Matrix-Bound Nanovesicles: The Effects of Isolation Method upon Yield, Purity, and Function.

Identification of matrix-bound nanovesicles (MBV) as ubiquitous components of the extracellular matrix (ECM) raises questions regarding their biologic functions and their potential theranostic application. Unlike liquid-phase extracellular vesicles (e.g., exosomes), MBV are tightly bound to the ECM, which makes their isolation and harvesting more challenging. The indiscriminate use of different methods to harvest MBV can alter or disrupt their structural and/or functional integrity. The objective of the present study was to compare the effect of various MBV harvesting methods upon yield, purity, and biologic activity. Combinations of four methods to solubilize the ECM (collagenase [COL], liberase [LIB], or proteinase K [PK] and nonenzymatic elution with potassium chloride) and four isolation methods (ultracentrifugation, ultrafiltration [UF], density barrier, and size exclusion chromatography [SEC]) were used to isolate MBV from urinary bladder-derived ECM. All combinations of solubilization and isolation methods allowed for the harvesting of MBV, however, distinct differences were noted. The highest yield, purity, cellular uptake, and biologic activity were seen with MBV isolated by a combination of liberase or collagenase followed by SEC. The combination of proteinase K and UF was shown to have detrimental effects on bioactivity. The results show the importance of selecting appropriate MBV harvesting methods for the characterization and evaluation of MBV and for analysis of their potential theranostic application. Impact statement Identification of matrix-bound nanovesicles (MBV) as ubiquitous components of the extracellular matrix (ECM) has raised questions regarding their biologic functions and their potential theranostic application. This study demonstrates that the harvesting methods used can result in samples with physical and biochemical properties that are unique to the isolation and solubilization methods used. Consequently, developing harvesting methods that minimize sample contamination with ECM remnants and/or solubilization agents will be essential in determining the theranostic potential of MBV in future studies.

[Predominant factors of neonatal encephalopathy: hypoxic and ischemia, a global problem]. / Factores predominantes de encefalopatía neonatal: hipoxia e isquemia, un problema global.

If a difficulty arises during birth, due to a maternal or fetal anomaly, acute or chronic, asphyxia of the fetal brain constitutes a greater risk, because it could result in the destruction of neurons and the possibility of evolving towards a Ischemic Hypoxic Encephalopathy with long -term sequelae. This review highlights the most recent scientific aspects but at the same time it offers an essential margin of knowledge regarding pathophysiology, diagnosis and treatment, as well as offering a perspective on the future of clinical care of ischemic hypoxic encephalopathy.

Si una dificultad sobreviene durante el nacimiento de un niño, por una anomalía materna o fetal, aguda o crónica, la asfixia del cerebro fetal constituye un riesgo mayor, porque ella podría dar como resultado la destrucción de las neuronas y la posibilidad de evolucionar hacia una encefalopatía hipóxico isquémica con secuelas a largo plazo. En esta revisión se resaltan los aspectos científicos más recientes pero a la vez se ofrece un margen de conocimiento imprescindible en cuant o a la patofisiología, diagnóstico y tratamiento, así como también se ofrece una perspectiva sobre el futuro de la atención clínica de la encefalopatía hipóxico isquémica.

Factores predominantes de encefalopatía neonatal: hipoxia e isquemia, un problema global / Predominant factors of neonatal encephalopathy: hypoxic and ischemia, a global problem

Si una dificultad sobreviene durante el nacimiento de un niño, por una anomalía materna o fetal, aguda o crónica, la asfixia del cerebro fetal constituye un riesgo mayor, porque ella podría dar como resultado la destrucción de las neuronas y la posibilidad de evolucionar hacia una encefalopatía hipóxico isquémica con secuelas a largo plazo. En esta revisión se resaltan los aspectos científicos más recientes pero a la vez se ofrece un margen de conocimiento imprescindible en cuanto a la patofisiología, diagnóstico y tratamiento, así como también se ofrece una perspectiva sobre el futuro de la atención clínica de la encefalopatía hipóxico isquémica.

If a difficulty arises during birth, due to a maternal or fetal anomaly, acute or chronic, asphyxia of the fetal brain constitutes a greater risk, because it could result in the destruction of neurons and the possibility of evolving towards a Ischemic Hypoxic Encephalopathy with long -term sequelae. This review highlights the most recent scientific aspects but at the same time it offers an essential margin of knowledge regarding pathophysiology, diagnosis and treatment, as well as offering a perspective on the future of clinical care of ischemic hypoxic encephalopathy.