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Changes in Morphology of Primary Muscle Cells Exposed to Di‐(2‐ethylhexyl) Phthalate
Author(s) -
Zenteno-Savín Tania,
Hernández-Almaraz Pablo,
Lugo-Lugo Orlando,
Gaxiola-Robles Ramón,
Vázquez-Medina José Pablo,
Ramírez-Jirano Luis Javier
Publication year - 2020
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2020.34.s1.04270
Subject(s) - phthalate , cytoplasm , andrology , cell , viability assay , chemistry , mitosis , biology , microbiology and biotechnology , medicine , biochemistry , organic chemistry
Di‐(2‐ethylhexyl) phthalate (DEHP) is used in plastic manufacture mainly to provide flexibility. Asthma, infertility, endocrine disruption and some types of cancer have been associated with environmental DEHP exposure in humans; however, the morphological, biochemical and physiological changes that lead to these pathologies have not been reported. The objective of this study was to analyze morphological changes in muscle cells exposed to DEHP. Abdominal muscle samples were collected from healthy women subjected to programmed cesarean section at Hospital de Especialidades Médicas de La Paz, in La Paz, Baja California Sur, México, and were subjected to cell culture at 35°C with 5% CO 2 and constant humidity. Primary smooth muscle cells (4.0 x 10 5 cells mL −1 ) from both species were exposed to 1000 μM DEHP for 13 days. Untreated controls reached 100% confluency, underwent constant mitotic divisions and had normal cell morphology throughout. In contrast, cells exposed to DEHP did not reach confluency beyond 85%, had significantly lower cell viability and total protein content at day 13 of exposure. Cells exposed to DEHP were distended and had granules in the cytoplasm, which were evident since day 2 of exposure and covered all the cytoplasm by day 13. In numerous cells, bright yellow refringent, round bodies were observed in the cytoplasm, with variable size and number among cells. These spheres were observed from day 2 of exposure and are presumed to be DEHP accumulated inside the cells. These intracellular DEHP deposits could be associated with the observed cell mortality and morphology changes and could potentially lead to the reported pathologies onset and progression. This research was conducted in accordance with institutional ethical standards and with the 1964 Declaration of Helsinki and later amendments; informed consent was obtained from all participants included in the study. The informed consent form and research protocol were approved by the Ethics Committee of Instituto Mexicano del Seguro Social. Support or Funding Information Consejo Nacional de Ciencia y Tecnología (CONACYT‐Fronteras 2016‐01‐2305)