Evaluation of in vivo angiogenetic effects of copper doped bioactive glass scaffolds in the AV loop model


effects of 3D scaffolds made from 45S5 bioactive glass (BG) doped with 1 wt. % copper ions in the arteriovenous loop model of the rat.

Materials and Methods: An arteriovenous loop was built in the groin of 10 rats and inserted in 1% copper doped 45S5 BG scaffolds and fibrin. The scaffold and the AV loop were inserted in Teflon isolation chambers and explanted 3 weeks after implantation. Afterwards the scaffolds were analyzed by Micro-CT and histology regarding vascularization. Results were compared to plain 45S5 BG-based scaffolds from a previous study.

Results: Micro-CT and histological evaluation showed consistent vascularization of the constructs. A tendency towards an increased vascularization in the copper doped BG group compared to plain BG constructs could be observed. However, therewas no significant difference in statistical analysis between both groups.

Conclusions: This study shows results that support an increased angiogenetic effect of 1% copper doped 45S5 BG compared to regular 45S5 BG scaffolds in the rat arteriovenous loop model although these tendencies are not backed by statistical evidence. Maybe higher copper doses could lead to a statistically significant angiogenetic effect.

If the inline PDF is not rendering correctly, you can download the PDF file here.

  • [1] Buehrer, G., Balzer, A., Arnold, I., Beier, J.P., Koerner, C., Bleiziffer, O., et al., Combination of BMP2 and MSCs significantly increases bone formation in the rat arterio-venous loop model. Tissue Eng Part A, 2015. 21(1–2): 96–105.

  • [2] Sasaki, J., Hashimoto, M., Yamaguchi, S., Itoh, Y., Yoshimoto, I., Matsumoto, T., et al., Fabrication of Biomimetic Bone Tissue Using Mesenchymal Stem Cell–Derived Three–Dimensional Constructs Incorporating Endothelial Cells. PLoS One, 2015. 10(6): e0129266.

  • [3] Quinlan, E., Lopez-Noriega, A., Thompson, E.M., Hibbitts, A., Cryan, S.A., and O’Brien, F.J., Controlled release of vascular endothelial growth factor from spray-dried alginate microparticles in collagen-hydroxyapatite scaffolds for promoting vascularization and bone repair. J Tissue Eng Regen Med, 2015.

  • [4] Gerhardt, L.-C. and Boccaccini, A.R., Bioactive Glass and Glass- Ceramic Scaffolds for Bone Tissue Engineering.Materials, 2010. 3(7): 3867.

  • [5] Gorustovich, A.A., Roether, J.A., and Boccaccini, A.R., Effect of bioactive glasses on angiogenesis: a review of in vitro and in vivo evidences. Tissue Eng Part B Rev, 2010. 16(2): 199–207.

  • [6] Hoppe, A., Guldal, N.S., and Boccaccini, A.R., A review of the biological response to ionic dissolution products from bioactive glasses and glass-ceramics. Biomaterials, 2011. 32(11): 2757– 2774.

  • [7] Habibovic, P. and Barralet, J.E., Bioinorganics and biomaterials: bone repair. Acta Biomater, 2011. 7(8): 3013–3026.

  • [8] Hoppe, A., Brandl, A., Bleiziffer, O., Arkudas, A., Horch, R.E., Jokic, B., et al., In vitro cell response to Co-containing 1,393 bioactive glass. Mater Sci Eng C Mater Biol Appl, 2015. 57: 157–163.

  • [9] Barralet, J., Gbureck, U., Habibovic, P., Vorndran, E., Gerard, C., and Doillon, C.J., Angiogenesis in calcium phosphate scaffolds by inorganic copper ion release. Tissue Eng Part A, 2009. 15(7): 1601–1609.

  • [10] Giavaresi, G., Torricelli, P., Fornasari, P.M., Giardino, R., Barbucci, R., and Leone, G., Blood vessel formation after soft-tissue implantation of hyaluronan-based hydrogel supplemented with copper ions. Biomaterials, 2005. 26(16): 3001–3008.

  • [11] Hu, G.F., Copper stimulates proliferation of human endothelial cells under culture. J Cell Biochem, 1998. 69(3): 326–335.

  • [12] Feng,W., Ye, F., Xue,W., Zhou, Z., and Kang, Y.J., Copper regulation of hypoxia-inducible factor-1 activity. Mol Pharmacol, 2009. 75(1): 174–182.

  • [13] Gerard, C., Bordeleau, L.J., Barralet, J., and Doillon, C.J., The stimulation of angiogenesis and collagen deposition by copper. Biomaterials, 2010. 31(5): 824–831.

  • [14] Zhang, Z., Qiu, L., Lin, C., Yang, H., Fu, H., Li, R., et al., Copperdependent and -independent hypoxia-inducible factor-1 regulation of gene expression. Metallomics, 2014. 6(10): 1889–1893.

  • [15] Rath, S.N., Brandl, A., Hiller, D., Hoppe, A., Gbureck, U., Horch, R.E., et al., Bioactive copper–doped glass scaffolds can stimulate endothelial cells in co–culture in combination with mesenchymal stem cells. PLoS One, 2014. 9(12): e113319.

  • [16] Sen, C.K., Khanna, S., Venojarvi, M., Trikha, P., Ellison, E.C., Hunt, T.K., et al., Copper-induced vascular endothelial growth factor expression and wound healing. Am J Physiol Heart Circ Physiol, 2002. 282(5): H1821–1827.

  • [17] Li, S., Xie, H., and Kang, Y.J., Copper stimulates growth of human umbilical vein endothelial cells in a vascular endothelial growth factor-independent pathway. Exp Biol Med (Maywood), 2012. 237(1): 77–82.

  • [18] Ewald, A., Kappel, C., Vorndran, E., Moseke, C., Gelinsky, M., and Gbureck, U., The effect of Cu(II)-loaded brushite scaffolds on growth and activity of osteoblastic cells. J Biomed Mater Res A, 2012. 100(9): 2392–2400.

  • [19] Hench, L.L., The story of Bioglass. J Mater SciMater Med, 2006. 17(11): 967–978.

  • [20] Wu, C., Zhou, Y., Xu, M., Han, P., Chen, L., Chang, J., et al., Copper–containing mesoporous bioactive glass scaffolds with multifunctional properties of angiogenesis capacity, osteostimulation and antibacterial activity. Biomaterials, 2013. 34(2): 422–433.

  • [21] Erol, O.O. and Spira, M., New capillary bed formation with a surgically constructed arteriovenous fistula. Surg Forum, 1979. 30: 530–531.

  • [22] Kneser, U., Polykandriotis, E., Ohnolz, J., Heidner, K., Grabinger, L., Euler, S., et al., Engineering of vascularized transplantable bone tissues: induction of axial vascularization in an osteoconductive matrix using an arteriovenous loop. Tissue Eng, 2006. 12(7): 1721–1731.

  • [23] Arkudas, A., Pryymachuk, G., Beier, J.P., Weigel, L., Korner, C., Singer, R.F., et al., Combination of extrinsic and intrinsic pathways significantly accelerates axial vascularization of bioartificial tissues. Plast Reconstr Surg, 2012. 129(1): 55e–65e.

  • [24] Arkudas, A., Balzer, A., Buehrer, G., Arnold, I., Hoppe, A., Detsch, R., et al., Evaluation of angiogenesis of bioactive glass in the arteriovenous loop model. Tissue Eng Part C Methods, 2013. 19(6): 479–486.

  • [25] Hoppe, A., Meszaros, R., Stahli, C., Romeis, S., Schmidt, J., Peukert, W., et al., In vitro reactivity of Cu doped 45S5 Bioglass

  • [ registered sign] derived scaffolds for bone tissue engineering. Journal of Materials Chemistry B, 2013. 1(41): 5659– 5674.

  • [26] Arkudas, A., Pryymachuk, G., Hoereth, T., Beier, J.P., Polykandriotis, E., Bleiziffer, O., et al., Dose-finding study of fibrin gelimmobilized vascular endothelial growth factor 165 and basic fibroblast growth factor in the arteriovenous loop rat model. Tissue Eng Part A, 2009. 15(9): 2501–2511.

  • [27] Kalal, Z.,Matas, J.&Mikolajczyk, K, 2010 IEEE Computer Society Conference on Computer Vision and Pattern Recognition. IEEE Conference on Computer Vision and Pattern Recognition (CVPR) 49—-56 (IEEE, 2010), 2010.

  • [28] Seber, G., SLAC Research Library | Community Pages. 1984.

  • [29] Laschke, M.W., Harder, Y., Amon, M.,Martin, I., Farhadi, J., Ring, A., et al., Angiogenesis in tissue engineering: breathing life into constructed tissue substitutes. Tissue Eng, 2006. 12(8): 2093– 2104.

  • [30] Horch, R.E., Beier, J.P., Kneser, U., and Arkudas, A., Successful human long–term application of in situ bone tissue engineering. J Cell Mol Med, 2014. 18(7): 1478–1485.

  • [31] Schmidt, V.J., Hilgert, J.G., Covi, J.M., Leibig, N.,Wietbrock, J.O., Arkudas, A., et al., Flow increase is decisive to initiate angiogenesis in veins exposed to altered hemodynamics. PLoS One, 2015. 10(1): e0117407.

  • [32] Weigand, A., Beier, J.P., Hess, A., Gerber, T., Arkudas, A., Horch, R.E., et al., Acceleration of vascularized bone tissue-engineered constructs in a large animal model combining intrinsic and extrinsic vascularization. Tissue Eng Part A, 2015. 21(9–10): 1680– 1694.

  • [33] Gerhardt, L.C., Widdows, K.L., Erol, M.M., Burch, C.W., Sanz– Herrera, J.A., Ochoa, I., et al., The pro-angiogenic properties of multi–functional bioactive glass composite scaffolds. Biomaterials, 2011. 32(17): 4096–4108.

  • [34] Leu, A., Stieger, S.M., Dayton, P., Ferrara, K.W., and Leach, J.K., Angiogenic response to bioactive glass promotes bone healing in an irradiated calvarial defect. Tissue Eng Part A, 2009. 15(4): 877–885.

  • [35] Yuan, Q., Bleiziffer, O., Boos, A.M., Sun, J.M., Brandl, A., Beier, J.P., et al., PHDs inhibitor DMOG promotes the vascularization process in the AV loop by HIF-1a up-regulation and the preliminary discussion on its kinetics in rat. Bmc Biotechnology, 2014. 14.

  • [36] Handel, M., Hammer, T.R., Nooeaid, P., Boccaccini, A.R., and Hoefer, D., 45S5-Bioglass((R))-based 3D-scaffolds seeded with human adipose tissue–derived stem cells induce in vivo vascularization in the CAM angiogenesis assay. Tissue Eng Part A, 2013. 19(23–24): 2703–2712.

  • [37] Zhong, Z., Gu, H., Peng, J., Wang, W., Johnstone, B.H., March, K.L., et al., GDNF secreted from adipose–derived stem cells stimulates VEGF–independent angiogenesis. Oncotarget, 2016. 7(24): 36829–36841.

  • [38] Yu, H.S., Pastor, S.A., Lam, K.W., and Yee, R.W., Ascorbateenhanced copper toxicity on bovine corneal endothelial cells in vitro. Curr Eye Res, 1990. 9(2): 177–182.

  • [39] Frangoulis, M., Georgiou, P., Chrisostomidis, C., Perrea, D., Dontas, I., Kavantzas, N., et al., Rat epigastric flap survival and VEGF expression after local copper application. Plast Reconstr Surg, 2007. 119(3): 837–843.

  • [40] Stahli, C., Muja, N., and Nazhat, S.N., Controlled copper ion release from phosphate–based glasses improves human umbilical vein endothelial cell survival in a reduced nutrient environment. Tissue Eng Part A, 2013. 19(3–4): 548–557.

  • [41] Stahli, C., James–Bhasin, M., Hoppe, A., Boccaccini, A.R., and Nazhat, S.N., Effect of ion release from Cu-doped 45S5 Bioglass( R) on 3D endothelial cell morphogenesis. Acta Biomater, 2015. 19: 15–22.

  • [42] Day, R.M., Boccaccini, A.R., Shurey, S., Roether, J.A., Forbes, A., Hench, L.L., et al., Assessment of polyglycolic acid mesh and bioactive glass for soft-tissue engineering scaffolds. Biomaterials, 2004. 25(27): 5857–5866.

  • [43] Arkudas, A., Beier, J.P., Pryymachuk, G., Hoereth, T., Bleiziffer, O., Polykandriotis, E., et al., Automatic quantitative micro– computed tomography evaluation of angiogenesis in an axially vascularized tissue–engineered bone construct. Tissue Eng Part C Methods, 2010. 16(6): 1503–1514.

  • [44] Rottensteiner, U., Sarker, B., Heusinger, D., Dafinova, D., Rath, S., Beier, J., et al., In vitro and in vivo Biocompatibility of Alginate Dialdehyde/Gelatin Hydrogels with and without Nanoscaled Bioactive Glass for Bone Tissue Engineering Applications. Materials, 2014. 7(3): 1957.


Journal + Issues

Biomedical Glasses is an international open access journal covering the field of glasses for biomedical applications. The aim of the journal is to provide a peer-reviewed forum for the publication of original research reports and authoritative review articles related to the development of biomedical glasses and their use in clinical applications.