Publicación: HYPOXIC PLACENTAL SMALL EXTRACELLULAR VESICLES DISRUPT THE BLOOD-BRAIN BARRIER IN THE FRONTAL AREA OF THE BRAIN
Fecha
2022
Título de la revista
ISSN de la revista
Título del volumen
Editor
PLACENTA
Resumen
OBJECTIVES: INVESTIGATE WHETHER SMALL EXTRACELLULAR VESICLES (PDSEVS) EXTRACTED FROM PLACENTAS CULTURED IN HYPOXIA DISRUPT THE BLOOD-BRAIN BARRIER (BBB) IN SELECTED AREAS OF THE BRAIN.
METHODS: PDSEVSWERE EXTRACTED FROM PLACENTAL EXPLANTS OF NORMAL PREGNANCY CULTURED IN NORMOXIA (21% O2) OR HYPOXIA (1% O2, 18 H). PDSEVS (100 MG OF PROTEIN) FROM NORMOXIA (N=6) OR HYPOXIA (N=6) WERE INJECTED IN NON-PREGNANT FEMALE C57BL/7 MICE. INJECTED MICE WERE EVALUATED USING THE RAPID MURINE COMA AND BEHAVIOR SCALE (RMSBS, 0-24 H), WHILE SYSTEMIC BLOOD PRESSURE WAS MEASURED BEFORE AND AFTER TREATMENT. MICE WERE INJECTED WITH EVAN S BLUE AND THEN EUTHANIZED TO EXTRACT THE BRAIN. MICE BRAINS WERE WEIGHED AND SECTIONED (100 MM SLICES) TO ANALYZE EVAN S BLUE EXTRAVASATION. ALSO, HUMAN CEREBRAL MICROVASCULAR ENDOTHELIAL CELLS (HCMEC/D3) WERE EXPOSED TO PDSEVS. BBB PARAMETERS INCLUDED ELECTRICAL RESISTANCE (TEER) AND CELL PERMEABILITY TO A HIGH-MOLECULAR-WEIGHT FLUORESCENT DYE. ALSO, THE AMOUNT OF CLAUDIN-5 IN THE CELL MEMBRANE WAS ANALYZED BY WESTERN BLOT. UPTAKE OF PDSEVS WAS ANALYZED USING PKH67.
RESULTS: MICE TREATED WITH HYPOXIC PDSEVS EXHIBITED NEUROLOGICAL DEFICITS (FROM 3 TO 24 H), AS INDICATED WITH A SIGNIFICANT REDUCTION IN THE SCORE FOR SPINAL REFLEXES AND MOTOR SKILLS, WHICH WAS ASSOCIATED WITH MORE SIGNIFICANT EXTRAVASATION OF EVAN S BLUE, MAINLY IN THE ANTERIOR PORTION OF THE BRAIN. THERE WERE NO SIGNIFICANT CHANGES IN SYSTOLIC/DIASTOLIC BLOOD PRESSURE IN ANY OF THE TREATED GROUPS. HYPOXIC PDSEVS GENERATED A MORE SIGNIFICANT REDUCTION IN THE TEER VALUES (P
