Effects of antithrombotic drugs fondaparinux and tinzaparin on in vitro proliferation and osteogenic and chondrogenic differentiation of bone‐derived mesenchymal stem cells

Abstract

An unexpected side effect of some classes of anticoagulants has been osteoporosis which may be, at least in part, related to deranged mesenchymal stem cell (MSC) function. The aim of the present study was to compare the effect of fondaparinux (FDP), a novel antithrombotic with a traditional widely used low molecular weight heparin, tinzaparin (TZP) on MSC proliferation and differentiation. MSCs were isolated from trabecular bone of 14 trauma patients by a collagenase-based digestion procedure and expanded in standard conditions until passage 3. Proliferation and differentiation of MSCs to chondrocytes and osteoblasts was assessed with or without the addition of FDP and TZP using standard in vitro assays and a broad range of drug concentrations. Flow cytometry was used for MSC phenotyping. In the age studied group (17-74 years old) the MSC frequency in collagenase-released fractions was 641/10 6 cells (range 110-2,158) and their growth characteristics were $4 days/population doubling. Cultures had a standard MSC phenotype (CD73þ, CD105þ, CD146þ, CD106þ, and CD166þ). Cell proliferation was assessed by both colony-forming unit-fibroblast (CFU-F) and colorimetric tetrazolium salt XTT assays. In both assays, MSC proliferation was inhibited by the addition of TZP, particularly at high concentrations. In contrast, FDP had no effect on MSC proliferation. Osteogenic differentiation and chondrogenic differentiation were not affected by the addition of either TZP or FDP. Whilst MSC proliferation, but not differentiation, is negatively affected by TZP, there was no evidence for adverse effects of FDP in this in vitro model system which argues well for its use in the orthopedic setting. ß

Human Embryonic Stem Cell-Derived Mesenchymal Stroma Cells (hES-MSCs) Engraft In Vivo and Support Hematopoiesis without Suppressing Immune Function: Implications for Off-The Shelf ES-MSC Therapies

“…However, considering the various CFU-F numbers reported previously for BM-MSCs [23], [40], hES-MP002.5 cells still showed reasonable CFU-F generation capability even in higher passages.…”

The mechanisms of nadroparin‐mediated inhibition of proliferation of two human lung cancer cell lines

“…These studies demonstrated that both osteoblast and MSC proliferation were inhibited by addition of LMWHs. In contrast, the pentasaccharide had no effect on osteoblast and MSC proliferation (44,45).…”

Heparin affects human bone marrow stromal cell fate: Promoting osteogenic and reducing adipogenic differentiation and conversion

“…Lanfer et al demonstrated multilineage differentiation of MSC (chondrogenic, adipogenic, and osteogenic) in aligned collagen matrices incorporating heparin [21]. The low molecular weight heparin tinzaparin (TZP) was reported to negatively affect hMSC proliferation but not osteogenic nor chondrogenic differentiation [22]. In addition, bone marrow-derived stromal cells grown on a heparin-treated surfaceshowed increased alkaline phosphatase (ALP) activity and mineralization [23].…”