Núcleo de Células-Tronco e Terapia Celular (NCT-TCA), Departamento de Clínica e CirurgiaVeterinárias, Escola de Veterinária, 154001UFMG, Belo Horizonte, Brazil.
Instituto de Ciências Biológicas, Departamento de Patologia Geral, 113014UFMG, Belo Horizonte, Brazil.
Hum Exp Toxicol. 2021 Dec;40(12_suppl):S414-S422. doi: 10.1177/09603271211045949. Epub 2021 Sep 25.
Thirteen female Wistar rats were divided into two groups: one treated with ethanol and the other of untreated. Four newborns from each mother were selected and weighed, measured, and evaluated for physical characteristics. From these neonates, chondrocytes were extracted from the articular cartilages of the femur and tibia, and cultivated in a chondrogenic medium at 37C and 5% CO. At 7, 14, and 21 days of cultivation, alkaline phosphatase activity tests, MTT conversion to formazan, and percentage area covered by cells per field were performed. At 21 days, the percentage of PAS areas in 3D cultures was performed, as well as the evaluation of gene transcript expression for aggrecan, SOX-9, collagen type II, collagen X, Runx-2, and VEGF by real-time RT-PCR. The means were compared by Student's t-test. The weight of the ethanol group neonates was significantly lower than that of the controls. Chondrocyte cultures from the ethanol group showed significantly higher AP activity, MTT conversion, and cell percentage. There was higher expression of collagen type II and lower expression of SOX-9 in the ethanol group. There was no difference in the percentage of PAS areas in pellets and in expression of aggrecan, collagen X, Runx-2, or VEGF between groups. In conclusion, prenatal exposure to ethanol alters the phenotype and activity of offspring chondrocytes, which may be mechanisms by which endochondral bone formation is compromised by maternal ethanol consumption.
十三只雌性 Wistar 大鼠被分为两组:一组用乙醇处理,另一组未处理。从每个母亲中选择四只新生仔鼠进行称重、测量,并评估其身体特征。从这些新生仔鼠中,提取股骨和胫骨关节软骨的软骨细胞,并在 37°C 和 5% CO 的软骨形成培养基中培养。在培养的第 7、14 和 21 天,进行碱性磷酸酶活性测试、MTT 转化为甲臜以及每个视野下细胞覆盖面积的百分比。在第 21 天,对 3D 培养物中的 PAS 区域百分比进行评估,并通过实时 RT-PCR 评估聚集蛋白聚糖、SOX-9、II 型胶原、X 型胶原、Runx-2 和 VEGF 的基因转录表达。通过学生 t 检验比较平均值。乙醇组新生仔鼠的体重明显低于对照组。乙醇组的软骨细胞培养物显示出明显更高的 AP 活性、MTT 转化和细胞百分比。乙醇组 II 型胶原表达升高,SOX-9 表达降低。在球囊中 PAS 区域的百分比和聚集蛋白聚糖、X 型胶原、Runx-2 或 VEGF 的表达方面,两组之间没有差异。总之,产前暴露于乙醇改变了后代软骨细胞的表型和活性,这可能是母体乙醇摄入损害软骨内骨形成的机制之一。