Abstract
The primary objective of this study was to evaluate in vitro responses of MLO-A5 osteogenic cells to two modifications of the bioactive glass 13-93. The modified glasses, which were designed for use as cell support scaffolds and contained added boron to form the glasses 13-93 B1 and 13-93 B3, were made to accelerate formation of a bioactive hydroxyapatite surface layer and possibly enhance tissue growth. Quantitative MTT cytotoxicity tests revealed no inhibition of growth of MLO-A5 cells incubated with 13-93 glass extracts up to 10 mg/ml, moderate inhibition of growth with 13-93 B1 glass extracts, and noticeable inhibition of growth with 13-93 B3 glass extracts. A morphology-based biocompatibility test was also performed and yielded qualitative assessments of the relative biocompatibilities of glass extracts that agree with those obtained by the quantitative MTT test. However, as a proof of concept experiment, when MLO-A5 cells were seeded onto 13-93 B3 scaffolds in a dynamic in vitro environment, cell proliferation occurred as evidenced by qualitative and quantitative MTT labeling of scaffolds. Together these results demonstrate the in vitro toxicity of released borate ion in static experiments; however borate ion release can be mitigated in a dynamic environment similar to the human body where microvasculature is present. Here we argue that despite toxicity in static environments, boron-containing 13-93 compositions may warrant further study for use in tissue engineering applications.
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Brown RF, Day DE, Day TE, Jung S, Rahaman MN, Fu Q. Growth and differentiation of osteoblastic cells on 13-93 bioactive glass fibers and scaffolds. Acta Biomater. 2008;4:387–96.
Fu Q, Rahaman MN, Bal SB, Brown RF, Day DE. Mechanical and in vitro performance of 13–93 bioactive glass scaffolds prepared by a polymer foam replication technique. Acta Biomater. 2008;4:1854–64.
Burg KJL, Porter S, Kellam JF. Biomaterial developments for bone tissue engineering. Biomaterials. 2000;21:2347–59.
Hutmacher DW. Scaffolds in tissue engineering bone and cartilage. Biomaterials. 2000;21:2529–43.
Kellomaki M, Niiranen H, Puumanen K, Ashamamakhi N, Waris T, Tormala P. Bioabsorbable scaffolds for guided bone regeneration and generation. Biomaterials. 2000;21:2495–505.
Du C, Cui FZ, Zhu XD, de Groot K. Three-dimensional nano-HAp/collagen matrix loading with osteogenic cells in organ culture. J Biomed Mater Res. 1999;44:407–15.
Griffith LG. Polymeric biomaterials. Acta Mater. 2000;48:263–77.
Goldstein SA, Patil PV, Moalli MR. Perspectives on tissue engineering of bone. Clin. Orthop. 1999;357S:S419–23.
Kneser U, Schaefer DJ, Munder B, Klemt C, Andree C, Stark GB. Tissue engineering of bone. Min. Invas. Ther. Alli. Tech. 2002;11:107–16.
Jones J, Ehrenfried L, Hench L. Optimising bioactive glass scaffolds for bone tissue engineering. Biomaterials. 2006;27:964–73.
Freyman TM, Yannas IV, Gibson LJ. Cellular materials as porous scaffolds for tissue engineering. Prog Mater Sci. 2001;45:273–82.
Hench LL, Wilson J. Surface-active biomaterials. Science. 1984;226:630–5.
Hench LL. Bioceramics. J Am Ceram Soc. 1998;81:1705–28.
Clupper DC, Mecholsky JJ, La Torre GP, Greenspan DC. Bioactivity of tape cast and sintered bioactive glass-ceramic in simulated body fluid. Biomaterials. 2002;23:2599–606.
Brink M. The influence of alkali and alkaline earths on the working range for bioactive glasses. J Biomed Mater Res. 1997;36:109–17.
Asikainen AJ, Hagstrom J, Sorsa T, Noponen J, Kellomaki M, Juuti H, Lindqvist C, Heitanen J, Suuronen R. Soft tissue reactions to bioactive glass 13–93 combined with chitosan. J Biomed Mater Res. 2006;83A:530–7.
Ruuttila P, Niiranen H, Kellomaki M, Tormala P, Konttinen YT, Hukkanen M. Characterization of human primary osteoblasts response on bioactive glass (BaG13–93) coated poly-L, DL-lactide (SR-PLA70) surface in vitro. J Biomed Mater Res. 2006;78 B:97–104.
Pirhonen E, Niiranen H, Niemela T, Brink M, Tormala P. Manufacturing, mechanical characterization, and in vitro performance of bioactive glass 13–93 fibers. J Biomed Mater Res. 2006;77:227–33.
Brink M, Turunen T, Happonen R-P, Yli-Urpo A. Compositional dependence of bioactivity of glasses in the system Na2O-K2O-MgOCaO-B2O3-P2O5-SiO2. J Biomed Mater Res. 1997;37:114–21.
Brink M, Yli-Urpo S, Yli-Urpo A.The resorption of a bioactive glass implanted into rat soft tissue. Presented at the 5th World Biomaterials Congress, Toronto, 48, 1996.
Modglin VC. In vitro evaluation of bioactive glass scaffolds and modified bioactive glasses with an osteogenic cell line. In: Biological Sciences, vol. (M.S. Rolla: Missouri University of Science and Technology, Rolla 2009).
Kato Y, Boskey A, Spevak L, Dallas M, Hori M, Bonewald LF. Establishment of an Osteoid Preosteocyte-like Cell MLO-A5 That Spontaneously Mineralizes in Culture. Bone and Min Res. 2001;16:1622–33.
Barragan-Adjemian C, Nicolella D, Dusevich V, Dallas MR, Eick JD, Bonewald LF. Mechanism by which MLO-A5 late osteoblasts/early osteocytes mineralize in culture: similarities with mineralization of lamellar bone. Calcif Tissue Int. 2006;79:340–53.
Zeitler T, Cormack A. Interaction of water with bioactive glass surfaces. J. Crystal Growth. 2006;294:96–102.
Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxic assays. J. Immun. Meth. 1983;65:55–63.
Gorustovich A, Lopez J, Guglielmotti M, Cabrini R. Biological performance of boron-modified bioactive glass particles implanted in rat tibia bone marrow. Biomed. Mat. 2006;3:100–5.
Dzondo-Gadet M, Mayap-Nzietchueng R, Hess K, Nabet P, Belleville F, Dousset B. Action of boron at the molecular level: effects on transcription and translation in an acellular system. Biol Trace Elem Res. 2002;85:23–33.
Vrouwenvelder WC, Groot CG, de Groot K. Better histology and biochemistry for osteoblasts cultured on titanium-doped bioactive glass: bioglass 45S5 compared with iron-, titanium-, fluorine- and boron-containing bioactive glasses. Biomaterials. 1994;15:97–106.
Burton JD. The MTT assay to evaluate chemosensitivity. Methods Mol Med. 2005;110:69–78.
Silver IA, Deas J, Erecinska M. Interactions of bioactive glasses with osteoblasts in vitro: effects of 45S5 Bioglass®, and 58S and 77S bioactive glasses on metabolism, intracellular ion concentrations and cell viability. Biomaterials. 2001;22:175–85.
Brown RF, Rahaman MN, Dwilewicz AB, Huang W, Day DE, Li Y, Bal BS. Effect of borate glass composition on its conversion to hydroxyapatite and on the proliferation of MC3T3-E1 cells. J Biomed Mater Res. 2008;88:392–400.
Richard M. Bioactive behavior of a borate glass. Ceramic Engineering, vol. (M.S. Rolla: University of Missouri-Rolla, 2000). p.140.
Heindel JJ, Price CJ, Schwetz BA. The developmental toxicity of boric acid in mice, rats, and rabbits. Environ Health Perspect. 1994;102:107–12.
Dolder J, Bancroft GN, Sikavitsas VI, Spauwen PHM, Jansen JA, Mikos AG. Flow perfusion culture of marrow stromal osteoblasts in titanium fiber mesh. J. Biomed. Mat. Res. 2002;64:235–41.
Smallwood CL, Lipscomb J, Swartout J, Teuschler L. Toxicological report of boron and compounds. In: Agency USEP, editor. Washington D.C.: EPA, 2004. p. 134.
Smallwood C. Boron in drinking-water, vol. 2. Geneva: World Health Organization; 2003.
Jung SB, Day DE, Brown RF, Bonewald LF. Potential toxicity of bioactive borate glasses in vitro and in vivo. ICACC, Daytona Beach, 2012 (Proceedings in press).
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This investigation was supported by funds from the Center for Bone and Tissue Repair and Regeneration at Missouri University of Science and Technology.
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Modglin, V.C., Brown, R.F., Jung, S.B. et al. Cytotoxicity assessment of modified bioactive glasses with MLO-A5 osteogenic cells in vitro. J Mater Sci: Mater Med 24, 1191–1199 (2013). https://doi.org/10.1007/s10856-013-4875-8
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DOI: https://doi.org/10.1007/s10856-013-4875-8