Hydroxychloroquine decreases human MSC‐derived osteoblast differentiation and mineralization in vitro

Abstract

We recently showed that patients with primary Sjögren Syndrome (pSS) have significantly higher bone mineral density (BMD) compared to healthy controls. The majority of those patients (69%) was using hydroxychloroquine (HCQ), which may have favourable effects on BMD. To study the direct effects of HCQ on human MSC‐derived osteoblast activity. Osteoblasts were cultured from human mesenchymal stromal cells (hMSCs). Cultures were treated with different HCQ doses (control, 1 and 5 µg/ml). Alkaline phosphatase activity and calcium measurements were performed to evaluate osteoblast differentiation and activity, respectively. Detailed microarray analysis was performed in 5 µg/ml HCQ‐treated cells and controls followed by qPCR validation. Additional cultures were performed using the cholesterol synthesis inhibitor simvastatin (SIM) to evaluate a potential mechanism of action. We showed that HCQ inhibits both MSC‐derived osteoblast differentiation and mineralization in vitro. Microarray analysis and additional PCR validation revealed a highly significant up‐regulation of the cholesterol biosynthesis, lysosomal and extracellular matrix pathways in the 5 µg/ml HCQ‐treated cells compared to controls. Besides, we demonstrated that 1 µM SIM also decreases MSC‐derived osteoblast differentiation and mineralization compared to controls. It appears that the positive effect of HCQ on BMD cannot be explained by a stimulating effect on the MSC‐derived osteoblast. The discrepancy between high BMD and decreased MSC‐derived osteoblast function due to HCQ treatment might be caused by systemic factors that stimulate bone formation and/or local factors that reduce bone resorption, which is lacking in cell cultures.

Zika virus infection perturbs osteoblast function

“…Human bone marrow-derived MSCs of two healthy donors (Donor 4266 and 3520, 33 and 20 year old males, respectively) were purchased from Lonza (PT-2501). MSCs were differentiated into osteoblasts as described previously 24 . Briefly, MSCs were cultured in αMEM medium (Gibco, Thermofisher) supplemented with 10% heat-inactivated fetal calf serum a (FCS, Sigma), 20 mM Hepes (Sigma), 100 U/mL penicillin (Lonza) and 100 µg/mL streptomycin (Thermofisher) and 1.8 mM CaCl 2 (Sigma) at 37 °C and 5% CO 2 in a humidified atmosphere.…”

“…RNA was extracted from ZIKV-infected and uninfected cells at different time points during osteoblast differentiation using TRIzol reagent (Thermo Fisher Scientific). RNA isolation, cDNA synthesis, and PCR reactions were performed as described previously 24 . Oligonucleotide primer pairs were designed to be either on exon boundaries or spanning at least one intron (Table 1 ).…”

The Association of Methotrexate, Sulfasalazine, and Hydroxychloroquine Use With Fracture in Postmenopausal Women With Rheumatoid Arthritis: Findings From the Women's Health Initiative

“…in vitro studies suggest that hydroxychloroquine both decreases mesenchymal-derived osteoblast differentiation and inhibits the formation of osteoclasts. (15,16) These potential conflicting actions on the skeleton from methotrexate, sulfasalazine, and hydroxychloroquine use make it important to study the association of these medications with osteoporosis.…”

Effect of 1,25-Dihydroxyvitamin D3 on Stem Cells from Human Apical Papilla: Adhesion, Spreading, Proliferation, and Osteogenic Differentiation

“…Mainly secreted by bone cells or chondrocytes, OCN is also a marker factor for osteogenic differentiation of stem cells, contributing to bone metabolism regulation [19]. Col I is one of the basic substances for bone formation and outer matrix mineralization [20][21][22]. The results of this study proved that 1,25-dihydroxyvitamin D 3 can positively promote the osteogenic differentiation of SCAPs, and the expression of related osteogenic differentiation markers increased by upregulating the transcription levels of Runx2, ALP, OCN, and Col I.…”