Alendronate Enhances Osteogenic Differentiation of Bone Marrow Stromal Cells: A Preliminary Study

Chonnam National University Hwasun Hospital Center for Joint Disease 160 Ilsimri, Hwasuneup, Hwasungun 519-809 Jeonnam Korea
Clinical Orthopaedics and Related Research (Impact Factor: 2.77). 12/2009; 467(12):3121-3128. DOI: 10.1007/s11999-008-0409-y


Alendronate inhibits osteoclastic activity. However, some studies suggest alendronate also has effects on osteoblast activity.
We hypothesized alendronate would enhance osteoblastic differentiation without causing cytotoxicity of the osteoblasts. We
evaluated the effect of alendronate on the osteogenic differentiation of mouse mesenchymal stem cells. D1 cells (multipotent
mouse mesenchymal stem cells) were cultured in osteogenic differentiation medium for 7days and then treated with alendronate
for 2days before being subjected to various tests using MTT assays, Alizarin Red, enzyme-linked immunosorbent assay, energy-dispersive
xray spectrophotometry, reverse transcriptase–polymerase chain reaction, confocal microscopy, and flow cytometric analysis.
D1 cells differentiated into osteoblasts in the presence of osteogenic differentiation medium as confirmed by positive Alizarin
Red S staining, increased alkaline phosphatase activity and osteocalcin mRNA expression, a calcium peak by energy-dispersive
xray spectrophotometry, and by positive immunofluorescence staining against CD44. Osteogenic differentiation was enhanced
after treatment with alendronate as confirmed by Alizarin Red S staining, elevated alkaline phosphatase activity and osteocalcin
mRNA expression, a greater calcium peak by energy-dispersive xray spectrophotometry, and by immunofluorescence staining against
CD44 by flow cytometric analysis. These data suggest alendronate enhances osteogenic differentiation when treated with mouse
mesenchymal stem cells in osteogenic differentiation medium.

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Available from: Hyung Keun Kim, Dec 16, 2013
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    • "In MG-63 osteoblastic cells, 1 nM to 100 µM Aln promoted cell proliferation and maturation [7], [8]. In human BMSCs, 10 nM Aln enhanced osteo-differentiation [9], [10]; however, the enhancement of osteo-differentiation required more than 14 days of treatment [9], [10]. In our previous study, 5 µM Aln sufficiently enhanced osteo-differentiation in human ADSCs [11]. "
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    ABSTRACT: Recent studies indicated that alendronate enhanced osteogenesis in osteoblasts and human bone marrow-derived stem cells. However, the time- and dose-dependent effects of Aln on ostegenic differentiation and cytotoxicity of hBMSCs remain undefined. In present study, we investigated the effective dose range and timing of hBMSCs. hBMSCs were treated with various Aln doses (1, 5 and 10 µM) according to the following groups: group A was treated with Aln during the first five days of bone medium, groups B, C and D were treated during the first, second, and final five days of osteo-induction medium and group E was treated throughout the entire experiment. The mineralization level and cytotoxicity were measured by quantified Alizarin Red S staining and MTT assay. In addition, the reversal effects of farnesyl pyrophosphate and geranylgeranyl pyrophosphate replenishment in group B were also investigated. The results showed that Aln treatment in groups A, B and E enhanced hBMSC mineralization in a dose-dependent manner, and the most pronounced effects were observed in groups B and E. The higher dose of Aln simultaneously enhanced mineralization and caused cytotoxicity in groups B, C and E. Replenishment of FPP or GGPP resulted in partial or complete reverse of the Aln-induced mineralization respectively. Furthermore, the addition of FPP or GGPP also eliminated the Aln-induced cytotoxicity. We demonstrated that hBMSCs are susceptible to 5 µM Aln during the initiation stage of osteogenic differentiation and that a 10 µM dose is cytotoxic.
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    • "Because of their strong affinity for bone and their inhibition of bone resorption, BPs have been widely used in clinical treatment of bone diseases with increased bone resorption, including Paget's disease, hypercalcemia of malignancy, fibrous dysplasia, and inflammation-related bone loss [3]–[7]. Recent studies indicated that BPs not only inhibit bone resorption induced by osteoclasts but also accelerate bone formation induced by osteoblasts, albeit with varying or conflicting effects, in relation to the concentration of BPs [1], [8]–[11]. In most studies, BPs were administered intravenously or orally. "
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    ABSTRACT: Bisphosphonates (BPs) have been widely used in clinical treatment of bone diseases with increased bone resorption because of their strong affinity for bone and their inhibition of bone resorption. Recently, there has been growing interest in their improvement of bone formation. However, the effect of local controlled delivery of BPs is unclear. We used polylactide acid-glycolic acid copolymer (PLGA) as a drug carrier to deliver various doses of the bisphosphonate zoledronate (Zol) into the distal femur of 8-week-old Sprague-Dawley rats. After 6 weeks, samples were harvested and analyzed by micro-CT and histology. The average bone mineral density and mineralized bone volume fraction were higher with medium- and high-dose PLGA-Zol (30 and 300 µg Zol, respectively) than control and low-dose Zol (3 µg PLGA-Zol; p<0.05). Local controlled delivery of Zol decreased the numbers of osteoclast and increased the numbers of osteoblast. Moreover, local controlled delivery of medium- and high-dose Zol accelerated the expression of bone-formation markers. PLGA used as a drug carrier for controlled delivery of Zol may promote local bone formation.
    Full-text · Article · Mar 2014 · PLoS ONE
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    • "BPs can increase OPG expression and decrease M-CSF expression; in consequence they might inhibit osteoclastogenesis [25]. Substantial evidence has accumulated that BPs modulate the proliferation and differentiation rates of osteoplastic cells, albeit with varying or conflicting effects, in relation to the concentration of BPs [5,27-31]. BPs can promote the growth and differentiation of osteoblasts at lower concentrations, ranging from 10-9 to 10-6 M but had inhibitory effects at >10-5 M [5]. "
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    ABSTRACT: It is now 40 years since bisphosphonates (BPs) were first used in the clinic. So, it is timely to provide a brief review of what we have learned about these agents in bone disease. BPs are bone-specific and have been classified into two major groups on the basis of their distinct molecular modes of action: amino-BPs and non-amino-BPs. The amino-BPs are more potent and they inhibit farnesyl pyrophosphate synthase (FPPS), a key enzyme of the mavalonate/cholesterol biosynthetic pathway, while the non-amino-BPs inhibit osteoclast activity, by incorporation into non-hydrolyzable analogs of ATP. Both amino-BPs and non-amino-BPs can protect osteoblasts and osteocytes against apoptosis. The BPs are widely used in the clinic to treat various diseases characterized by excessive bone resorption, including osteoporosis, myeloma, bone metastasis, Legg-Perthes disease, malignant hyperparathyroidism, and other conditions featuring bone fragility. This review provides insights into some of the adverse effects of BPs, such as gastric irritation, osteonecrosis of the jaw, atypical femoral fractures, esophageal cancer, atrial fibrillation, and ocular inflammation. In conclusion, this review covers the biochemical and molecular mechanisms of action of BPs in bone, particularly the discovery that BPs have direct anti-apoptotic effects on osteoblasts and osteocytes, and the current situation of BP use in the clinic.
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