Doxorubicin Activity Is Modulated by Traditional Herbal Extracts in a 2D and 3D Multicellular Sphere Model of Leukemia.

The modulation of the tumor microenvironment by natural products may play a significant role in the response of tumor cells to chemotherapy. In this study, we evaluated the effect of extracts derived from P2Et (Caesalpinia spinosa) and Anamú-SC (Petiveria alliacea) plants, previously studied by our group, on the viability and ROS levels in the K562 cell line (Pgp- and Pgp+), endothelial cells (ECs, Eahy.926 cell line) and mesenchymal stem cells (MSC) cultured in 2D and 3D. The results show that: (a) the two botanical extracts are selective on tumor cells compared to doxorubicin (DX), (b) cytotoxicity is independent of the modulation of intracellular ROS for plant extracts, unlike DX, (c) the interaction with DX can be influenced by chemical complexity and the expression of Pgp, (d) the 3D culture shows a greater sensitivity of the tumor cells to chemotherapy, in co-treatment with the extracts. In conclusion, the effect of the extracts on the viability of leukemia cells was modified in multicellular spheroids with MSC and EC, suggesting that the in vitro evaluation of these interactions can contribute to the comprehension of the pharmacodynamics of the botanical drugs.

Revisión narrativa: unidad de ortogeriatría, la propuesta para reducir los desenlaces adversos en adultos mayores con fracturas de cadera por fragilidad / Narrative review: Orthogeriatrics unit, the proposal to reduce adverse outcomes in older adults with fragility fractures

Introducción Las fracturas de cadera son un problema de salud pública debido a su asociación con altas tasas de mortalidad a un año, deterioro funcional y empeoramiento de la calidad de vida. Un enfoque multidisciplinario mejora los resultados de los pacientes adultos mayores con fracturas por fragilidad. Objetivo Realizar una revisión narrativa para evaluar la evidencia de las unidades de ortogeriatría como estrategia para reducir desenlaces adversos en adultos mayores con fracturas de cadera por fragilidad. Materiales y métodos Se realizó una búsqueda de la literatura en las siguientes bases de datos: PubMed,Cochrane y Scielo, con los términos: "hip fracture" or "fragility fracture" AND "multidisciplinary team" or "Geriatric co management" or AND "orthogeriatric" or "ortho-geriatric". Resultados El manejo por ortogeriatría reduce el tiempo de evaluación pre-quirúrgico, complicaciones postoperatorias y la estancia hospitalaria. La valoración interdisciplinaria ha demostrado ser eficaz para disminuir el deterioro funcional, la mortalidad intrahospitalaria, la mortalidad a los 30 días y el riesgo de institucionalización en comparación con otros modelos de atención. Estos factores impactan sobre la optimización de recursos disminuyendo los costos en salud. Conclusión Los servicios de ortogeriatría son el modelo ideal para tratar las fracturas de cadera por fragilidad en adultos mayores. En Colombia se deben implementar más unidades de ortogeriatría para mejorar la atención intrahospitalaria, crear programas de seguimiento y rehabilitación.

Introduction Hip fracture is a public health problem due to its association with high mortality rates at one year, functional decline and worsening of quality of life. A multidisciplinary approach improves outcomes for elderly patients with fragility fracture. Objective To carry out a narrative review to evaluate the evidence of orthogeriatric units as a strategy to reduce adverse outcomes in older adults with fragility hip fractures. Material and Methods A literature search was carried out in the following databases: PubMed, Cochrane and Scielo, with the terms: "hip fracture" or "fragility fracture" AND "multidisciplinary team" or "Geriatric co management" or AND "orthogeriatric" or "ortho-geriatric". Results Care by orthogeriatrics reduces pre-operative surgical time, postoperative complications and hospital stay. Interdisciplinary assessment has been shown to be effective in reducing functional decline, in-hospital mortality, 30-day mortality, and the risk of institutionalization compared to other models of care. These factors impact on the optimization of resources, reducing health costs. Conclusion Orthogeriatric services are the ideal model to treat fragility fractures in older adults. In Colombia, more orthogeriatric units should be implemented to improve in-hospital care, create follow-up and rehabilitation programs.

3D Multicellular Spheroid for the Study of Human Hematopoietic Stem Cells: Synergistic Effect Between Oxygen Levels, Mesenchymal Stromal Cells and Endothelial Cells.

INTRODUCTION: The human bone marrow microenvironment is composed of biological, chemical and physical factors that act in a synergistic way to modulate hematopoietic stem cell biology, such as mesenchymal stromal cells (MSCs), endothelial cells (ECs) and low oxygen levels; however, it is difficult to mimic this human microenvironment in vitro. METHODS: In this work, we developed 3D multicellular spheroid (3D-MS) for the study of human hematopoietic stem cells (HSCs) with some components of perivascular niche. HSCs were isolated from umbilical cord blood, MSCs were isolated from human bone marrow and a microvasculature EC line (CC-2811, Lonza®) was used. For the formation of a 3D structure, a magnetic levitation culture system was used. Cultures were maintained in 21%, 3% and 1% O2 for 15 days. Culture volume, sphericity index and cell viability were determined. Also, human HSC proliferation, phenotype and production of reactive oxygen species were evaluated. RESULTS: After 15 days, 3D-MS exhibited viability greater than 80%. Histology results showed structures without necrotic centers, and higher cellular proliferation with 3% O2. An increase in the expression of the CD34 antigen and other hematopoietic antigens were observed to 1% O2 with MSCs plus ECs and low ROS levels. CONCLUSION: These findings suggest that 3D-MS formed by MSCs, ECs and HSCs exposed to low concentrations of oxygen (1-3% O2) modulate human HSC behavior and mimics some features of the perivascular niche, which could reduce the use of animal models and deepen the relationship between the microenvironment of HSC and human hematological diseases development.

TRPM8 Channel Promotes the Osteogenic Differentiation in Human Bone Marrow Mesenchymal Stem Cells.

Various families of ion channels have been characterized in mesenchymal stem cells (MSCs), including some members of transient receptor potential (TRP) channels family. TRP channels are involved in critical cellular processes as differentiation and cell proliferation. Here, we analyzed the expression of TRPM8 channel in human bone marrow MSCs (hBM-MSCs), and its relation with osteogenic differentiation. Patch-clamp recordings showed that hBM-MSCs expressed outwardly rectifying currents which were increased by exposure to 500 µM menthol and were partially inhibited by 10 µM of BCTC, a TRPM8 channels antagonist. Additionally, we have found the expression of TRPM8 by RT-PCR and western blot. We also explored the TRPM8 localization in hBM-MSCs by immunofluorescence using confocal microscopy. Remarkably, hBM-MSCs treatment with 100 µM of menthol or 10 µM of icilin, TRPM8 agonists, increases osteogenic differentiation. Conversely, 20 µM of BCTC, induced a decrease of osteogenic differentiation. These results suggest that TRPM8 channels are functionally active in hBM-MSCs and have a role in cell differentiation.

Comportamiento de reingreso a 30 días en un centro de reemplazos articulares. Estudio de cohorte longitudinal / 30 days re-admission in an articular Joint Arthroplasty center. A cohort Study

Introducción Los reemplazos articulares son calificados como procedimientos seguros y exitosos para el tratamiento de la osteoartrosis severa de cadera y rodilla alrededor del mundo. Las tasas de reingreso en los primeros 30 días y las complicaciones se han descrito como una medida para evaluar estándares de calidad. En Colombia no se conoce el comportamiento de éstos estándares. Este estudio pretende aportar información de un centro especializado en reemplazos articulares en Bogotá DC, Colombia, Suramérica. Materiales & Métodos Se realizó un estudio observacional prospectivo de una cohorte de 1.126 pacientes con 626 reemplazos totales de cadera (RTC) y 500 de rodilla (RTR), operados entre los años 2011-2015 en el Hospital Universitario San Ignacio de Bogotá DC, Colombia. Se reportan las tasas de reingreso a 30 días y las complicaciones. Se analiza mediante un modelo de regresión las asociaciones entre variables sociodemográficas y clínicas con el reingreso y las complicaciones. Resultados El 71% de los pacientes fueron mujeres, el promedio de edad de 63 años en los hombres y 65 años en mujeres. La estancia hospitalaria promedio fue de 3,5 días (DE:0,81-6,29). La tasa de reingreso a 30 días por cualquier causa fue 10,8% y la tasa de complicaciones asociadas al procedimiento fue 5,8%, se requirió reintervención quirúrgica en 1,5% de los pacientes. Se encontró asociación estadísticamente significativa entre el índice de Charlson mayor a 2 (OR=2,33[IC95% 1,3-4,18], p=0.004) y el reingreso a 30 días. La principal complicación en RTC fue luxación (1,4%) y en RTR infección de sitio quirúrgico (4%). Se encontró una asociación significativa entre complicaciones a 30 días con los días de hospitalización (OR=1,17[IC95% 1,09-1,25], p=0,00); y con el cirujano 4 en RTR (OR=2,28[IC95% 1,29-4,01], p=0,04); las demás variables no mostraron diferencias significativas. No se aprecian grandes diferencias con lo reportado en la literatura, asociación con estancia hospitalaria y experiencia del cirujano. Discusión La experiencia del cirujano y la aplicación de guías clínicas que optimicen los procesos intrahospitalarios y regulen los días de estancia hospitalaria, se relacionan con un mejor desenlace en la evolución de los pacientes en cuanto al número de complicaciones y reingresos a 30 días.

Background Joint replacements are always defined as safe and successful procedures for the treatment of severe osteoarthritis of the hip and knee around the world. Rates of readmission within initial post surgical 30 days and complications are described as a measure to assess quality standards. In Colombia the behavior of the standards are mainly unknown. Aim of study is to provide information from a specialized in joint replacements center in Colombia. Methods A prospective observational study was carried out on a cohort of 1,126 patients with 626 total hip replacements (THR) and 500 knee replacements (TKR), who underwent surgery between 2011-2015 at a University Hospital in Bogotá, Colombia. 30-day readmission rates and complications were reported. The associations between sociodemographic and clinical variables with readmission and complications were analyzed using a regression model. Results 71% of the patients were women, the average age was 63 years for men and 65 years for women. The mean hospital stay was 3.5 days (SD: 0.81-6.29). The 30-day readmission rate for any cause was 10.8% and the rate of complications associated with the procedure was 5.8%. Additional surgical procedures were required in 1.5% of the patients. A statistically significant association was found between the Charlson index greater than 2 (OR=2.33 [95% CI 1.3-4.18], p=0.004) and readmission at 30 days. The main complication in THR was dislocation (1.4%) and surgical site infection (4%) in TKR. A significant association was found between complications at 30 days and days of hospitalization (OR=1.17 [95% CI 1.09-1.25], p=0.00); and with surgeon 4 in TKR (OR=2.28 [95% CI 1.29-4.01], p=0.04); the other variables did not show significant differences. No differences were found with what is reported in the literature, association with hospital stay and experience of the surgeon. Discussion The surgeon's experience and the application of clinical guidelines that optimize in-hospital processes and regulate the days of hospital stay are related to a better outcome in the evolution of patients in terms of the number of complications and readmissions after 30 days.

Generation of Organotypic Multicellular Spheres by Magnetic Levitation: Model for the Study of Human Hematopoietic Stem Cells Microenvironment.

BACKGROUND AND OBJECTIVE: The characteristics of human hematopoietic stem cells are conditioned by the microenvironment of the bone marrow, where they interact with other cell populations, such as mesenchymal stem cells and endothelial cells; however, the study of this microenvironment is complex. The objective of this work was to develop a 3D culture system by magnetic levitation that imitates the microenvironment of human HSC. METHODS AND RESULTS: Human bone marrow-mesenchymal stem cells, umbilical cord blood-hematopoietic stem cells and a non-tumoral endothelial cell line (CC2811, LonzaⓇ) were used to develop organotypic multicellular spheres by the magnetic levitation method. We obtained viable structures with an average sphericity index greater than 0.6, an average volume of 0.5 mm3 and a percentage of aggregation greater than 70%. Histological studies of the organotypic multicellular spheres used hematoxylin and eosin stains, and an evaluation of vimentin expression by means of immunohistochemistry demonstrated an organized internal structure without picnotic cells and a high expression of vimentin. The functional capacity of human hematopoietic stem cells after organotypic multicellular spheres culture was evaluated by multipotency tests, and it was demonstrated that 3D structures without exogenous Flt3L are autonomous in the maintenance of multipotency of human hematopoietic stem cells. CONCLUSIONS: We developed organotypic multicellular spheres from normal human cells that mimic the microenvironment of the human hematopoietic stem cells. These structures are the prototype for the development of complex organoids that allow the further study of the biology of normal human stem cells and their potential in regenerative medicine.

Impact of donor characteristics on the quality of bone marrow as a source of mesenchymal stromal cells.

In recent years, the therapeutic use of mesenchymal stromal cells (MSC) has generated a valuable number of scientific studies that delve into their biological characteristics and their potential in regenerative medicine; however, the impact of the clinical characteristics of tissue donors, from which these cells are isolated, on their potential in applied clinical research is not yet clear. The objective of this study was to evaluate the impact of the clinical characteristics of bone marrow donors on the quality of this tissue as a source of MSC for therapeutic use. Human MSC were isolated, characterized and cultured (according to ISCT criteria) from bone marrow samples from volunteer donors (n = 70) attending the Department of Orthopedics and Traumatology of the Hospital Universitario San Ignacio (Bogota, Colombia) for surgery of prosthetic hip replacement that agreed to participate voluntarily in the study. Donor data such as age, gender, weight, smoker and type of anesthesia used during the surgical procedure were recorded, and the impact of these characteristics on the volume of tissue collection, mononuclear cell count and confluence time of cells with fibroblastoid morphology was evaluated. Correlation coefficients between quantitative variables were calculated with Spearman's correlation test, and the association between qualitative and quantitative variables was evaluated with biserial correlation coefficient. A significant correlation was observed between the age of the donors and the time necessary to obtain confluent cells in vitro (r = 0.2489, P = 0.0377); similarly, the correlation between the volume of bone marrow collected and the number of mononuclear cells obtained was significant (r = 0.7101, P = 0.0001). Although a negative correlation tendency was observed between the mononuclear cell count and the confluence time, this was not significant (r = -0.2041, P = 0.0950). No significant associations were observed between gender, smoking status or type of anesthesia and the expansion characteristics of human mesenchymal stromal cells. Bone marrow donor age and the tissue collection volume impact the time of obtaining MSC in vitro and the mononuclear cell count with which the culture starts. These conditions must be considered when the bone marrow is selected as the tissue for obtaining MSC.