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In vitro degradation of four magnesium–zinc–strontium alloys and their cytocompatibility with human embryonic stem cells

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Abstract

Magnesium alloys have attracted great interest for medical applications due to their unique biodegradable capability and desirable mechanical properties. When designed for medical applications, these alloys must have suitable degradation properties, i.e., their degradation rate should not exceed the rate at which the degradation products can be excreted from the body. Cellular responses and tissue integration around the Mg-based implants are critical for clinical success. Four magnesium–zinc–strontium (ZSr41) alloys were developed in this study. The degradation properties of the ZSr41 alloys and their cytocompatibility were studied using an in vitro human embryonic stem cell (hESC) model due to the greater sensitivity of hESCs to known toxicants which allows to potentially detect toxicological effects of new biomaterials at an early stage. Four distinct ZSr41 alloys with 4 wt% zinc and a series of strontium compositions (0.15, 0.5, 1, and 1.5 wt% Sr) were produced through metallurgical processing. Their degradation was characterized by measuring total mass loss of samples and pH change in the cell culture media. The concentration of Mg ions released from ZSr41 alloy into the cell culture media was analyzed using inductively coupled plasma atomic emission spectroscopy. Surface microstructure and composition before and after culturing with hESCs were characterized using field emission scanning electron microscopy and energy dispersive X-ray spectroscopy. Pure Mg was used as a control during cell culture studies. Results indicated that the Mg–Zn–Sr alloy with 0.15 wt% Sr provided slower degradation and improved cytocompatibility as compared with pure Mg control.

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References

  1. Witte F, Hort N, Vogt C, Cohen S, Kainer KU, Willumeit R, et al. Degradable biomaterials based on magnesium corrosion. Curr Opin Solid State Mater Sci. 2008;12:63–72.

    Article  CAS  Google Scholar 

  2. Staiger MP, Pietak AM, Huadmai J, Dias G. Magnesium and its alloys as orthopedic biomaterials: a review. Biomaterials. 2006;27:1728–34.

    Article  CAS  Google Scholar 

  3. Liu H. The effects of surface and biomolecules on magnesium degradation and mesenchymal stem cell adhesion. J Biomed Mater Res A. 2011;99:249–60.

    Google Scholar 

  4. Breen DJ, Stoker DJ. Titanium lines: a manifestation of metallosis and tissue response to titanium alloy megaprostheses at the knee. Clin Radiol. 1993;47:274–7.

    Article  CAS  Google Scholar 

  5. Beuf O, Briguet A, Lissac M, Davis R. Magnetic resonance imaging for the determination of magnetic susceptibility of materials. J Magn Reson B. 1996;112:111–8.

    Article  CAS  Google Scholar 

  6. Ludeke KM, Roschmann P, Tischler R. Susceptibility artefacts in NMR imaging. Magn Reson Imaging. 1985;3:329–43.

    Article  CAS  Google Scholar 

  7. Petersilge CA, Lewin JS, Duerk JL, Yoo JU, Ghaneyem AJ. Optimizing imaging parameters for MR evaluation of the spine with titanium pedicle screws. Am J Roentgenol. 1996;166:1213–8.

    Article  CAS  Google Scholar 

  8. Tran N, Webster TJ. Nanotechnology for bone materials. Wiley Interdiscip Rev Nanomed Nanobiotechnol. 2009;1:336–51.

    Article  CAS  Google Scholar 

  9. Lembeck B, Wulker N. Severe cartilage damage by broken poly-l-lactic acid (PLLA) interference screw after ACL reconstruction. Knee Surg Sports Traumatol Arthrosc. 2005;13:283–6.

    Article  Google Scholar 

  10. Smith CA, Tennent TD, Pearson SE, Beach WR. Fracture of Bilok interference screws on insertion during anterior cruciate ligament reconstruction. Arthroscopy. 2003;19:E115–7.

    Article  Google Scholar 

  11. Baums MH, Zelle BA, Schultz W, Ernstberger T, Klinger HM. Intraarticular migration of a broken biodegradable interference screw after anterior cruciate ligament reconstruction. Knee Surg Sports Traumatol Arthrosc. 2006;14:865–8.

    Article  CAS  Google Scholar 

  12. Shafer BL, Simonian PT. Broken poly-l-lactic acid interference screw after ligament reconstruction. Arthroscopy. 2002;18:E35.

    Google Scholar 

  13. Sidhu DS, Wroble RR. Intraarticular migration of a femoral interference fit screw. A complication of anterior cruciate ligament reconstruction. Am J Sports Med. 1997;25:268–71.

    Article  CAS  Google Scholar 

  14. Takizawa T, Akizuki S, Horiuchi H, Yasukawa Y. Foreign body gonitis caused by a broken poly-l-lactic acid screw. Arthroscopy. 1998;14:329–30.

    Article  CAS  Google Scholar 

  15. Bostman O, Hirvensalo E, Makinen J, Rokkanen P. Foreign-body reactions to fracture fixation implants of biodegradable synthetic polymers. J Bone Joint Surg Br. 1990;72:592–6.

    CAS  Google Scholar 

  16. Kwak JH, Sim JA, Kim SH, Lee KC, Lee BK. Delayed intra-articular inflammatory reaction due to poly-l-lactide bioabsorbable interference screw used in anterior cruciate ligament reconstruction. Arthroscopy. 2008;24:243–6.

    Article  Google Scholar 

  17. Walton M, Cotton NJ. Long-term in vivo degradation of poly-l-lactide (PLLA) in bone. J Biomater Appl. 2007;21:395–411.

    Article  CAS  Google Scholar 

  18. Beevers DJ. Metal vs bioabsorbable interference screws: initial fixation. Proc Inst Mech Eng H. 2003;217:59–75.

    CAS  Google Scholar 

  19. Witte F. The history of biodegradable magnesium implants: a review. Acta Biomater. 2010;6:1680–92.

    Article  CAS  Google Scholar 

  20. Johnson I, Perchy D, Liu H. In vitro evaluation of the surface effects on magnesium–yttrium alloy degradation and mesenchymal stem cell adhesion. J Biomed Mater Res A. 2012;100:477–85.

    Google Scholar 

  21. Guan RG, Johnson I, Cui T, Zhao T, Zhao ZY, Li X, et al. Electrodeposition of hydroxyapatite coating on Mg-4.0Zn-1.0Ca-0.6Zr alloy and in vitro evaluation of degradation, hemolysis, and cytotoxicity. J Biomed Mater Res A. 2012;100:999–1015.

    Google Scholar 

  22. Cipriano A, Guan RG, Cui T, Zhao ZY, Garcia S, Johnson I, Liu H. In vitro degradation and cytocompatibility of magnesium-zinc-strontium alloys with human embryonic stem cells. 34th Annual International IEEE EMBS Conference Proceeding, 28 August–1 September, 2012, San Diego, CA, USA.

  23. Borkar H, Hoseini M, Pekguleryuz M. Effect of strontium on flow behavior and texture evolution during the hot deformation of Mg–1 wt% Mn alloy. Mater Sci Eng A. 2012;537:49–57.

    Article  CAS  Google Scholar 

  24. Xu L, Zhang E, Yin D, Zeng S, Yang K. In vitro corrosion behaviour of Mg alloys in a phosphate buffered solution for bone implant application. J Mater Sci. 2008;19:1017–25.

    Google Scholar 

  25. Maeng DY, Kim TS, Lee JH, Hong SJ, Seo SK, Chun BS. Microstructure and strength of rapidly solidified and extruded Mg–Zn alloys. Scripta Mater. 2000;43:385–9.

    Article  CAS  Google Scholar 

  26. Somekawa H, Osawa Y, Mukai T. Effect of solid-solution strengthening on fracture toughness in extruded Mg–Zn alloys. Scripta Mater. 2006;55:593–6.

    Article  CAS  Google Scholar 

  27. Gao X, Nie JF. Characterization of strengthening precipitate phases in a Mg–Zn alloy. Scripta Mater. 2007;56:645–8.

    Article  CAS  Google Scholar 

  28. Coleman JE. Zinc proteins: enzymes, storage proteins, transcription factors, and replication proteins. Annu Rev Biochem. 1992;61:897–946.

    Article  CAS  Google Scholar 

  29. Gu XN, Xie XH, Li N, Zheng YF, Qin L. In vitro and in vivo studies on a Mg–Sr binary alloy system developed as a new kind of biodegradable metal. Acta Biomater. 2012;8:2360.

    Article  CAS  Google Scholar 

  30. Dahl SG, Allain P, Marie PJ, Mauras Y, Boivin G, Ammann P, et al. Incorporation and distribution of strontium in bone. Bone. 2001;28:446–53.

    Article  CAS  Google Scholar 

  31. Marie PJ, Ammann P, Boivin G, Rey C. Mechanisms of action and therapeutic potential of strontium in bone. Calcif Tissue Int. 2001;69:121–9.

    Article  CAS  Google Scholar 

  32. Meunier PJ, Roux C, Seeman E, Ortolani S, Badurski JE, Spector TD, et al. The effects of strontium ranelate on the risk of vertebral fracture in women with postmenopausal osteoporosis. N Engl J Med. 2004;350:459–68.

    Article  CAS  Google Scholar 

  33. Laschinski G, Vogel R, Spielmann H. Cytotoxicity test using blastocyst-derived euploid embryonal stem cells: a new approach to in vitro teratogenesis screening. Reprod Toxicol. 1991;5:57–64.

    Article  CAS  Google Scholar 

  34. Talbot P, Lin S. Mouse and human embryonic stem cells: can they improve human health by preventing disease? Curr Top Med Chem. 2011;11:1638–52.

    Article  CAS  Google Scholar 

  35. Seiler AE, Spielmann H. The validated embryonic stem cell test to predict embryotoxicity in vitro. Nat Protoc. 2011;6:961–78.

    Article  CAS  Google Scholar 

  36. West PR, Weir AM, Smith AM, Donley ELR, Cezar GG. Predicting human developmental toxicity of pharmaceuticals using human embryonic stem cells and metabolomics. Toxicol Appl Pharmacol. 2010;247:18–27.

    Article  CAS  Google Scholar 

  37. Davila JC, Cezar GG, Thiede M, Strom S, Miki T, Trosko J. Use and application of stem cells in toxicology. Toxicol Sci. 2004;79:214–23.

    Article  CAS  Google Scholar 

  38. Adler S, Pellizzer C, Hareng L, Hartung T, Bremer S. First steps in establishing a developmental toxicity test method based on human embryonic stem cells. Toxicol In Vitro. 2008;22:200–11.

    Article  CAS  Google Scholar 

  39. Eiges R, Schuldiner M, Drukker M, Yanuka O, Itskovitz-Eldor J, Benvenisty N. Establishment of human embryonic stem cell-transfected clones carrying a marker for undifferentiated cells. Curr Biol. 2001;11:514–8.

    Article  CAS  Google Scholar 

  40. Hidaka K, Lee JK, Kim HS, Ihm CH, Iio A, Ogawa M, et al. Chamber-specific differentiation of Nkx2.5-positive cardiac precursor cells from murine embryonic stem cells. FASEB J. 2003;17:740–2.

    CAS  Google Scholar 

  41. Xin Y, Huo K, Tao H, Tang G, Chu PK. Influence of aggressive ions on the degradation behavior of biomedical magnesium alloy in physiological environment. Acta Biomater. 2008;4:2008–15.

    Article  CAS  Google Scholar 

  42. Rettig R, Virtanen S. Composition of corrosion layers on a magnesium rare-earth alloy in simulated body fluids. J Biomed Mater Res. 2009;88:359–69.

    Article  Google Scholar 

  43. Zainal Abidin NI, Martin D, Atrens A. Corrosion of high purity Mg, AZ91, ZE41 and Mg2Zn0·2Mn in Hank’s solution at room temperature. Corros Sci. 2011;53:862–72.

    Article  Google Scholar 

  44. Yang M, Pan F, Cheng R, Tang A. Effect of Mg–10Sr master alloy on grain refinement of AZ31 magnesium alloy. Mater Sci Eng A. 2008;491:440–5.

    Article  Google Scholar 

  45. Noronha JL, Matuschak GM. Magnesium in critical illness: metabolism, assessment, and treatment. Intensive Care Med. 2002;28:667–79.

    Article  Google Scholar 

  46. Fosmire GJ. Zinc toxicity. Am J Clin Nutr. 1990;51:225–7.

    CAS  Google Scholar 

  47. Feyerabend F, Fischer J, Holtz J, Witte F, Willumeit R, Drücker H, et al. Evaluation of short-term effects of rare earth and other elements used in magnesium alloys on primary cells and cell lines. Acta Biomater. 2010;6:1834–42.

    Article  CAS  Google Scholar 

  48. Winek CL, Buehler EV. Intravenous toxicity of zinc pyridinethione and several zinc salts. Toxicol Appl Pharmacol. 1966;9:269–73.

    Article  CAS  Google Scholar 

  49. Kroes R, den Tonkelaar EM, Minderhoud A, Speijers GJ, Vonk-Visser DM, Berkvens JM, et al. Short-term toxicity of strontium chloride in rats. Toxicology. 1977;7:11–21.

    Article  CAS  Google Scholar 

  50. Pizzoferrato A, Ciapetti G, Stea S, Cenni E, Arciola CR, Granchi D, et al. Cell culture methods for testing biocompatibility. Clin Mater. 1994;15:173–90.

    Article  CAS  Google Scholar 

  51. Zhang Z, Couture A, Luo A. An investigation of the properties of Mg–Zn–Al alloys. Scripta Materialia. 1998;39(1):45–53.

    Article  CAS  Google Scholar 

  52. Liu SF, Liu LY, Kang LG. Refinement role of electromagnetic stirring and strontium in AZ91 magnesium alloy. J Alloy Compd. 2008;450:546–50.

    Article  CAS  Google Scholar 

  53. Ziats NP, Miller KM, Anderson JM. In vitro and in vivo interactions of cells with biomaterials. Biomaterials. 1988;9:5–13.

    Article  CAS  Google Scholar 

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Acknowledgments

The authors would like to thank the U.S.A. NSF BRIGE award (CBET 1125801), Burroughs Welcome Fund (1011235), Hellman Fellowship, and the University of California Regents Faculty Fellowship for financial support. The authors also thank National Natural Science Foundation of China (Grant 51034002, 50974038 and 51074049) and the Fok Ying Tong Education Foundation (132002) for financial support. The authors thank the Central Facility for Advanced Microscopy and Microanalysis (CFAMM) and the Stem Cell Center (Drs. Prudence Talbot and Duncan Liew) at the University of California, Riverside.

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Correspondence to Huinan Liu.

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Cipriano, A.F., Zhao, T., Johnson, I. et al. In vitro degradation of four magnesium–zinc–strontium alloys and their cytocompatibility with human embryonic stem cells. J Mater Sci: Mater Med 24, 989–1003 (2013). https://doi.org/10.1007/s10856-013-4853-1

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