3D-printed Biomimetic Bioactive Glass Scaffolds for Bone Regeneration in Rat Calvarial Defects
The pore geometry of scaffold intended for the use in the bone repair or replacement is one of the most important parameters in bone tissue engineering. It affects not only the mechanical properties of the scaffold but also the amount of bone regeneration after implantation. Scaffolds with five different architectures (cubic, spherical, x, gyroid, and diamond) at different porosities were fabricated with bioactive borate glass using the selective laser sintering (SLS) process. The compressive strength of scaffolds with porosities ranging from 60% to 30% varied from 1.7 to 15.5 MPa. The scaffold’s compressive strength decreased significantly (up to 90%) after 1-week immersion in simulated body fluids. Degradation of scaffolds is dependent on porosity, in which the scaffold with the largest surface area has the largest reduction in strength. Scaffolds with traditional cubic architecture and biomimetic diamond architecture were implanted in 4.6 mm diameter fullthickness rat calvarial defects for 6 weeks to evaluate the bone regeneration with or without bone morphogenetic protein 2 (BMP-2). Histological analysis indicated no significant difference in bone formation in the defects treated with the two different architectures. However, the defects treated with the diamond architecture scaffolds had more fibrous tissue formation and thus have the potential for faster bone formation. Overall, the results indicated that borate glass scaffolds fabricated using the SLS process have the potential for bone repair and the addition of BMP-2 significantly improves bone regeneration.
1. Jones JR, 2013, Review of Bioactive Glass: From Hench to Hybrids. Acta Biomater, 9:4457–86.
2. Hench LL, 2006, The Story of Bioglass®. J Mater Sci Mater Med, 17:967–78.
3. Greenspan D, 2019, Bioglass at 50-A Look at Larry Hench’s Legacy and Bioactive Materials. Biomed Glas, 5:178–84. DOI: 10.1515/bglass-2019-0014.
4. Rahaman MN, Day DE, Bal BS, et al., 2011, Bioactive Glass in Tissue Engineering. Acta Biomater, 7:2355–73.
5. Fu Q, Rahaman MN, Fu H, et al., 2010, Silicate, Borosilicate, and Borate Bioactive Glass Scaffolds with Controllable Degradation Rate for Bone Tissue Engineering Applications. I. Preparation and In Vitro Degradation. J Biomed Mater Res Part A, 95A:164–71. DOI: 10.1002/jbm.a.32824.
6. Jung S, Day D, 2009, Conversion Kinetics of Silicate, Borosilicate, and Borate Bioactive Glasses to Hydroxyapatite. Phys Chem Glas, 50:85–8.
7. Balasubramanian P, Kolzow J, Chen RR, et al., 2018, Boron-containing Bioactive Glasses in Bone and Soft Tissue Engineering. J Eur Ceram Soc, 38:855–69.
8. Yuan S, Shen F, Chua CK, et al., 2019, Polymeric Composites for Powder-based Additive Manufacturing: Materials and Applications. Prog Polym Sci, 91:141–68.
9. Ng WL, Lee JM, Zhou M, et al., 2020, Vat Polymerization based Bioprinting Process, Materials, Applications and Regulatory Challenges IOP Science. Biofabrication, 12:022001. DOI: 10.1088/1758-5090/ab6034.
10. Goh GD, Yap YL, Tan HK, et al., 2020, Process Structure Properties in Polymer Additive Manufacturing via Material Extrusion: A Review. Crit Rev Solid State Mater Sci, 45:113–33. DOI: 10.1080/10408436.2018.1549977.
11. Cai S, Xi J, 2008, A Control Approach for Pore Size Distribution in the Bone Scaffold Based on the Hexahedral Mesh Refinement. CAD Comput Aided Des, 40:1040–50. DOI: 10.1016/j.cad.2008.09.004.
12. Melchels FP, Bertoldi K, Gabbrielli R, et al., 2010, Mathematically Defined Tissue Engineering Scaffold Architectures Prepared by Stereolithography. Biomaterials, 31:6909–16. DOI: 10.1016/j.biomaterials.2010.05.068.
13. Challis VJ, Roberts AP, Grotowski JF, et al., 2010, Prototypes for Bone Implant Scaffolds Designed via Topology Optimization and Manufactured by Solid Freeform Fabrication. Adv Eng Mater, 12:1106–10. DOI: 10.1002/adem.201000154.
14. Feng J, Fu J, Li Z, et al., 2018, A Review of the Design Methods of Complex Topology Structures for 3D Printing. Vis Comput Ind Biomed Art, 1:5.
15. Wang G, Shen L, Zhao J, et al., 2018, Design and Compressive Behavior of Controllable Irregular Porous Scaffolds: Based on Voronoi-Tessellation and for Additive Manufacturing. ACS Biomater Sci Eng, 4:719–27. DOI: 10.1021/acsbiomaterials.7b00916.
16. Ng WL, Chua CK, Shen YF, et al., 2019, Print Me an Organ! Why We Are Not There Yet. Prog Polym Sci, 97:101145. DOI: 10.1016/j.progpolymsci.2019.101145.
17. Sing SL, Yeong WY, Wiria FE, et al., 2017, Direct Selective Laser Sintering and Melting of Ceramics: A Review. Rapid Prototyp J, 23:611–23. DOI: 10.1108/rpj-11-2015-0178.
18. Datsiou KC, Saleh E, Spirrett F, et al., 2019, Additive Manufacturing of Glass with Laser Powder Bed Fusion. J Am Ceram Soc, 102:4410–4. DOI: 10.1111/jace.16440.
19. Yves-Christian H, et al., 2010, Net Shaped High Performance Oxide Ceramic Parts by Selective Laser Melting. In: Physics Procedia. Vol. 5. Elsevier B.V., Berlin, pp. 587–94. DOI: 10.1016/j.phpro.2010.08.086.
20. Verga F, Mario B, Laura C, et al., 2020, Laser-based Powder Bed Fusion of Alumina Toughened Zirconia. Addit Manuf, 31:100959. DOI: 10.1016/j.addma.2019.100959.
21. Kolan KC, Leu MC, Hilmas GE, et al., 2011, Fabrication of 13-93 Bioactive Glass Scaffolds for Bone Tissue Engineering Using Indirect Selective Laser Sintering. Biofabrication, 3:025004. DOI: 10.1088/1758-5082/3/2/025004.
22. Goodridge RD, Dalgarno KW, Wood DJ, et al., 2006, Indirect Selective Laser Sintering of an Apatite-mullite Glass-Ceramic for Potential Use in Bone Replacement Applications. Proc Inst Mech Eng Part H J Eng Med, 220:57–68. DOI:10.1243/095441105x69051.
23. Van Bael S, Chai YC, Truscello S, et al., 2012, The Effect of Pore Geometry on the In Vitro Biological Behavior of Human Periosteum-Derived Cells Seeded on Selective Laser-melted Ti6Al4V Bone Scaffolds. Acta Biomater, 8:2824–34. DOI: 10.1016/j.actbio.2012.04.001.
24. Zadpoor AA, 2015, Bone Tissue Regeneration: The Role of Scaffold Geometry. Biomater Sci, 3:231–45. DOI: 10.1039/c4bm00291a.
25. Ouyang P, Dong H, He X, et al., 2019, Hydromechanical Mechanism behind the Effect of Pore Size of Porous Titanium Scaffolds on Osteoblast Response and Bone Ingrowth. Mater Des, 183:108151. doi.org/10.1016/j.matdes.2019.108151.
26. Gariboldi MI, Best SM, 2015, Effect of Ceramic Scaffold Architectural Parameters on Biological Response. Front Bioeng Biotechnol, 3:151. DOI: 10.3389/fbioe.2015.00151.
27. Roosa SM, Kemppainen JM, Moffitt EN, et al., 2010, The Pore Size of Polycaprolactone Scaffolds has Limited Influence on Bone Regeneration in an In Vivo Model. J Biomed Mater Res Part A, 92:359–68. DOI: 10.1002/jbm.a.32381.
28. Perez RA, Mestres G, 2016, Role of Pore Size and Morphology in Musculo-skeletal Tissue Regeneration. Mater Sci Eng C, 61:922–39. DOI: 10.1016/j.msec.2015.12.087.
29. Lin Y, Liu X, Xiao W, et al., 2015, Long-term Bone Regeneration, Mineralization and Angiogenesis in Rat Calvarial Defects Implanted with Strong Porous Bioactive Glass (13-93) Scaffolds. J Non Cryst Solids, 432:4–13. DOI: 10.1016/j.jnoncrysol.2015.04.008.
30. Bi L, Jung S, Day D, et al., 2012, Evaluation of Bone Regeneration, Angiogenesis, and Hydroxyapatite Conversion in Critical-sized Rat Calvarial Defects Implanted with Bioactive Glass Scaffolds. J Biomed Mater Res Part A, 100A:3267–75. DOI: 10.1002/jbm.a.34272.
31. Bidan CM, Kommareddy KP, Rumpler M, et al., 2013, Geometry as a Factor for Tissue Growth: Towards Shape Optimization of Tissue Engineering Scaffolds. Adv Healthc Mater, 2:186–94. DOI: 10.1002/adhm.201200159.
32. Bidan CM, Kommareddy KP, Rumpler M, et al., 2012, How Linear Tension Converts to Curvature: Geometric Control of Bone Tissue Growth. PLoS One, 7:e36336. DOI: 10.1371/journal.pone.0036336.
33. Rumpler M, Woesz A, Dunlop JW, et al., 2008, The Effect of Geometry on Three-dimensional Tissue Growth. J R Soc Interface, 5:1173–80.
34. Kolan KC, Thomas A, Leu MC, et al., 2015, In Vitro Assessment of Laser Sintered Bioactive Glass Scaffolds with Different Pore Geometries. Rapid Prototyp J, 21:152–8. DOI:10.1108/rpj-12-2014-0175.
35. Kolan KC, Leu MC, Hilmas GE, et al., 2012, Effect of Material, Process Parameters, and Simulated Body Fluids on Mechanical Properties of 13-93 Bioactive Glass Porous Constructs Made by Selective Laser Sintering. J Mech Behav Biomed Mater, 13:14–24. DOI: 10.1016/j.jmbbm.2012.04.001.
36. Kokubo T, Takadama H, 2006, How Useful is SBF in Predicting In Vivo Bone Bioactivity? Biomaterials, 27:2907–15. DOI: 10.1016/j.biomaterials.2006.01.017.
37. Schindelin J, Arganda-Carreras I, Frise E, et al., 2012, Fiji:An Open-source Platform for Biological-image Analysis. Nat Methods, 9:676–82. DOI: 10.1038/nmeth.2019.
38. Melchels FP, Barradas AM, van Blitterswijk CA, et al., 2010, Effects of the Architecture of Tissue Engineering Scaffolds on Cell Seeding and Culturing. Acta Biomater, 6:4208–17. DOI: 10.1016/j.actbio.2010.06.012.
39. Carter DR, Hayes WC, 1976, Bone Compressive Strength: The Influence of Density and Strain Rate. Science, 194:1174–6. DOI: 10.1126/science.996549.
40. Wu D, Isaksson P, Ferguson SJ, et al., 2018, Young’s Modulus of Trabecular Bone at the Tissue Level: A Review. Acta Biomater, 78:1–12. DOI: 10.1016/j.actbio.2018.08.001.
41. Freitas GP, Lopes HB, Souza AT, et al., 2019, Cell Therapy: Effect of Locally Injected Mesenchymal Stromal Cells Derived from Bone Marrow or Adipose Tissue on Bone Regeneration of Rat Calvarial Defects. Sci Rep, 9:13476. DOI: 10.1038/s41598-019-50067-6.
42. Gibson LJ, Ashby MF, 1982, The Mechanics of Three-Dimensional Cellular Materials. Proc R Soc A Math Phys Sci, 382:43–59.
43. Ryshkewitch E, 1953, Compression Strength of Porous Sintered Alumina and Zirconia. J Am Ceram Soc, 36:65–8.
44. Duckworth W, 1953, Discussion of Ryshkewitch Paper. J Am Ceram Soc, 36:68.
45. Rice RW, 1996, Evaluation and Extension of Physical Property-porosity Models Based on Minimum Solid Area. J Mater Sci, 31:102–18.
46. Hattiangadi A, Bandyopadhyay A, 2000, Strength Degradation of Nonrandom Porous Ceramic Structures under Uniaxial Compressive Loading. J Am Ceram Soc, 83:2730–6. DOI: 10.1111/j.1151-2916.2000.tb01624.x.
47. Rice RW, 1993, Comparison of Stress Concentration Versus Minimum Solid Area Based Mechanical Property-porosity Relations. J Mater Sci, 28:2187–90. DOI: 10.1007/bf00367582.
48. Deliormanl AM, 2012, In Vitro Assessment of Degradation and Bioactivity of Robocast Bioactive Glass Scaffolds in Simulated Body Fluid. Ceram Int, 38:6435–44. DOI: 10.1016/j.ceramint.2012.05.019.
49. Deliormanli AM, Rahaman MN, 2012, Direct-write Assembly of Silicate and Borate Bioactive Glass Scaffolds for Bone Repair. J Eur Ceram Soc, 32:3637–46. DOI: 10.1016/j.jeurceramsoc.2012.05.005.
50. Kolan KC, Semon J, Bromet B, et al., 2019, Bioprinting with Human Stem Cells-laden Alginate-gelatin Bioink and Bioactive Glass for Tissue Engineering. Int J Bioprint, 5:3. DOI: 10.18063/ijb.v5i2.2.204.
51. Murphy C, Kolan K, Li W, et al., 2017, 3D Bioprinting of Stem Cells and Polymer/Bioactive Glass Composite Scaffolds for Tissue Engineering. Int J Bioprinting, 3:54–64. DOI: 10.18063/ijb.2017.01.005.
52. Hustedt JW, Blizzard DJ, 2018, The Controversy Surrounding Bone Morphogenetic Proteins in the Spine: A Review of Current Research. In: Getting to Good: Research Integrity in the Biomedical Sciences. Vol. 87. Springer International Publishing, Basel, Switzerland, pp. 9–22.
53. Carragee EJ, Chu G, Rohatgi R, et al., 2013, Cancer Risk After Use of Recombinant Bone Morphogenetic Protein-2 for Spinal Arthrodesis. J Bone Joint Surg Am, 95:1537–45. DOI: 10.1016/j.spinee.2013.11.026.
54. Injamuri S, Rahaman MN, Shen Y, et al., 2020, Relaxin Enhances Bone Regeneration with BMP-2-Loaded Hydroxyapatite Microspheres. J Biomed Mater Res Part A, 108:1231–42. DOI: 10.1002/jbm.a.36897.
55. Gu Y, Bal B, Rahaman N, et al., 2015, In Vivo Evaluation of Scaffolds with a Grid-Like Microstructure Composed of a Mixture of Silicate (13-93) and Borate (13-93B3) Bioactive Glasses. John Wiley and Sons, Inc., New York, pp. 53–64. DOI: 10.1002/9781119040392.ch6.
56. Karageorgiou V, Kaplan D, 2005, Porosity of 3D Biomaterial Scaffolds and Osteogenesis. Biomaterials, 26:5474–91. DOI: 10.1016/j.biomaterials.2005.02.002.
57. Sopyan I, Gunawan, 2013, Development of Porous Calcium Phosphate Bioceramics for Bone Implant Applications: A Review. Recent Patents Mater Sci, 6:238–52. DOI: 10.2174/18744648113069990012.
58. Gu Y, Huang W, Rahaman MN, et al., 2013, Bone Regeneration in Rat Calvarial Defects Implanted with Fibrous Scaffolds Composed of a Mixture of Silicate and Borate Bioactive Glasses. Acta Biomater, 9:9126–36. DOI: 10.1016/j.actbio.2013.06.039.
59. Bi L, Zobell B, Liu X, et al., 2014, Healing of Critical-size Segmental Defects in Rat Femora Using Strong Porous Bioactive Glass Scaffolds. Mater Sci Eng C, 42:816–24. DOI: 10.1016/j.msec.2014.06.022.
60. Wang H, Zhao S, Xiao W, et al., 2015, Three-dimensional Zinc Incorporated Borosilicate Bioactive Glass Scaffolds for Rodent Critical-sized Calvarial Defects Repair and Regeneration. Colloids Surfaces B Biointerfaces, 130:149–56. DOI: 10.1016/j.colsurfb.2015.03.053.
61. Wang H, Zhao S, Zhou, J, et al., 2014, Evaluation of Borate Bioactive Glass Scaffolds as a Controlled Delivery System for Copper Ions in Stimulating Osteogenesis and Angiogenesis in Bone Healing. J Mater Chem B, 2:8547–57. DOI: 10.1039/c4tb01355g.
62. Hart NH, Nimphius S, Rantalainen T, et al., 2017, Mechanical Basis of Bone Strength: Influence of Bone Material, Bone Structure and Muscle Action. J Musculoskelet Neuronal Interact, 17:114–39.