Acetylsalicylic Acid Inhibits Proliferation of Human Bone Marrow Stromal Cells and Matrix Mineralization

Abstract

Acetylsalicylic acid (ASA) and other non-steroidal anti-inflammatory drugs have been shown to potentially inhibit bone healing and bone formation in both animal and clinical studies. Due to the extensive diffusion of ASA-based long-term therapies, the implications of such a side-effect are of interest in all types of bone surgery, including bone grafting procedures and dental implant placement. In this study, we investigate the effect of ASA at therapeutic concentrations on the proliferation and osteogenic differentiation of human bone marrow stromal cells (BMSCs). Primary cultures of BMSCs were isolated and expanded. Their proliferation in response to ASA 50, 100 and 200 Jlg/ml was evaluated by MTT assay and 3H-thymidine incorporation. Cell cycle machinery was also investigated by FACS and analysis of inhibitors of cyclin-dependent kinases (CDKIs). ASA inhibited BMSC proliferation and DNA synthesis in a dose-dependent manner down to 60% of control (ASA 200 ug/ml) at 72 h. Cell cycle analysis showed a decrease ofBMSCs in the Sand G2/M phases with a concomitant accumulation in GO/ 1 in ASA treated cells. The finding was associated to increased levels of some CDKIs, namely p27 Kipl and p21 Cipl, whereas ASA did not affected p16Ink4A level at any ofthe concentrations employed. The matrix mineralization, that represents the major feature of the osteogenic commitment, was assessed by a specific staining procedure (von Kossa) and by calcium content determlnation.Both the methods demonstrated an extensive reduction (>90%) of extracellular calcification at 200 Jlg/ml ASA. On the basis of our results, we can hypothesize that the widely reported inhibition of bone healing by ASA might be sustained both by a direct anti-proliferative effect on BMSCs and by an alteration ofthe extracellular calcification.Aspirin (acetylsalicylic acid, ASA) represents one of the most widely prescribed drugs worldwide, and is the forefather of the entire class of non-steroidal anti-inflammatory drugs (NSAIDs). Originally employed exclusively for relieving pain, fever and inflammation, ASA further expanded its indications to the treatment of the chronic musculoskeletal conditions and the primary/secondary prophylaxis of stroke and myocardial infarction (1-2).A number of studies have shown that NSAIDs

Effect of Aspirin on Bone Healing in a Rabbit Ulnar Osteotomy Model

“…None of these studies examined the effect of aspirin on bone healing within a dosing range equivalent to that commonly used in humans (<1 g per dose). The dose-dependent effect of aspirin on bone healing was also examined in an in vitro study 23 . That study showed aspirin to have a dose-dependent inhibitory effect on human bone-marrow stromal cell proliferation and matrix mineralization.…”

“…Previous animal studies have shown aspirin to inhibit bone healing at high doses [19][20][21][22][23] . To our knowledge, our study is the first to demonstrate inhibition of bone healing by aspirin in an animal model within a dosing range equivalent to that typically employed in humans.…”

“…The effect of aspirin on bone healing was examined in four animal studies and a human in vitro study [19][20][21][22][23] . All of these studies demonstrated an inhibitory effect of aspirin on bone healing.…”

An Oligodeoxynucleotide That Induces Differentiation of Bone Marrow Mesenchymal Stem Cells to Osteoblasts in Vitro and Reduces Alveolar Bone Loss in Rats with Periodontitis

“…Proliferation of BMSCs was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay as previously described [41]. Briefly, 5 × 10 3 cells/well in 200 μL DMEM containing 10% fetal calf serum were inoculated into 96-well plates and cultured with medium containing various ODNs at a final concentration of 1 μg/mL.…”

Mast Cells and Arachidonic Acid Cascade in Inflammation

“…These cells are recruited and activated either by mast cellassociated chemotactic non-specific factors such as LTB4, PAF, the eosinophil chemotactic factor of anaphylaxis (ECF-A), and chemotactic specific factors: chemokines (44)(45)(46). These compounds also participate in the late phase reactions six hours subsequent to the exposure to the allergen (47).…”