Inhibition of the osteogenic differentiation of mesenchymal stem cells derived from the offspring of rats treated with caffeine during pregnancy and lactation

Abstract

Caffeine is an alkaloid that is widely consumed due to its presence in drugs, coffee, tea, and chocolate. This compound passes to offspring through the placenta and milk; can cause teratogenic mutations; and reduces the formation, growth, and mass of bone. Because mesenchymal stem cells (MSCs) are responsible for generating the entire skeleton, we hypothesized that these cells are targets of caffeine. This study evaluated the osteogenic differentiation of MSCs derived from the offspring of rats treated with caffeine during pregnancy and lactation. Twenty-four adult Wistar rats were randomly divided into four groups, including one control group and three experimental groups treated with 25, 50, or 100 mg/kg of caffeine. At weaning, three 21-day-old pups from each dam in each group were euthanized for extraction of bone marrow cells for in vitro tests. Caffeine doses of 50 and 100 mg/kg significantly reduced the activity of alkaline phosphatase at 7, 14, and 21 days and the expression of collagen I at 21 days. However, the expression of gene transcripts for alkaline phosphatase, Runx-2, and bone sialoprotein, as well as the synthesis of mineralization nodules, decreased significantly in all groups treated with caffeine. The expression of osteocalcin was significantly reduced only in the group treated with 50 mg/kg caffeine. The caffeine that passes from the mother to the offspring during pregnancy and lactation reduces the osteogenic differentiation of MSCs. We propose that this reduction in the osteogenic potential of MSCs may be involved in the pathogenesis of osteopenia resulting from caffeine consumption.

Direct inhibitory effect of caffeine on viability, synthesis activity and gene expression in cultures of chondrocytes extracted from the articular cartilage of rats

“…In MSCs, high concentrations of caffeine suppresses cell differentiation, decreased mineralization and alkaline phosphatase activityregulating genes, such as osteocalcin, osteoprotegerin and Runx-2 (Su et al, 2013). The inhibitory effect of caffeine on the osteogenic differentiation of MSCs was also observed in a study that investigated the MSCs of the offspring of rats given caffeine during pregnancy and lactation (Reis et al, 2016).…”

“…We analyzed the percentage of chondrocytes in the sections stained with Hematoxylin and eosin (HE) and the percentage of the chondrogenic matrix stained with safranin O, alcian blue and PAS (Sigma, St. Louis, MO, USA). The analysis was performed using a 121-point graticule coupled to an optical microscope with a 10X objective in 50 microscopic fields (Reis et al, 2016). Six repetitions of the analysis were performed per group.…”

“…Caffeine is a substance that can alter bone growth (Reis et al, 2014). In addition, as caffeine is widely consumed by the human population, it can be conveyed by the placenta and via transmammary passage, interfering with the formation and endochondral bone growth of offspring (Reissig et al, 2009). There are several reports of the effects of caffeine on bone tissue and its cells (Tassinari et al, 1991;Tsuang et al, 2006;Reis et al, 2014), but there are no studies reporting its effect on chondrocytes, despite the knowledge that a mother's caffeine consumption during pregnancy and lactation affects the growth of the offspring's cartilage (Reis et al, 2014).…”

“…The effect of caffeine in osteoblasts and MSCs may explain some of the mechanisms by which it causes osteopenia but does not provide information on the genesis of the growth changes caused by this alkaloid (Tassinari et al, 1991;Rapuri et al, 2007;Reis et al, 2016), as the effects of caffeine on chondrocyte viability and synthesis activity have not been studied.…”

The Solanum glaucophyllum Desf. extract reduces mineralized matrix synthesis in osteogenically differentiated rat mesenchymal stem cells in vitro

“…In order to study the effects of S. glaucophyllum Desf. on MSCs submitted to the osteogenic differentiation process, the plant extract was added to the culture medium together with factors known to promote osteogenic differentiation (Ocarino et al, 2008; Pittenger et al, 1999; Reis et al, 2016). S. glaucophyllum Desf.…”