A pilot study of circulating levels of TGF-ß1 and TGF-ß2 as biomarkers of bone healing in patients with non-hypertrophic pseudoarthrosis of long bones.

BACKGROUND: Pseudoarthrosis or non-union is a complication with an incidence of 5-10% of bone fractures, most frequently located in the diaphysis of long bones. The management of this complication is addressed by means of complex surgical procedures and is a concern for orthopaedic and trauma surgeons nowadays. The use of biomarkers for diagnosing patients at risk of non-union would help us to establish special measures for early corrective treatment. METHODS: Prospective exploratory pilot study with a cohort of 20 patients diagnosed of non-hypertrophic pseudoarthrosis of long bones who were treated surgically with either autologous bone graft or a Tissue Engineering Product composed of bone marrow-derived Mesenchymal Stromal Cells. Patients were followed for 12 months and plasma blood samples were obtained to determine circulating levels of Transforming Growth Factor Beta 1 and Beta 2 (TGF-ß1 and TGF-ß2, respectively) at inclusion, and at 1 week, 2 weeks, and months 1, 2, 3, 6 and 12 after surgery. Radiological bone healing was evaluated by the Tomographic Union Score (TUS). RESULTS: Basal levels of TGF-ß1 and TGF-ß2 were determined in the twenty patients (26,702 ± 14,537 pg/mL and 307.8 ± 83.1 pg/mL, respectively). Three of them withdrew from the study, so complete follow-up was conducted on 17 patients (9 successfully healed vs. 8 that did not heal). Statistically significant differences between the bone healing group and the non-union group were found at month 12 for both TGF-ß1 (p = 0.005) and TGF-ß2 (p = 0.02). CONCLUSIONS: TGF-ß1 and TGF-ß2 are biomarkers that correlate with clinical evidence of bone regeneration and may be used to monitor patients, although early predictive value after intervention needs to be further studied in combination with other molecules.

Evaluation of a cell-based osteogenic formulation compliant with good manufacturing practice for use in tissue engineering.

Proper bony tissue regeneration requires mechanical stabilization, an osteogenic biological activity and appropriate scaffolds. The latter two elements can be combined in a hydrogel format for effective delivery, so it can readily adapt to the architecture of the defect. We evaluated a Good Manufacturing Practice-compliant formulation composed of bone marrow-derived mesenchymal stromal cells in combination with bone particles (Ø = 0.25 to 1 µm) and fibrin, which can be readily translated into the clinical setting for the treatment of bone defects, as an alternative to bone tissue autografts. Remarkably, cells survived with unaltered phenotype (CD73+, CD90+, CD105+, CD31-, CD45-) and retained their osteogenic capacity up to 48 h after being combined with hydrogel and bone particles, thus demonstrating the stability of their identity and potency. Moreover, in a subchronic toxicity in vivo study, no toxicity was observed upon subcutaneous administration in athymic mice and signs of osteogenesis and vascularization were detected 2 months after administration. The preclinical data gathered in the present work, in compliance with current quality and regulatory requirements, demonstrated the feasibility of formulating an osteogenic cell-based tissue engineering product with a defined profile including identity, purity and potency (in vitro and in vivo), and the stability of these attributes, which complements the preclinical package required prior to move towards its use of prior to its clinical use.

Effect of Allogeneic Cell-Based Tissue-Engineered Treatments in a Sheep Osteonecrosis Model.

Osteonecrosis of the femoral head (ONFH) is defined as a tissue disorder and successive subchondral bone collapse resulting from an ischemic process, which may progress to hip osteoarthritis. Cell therapy with multipotent bone marrow mesenchymal stromal cells (BM-MSC) of autologous origin appears to be safe and has shown regenerative potential in previous preclinical and clinical studies. The use of allogeneic cells is far more challenging, but may be a promising alternative to use of autologous cells. Moreover, an optimized dosage of cells from an allogeneic source is needed to obtain off-the-shelf tissue engineering products (TEPs). The purpose of this study was to evaluate the efficacy of a TEP composed of undifferentiated ex vivo expanded BM-MSC of allogeneic origin, combined with bone matrix particles in variable doses. A comparative analysis of TEP's bone regenerative properties against its autologous counterpart was performed in an early-stage ONFH preclinical model in mature sheep. Allogeneic BM-MSC groups demonstrated bone regeneration capacity in osteonecrotic lesions equivalent to autologous BM-MSC groups 6 weeks after treatment. Likewise, stimulation of bone regeneration by a low cell dose of 0.5 × 106 BM-MSC/cm3 was equivalent to that of a high cell dose, 5 × 106 BM-MSC/cm3. Neither local nor systemic immunological reactions nor tumorigenesis were reported, strengthening the safety profile of allogeneic BM-MSC therapy in this model. Our results suggest that low-dose allogeneic BM-MSC is sufficient to promote bone regeneration in femoral head osteonecrotic lesions, and should be considered in translation of new allogeneic cell-based TEPs to human clinics. Impact statement Cell therapy and tissue engineering hold promise as novel regenerative therapies for musculoskeletal diseases, and particularly in bone regeneration strategies. In this article, we report the evaluation of the efficacy of an allogeneic cell-based tissue engineering product (TEP) in an early-stage osteonecrosis of the femoral head preclinical model in skeletally mature sheep. Moreover, we demonstrate its bone regeneration capacity and safety in vivo and its equivalence to autologous counterparts. These findings have important implications for the translation of new allogeneic cell-based TEPs to human clinics.

Emotional Processing and Personality as Predictors of Obsessive-compulsive Symptoms in College Students.

The categorical definition of obsessive-compulsive disorder, and exclusive focus on thoughts and behaviors, have constrained the study and treatment of its symptoms. The present study's aim was to search for relationships among emotional processing dimensions, five major personality dimensions, and self-perceived obsessive-compulsive symptoms. The participants were 100 college students, and the questionnaires used were a selection of images from the International Affective Picture System (IAPS), the Self-assessment Manikin (SAM), the Dimensional Obsessive-Compulsive Scale (DOCS), and the NEO-FFI. We found differences in emotional processing dimensions between participants with high and low DOCS scores, grouped according to sex (d = .56); and evidence that the neuroticism and agreeableness dimensions predict self-perceived obsessive-compulsive symptoms. Emotional processing dimensions and personality are considered useful to comprehending obsessive-compulsive symptoms, which lends support to dimensional models of OC symptomatology, as well as planning and developing psychological interventions.

Levels of IL-17F and IL-33 correlate with HLA-DR activation in clinical-grade human bone marrow-derived multipotent mesenchymal stromal cell expansion cultures.

BACKGROUND AIMS: Multipotent mesenchymal stromal cell (MSC)-based medicines are extensively investigated for use in regenerative medicine and immunotherapy applications. The International Society for Cell and Gene Therapy (ISCT) proposed a panel of cell surface molecules for MSC identification that includes human leukocyte antigen (HLA)-DR as a negative marker. However, its expression is largely unpredictable despite production under tightly controlled conditions and compliance with current Good Manufacturing Practices. Herein, we report the frequency of HLA-DR expression in 81 batches of clinical grade bone marrow (BM)-derived MSCs and investigated its impact on cell attributes and culture environment. METHODS: The levels of 15 cytokines (interleukin [IL]-1ß, IL-4, IL-6, IL-10, IL-17A, IL-17F, IL-21, IL-22, IL-23, IL-25, IL-31, IL-33, interferon-γ, soluble CD40 ligand and tumor necrosis factor-α) were determined in sera supplements and supernatants of BM-MSC cultures. Identity, multipotentiality and immunopotency assays were performed on high (>20% of cells) and low (≤20% of cells) HLA-DR+ cultures. RESULTS: A correlation was found between HLA-DR expression and levels of IL-17F and IL-33. Expression of HLA-DR did neither affect MSC identity, in vitro tri-lineage differentiation potential (into osteogenic, chondrogenic and adipogenic lineages), nor their ability to inhibit the proliferation of stimulated lymphocytes. DISCUSSION: Out of 81 batches of BM-MSCs for autologous use analyzed, only three batches would have passed the ISCT criteria (<2%), whereas 60.5% of batches were compliant with low HLA-DR values (≤20%). Although a cause-effect relationship cannot be drawn, we have provided a better understanding of signaling events and cellular responses in expansion culture conditions relating with HLA-DR expression.

Emotional processing and personality as predictors of obsessive-compulsive symptoms in college students

The categorical definition of obsessive-compulsive disorder, and exclusive focus on thoughts and behaviors, have constrained the study and treatment of its symptoms. The present study's aim was to search for relationships among emotional processing dimensions, five major personality dimensions, and self-perceived obsessive-compulsive symptoms. The participants were 100 college students, and the questionnaires used were a selection of images from the International Affective Picture System (IAPS), the Self-assessment Manikin (SAM), the Dimensional Obsessive-Compulsive Scale (DOCS), and the NEO-FFI. We found differences in emotional processing dimensions between participants with high and low DOCS scores, grouped according to sex (d = .56); and evidence that the neuroticism and agreeableness dimensions predict self-perceived obsessive-compulsive symptoms. Emotional processing dimensions and personality are considered useful to comprehending obsessive-compulsive symptoms, which lends support to dimensional models of OC symptomatology, as well as planning and developing psychological interventions

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Derivation of Multipotent Mesenchymal Stromal Cells from Ovine Bone Marrow.

In the field of orthopedics, translational research of novel therapeutic approaches involves the use of large animal models (such as sheep, goat, pig, dog, and horse) due to the similarities with humans in weight, size, joint structure, and bone/cartilage healing mechanisms. Particularly in the development of cell-based therapies, the lack of manageable immunocompromised preclinical large animal models prevents the use of human cells, which makes it necessary to produce equivalent homologous cell types for the study of their pharmacodynamics, pharmacokinetics, and toxicology. The methods described herein allow for the isolation, expansion, manipulation, and characterization of fibroblastic-like ovine bone marrow-derived multipotent mesenchymal stromal cells (BM-MSC) that, similar to human BM-MSC, adhere to standard plastic surfaces; express specific surface markers such as CD44, CD90, CD140a, CD105, and CD166; and display trilineage differentiation potential in vitro. Homogeneous cell cultures result from a 3-week bioprocess yielding cell densities in the range of 2-4 × 104 MSC/cm2 at passage 2, which corresponds to ∼8 cumulative population doublings. Large quantities of BM-MSC resulting from following this methodology can be readily used in proof of efficacy and safety studies in the preclinical development stage. © 2018 by John Wiley & Sons, Inc.

A reproducible method for the isolation and expansion of ovine mesenchymal stromal cells from bone marrow for use in regenerative medicine preclinical studies.

The use of multipotent mesenchymal stromal cells (MSCs) as candidate medicines for treating a variety of pathologies is based on their qualities as either progenitors for the regeneration of damaged tissue or producers of a number of molecules with pharmacological properties. Preclinical product development programmes include the use of well characterized cell populations for proof of efficacy and safety studies before testing in humans. In the field of orthopaedics, an increasing number of translational studies use sheep as an in vivo test system because of the similarities with humans in size and musculoskeletal architecture. However, robust and reproducible methods for the isolation, expansion, manipulation and characterization of ovine MSCs have not yet been standardised. The present study describes a method for isolation and expansion of fibroblastic-like, adherent ovine MSCs that express CD44, CD90, CD140a, CD105 and CD166, and display trilineage differentiation potential. The 3-week bioprocess proposed here typically yielded cell densities of 1.4 × 104 MSCs/cm2 at passage 2, with an expansion factor of 37.8 and approximately eight cumulative population doublings. The osteogenic potential of MSCs derived following this methodology was further evaluated in vivo in a translational model of osteonecrosis of the femoral head, in which the persistence of grafted cells in the host tissue and their lineage commitment into osteoblasts and osteocytes was demonstrated by tracking enhanced green fluorescent protein-labelled cells. Copyright © 2016 John Wiley & Sons, Ltd.

Dengue seroprevalence and risk factors for past and recent viral transmission in Venezuela: a comprehensive community-based study.

Dengue transmission in Venezuela has become perennial and a major public health problem. The increase in frequency and magnitude of recent epidemics prompted a comprehensive community-based cross-sectional study of 2,014 individuals in high-incidence neighborhoods of Maracay, Venezuela. We found a high seroprevalence (77.4%), with 10% of people experiencing recent infections. Multivariate logistic regression analysis showed that poverty-related socioeconomic factors (place and duration of residence, crowding, household size, and living in a shack) and factors/constraints related to intradomiciliary potential mosquito breeding sites (storing water and used tires) were linked with a greater risk of acquiring a dengue infection. Our results also suggest that transmission occurs mainly at home. The combination of increasingly crowded living conditions, growing population density, precarious homes, and water storage issues caused by enduring problems in public services in Maracay are the most likely factors that determine the permanent dengue transmission and the failure of vector control programs.

Georreferenciación de pacientes diagnosticados con Fibrosis Quistica en la unidad proyecto Aragua. Universidad de Carabobo, período junio 1993-junio 2011, Venezuela / Georeferencng off patients with cystic fibrosis diagnosed i the project arragua unit - carabobo university, period june 1993 - june 2011, Venezuela.

Los Sistemas de Información Geográfica son definidos, como conjunto de tecnologías, métodos y procedimientos informaticos, que pueden ser utilizados para conocer la distribución de patologías como la Fibrosis Quística, que es una enfermedad de etiología genética, con un patrón de herencia autosómica recesiva, que aunque tiene una baja incidencia, sus complicaciones son severas, requiriendo de tratamiento diario y frecuentes hospitalizaciones, lo que representa un alto costo para la sociedad. En Venezuela no se conoce de forma precisa la distribución espacial de esta patología, esto motivo a la realización de una investigación de tipo descriptiva, documental y retrospectiva, que permitió desarrollar un modelo georreferencial con 69 pacientes diganosticados con Fibrosis Quística en la Unidad Proyecto Aragua (UPA), durante el periodo Junio 1993- Junio 2011. Los resultados obtenidos evidenciaron que los lugares de procedencia más frecuentes fueron: Distrito Capital, Aragua y Carabobo, igualmente facilitó la identificación de los patrones de distribución georreferencial de dichos pacientes y sus familiares hasta el segundo grado de consanguinidad y se presenta la posible migración o rutas genética de los genes de Fibrosis Quistica.

Geographic Information Systems are defined as a set of computer technology, methods and procedures used in the handling and analysis of geographically referenced data. They can be used to map the distribution of diseases such as Cystic Fibrosis is a disease of genetic etiology with an autosomal recessive inheritance pattern, although it has a low incidence, complications of this disease are very severe and require treatment daily and frequent hospitalizations, representing a high cost to society. In Venezuela it is not known precisely the spatial distribution of this disease, which is why there was a descriptive research, documentary, retrospective, which allowed developing a geospatial model with 69 CF patients who were diagnosed in the Project Aragua Unit (UPA) during the period June 1993 - June 2011, resulting in the most common places of origin were: Capital District, Aragua and Carabobo; it also facilitated the identification of the pattient of distribution of georreferencial such patient and their relatives up to the second degree of consaguinity and presents the possible migration or “path” genetics of the genes for Cystic Fibrosis.

Integración de plataformas tecnológicas por productos, entre la Universidad de Carabobo y el Sistema Público Nacional de Salud, mediante la estrategia de comunidades virtuales / Integration of technology platforms for products from the Carabobo University and National Public health system, through Virtual Communities Strategy

Esta investigación, propone la integración sistemática de las plataformas tecnológicas de la Universidad de Carabobo, en su Facultad de Ciencias de la Salud-Sede Aragua, con la del Sistema Público Nacional de Salud, a través del desarrollo de productos de las Nuevas Tecnologías de Información y Comunicación, en este caso, haciendo uso de la estrategia de comunidades virtuales. Las comunidades virtuales, constituyen espacios propicios de desarrollo de las redes sociales, que con la llegada de la Internet permiten su creación y progreso más allá del contexto físico. Se empleó la modalidad de proyecto factible, para el diseño de un prototipo de curso on line, considerado como experiencia piloto, para la enseñanza de la estrategia de comunidades virtuales, Grupos Yahoo - Facebook, como recurso de mediación de experiencias de aprendizaje en las Pasantías de Ejercicio Profesional en Ciencias de la Salud. Fundamentan la propuesta, principios de convergencia tecnológica, de encuentro de saberes, la necesidad de construcción de la ciudadanía digital en salud, la búsqueda de representaciones sociales de la tecnología, la inclusión digital de las comunidades para su autonomía y bienestar, implicando también el desarrollo de espacios virtuales de emancipación, que finalmente asienten una tecnología supeditada a lo humano, a la justicia social.

The research proposes the systematic integration of technology platforms at the Carabobo University, in its Health Sciences Faculty, Aragua headquarters with the National Public Health System, through out development by informational and communication technologies products. Also virtual community strategy was involved. Virtual communities constitute appropriate spaces for the development of social networks, that with the advent of the Internet allows the creation and development beyond the physical context. This approach used the feasible project modality. Here, it was developed an online course for teaching virtual communities strategy, as a learning mediation resource experiences in the Professional Internships Health Sciences Practice. Characterize the proposed system, the principles of technological convergence, the coming together of knowledge, the construction of digital citizenship and health technology search social representations, digital community inclusion destiny to their autonomy and welfare, virtual emancipation spaces development, finally settled all technology subject to the humanity and social justice.

Fenotipo-Genotipo de Pacientes con Fibrosis Quística: Unidad Proyecto Aragua 1997-2007 / Phenotype - Genotype of Cystic Fibrosis Patients: Diagnosed at Unidad Proyecto Aragua 1997-2007

La Fibrosis Quística (FQ) es una enfermedad autosómica recesiva multisistémica de evolución crónica progresiva y letal. Varios estudios de correlación fenotipo-genotipo han identificado mutaciones relacionadas con formas clínicas severas y otras leves, esta asociación aún no ha sido investigada en los pacientes FQ diagnosticados en la Unidad Proyecto Aragua (UPA) de la Universidad de Carabobo Sede Aragua. Por tal motivo esta investigación es realizada a partir de la revisión de diez historias clínicas de pacientes con diagnóstico de FQ realizado en la UPA y atendidos en el Hospital Central de Maracay durante el período 1997-2007. El objetivo del trabajo fue determinar las características fenotípicas y relacionarlas con el genotipo. Los genotipos presentes fueron DeltaF508/Otra (33,3%), G542X/Otra (11,5%), Otra/Otra (55,6%); se formaron dos grupos según el genotipo: El Nº1, integrado por individuos con genotipo compuesto, donde uno de los alelos corresponde a una mutación severa; y en el Nº 2 se incluyeron los pacientes con mutaciones no identificadas y un paciente fallecido antes de investigar el genotipo. El promedio de la edad al diagnóstico es 1,92 años en el grupo 1 y 4,26 años en el grupo 2. La insuficiencia pancreática (IP) y las complicaciones digestivas son más severas en el grupo 1; los pacientes del grupo 2 presentan mayor heterogeneidad clínica. Se concluye que el genotipo es un buen predictor de la función exocrina pancreática ya que todos los pacientes con IP, tienen al menos una mutación severa. Se manifiesta la necesidad de indagar otras mutaciones y sugerimos un diagnóstico precoz y selectivo, que ayude a prevenir las complicaciones de esta patología y mejorar la calidad de vida de pacientes y sus familias.

Cystic fibrosis is an autosomal recessive and multisystemic disease of chronic progressive and lethal evolution. Several studies of fenotype-genotype correlations have identified mutations related to clinical severe and mild forms, this association still has not been investigated in the FQ patients diagnosed in the Unidad Proyecto Aragua (UPA) of Universidad de Carabobo Sede Aragua (University of Carabobo, Aragua Campus). This is a descriptive and retrospective investigation carried out from the review of ten patients’ case histories with FQ diagnosed in the UPA and attended at the Central Hospital of Maracay during the period 1997-2007. The aim of this work was to determine the phenotypic characteristics and to relate them to the genotype. The genotypes found were DeltaF508/Other (33.3%), G542X/Other (11.5%), Other / Other (55.6%); the patients were classified according to genotype in two groups: Group Nº 1 integrated by individuals with composed genotype, where one of the allele corresponds to a severe mutation; and in group Nº 2 were included the patients with not identified mutations and a deceased patient before investigating the genotype. The mean age at diagnosis was 1.92 years in group 1 and of 4.26 years in group 2. The pancreatic insufficiency (IP) and the digestive complications are severe in group 1; the patients of group 2 showed major clinical heterogeneity. It is concluded that the genotype is a good predictor of the exocrine pancreatic function, since all the patients with IP, have at least a severe mutation. It demonstrates the need to investigate other mutations and we suggest a selective and early diagnosis, to help prevent the complications of this pathology and improve the quality of patients’ life and his families.