title: Osteoanabolic effect of alendronate and zoledronate on bone marrow stromal cells (BMSCs) isolated from aged female osteoporotic patients and its implications for their mode of action in the treatment of age-related bone loss.
abstract: CONCLUSIONS
  In the present study, we evaluated the potential for aminobisphosphonates to enhance the development of bone-forming osteoblasts from progenitor cells isolated from aged female osteoporotic patients. The aminobisphosphonates tested significantly enhanced osteoblast formation and thus lend further insights into their possible mode of action in the treatment of osteoporosis.
  
  
  BACKGROUND
  The primary aim of this study was to evaluate the influence of aminobisphosphonates on the osteogenesis of human bone marrow stromal cells (hBMSCs) and mineralization of differentiating bone-forming cells isolated from osteoporotic patients.
  
  
  METHODS
  The influence of aminobisphosphonate treatment on hBMSC osteogenesis was assessed by the quantitative measurement of alkaline phosphatase (ALP) activity, in addition to quantitative reverse transcription polymerase chain reaction and Western blot analysis of known osteogenic markers. Mineralized matrix formation by hBMSC-derived osteoblasts was visualized and quantified using Alizarin red staining.
  
  
  RESULTS
  hBMSC cultures treated with osteogenic medium supplemented with zoledronate demonstrated a significant increase in Alizarin red staining after 3 weeks as compared to cells cultured in osteogenic medium alone. Similarly, cultures of differentiating hBMSCs isolated from patients receiving alendronate treatment also demonstrated an increased propensity for mineralization, even in the absence of further in vitro stimulation by zoledronate. The stimulatory effects of aminobisphosphonate treatment on hBMSC-derived osteoblast-mediated mineralization were independent of any alterations in ALP activity, although significant decreases in the expression levels of osteopontin (SPP1) were evident in hBMSCs following exposure to aminobisphosphonates. Further analysis including Western blotting and loss-of-function studies revealed osteopontin as having a negative influence on the mineralization of differentiating osteoporotic bone-forming cells.
  
  
  CONCLUSIONS
  The results presented here demonstrate for the first time that aminobisphosphonate treatment of osteoporotic hBMSCs enhances their capacity for osteoblast formation and subsequent mineral deposition, thus supporting the concept of aminobisphosphonates as having an osteoanabolic effect in osteoporosis.
authors: Department of Trauma Surgery and Sports Medicine, Innsbruck Medical University, Innsbruck, Austria.
authors: R A
authors: RA
authors: Lindtner
authors: A N
authors: AN
authors: Tiaden
authors: K
authors: K
authors: Genelin
authors: H L
authors: HL
authors: Ebner
authors: C
authors: C
authors: Manzl
authors: M
authors: M
authors: Klawitter
authors: I
authors: I
authors: Sitte
authors: B
authors: B
authors: von Rechenberg
authors: M
authors: M
authors: Blauth
authors: P J
authors: PJ
authors: Richards
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D019386:Alendronate / Q000494:pharmacology* / Q000627:therapeutic use; D000469:Alkaline Phosphatase / Q000378:metabolism; D015519:Bone Density / Q000187:drug effects; D050071:Bone Density Conservation Agents / Q000494:pharmacology* / Q000627:therapeutic use; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D004164:Diphosphonates / Q000494:pharmacology*; D005260:Female; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology*; D059630:Mesenchymal Stem Cells / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D010012:Osteogenesis / Q000187:drug effects; D053495:Osteopontin / Q000502:physiology; D015663:Osteoporosis, Postmenopausal / Q000188:drug therapy / Q000473:pathology* / Q000503:physiopathology; D000077211:Zoledronic Acid
chemical_list: D050071:Bone Density Conservation Agents; D004164:Diphosphonates; D007093:Imidazoles; D053495:Osteopontin; D000077211:Zoledronic Acid; D000469:Alkaline Phosphatase; D019386:Alendronate
references: Clin Endocrinol (Oxf). 2010 Feb;72(2):161-8
references: 19302584
references: Bone. 2009 May;44(5):858-64
references: 19442618
references: J Bone Miner Res. 2005 Mar;20(3):419-27
references: 15746986
references: J Clin Invest. 1996 Apr 1;97(7):1732-40
references: 8601639
references: Basic Clin Pharmacol Toxicol. 2005 Dec;97(6):382-91
references: 16364054
references: Biomaterials. 2005 Dec;26(34):6941-9
references: 16009417
references: Bone. 2011 Jul;49(1):34-41
references: 21111853
references: J Biol Chem. 2010 Aug 13;285(33):25103-8
references: 20501658
references: Biomaterials. 2010 Nov;31(33):8674-83
references: 20719378
references: Methods Mol Biol. 2008;449:3-25
references: 18370080
references: Aging Cell. 2008 Jan;7(1):23-31
references: 18028256
references: J Orthop Res. 2010 Jun;28(6):798-804
references: 20014309
references: N Engl J Med. 2004 Mar 18;350(12):1189-99
references: 15028823
references: J Biol Chem. 2007 May 25;282(21):15872-83
references: 17383965
references: J Clin Invest. 1999 Nov;104(10):1363-74
references: 10562298
references: PLoS One. 2012;7(9):e45142
references: 23028809
references: Bone. 2011 Jul;49(1):122-7
references: 20736091
references: Nat Med. 2007 Jul;13(7):791-801
references: 17618270
references: Bone. 2011 Jul;49(1):82-8
references: 21349354
references: Bone Miner. 1989 Apr;6(1):33-43
references: 2526673
references: J Bone Miner Res. 2010 Nov;25(11):2267-94
references: 20842676
references: J Cell Biochem. 2005 Jan 1;94(1):50-7
references: 15517602
references: Science. 1999 Apr 2;284(5411):143-7
references: 10102814
references: Bone. 1998 May;22(5):455-61
references: 9600778
references: J Clin Invest. 1989 Jun;83(6):1930-5
references: 2524504
references: Stem Cells. 2012 Oct;30(10):2271-82
references: 22865667
references: Endocrinol Metab Clin North Am. 2012 Sep;41(3):487-506
references: 22877426
references: Injury. 2008 Feb;39(2):224-31
references: 18222447
references: J Bone Miner Res. 2006 Sep;21(9):1377-86
references: 16939396
references: Osteoporos Int. 2005 Oct;16(10):1291-8
references: 15986101
references: Endocrinology. 2008 Mar;149(3):1302-13
references: 18079194
references: J Bone Miner Res. 1998 Nov;13(11):1721-9
references: 9797481
references: Cancer Res. 2000 Nov 1;60(21):6001-7
references: 11085520
references: J Cell Biochem. 2011 May;112(5):1229-42
references: 21465521
references: J Bone Miner Res. 1995 Oct;10(10):1478-87
references: 8686503
references: Bone. 2011 Jul;49(1):2-19
references: 21555003
references: Calcif Tissue Int. 2002 Aug;71(2):145-54
references: 12073157
references: Lancet. 2011 Apr 9;377(9773):1276-87
references: 21450337
references: J Orthop Res. 2009 Aug;27(8):1067-73
references: 19215023
references: Biomaterials. 2004 Aug;25(18):4105-15
references: 15046901
references: Arthritis Rheum. 2009 Nov;60(11):3356-65
references: 19877060
references: N Engl J Med. 2006 May 25;354(21):2250-61
references: 16723616
references: Nature. 1997 Nov 6;390(6655):45-51
references: 9363890
references: J Clin Invest. 1999 Aug;104(3):229-37
references: 10430604
references: J Bone Joint Surg Br. 2012 Mar;94(3):385-90
references: 22371548
references: J Bone Miner Res. 2007 Oct;22(10):1603-11
references: 17605634
references: Proc Natl Acad Sci U S A. 2003 May 13;100(10):5840-5
references: 12732718
references: J Cell Biochem. 1999 Dec 1;75(3):414-23
references: 10536365
references: J Cell Biochem. 2000 Sep 14;79(4):557-65
references: 10996846