Indirect 3D printing CDHA scaffolds with hierarchical porous structure to promote osteoinductivity and bone regeneration

doi:10.1016/j.jmst.2024.04.032

Abstract

Abstract: Hierarchical porous structure, which include macropores, minor pores, and micropores in scaffolds, are essential in the multiple biological functions of bone repair and regeneration. In this study, patient-customized calcium-deficient hydroxyapatite (CDHA) scaffolds with three-level hierarchical porous structure were fabricated by indirect 3D printing technology and particulate leaching method. The sacrificial template scaffolds were fabricated using a photo-curing 3D printer, which provided a prerequisite for the integral structure and interconnected macropores of CDHA scaffolds. Additionally, 20 wt% pore former was incorporated into the slurry to enhance the content of smaller pores within the CDHA-2 scaffolds, and then the CDHA-2 scaffolds were sintered to remove the sacrificial template scaffolds and pore former. The obtained CDHA-2 scaffolds exhibited interconnected macropores (300-400 μm), minor pores (∼10-100 μm), and micropores (< 10 μm) distributed throughout the scaffolds, which could promote bone tissue ingrowth, increase surface roughness, and enhance protein adsorption of scaffolds. In vitro studies identified that CDHA-2 scaffolds had nanocrystal grains, high specific surface area, and outstanding protein adsorption capacity, which could provide a microenvironment for cell adhesion, spreading, and proliferation. In addition, the murine intramuscular implantation experiment suggested that CDHA-2 scaffolds exhibited excellent osteoinductivity and were superior to traditional BCP ceramics under conditions without the addition of live cells and exogenous growth factors. The rabbit calvarial defect repair results indicated that CDHA-2 scaffolds could enhance in situ bone regeneration. In conclusion, these findings demonstrated that the hierarchical porous structure of CDHA scaffolds was a pivotal factor in modulating osteoinductivity and bone regeneration, and CDHA-2 scaffolds were potential candidates for bone regeneration.

Key words: Calcium-deficient hydroxyapatite, Indirect 3D printing technology, Hierarchical porous structure, Osteoinductivity, Bone regeneration

Wenling Dai, Shikui Li, Hengxing Jia, Xingchen Zhao, Chenxin Liu, Changchun Zhou, Yumei Xiao, Likun Guo, Yujiang Fan, Xingdong Zhang. Indirect 3D printing CDHA scaffolds with hierarchical porous structure to promote osteoinductivity and bone regeneration[J]. J. Mater. Sci. Technol., 2025, 207: 295-307.

References

[1] A. Mostafavi, T. Abdullah, C.S. Russell, E. Mostafavi, T.J. Williams, N. Salah, A. Alshahrie, S. Harris, S.M.M.Basri, Y.K. Mishra, T.J. Webster, A. Memic, A.Tamayol, Acta Biomater. 127(2021) 313-326.
[2] R. Dimitriou, E. Jones, D. McGonagle, P.V. Giannoudis, BMC Med. 9(2011) 66.
[3] J.J. Li, M. Ebied, J. Xu, H. Zreiqat, Adv Healthc. Mater. 7(2018) e1701061.
[4] T.W. Bauer, G.F. Muschler, Bone Graft Mater. Clin. Orthop. Relat. R. 371(2000) 10-27.
[5] M. Lopez-Alvarez, S. Perez-Davila, C. Rodriguez-Valencia, P. Gonzalez, J. Serra, Biomed. Mater. 11(2016) 035011.
[6] Y. Ma, T. Wang, J. Liao, H. Gu, X. Lin, Q. Jiang, M.K. Bulsara, M. Zheng, Q. Zheng, Stem. Cell. Res. Ther. 5(2014) 115.
[7] W. Wang, K.W.K.Yeung, Bioact. Mater. 2(2017) 224-247.
[8] A. Oryan, S. Alidadi, A. Moshiri, N. Maffulli, J. Orthop. Surg.Res. 9(2014) 18.
[9] H. Liang, Y. Yang, D. Xie, L. Li, N. Mao, C. Wang, Z. Tian, Q. Jiang, L. Shen, J. Mater. Sci.Technol. 35(2019) 1284-1297.
[10] T. Yang, Y. Hu, C. Wang, B.P. Binks, ACS Appl. Mater. Interfaces 9 (2017) 22950-22958.
[11] J.S. Lee, H. Nah, D. Lee, S.H. An, W.K. Ko, S.J. Lee, S.Y. Lee, K.M. Park, J.B. Lee, H.J. Yi, I.K. Kwon, K.S. Choi, D.N. Heo, Mater. Des. 219(2022) 110801.
[12] M. Mirkhalaf, Y. Men, R. Wang, Y. No, H. Zreiqat, Acta Biomater. 156(2023) 110-124.
[13] Z.H.E. Wang, Z. Tang, F. Qing, Y. Hong, X. Zhang, Nano 07 (2012) 1230004.
[14] Z. Tang, X. Li, Y. Tan, H. Fan, X. Zhang, Regen. Biomater. 5(2018) 43-59.
[15] M. Vallet-Regí, E. Ruiz-Hernández, Adv. Mater. 23(2011) 5177-5218.
[16] P. Habibovic, T.M. Sees, M.A. van den Doel, C.A. van Blitterswijk, K. de Groot, J. Biomed. Mater. Res. Part A 77A (2006) 747-762.
[17] X. Li, Y. Deng, X. Chen, Y. Xiao, Y. Fan, X. Zhang, Mater. Lett. 185(2016) 428-431.
[18] Y. Tang, J. Wang, Q. Cao, F. Chen, M. Wang, Y. Wu, X. Chen, X. Zhu, X. Zhang, Biomater. Adv. 140(2022) 213030.
[19] X. Li, T. Song, X. Chen, M. Wang, X. Yang, Y. Xiao, X. Zhang, ACS Appl. Mater. Interfaces 11 (2019) 3722-3736.
[20] J. Wang, Y. Tang, Q. Cao, Y. Wu, Y. Wang, B. Yuan, X. Li, Y. Zhou, X. Chen, X. Zhu, C. Tu, X. Zhang, Regen. Biomater. 9 (2022) rbac005.
[21] B. Zhang, X. Pei, P. Song, H. Sun, H. Li, Y. Fan, Q. Jiang, C. Zhou, X. Zhang, Compos. Pt. B-Eng. 155(2018) 112-121.
[22] J. Wang, Y. Chen, X. Zhu, T. Yuan, Y. Tan, Y. Fan, X. Zhang, J. Biomed. Mater. Res. Part A 102 (2014) 4234-4243.
[23] S. Goenka, R. Balu, T.S.S.Kumar, J. Mech. Behav. Biomed. Mater. 7(2012) 69-76.
[24] S.V. Dorozhkin, Acta Biomater. 6(2010) 715-734.
[25] S.C. Liou, S.Y. Chen, H.Y. Lee, J.S. Bow, Biomaterials 25 (2004) 189-196.
[26] M.A. Brennan, D.S. Monahan, B. Brulin, S. Gallinetti, P. Humbert, C. Tringides, C. Canal, M.P. Ginebra, P. Layrolle, Acta Biomater. 135(2021) 689-704.
[27] B. Li, X. Chen, B. Guo, X. Wang, H. Fan, X. Zhang, Acta Biomater. 5(2009) 134-143.
[28] A. Barba, A. Diez-Escudero, Y. Maazouz, K. Rappe, M. Espanol, E.B. Montufar, M. Bonany, J.M. Sadowska, J. Guillem-Marti, C. Ohman-Magi, C. Persson, M.C. Manzanares, J. Franch, M.P. Ginebra, ACS Appl. Mater. Interfaces 9 (2017) 41722-41736.
[29] A. Barba, A. Diez-Escudero, M. Espanol, M. Bonany, J.M. Sadowska, J. Guillem- Marti, C. Ohman-Magi, C. Persson, M.C. Manzanares, J. Franch, M.P. Ginebra, ACS Appl. Mater. Interfaces 11 (2019) 8818-8830.
[30] P. Kasten, R. Luginbuhl, M. van Griensven, T. Barkhausen, C. Krettek, M. Bohner, U. Bosch, Biomaterials 24 (2003) 2593-2603.
[31] M. Alizadeh-Osgouei, Y. Li, C. Wen, Bioact. Mater. 4(2019) 22-36.
[32] M.A .A. Ansari, A .A. Golebiowska, M. Dash, P. Kumar, P.K. Jain, S.P. Nukavarapu, S. Ramakrishna, H.S. Nanda, Biomater. Sci. 10(2022) 2789-2816.
[33] C. Wang, W. Huang, Y. Zhou, L. He, Z. He, Z. Chen, X. He, S. Tian, J. Liao, B. Lu, Y. Wei, M. Wang, Bioact. Mater. 5(2020) 82-91.
[34] X. Pei, L. Ma, B. Zhang, J. Sun, Y. Sun, Y. Fan, Z. Gou, C. Zhou, X. Zhang, Biofabrication 9 (2017) 045008.
[35] Y. Hong, H. Fan, B. Li, B. Guo, M. Liu, X. Zhang, Mater. Sci. Eng. R-Rep. 70(2010) 225-242.
[36] A. Barba, Y. Maazouz, A. Diez-Escudero, K. Rappe, M. Espanol, E.B. Montufar, C. Öhman-Mägi, C. Persson, P. Fontecha, M.C. Manzanares, J. Franch, M.P. Ginebra, Acta Biomater. 79(2018) 135-147.
[37] X. Chen, M. Wang, F. Chen, J. Wang, X. Li, J. Liang, Y. Fan, Y. Xiao, X. Zhang, Acta Biomater. 103(2020) 318-332.
[38] S. Hendrikx, C. Kascholke, T. Flath, D. Schumann, M. Gressenbuch, F.P. Schulze, M.C. Hacker, M. Schulz-Siegmund, Acta Biomater. 35(2016) 318-329.
[39] J.W. Cai, B.C. Zhang, M.H. Zhang, Y.J. Wen, X.H. Qu, J. Cent. South.Univ. 28(2021) 983-1002.
[40] P. Habibovic, H. Yuan, C.M. van der Valk, G. Meijer, C.A. van Blitterswijk, K. de Groot, Biomaterials 26 (2005) 3565-3575.
[41] Zhou. Changchun, Li. Xiangfeng, Cheng. Junqiu, Fan. Hongsong, Zhang. Xingdong, Bioactive Ceramics and Metals for Regenerative Engineering, Regenerative Engineering, CRC Press, Boca Raton, USA, 2018, pp.32-46.
[42] K. Hurle, J. Neubauer, M. Bohner, N. Doebelin, F. Goetz-Neunhoeffer, Acta Biomater. 10(2014) 3931-3941.
[43] S.V. Dorozhkin, M. Epple, Angew. Chem. Int. Ed. 41(2002) 3130-3146.
[44] K. Ishikawa, P. Ducheyne, S. Radin, J. Mater. Sci.Mater. Med. 4(1993) 165-168.
[45] L. Lu, A.G. Mikos, MRS Bull. 21(1996) 28-32.
[46] X.P. Tan, Y.J. Tan, C.S.L.Chow, S.B. Tor, W.Y. Yeong, Mater. Sci. Eng. C-Mater. Biol. Appl. 76(2017) 1328-1343.
[47] J.M. Sadowska, F. Wei, J. Guo, J. Guillem-Marti, M.P. Ginebra, Y. Xiao, Biomaterials 181 (2018) 318-332.
[48] D.A. Puleo, A. Nanci, Biomaterials 20 (1999) 2311-2321.
[49] C. Feng, Y. Wu, Q. Li, T. He, Q. Cao, X. Li, Y. Xiao, J. Lin, X. Zhu, X. Zhang, Adv. Funct. Mater. 32(2022) 2204974.
[50] X.D. Zhu, H.J. Zhang, H.S. Fan, W. Li, X.D. Zhang, Acta Biomater. 6(2010) 1536-1541.
[51] X.D. Zhu, H.S. Fan, Y.M. Xiao, D.X. Li, H.J. Zhang, T. Luxbacher, X.D. Zhang, Acta Biomater. 5(2009) 1311-1318.
[52] L. Stipniece, V. Stepanova, I. Narkevica, K. Salma-Ancane, A.R. Boyd, J. Eur. Ceram.Soc. 38(2018) 761-768.
[53] J. Wang, Y. Su, L. Xu, D. Li, Ceram. Int. 46(2020) 4801-4812.
[54] F. Tamimi, J. Torres, K. Al-Abedalla, E. Lopez-Cabarcos, M.H. Alkhraisat, D.C. Bassett, U. Gbureck, J.E. Barralet, Biomaterials 35 (2014) 5436-5445.
[55] S. Cazalbou, D. Eichert, X. Ranz, C. Drouet, C. Combes, M.F. Harmand, C. Rey, J. Mater. Sci.: Mater. Med. 16(2005) 405-409.
[56] J.M. Sadowska, J. Guillem-Marti, E.B. Montufar, M. Espanol, M.P. Ginebra, Tissue Eng. Part A 23 (2017) 1297-1309.
[57] J.M. Sadowska, J. Guillem-Marti, M. Espanol, C. Stähli, N. Döbelin, M.P. Ginebra, Acta Biomater. 76(2018) 319-332.
[58] O. Chan, M.J. Coathup, A. Nesbitt, C.Y. Ho, K.A. Hing, T. Buckland, C. Campion, G.W. Blunn, Acta Biomater. 8(2012) 2788-2794.
[59] J. Zhang, X. Luo, D. Barbieri, A.M.C.Barradas, J.D.de Bruijn, C.A. van Blitterswijk, H. Yuan, Acta Biomater. 10(2014) 3254-3263.
[60] Y.C. Chai, S.J. Roberts, J. Schrooten, F.P. Luyten, Tissue Eng. Part A 17 (2011) 1083-1097.
[61] M. Kroeger, Trends Biotechnol. 24(2006) 343-346.
[62] A. Salerno, D. Guarnieri, M. Iannone, S. Zeppetelli, P.A. Netti, Tissue Eng. Part A 16 (2010) 2661-2673.
[63] H. Yuan, Z. Yang, Y. Li, X. Zhang, J.D.De Bruijn, K.De Groot, J. Mater. Sci.: Mater. Med. 9(1998) 723-726.
[64] L.E. Rustom, T. Boudou, B.W. Nemke, Y. Lu, D.J. Hoelzle, M.D. Markel, C. Picart, A.J.Wagoner Johnson, ACS Biomater. Sci. Eng. 3(2017) 2768-2778.
[65] L.E. Rustom, T. Boudou, S. Lou, I. Pignot-Paintrand, B.W. Nemke, Y. Lu, M.D. Markel, C. Picart, A.J.Wagoner Johnson, Acta Biomater. 44(2016) 144-154.
[66] L.E. Rustom, M.J. Poellmann, A.J.Wagoner Johnson, Acta Biomater. 83(2019) 435-455.
[67] S.J. Polak, L.E. Rustom, G.M. Genin, M. Talcott, A.J.Wagoner Johnson, Acta Biomater. 9(2013) 7977-7986 .