell proliferation and migration of periodontal cells; however, this effect was recovered in the presence of leucocyte- and platelet-rich fibrin conditioned media. CONCLUSION: Leucocyte- and platelet-rich fibrin derived from smokers could be an autologous source of biomolecules to stimulate cell biological activities involved in wound healing in smokers who have difficulties in ceasing this habit. Clinical trials are required to evaluate the impact of leucocyte- and platelet-rich fibrin on healing responses in smokers.

Diabetes alters the involvement of myofibroblasts during periodontal wound healing.

OBJECTIVES: Myofibroblasts constitute a specific cell phenotype involved in connective tissue healing. Diabetes alters the wound healing response. However, it is not clear whether diabetes modifies the involvement of myofibroblasts in periodontal wounds. MATERIALS AND METHODS: Type I diabetes was induced in rats through streptozotocin injection, and periodontal wounds were performed. Wound healing was evaluated histologically at 2, 5, 7, and 15 days by measuring epithelial migration, neutrophil infiltration, and collagen and biofilm formation. Distribution of myofibroblasts was evaluated through immunofluorescence for α-smooth muscle actin. Data analyses were performed using the Shapiro-Wilk, ANOVA, or Kruskal-Wallis tests. RESULTS: Diabetic wounds were characterized by delayed epithelial closure, increased neutrophil infiltration, biofilm formation, and reduced collagen formation. Quantification of the myofibroblasts showed a significant reduction at 5 and 7 days in wounds of diabetic rats and an increase at 15 days when compared to wounds of non-diabetic rats. CONCLUSIONS: Diabetic wound healing was associated with decreased epithelial and connective tissue healing, increased levels of inflammation, and biofilm formation. Myofibroblast differentiation was delayed in diabetic periodontal wounds at early time points. However, myofibroblasts persisted at later time points of healing. The present study suggests that diabetes alters the involvement of myofibroblasts during periodontal wound healing.

Uncoupled inflammatory, proliferative, and cytoskeletal responses in senescent human gingival fibroblasts.

BACKGROUND AND OBJECTIVE: Aging is characterized by a decline in tissue structure and function that may be explained by the development of cellular senescence. However, the acquisition of specific phenotypic responses in senescent gingival fibroblasts is still poorly understood. Here, we have analyzed whether proliferation of primary cultures of human gingival fibroblasts may affect different cell functions relevant to cellular senescence and tissue deterioration. METHODS: Human gingival fibroblasts from five young donors were expanded until cellular senescence was achieved. Cellular senescence was evaluated by determining modifications in cell size, cell proliferation, p16 and p21 mRNA levels, H2Ax phosphorylation, cell viability, and senescence-associated beta-galactosidase staining. Inflammation was evaluated by analyzing the secretion of cytokines and nuclear translocation of NF-κB. Collagen remodeling was evaluated using a collagen gel contraction assay. Immunofluorescence and confocal microscopy were used to determine changes in the localization of the cytoskeletal proteins. Data analysis was performed to identify changes between cultures of the same donor at early and late passages using the paired sample t test or the Wilcoxon matched-pairs signed-rank test. RESULTS: Late passage cells showed changes compatible with cellular senescence that included increased cell size, reduced cell proliferation, staining for SA-beta gal, phosphorylated H2Ax, and increased p16 and p21 mRNA levels. Late passage cells showed a decrease in collagen contraction and reduced co-localization between the cytoskeletal proteins actin and vinculin. Importantly, late passage cells neither demonstrated changes in the secretion of inflammatory cytokines nor NF-κB activation. CONCLUSION: Our results imply that cytoskeletal changes and inhibition of cell proliferation represent early modifications in the structure and function of senescent gingival fibroblasts that are not coupled with the acquisition of an inflammatory phenotype. Further studies are needed to clarify the impact of different senescence stages during aging of the periodontium.

Role of Fibroblast Populations in Periodontal Wound Healing and Tissue Remodeling.

After injury to periodontal tissues, a sequentially phased healing response is initiated that enables wound closure and partial restoration of tissue structure and function. Wound closure in periodontal tissues involves the tightly regulated coordination of resident cells in epithelial and connective tissue compartments. Multiple cell populations in these compartments synergize their metabolic activities to reestablish a mucosal seal that involves the underlying periodontal connective tissues and the attachment of these tissues to the tooth surface. The formation of an impermeable seal around the circumference of the tooth is of particular significance in oral health since colonization of tooth surfaces by pathogenic biofilms promotes inflammation, which can contribute to periodontal tissue degradation and tooth loss. The reformation of periodontal tissue structures in the healing response centrally involves fibroblasts, which synthesize and organize the collagen fibers that link alveolar bone and gingiva to the cementum covering the tooth root. The synthesis and remodeling of nascent collagen matrices are of fundamental importance for the reestablishment of a functional periodontium and are mediated by diverse, multi-functional fibroblast populations that reside within the connective tissues of gingiva and periodontal ligament. Notably, after gingival wounding, a fibroblast sub-type (myofibroblast) arises, which is centrally involved in collagen synthesis and fibrillar remodeling. While myofibroblasts are not usually seen in healthy, mature connective tissues, their formation is enhanced by wound-healing cytokines. The formation of myofibroblasts is also modulated by the stiffness of the extracellular matrix, which is mechanosensed by resident precursor cells in the gingival connective tissue microenvironment. Here, we consider the cellular origins and the factors that control the differentiation and matrix remodeling functions of periodontal fibroblasts. An improved understanding of the regulation and function of periodontal fibroblasts will be critical for the development of new therapies to optimize the restoration of periodontal structure and function after wounding.

Comparative effect of platelet-rich plasma, platelet-poor plasma, and fetal bovine serum on the proliferative response of periodontal ligament cell subpopulations.

OBJECTIVES: Cell-based therapies involve the need to expand cell cultures ex vivo for their subsequent implantation in an autologous manner. An important limitation regarding this technology is the use of fetal bovine serum (FBS) that has critical safety limitations. Platelet-derived fractions represent an autologous source of growth factors that may be used for the expansion of these cell cultures. Periodontal ligament (PDL) cells comprise a heterogeneous cell population that may not necessarily respond in a uniform manner to proliferative stimuli. The aim of this study was to evaluate the ability of two platelet-derived fractions, platelet-rich plasma (PRP) and platelet-poor plasma (PPP) and FBS on the proliferative response of different subpopulations of PDL cell cultures. MATERIALS AND METHODS: PDL cells were characterized and then exposed to PRP, PPP, or FBS during 2, 5, or 14 days to analyze cell proliferation and clonogenic capability. Cell proliferation was evaluated through immunofluorescence for Ki67 and by tracing carboxyfluorescein diacetate succinimidyl ester (CFSE) dye in combination with mesenchymal stem cell markers using flow cytometry. RESULTS: Both PRP and PPP stimulated PDL cell proliferation and their clonogenic ability. We found a significant increase of CD73- and CD90-positive cells after PRP or PPP treatment, compared to FBS. Otherwise, no differences were found regarding the response of CD146-or CD105-positive cells when stimulated with PRP, PPP, or FBS. CONCLUSION: PRP and PPP can stimulate the proliferation and clonogenicity of PDL cell populations including cells positive for CD90 and CD73 markers. CLINICAL RELEVANCE: These findings may have implications for future therapies aiming to stimulate periodontal regeneration using autologous growth factors.

[Chikungunya, emerging disease in Latin America. Description of the first cases in Chile]. / Chikungunya, enfermedad emergente en América Latina. Descripción de los primeros casos en Chile.

BACKGROUND: Chikungunya (CHIK) was introduced in The Americas in 2013, spreading rapidly. In 2014, the first imported case was diagnosed in Chile. AIM: To identify patients with clinical suspicion of CHIK and describe their clinical and laboratory characteristics. PATIENTS AND METHODS: Patients with suspected CHIK were enrolled. All were confirmed by PCR, IgM or IgG CHIK. A structured survey was applied, which included demographic questions, travel characteristics, clinical manifestations, and laboratory results. RESULTS: 21 patients were enrolled and CHIK was confirmed in 16, who were further analyzed; 12 were female (75%), average age 39 years (27-52). The Caribbean and South Americawere the most frequent sites of exposure. In 63%, the initial symptom was arthralgia. Most frequent symptoms were myalgias, malaise (both 100%), fever, and polyarthralgia (both 94%). The median duration of arthralgias was 90 days (3-262); in 53% arthralgias lasted ≥ 3 months. Main joints involved were ankles, hands, and wrists; 87% reported invalidating pain. Arthritis lasted longer in men than in women (p < 0.001). 38% of patients presented lymphopenia and one patient mild thrombocytopenia. Two patients required hospitalization, one with severe headaches, the other with acute pyelonephritis. CONCLUSIONS: Chikungunya should be suspected in returning travelers presenting with fever and severe polyarthralgia. Travelers to endemic areas should apply prevention measures to avoid mosquito bites.

Chikungunya, enfermedad emergente en América Latina. Descripción de los primeros casos en Chile / Chikungunya, emerging disease in Latin America. Description of the first cases in Chile

Resumen Introducción: Chikungunya (CHIK) se introduce en América el año 2013 diseminándose rápidamente. En 2014, se diagnosticó el primer caso importado en Chile. Objetivos: Identificar pacientes con sospecha clínica de CHIK. Describir sus características clínicas y laboratorio. Pacientes y Métodos: Se enrolaron pacientes con sospecha de CHIK. Se confirmó mediante reacción de polimerasa en cadena (RPC), IgM o IgG CHIKV. Se aplicó encuesta con preguntas demográficas, características del viaje, manifestaciones clínicas y laboratorio a pacientes y médicos tratantes. Resultados: Se enrolaron 21 pacientes, confirmando CHIK en 16 que se analizaron; 12 mujeres (75%), promedio edad 39 años (27-52). Exposición más frecuente fue el Caribe y Sudamérica. El síntoma inicial fue artralgia en 63%. Los síntomas más frecuentes fueron mialgias y malestar general (100%), fiebre y poliartralgia (94%). La mediana de duración de artralgias 90 días (3-262 días) y en 53% > 3 meses. Las articulaciones más comprometidas fueron tobillos, manos y muñecas, 87% con dolor invalidante. La artritis duró más en hombres que en mujeres (p < 0,001). El 38% de pacientes presentó linfopenia y un paciente trombocitopenia leve. Hubo dos hospitalizaciones por cefalea y pielonefritis aguda. Conclusiones: Chikungunya debe sospecharse en viajeros que regresan febriles y con poliartralgias intensas. Medidas de prevención deben ser indicadas a viajeros a zonas de riesgo.

Background: Chikungunya (CHIK) was introduced in The Americas in 2013, spreading rapidly. In 2014, the first imported case was diagnosed in Chile. Aim: To identify patients with clinical suspicion of CHIK and describe their clinical and laboratory characteristics. Patients and Methods: Patients with suspected CHIK were enrolled. All were confirmed by PCR, IgM or IgG CHIK. A structured survey was applied, which included demographic questions, travel characteristics, clinical manifestations, and laboratory results. Results: 21 patients were enrolled and CHIK was confirmed in 16, who were further analyzed; 12 were female (75%), average age 39 years (27-52). The Caribbean and South Americawere the most frequent sites of exposure. In 63%, the initial symptom was arthralgia. Most frequent symptoms were myalgias, malaise (both 100%), fever, and polyarthralgia (both 94%). The median duration of arthralgias was 90 days (3-262); in 53% arthralgias lasted ≥ 3 months. Main joints involved were ankles, hands, and wrists; 87% reported invalidating pain. Arthritis lasted longer in men than in women (p < 0.001). 38% of patients presented lymphopenia and one patient mild thrombocytopenia. Two patients required hospitalization, one with severe headaches, the other with acute pyelonephritis. Conclusions: Chikungunya should be suspected in returning travelers presenting with fever and severe polyarthralgia. Travelers to endemic areas should apply prevention measures to avoid mosquito bites.

Glycated Collagen Stimulates Differentiation of Gingival Myofibroblasts.

BACKGROUND: Glucose-derived metabolites may alter the structure and biologic properties of important proteins in periodontium, such as collagens. As a consequence, it is possible that collagen-binding cells may change their phenotypic traits. Although the glucose-derived product methylglyoxal (MGO) has been detected in periodontal lesions, the precise effect of collagen glycation on gingival connective tissue biology is not fully understood. The present study evaluates whether collagen glycation by MGO may affect phenotypic properties and remodeling capacity of human gingival fibroblasts (HGFs). METHODS: Primary cultures of HGFs were grown on Type I collagen matrices previously treated with MGO. Cell cultures were tested for cell viability, apoptosis, α-smooth muscle actin (SMA), fibronectin (FN) production, and collagen remodeling. Mechanical properties and morphology of MGO-treated collagen gels were evaluated using rheometry and atomic force microscopy. Statistical analysis was performed by Kruskal-Wallis and Mann-Whitney U tests. RESULTS: MGO-treated collagen did not affect cell viability or apoptosis. In addition, MGO did not induce significant changes in morphology or mechanical properties of the collagen matrix. However, MGO-treated collagen stimulated an increase in the myofibroblast marker α-SMA, production and assembly of FN, and contraction of collagen matrices. Moreover, use of a triple-helical peptide that reconstitutes the collagen-binding domain for integrins GFOGER reverted the assembly of FN induced by MGO-treated collagen. CONCLUSIONS: The present study shows that collagen glycation by MGO stimulates differentiation of myofibroblasts and production and assembly of FN. These responses may alter the homeostatic balance and wound-healing response of gingival connective tissues affected by diabetes mellitus or aging.

Platelet-Poor and Platelet-Rich Plasma Stimulate Bone Lineage Differentiation in Periodontal Ligament Stem Cells.

BACKGROUND: Plasma-derived fractions have been used as an autologous source of growth factors; however, limited knowledge concerning their biologic effects has hampered their clinical application. In this study, the authors analyze the content and specific effect of both platelet-rich plasma (PRP) and platelet-poor plasma (PPP) on osteoblastic differentiation using primary cultures of human periodontal ligament stem cells (HPLSCs). METHODS: The authors evaluated the growth factor content of PRP and PPP using a proteome profiler array and enzyme-linked immunosorbent assay. HPLSCs were characterized by flow cytometry and differentiation assays. The effect of PRP and PPP on HPLSC bone differentiation was analyzed by quantifying calcium deposition after 14 and 21 days of treatment. RESULTS: Albeit at different concentrations, the two fractions had similar profiles of growth factors, the most representative being platelet-derived growth factor (PDGF) isoforms (PDGF-AA, -BB, and -AB), insulin-like growth factor binding protein (IGFBP)-2, and IGFBP-6. Both formulations exerted a comparable stimulus on osteoblastic differentiation even at low doses (2.5%), increasing calcium deposits in HPLSCs. CONCLUSIONS: PRP and PPP showed a similar protein profile and exerted comparable effects on bone differentiation. Further studies are needed to characterize and compare the effects of PPP and PRP on bone healing in vivo.

The influence of platelet-derived products on angiogenesis and tissue repair: a concise update.

Platelet degranulation allows the release of a large amount of soluble mediators, is an essential step for wound healing initiation, and stimulates clotting, and angiogenesis. The latter process is one of the most critical biological events observed during tissue repair, increasing the growth of blood vessels in the maturing wound. Angiogenesis requires the action of a variety of growth factors that act in an appropriate physiological ratio to assure functional blood vessel restoration. Platelets release main regulators of angiogenesis: Vascular Endothelial Growth Factors (VEGFs), basic fibroblast growth factor (FGF-2), and Platelet derived growth factors (PDGFs), among others. In order to stimulate tissue repair, platelet derived fractions have been used as an autologous source of growth factors and biomolecules, namely Platelet Rich Plasma (PRP), Platelet Poor Plasma (PPP), and Platelet Rich Fibrin (PRF). The continuous release of these growth factors has been proposed to promote angiogenesis both in vitro and in vivo. Considering the existence of clinical trials currently evaluating the efficacy of autologous PRP, the present review analyses fundamental questions regarding the putative role of platelet derived fractions as regulators of angiogenesis and evaluates the possible clinical implications of these formulations.

Synthesis of nanostructured porous silica coatings on titanium and their cell adhesive and osteogenic differentiation properties.

Nanostructured porous silica coatings were synthesized on titanium by the combined sol-gel and evaporation-induced self-assembly process. The silica-coating structures were characterized by X-ray diffraction, transmission electron microscopy, scanning electron microscopy, and nitrogen sorptometry. The effect of the nanoporous surface on apatite formation in simulated body fluid, protein adsorption, osteoblast cell adhesion behavior, and osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs) is reported. Silica coatings with highly ordered sub-10 nm porosity accelerate early osteoblast adhesive response, a favorable cell response that is attributed to an indirect effect due to the high protein adsorption observed on the large-specific surface area of the nanoporous coating but is also probably due to direct mechanical stimulus from the nanostructured topography. The nanoporous silica coatings, particularly those doped with calcium and phosphate, also promote the osteogenic differentiation of hBMSCs with spontaneous mineral nodule formation in basal conditions. The bioactive surface properties exhibited by the nanostructured porous silica coatings make these materials a promising alternative to improve the osseointegration properties of titanium dental implants and could have future impact on the nanoscale design of implant surfaces.

La vacuna polio oral en lactantes no interfiere con la detección de enterovirus en sangre / Oral polio vaccine in infants does not interfere in detection of enterovirus in blood

Introduction: There is not known if a viraemia post-oral polio vaccine (OPV) is detectable by modern molecular techniques. Such viraemia could affect the performance of the real time-polymerase chain reaction (PCR) for non polio enterovirus (EV) detection, technique of growing clinical use for the study of febrile infants. Objective: To determine viraemia post-first dose of OPV in healthy infants, by molecular techniques. Patients and Methods: 50 infants less than three months without previous VPO were randomized in 5 groups: a control group with pre-vaccination blood sample (BS), group 1 BS at day 2, group 2 BS at day 4, group 3, BS at day 6 and group 4, BS at day 8 post-vaccination. Conventional and specific PCR for poliovirus and real time PCR for non polio EV were performed in BS and in OPV samples. Results: No genetic material of poliovirus was detected in any infant, while in 9 of them (18%) non polio EV was identified. Real time PCR for EV did not amplify poliovirus from OPV samples. Discussion: Results suggest that no post VPO viraemia detectable by molecular methods exists. Considering that real time PCR for EV does not allow to identify polio virus, no false positives of the test are expected as a result of a recent VPO vaccination. We documented presence of non polio EV in blood of healthy asymptomatic infants.

Introducción: No existen estudios que indiquen si la vacuna polio oral (VPO) produce viremia detectable mediante métodos moleculares. Una eventual viremia podría afectar el rendimiento de la RPC tiempo real para detectar enterovirus (EV) no polio, examen de creciente uso clínico en lactantes pequeños con fiebre sin foco. Objetivo: Determinar viremia post VPO en lactantes sanos, por métodos moleculares. Métodos: 50 menores de 3 meses, al momento de recibir su primera VPO se distribuyeron en forma aleatoria en 5 grupos: control, muestra de sangre pre-vacunación; grupo 1, muestra al 2° día; grupo 2, al 4° día; grupo 3, al 6° día y grupo 4, al 8° día post-vacunación. Se realizó RPC convencional específica para virus polio y RPC tiempo real para EV no polio en las muestras de sangre y en muestras de VPO. Resultados: No se identificó presencia de material genético de virus polio en lactante alguno, mientras que en 9 (18%) se identificó presencia de EV no polio. La RPC tiempo real para EV no polio no amplificó material genético a partir de las muestras de VPO. Discusión: Los resultados sugieren que no existe viremia post-VPO detectable por métodos moleculares. Considerando que la RPC tiempo real de EV no polio de uso clínico no permite identificar la presencia de virus polio, estos hallazgos indican que no existirán falsos positivos de este examen como resultado de una vacunación VPO reciente. Adicionalmente se documentó presencia de EV no polio en sangre de lactantes asintomáticos.

Proliferation of murine midbrain neural stem cells depends upon an endogenous sonic hedgehog (Shh) source.

The Sonic Hedgehog (Shh) pathway is responsible for critical patterning events early in development and for regulating the delicate balance between proliferation and differentiation in the developing and adult vertebrate brain. Currently, our knowledge of the potential role of Shh in regulating neural stem cells (NSC) is largely derived from analyses of the mammalian forebrain, but for dorsal midbrain development it is mostly unknown. For a detailed understanding of the role of Shh pathway for midbrain development in vivo, we took advantage of mouse embryos with cell autonomously activated Hedgehog (Hh) signaling in a conditional Patched 1 (Ptc1) mutant mouse model. This animal model shows an extensive embryonic tectal hypertrophy as a result of Hh pathway activation. In order to reveal the cellular and molecular origin of this in vivo phenotype, we established a novel culture system to evaluate neurospheres (nsps) viability, proliferation and differentiation. By recreating the three-dimensional (3-D) microenvironment we highlight the pivotal role of endogenous Shh in maintaining the stem cell potential of tectal radial glial cells (RGC) and progenitors by modulating their Ptc1 expression. We demonstrate that during late embryogenesis Shh enhances proliferation of NSC, whereas blockage of endogenous Shh signaling using cyclopamine, a potent Hh pathway inhibitor, produces the opposite effect. We propose that canonical Shh signaling plays a central role in the control of NSC behavior in the developing dorsal midbrain by acting as a niche factor by partially mediating the response of NSC to epidermal growth factor (EGF) and fibroblast growth factor (FGF) signaling. We conclude that endogenous Shh signaling is a critical mechanism regulating the proliferation of stem cell lineages in the embryonic dorsal tissue.

[Oral polio vaccine in infants does not interfere in detection of enterovirus in blood]. / La vacuna polio oral en lactantes no interfiere con la detección de enterovirus en sangre.

INTRODUCTION: There is not known if a viraemia post-oral polio vaccine (OPV) is detectable by modern molecular techniques. Such viraemia could affect the performance of the real time-polymerase chain reaction (PCR) for non polio enterovirus (EV) detection, technique of growing clinical use for the study of febrile infants. OBJECTIVE: To determine viraemia post-first dose of OPV in healthy infants, by molecular techniques. PATIENTS AND METHODS: 50 infants less than three months without previous VPO were randomized in 5 groups: a control group with pre-vaccination blood sample (BS), group 1 BS at day 2, group 2 BS at day 4, group 3, BS at day 6 and group 4, BS at day 8 post-vaccination. Conventional and specific PCR for poliovirus and real time PCR for non polio EV were performed in BS and in OPV samples. RESULTS: No genetic material of poliovirus was detected in any infant, while in 9 of them (18%) non polio EV was identified. Real time PCR for EV did not amplify poliovirus from OPV samples. DISCUSSION: Results suggest that no post VPO viraemia detectable by molecular methods exists. Considering that real time PCR for EV does not allow to identify polio virus, no false positives of the test are expected as a result of a recent VPO vaccination. We documented presence of non polio EV in blood of healthy asymptomatic infants.