Guiding the osteogenic fate of mouse and human mesenchymal stem cells through feedback system control.

Stem cell-based disease modeling presents unique opportunities for mechanistic elucidation and therapeutic targeting. The stable induction of fate-specific differentiation is an essential prerequisite for stem cell-based strategy. Bone morphogenetic protein 2 (BMP-2) initiates receptor-regulated Smad phosphorylation, leading to the osteogenic differentiation of mesenchymal stromal/stem cells (MSC) in vitro; however, it requires supra-physiological concentrations, presenting a bottleneck problem for large-scale drug screening. Here, we report the use of a double-objective feedback system control (FSC) with a differential evolution (DE) algorithm to identify osteogenic cocktails of extrinsic factors. Cocktails containing significantly reduced doses of BMP-2 in combination with physiologically relevant doses of dexamethasone, ascorbic acid, beta-glycerophosphate, heparin, retinoic acid and vitamin D achieved accelerated in vitro mineralization of mouse and human MSC. These results provide insight into constructive approaches of FSC to determine the applicable functional and physiological environment for MSC in disease modeling, drug screening and tissue engineering

A novel method to optimize autologous adipose tissue recovery with extracellular matrix preservation

This work aims to characterize a new method to recover low-manipulated human adipose tissue, enriched with adipose tissue-derived mesenchymal stem cells (ATD-MSCs) for autologous use in regenerative medicine applications. Lipoaspirated fat collected from patients was processed through Lipocell, a Class II-a medical device for dialysis of adipose tissue, by varying filter sizes and washing solutions. ATD-MSC yield was measured with flow cytometry after stromal vascular fraction (SVF) isolation in fresh and cultured samples. Purification from oil and blood was measured after centrifugation with spectrophotometer analysis. Extracellular matrix preservation was assessed through hematoxylin and eosin (H&E) staining and biochemical assay for total collagen, type-2 collagen, and glycosaminoglycans (GAGs) quantification. Flow cytometry showed a two-fold increase of ATD-MSC yield in treated samples in comparison with untreated lipoaspirate; no differences where reported when varying filter size. The association of dialysis and washing thoroughly removed blood and oil from samples. Tissue architecture and extracellular matrix integrity were unaltered after Lipocell processing. Dialysis procedure associated with Ringer’s lactate preserves the proliferation ability of ATD-MSCs in cell culture. The characterization of the product showed that Lipocell is an efficient method for purifying the tissue from undesired byproducts and preserving ATD-MSC vitality and extracellular matrix (ECM) integrity, resulting in a promising tool for regenerative medicine applications

A novel method to optimize autologous adipose tissue recovery with extracellular matrix preservation

This work aims to characterize a new method to recover low-manipulated human adipose tissue, enriched with adipose tissue-derived mesenchymal stem cells (ATD-MSCs) for autologous use in regenerative medicine applications. Lipoaspirated fat collected from patients was processed through Lipocell, a Class II-a medical device for dialysis of adipose tissue, by varying filter sizes and washing solutions. ATD-MSC yield was measured with flow cytometry after stromal vascular fraction (SVF) isolation in fresh and cultured samples. Purification from oil and blood was measured after centrifugationwith spectrophotometer analysis. Extracellularmatrix preservationwas assessed through hematoxylin and eosin (H&E) staining and biochemical assay for total collagen, type-2 collagen, and glycosaminoglycans (GAGs) quantification. Flow cytometry showed a two-fold increase of ATD-MSC yield in treated samples in comparisonwith untreated lipoaspirate; no differenceswhere reportedwhen varying filter size. The association of dialysis and washing thoroughly removed blood and oil from samples. Tissue architecture and extracellular matrix integrity were unaltered after Lipocell processing. Dialysis procedure associated with Ringer's lactate preserves the proliferation ability of ATD-MSCs in cell culture. The characterization of the product showed that Lipocell is an efficient method for purifying the tissue from undesired byproducts and preserving ATD-MSC vitality and extracellular matrix (ECM) integrity, resulting in a promising tool for regenerative medicine applications

Isolation and characterization of buccal fat pad and dental pulp mscs from the same donor

Mesenchymal stem cells (MSCs) can be harvested from different sites in the oral cavity, representing a reservoir of cells useful for regenerative purposes. As direct comparisons between at least two types of MSCs deriving from the same patient are surprisingly rare in scientific literature, we isolated and investigated the osteoinductive potential of dental pulp stem cells (DPSCs) and buccal fat pad stem cells (BFPSCs). MSCs were isolated from the third molar dental pulp and buccal fat pads of 12 patients. The number of viable cells was quantified through manual count. Proliferation and osteodifferentiation assays, flow cytometry analysis of cell phenotypes, and osteocalcin release in vitro were performed. The isolation of BFPSCs and DPSCs was successful in 7 out of 12 (58%) and 3 out of 12 (25%) of retrieved samples, respectively. The yield of cells expressing typical stem cell markers and the level of proliferation were higher in BFPSCs than in DPSCs. Both BFP-SCs and DPSCs differentiated into osteoblast-like cells and were able to release a mineralized matrix. The release of osteocalcin, albeit greater for BFPSCs, did not show any significant difference between BFPSCs and DPSCs. The yield of MSCs depends on their site of origin as well as on the protocol adopted for their isolation. Our data show that BFP is a valuable source for the derivation of MSCs that can be used for regenerative treatments

Influence of chitosan on the mechanical and biological properties of HDPE for biomedical applications

High density polyethylene (HDPE) is widely used in biomedical field, except when strong cell-material interactions and high mechanical properties are required. To address this pitfall, two kinds of chitosan in different amounts were used as filler in the present research. Composites were prepared by melt extrusion process and their microstructural, thermal and mechanical properties were widely investigated. Also roughness and wettability were studied, as features of paramount importance in dictating cell response. Both types of chitosan endowed HDPE with higher Young modulus and lower elongation at break. Interestingly, fibroblast adhesion and viability were enhanced when a low amount of filler was used. The interaction of HDPE/chitosan composites with biological environment was investigated for the first time in order to assess the feasibility of these composites as materials for biomedical application

Endothelial heme dynamics drive cancer cell metabolism by shaping the tumor microenvironment

The crosstalk among cancer cells (CCs) and stromal cells within the tumor microenvironment (TME) has a prominent role in cancer progression. The significance of endothelial cells (ECs) in this scenario relies on multiple vascular functions. By forming new blood vessels, ECs support tumor growth. In addition to their angiogenic properties, tumor-associated ECs (TECs) establish a unique vascular niche that actively modulates cancer development by shuttling a selected pattern of factors and metabolites to the CC. The profile of secreted metabolites is strictly dependent on the metabolic status of the cell, which is markedly perturbed in TECs. Recent evidence highlights the involvement of heme metabolism in the regulation of energy metabolism in TECs. The present study shows that interfering with endothelial heme metabolism by targeting the cell membrane heme exporter Feline Leukemia Virus subgroup C Receptor 1a (FLVCR1a) in TECs, resulted in enhanced fatty acid oxidation (FAO). Moreover, FAO-derived acetyl-CoA was partly consumed through ketogenesis, resulting in ketone bodies (KBs) accumulation in FLVCR1a-deficient TECs. Finally, the results from this study also demonstrate that TECs-derived KBs can be secreted in the extracellular environment, inducing a metabolic rewiring in the CC. Taken together, these data may contribute to finding new metabolic vulnerabilities for cancer therapy

Tercer reporte de eventos adversos con tratamientos biológicos en Argentina

Introducción: BIOBADASAR ((Registro Argentino de Eventos Adversos con Tratamientos Biológicos en Reumatología) comenzó en agosto de 2010. La importancia de este registro es mostrar datos locales que, probablemente, puedan diferir de otros registros. El objetivo es comunicar los resultados del tercer reporte de BIOBADASAR. Métodos: Todos los pacientes con enfermedades reumáticas que requirieron tratamiento con agentes biológicos y pacientes controles sin estos tratamientos fueron incluidos en la base de datos provenientes de 32 centros participando a lo largo de la Argentina. Tres áreas de datos son analizados: características de los pacientes, tratamientos y eventos adversos. El inicio de la carga fue agosto de 2010 y cerró para este análisis en agosto de 2013. Se utilizó software Infostat para el análisis estadístico. Se calcularon porcentajes, riesgos relativos e incidencia persona/año para los eventos adversos. Resultados: Se incorporaron 2356 pacientes (1277 casos y 1079 controles) utilizando 2940 tratamientos. 1862 mujeres (79%) y 494 hombres (21%). La edad promedio fue 54 años (1-90). 1107 pacientes (58%) fueron tratados con agentes biológicos (casos) y 802 (42%) fueron controles. 1829 pacientes tenían artritis reumatoidea y 218 artritis psoriásica entre los diagnósticos principales. El tiempo promedio de la enfermedad fue 9,81 años para los controles y 12,59 para los casos. El biológico más utilizado fue el etanercept (46,7% de los tratamientos) con una mediana de sobrevida al tratamiento de 31 meses seguido de adalimumab con 23,6% de los tratamientos y una mediana de sobrevida de 26,4 meses. La causa más frecuente de interrupción de tratamientos para los casos fue ineficacia (30%) seguido por eventos adversos (26%). La incidencia de eventos adversos serios fue 37/1000 pacientes año en el grupo biológico vs. 5/1000 pacientes año en el grupo control (RR 7,44; IC 5,17-10,7; p<0,05). El evento adverso más frecuente fue la infección con un RR 1,66 (IC 1,38-2,0; p<0,05). Dentro de las infecciones, la neumonía tuvo un RR de 29,31 (IC 9,21-93,25; p<0,05), herpes zoster RR 4,12 (IC 2,08-8,15; p<0,05) e infecciones cutáneas con un RR 7,21 (IC 3,89-13,36; p<0,05). La tuberculosis tuvo un RR de 4,53 (IC 0,472-43,56; sin significancia). Las enfermedades neoplásicas tuvieron un RR 2,82 (IC 1,61-4,95; p<0,05). Conclusiones: Éste es el tercer reporte de BIOBADASAR mostrando la realidad de los tratamientos biológicos en Argentina. Los pacientes de Latinoamérica podrían mostrar diferencias con respecto a otros países utilizando los mismos tratamientos debido a enfermedades regionales, esquemas diferentes de vacunación o tolerancia a patógenos. Podría haber sesgos en el registro que serán minimizados con el control estricto y el tiempo del registro