In vitro Evaluation of a 20% Bioglass-Containing 3D printable PLA Composite for Bone Tissue Engineering
DOI:
https://doi.org/10.18063/ijb.v8i4.602Keywords:
Bone tissue engineering, Composite, Polylactic acid, Bioglass, Osteoconductive, OsteoinductiveAbstract
Three-dimensional (3D) printing is considered a key technology in the production of customized scaffolds for bone tissue engineering. In a previous work, we developed a 3D printable, osteoconductive, hierarchical organized scaffold system. The scaffold material should be osteoinductive. Polylactic acid (PLA) (polymer)/Bioglass (BG) (mineral/ion source) composite materials are promising. Previous studies of PLA/BG composites never exceed BG fractions of 10%, as increase of bioactive BG component negatively affects the printability of the composite material. Here, we test a novel, 3D printable PLA/ BG composite with BG fractions up to 20% for its biological activity in vitro. PLA/BG filaments suitable for microstructure 3D printing were spun and the effect of different BG contents (5%, 10%, and 20%) in this material on mesenchymal stem cell (MSC) activity was tested in vitro. Our results showed that all tested composites are biocompatible. MSC cell adherence and metabolic activity increase with increasing BG content. The presence of BG component in scaffold has only slight effect on osteogenic gene expression, but it has significant suppressive effect on the expression of inflammatory genes in MSC. In addition, the material did not provoke any significant inflammatory response in whole-blood stimulation assay. The results show that by increasing the BG content, the bioactivity can be further enhanced.
References
Söhling N, Neijhoft J, Nienhaus V, et al., 2020, 3D-Printing of Hierarchically Designed and Osteoconductive Bone Tissue Engineering Scaffolds. Materials (Basel), 13:1836. https://doi.org/10.3390/ma13081836
Grémare A, Guduric V, Bareille R, et al., 2018, Characterization of Printed PLA Scaffolds for Bone Tissue Engineering. J Biomed Mater Res Part A, 106:887–94. https://doi.org/10.1002/jbm.a.36289
Lam CX, Hutmacher DW, Schantz JT, et al., 2009, Evaluation of Polycaprolactone Scaffold Degradation for 6 Months In Vitro and In Vivo. J Biomed Mater Res A, 90:906–19. https://doi.org/10.1002/jbm.a.32052
Shrivats AR, McDermott MC, Hollinger JO, 2014, Bone Tissue Engineering: State of the Union. Drug Discov Today, 19:781–6. https://doi.org/10.1016/j.drudis.2014.04.010
Mehrpouya M, Vahabi H, Barletta M, et al., 2021, Additive Manufacturing of Polyhydroxyalkanoates (PHAs) Biopolymers: Materials, Printing Techniques, and Applications. Mater Sci Eng C, 127:112216. https://doi.org/10.1016/j.msec.2021.112216
Kalia VC, Singh Patel SK, Shanmugam R, et al., 2021, Polyhydroxyalkanoates: Trends and Advances Toward Biotechnological Applications. Bioresour Technol, 326:124737. https://doi.org/10.1016/j.biortech.2021.124737
Eldesoqi K, Seebach C, Ngoc CN, et al., 2013, High Calcium Bioglass Enhances Differentiation and Survival of Endothelial Progenitor Cells, Inducing Early Vascularization in Critical Size Bone Defects. PLoS One, 8:e79058. https://doi.org/10.1371/journal.pone.0079058
Zhang J, Liu W, Schnitzler V, et al., 2014, Calcium Phosphate Cements for Bone Substitution: Chemistry, Handling and Mechanical Properties. Acta Biomater, 10:1035–49. https://doi.org/10.1016/j.actbio.2013.11.001
Henriksen SS, Ding M, Juhl MV, et al., 2011, Mechanical Strength of Ceramic Scaffolds Reinforced with Biopolymers is Comparable to that of Human Bone. J Mater Sci Mater Med, 22:1111–8. https://doi.org/10.1007/s10856-011-4290-y
Wu C, Fan W, Zhou Y, et al., 2012, 3D-Printing of Highly Uniform CaSiO3 Ceramic Scaffolds: Preparation, Characterization and In Vivo Osteogenesis. J Mater Chem, 22:12288–95. https://doi.org/10.1039/c2jm30566f
Trombetta R, Inzana JA, Schwarz EM, et al., 2017, 3D Printing of Calcium Phosphate Ceramics for Bone Tissue Engineering and Drug Delivery. Ann Biomed Eng, 45:23–44. https://doi.org/10.1007/s10439-016-1678-3
Hwang KS, Choi JW, Kim JH, et al., 2017, Comparative Efficacies of Collagen-Based 3D Printed PCL/PLGA/β-TCP Composite Block Bone Grafts and Biphasic Calcium Phosphate Bone Substitute for Bone Regeneration. Materials (Basel), 10:421. https://doi.org/10.3390/ma10040421
Pei F, Ping W, Chengde G, et al., 2018, A Multimaterial Scaffold with Tunable Properties: Toward Bone Tissue Repair. Adv Sci, 5:1700817. https://doi.org/10.1002/advs.201700817
Nyberg E, Rindone A, Dorafshar A, et al., 2017, Comparison of 3D-Printed Poly-ϵ-Caprolactone Scaffolds Functionalized with Tricalcium Phosphate, Hydroxyapatite, Bio-Oss, or Decellularized Bone Matrix. Tissue Eng Part A, 23:503–14. https://doi.org/10.1089/ten.tea.2016.0418
Scaffaro R, Lopresti F, Botta L, et al., 2016, Integration of PCL and PLA in a Monolithic Porous Scaffold for Interface Tissue Engineering. J Mech Behav Biomed Mater, 63:303–13. https://doi.org/10.1016/j.jmbbm.2016.06.021
Poh PS, Chhaya MP, Wunner FM, et al., 2016, Polylactides in Additive Biomanufacturing. Adv Drug Deliv Rev, 107:228–46. https://doi.org/10.1016/j.addr.2016.07.006
Ojansivu M, Wang X, Hyväri L, et al., 2018, Bioactive Glass Induced Osteogenic Differentiation of Human Adipose Stem Cells is Dependent on Cell Attachment Mechanism and Mitogen-Activated Protein Kinases. Eur Cells Mater, 35:54–72. https://doi.org/10.22203/eCM.v035a05
El-Rashidy AA, Roether JA, Harhaus L, et al., 2017, Regenerating Bone with Bioactive Glass Scaffolds: A Review of In Vivo Studies in Bone Defect Models. Acta Biomater, 62:1–28. https://doi.org/10.1016/j.actbio.2017.08.030
Popa AC, Stan GE, Husanu MA, et al., 2017, Bioglass Implant-Coating Interactions in Synthetic Physiological Fluids with Varying Degrees of Biomimicry. Int J Nanomedicine, 12:683–707. https://doi.org/10.2147/IJN.S123236
Saboori A, Rabiee M, Moztarzadeh F, et al., 2009, Synthesis, Characterization and In Vitro Bioactivity of Sol-Gel-Derived SiO2-CaO-P2O5-MgO Bioglass. Mater Sci Eng C, 29:335–40. https://doi.org/10.1016/j.msec.2008.07.004
Westhauser F, Karadjian M, Essers C, et al., 2019, Osteogenic Differentiation of Mesenchymal Stem Cells is Enhanced in a 45S5-Supplemented β-TCP Composite Scaffold: An In-Vitro Comparison of Vitoss and Vitoss BA. PLoS One, 14:1–18. https://doi.org/10.1371/journal.pone.0212799
Al Malat T, Glombitza M, Dahmen J, et al., 2018, The Use of Bioactive Glass S53P4 as Bone Graft Substitute in the Treatment of Chronic Osteomyelitis and Infected Non-Unions a Retrospective Study of 50 Patients Anwendung von Bioglas S53P4 als Knochenersatzmaterial Bei Chronischer Osteomyelitis Und Infe. Z Orthop Unfall, 156:152–159. https://doi.org/10.1055/s-0043-124377
Drago L, Toscano M, Bottagisio M, 2018, Recent Evidence on Bioactive Glass Antimicrobial and Antibiofilm Activity: A Mini-Review. Materials (Basel), 11:1–11. https://doi.org/10.3390/ma11020326
Lyyra I, Leino K, Hukka T, et al., 2021, Impact of Glass Composition on Hydrolytic Degradation of Polylactide/Bioactive Glass Composites. Materials (Basel), 14:1–20. https://doi.org/10.3390/ma14030667
Ng WL, Chua CK, Shen YF, 2019, Print Me An Organ! Why We Are Not There Yet. Prog Polym Sci, 97:101145. https://doi.org/10.1016/j.progpolymsci.2019.101145
Yang Y, Wang G, Liang H, et al., 2019, Additive Manufacturing of Bone Scaffolds. Int J Bioprint, 5:1–25. https://doi.org/10.18063/IJB.v5i1.148
Qu H, 2020, Additive Manufacturing for Bone Tissue Engineering Scaffolds. Mater Today Commun, 24:101024. https://doi.org/10.1016/j.mtcomm.2020.101024
Vergnol G, Ginsac N, Rivory P, et al., 2016, In Vitro and In Vivo Evaluation of a Polylactic Acid-Bioactive Glass Composite for Bone Fixation Devices. J Biomed Mater Res B Appl Biomater, 104:180–91. https://doi.org/10.1002/jbm.b.33364
Maquet V, Boccaccini AR, Pravata L, et al., 2004, Porous poly(α-hydroxyacid)/Bioglass® Composite Scaffolds for Bone Tissue Engineering. I: Preparation and In Vitro Characterisation. Biomaterials, 25:4185–94. https://doi.org/10.1016/j.biomaterials.2003.10.082
Estrada SA, Armendáriz IO, García AT, et al., 2017, Evaluation of In Vitro Bioactivity of 45S5 Bioactive Glass/Poly Lactic Acid Scaffolds Produced by 3D Printing. Int J Compos Mater, 7:144–9. https://doi.org/10.5923/j.cmaterials.20170705.03
Alksne M, Kalvaityte M, Simoliunas E, et al., 2020, In Vitro Comparison of 3d Printed Polylactic Acid/Hydroxyapatite and Polylactic Acid/Bioglass Composite Scaffolds: Insights Into Materials for Bone Regeneration. J Mech Behav Biomed Mater, 104:103641. https://doi.org/10.1016/j.jmbbm.2020.103641
Distler T, Fournier N, Grünewald A, et al., 2020, Polymer-Bioactive Glass Composite Filaments for 3D Scaffold Manufacturing by Fused Deposition Modeling: Fabrication and Characterization. Front Bioeng Biotechnol, 8:1–17. https://doi.org/10.3389/fbioe.2020.00552
Ginsac N, Chenal JM, Meille S, et al., 2011, Crystallization Processes at the Surface of Polylactic Acid-Bioactive Glass Composites During Immersion in Simulated Body Fluid. J Biomed Mater Res Part B Appl Biomater, 99B:412–9. https://doi.org/10.1002/jbm.b.31913
Henrich D, Verboket RR, Schaible A, et al., 2015, Characterization of Bone Marrow Mononuclear Cells on Biomaterials for Bone Tissue Engineering In Vitro. Biomed Res Int, 2015:762407. https://doi.org/10.1155/2015/762407
Seebach C, Henrich D, Tewksbury R, et al., 2007, Number and Proliferative Capacity of Human Mesenchymal Stem Cells are Modulated Positively in Multiple Trauma Patients and Negatively in Atrophic Nonunions. Calcif Tissue Int, 80:294–300. https://doi.org/10.1007/s00223-007-9020-6
Oliveira KM, Leppik L, Keswani K, et al., 2020, Electrical Stimulation Decreases Dental Pulp Stem Cell Osteo-/Odontogenic Differentiation. Biores Open Access, 9:162–73. https://doi.org/10.1089/biores.2020.0002
Schätzlein E, Kicker C, Söhling N, et al., (2022) 3D-printed PLA bioglass scaffolds with controllable calcium release and MSC adhesion for bone tissue engineering. Polymers, 14:2389. https://doi.org/10.3390/polym14122389
Yang J, Shi G, Bei J, et al., 2002, Fabrication and Surface Modification of Macroporous Poly(L-Lactic Acid) and Poly(LLactic-Co-Glycolic Acid) (70/30) Cell Scaffolds for Human Skin Fibroblast Cell Culture. J Biomed Mater Res, 62:438–46. https://doi.org/10.1002/jbm.10318
Johari N, Fathi MH, Golozar MA, et al., 2012, Poly(ECaprolactone)/Nano Fluoridated Hydroxyapatite Scaffolds for Bone Tissue Engineering: In Vitro Degradation and Biocompatibility Study. J Mater Sci Mater Med, 23:763–70. https://doi.org/10.1007/s10856-011-4528-8
Livak KJ, Schmittgen TD, 2001, Analysis of Relative Gene Expression Data Using Real- Time Quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods, 408:402–8. https://doi.org/10.1006/meth.2001.1262
Wutzler S, Maier M, Lehnert M, et al., 2009, Suppression and Recovery of Lps-Stimulated Monocyte Activity after Trauma is Correlated with Increasing Injury Severity: A Prospective Clinical Study. J Trauma, 66:1273–80. https://doi.org/10.1097/TA.0b013e3181968054
Eldesoqi K, Henrich D, El-Kady AM, et al., 2014, Safety Evaluation of a Bioglass-Polylactic Acid Composite Scaffold Seeded with Progenitor Cells in a Rat Skull Critical-Size Bone Defect. PLoS One, 9:e87642. https://doi.org/10.1371/journal.pone.0087642
Azizi L, Turkki P, Huynh N, et al., 2021, Surface Modification of Bioactive Glass Promotes Cell Attachment and Spreading. ACS Omega, 6:22635–42. https://doi.org/10.1021/acsomega.1c02669
Vissers CA, Harvestine JN, Leach JK, 2015, Pore Size Regulates Mesenchymal Stem Cell Response to Bioglass-Loaded Composite Scaffolds. J Mater Chem B, 3:8650–8. https://doi.org/10.1039/c5tb00947b
Aguirre A, González A, Navarro M, et al., 2012, Control of Microenvironmental Cues with a Smart Biomaterial Composite Promotes Endothelial Progenitor Cell Angiogenesis. Eur Cells Mater, 24:90–106. https://doi.org/10.22203/eCM.v024a07
Kulterer B, Friedl G, Jandrositz A, et al., 2007, Gene Expression Profiling of Human Mesenchymal Stem Cells Derived from Bone Marrow During Expansion and Osteoblast Differentiation. BMC Genomics, 8:1–15. https://doi.org/10.1186/1471-2164-8-70
Schroeder TM, Jensen ED, Westendorf JJ, 2005, Runx2: A Master Organizer of Gene Transcription in Developing and Maturing Osteoblasts. Birth Defects Res Part C Embryo Today Rev, 75:213–25. https://doi.org/10.1002/bdrc.20043
Li X, Yi W, Jin A, et al., 2015, Effects of Sequentially Released BMP-2 and BMP-7 from PELA Microcapsule-Based Scaffolds on the Bone Regeneration. Am J Transl Res, 7:1417–28.
Bouyer M, Guillot R, Lavaud J, et al., 2016, Surface Delivery of Tunable Doses of Bmp-2 from an Adaptable Polymeric Scaffold Induces Volumetric Bone Regeneration. Biomaterials, 104:168–81. https://doi.org/10.1016/j.biomaterials.2016.06.001
Yan H, Wu M, Yuan Y, et al., 2014, Priming of Toll-like Receptor 4 Pathway In Mesenchymal Stem Cells Increases Expression of B Cell Activating Factor. Biochem Biophys Res Commun, 448:212–17. https://doi.org/10.1016/j.bbrc.2014.04.097
Dorronsoro A, Lang V, Ferrin I, et al., 2020, Intracellular Role of IL-6 in Mesenchymal Stromal Cell Immunosuppression and Proliferation. Sci Rep, 10:1–12. https://doi.org/10.1038/s41598-020-78864-4
Pricola KL, Kuhn NZ, Haleem-Smith H, et al., 2009, Interleukin-6 Maintains Bone Marrow-Derived Mesenchymal Stem Cell Stemness by an ERK1/2-Dependent Mechanism. J Cell Biochem, 108:577–88. https://doi.org/10.1002/jcb.22289
Semba T, Sammons R, Wang X, et al., 2020, Concise Review: JNK Signaling in Stem Cell Self-Renewal and Differentiation. Int J Mol Sci, 21:1–19. https://doi.org/10.3390/ijms21072613
Yue C, Guo Z, Luo Y, et al., 2020, C-Jun Overexpression Accelerates Wound Healing in Diabetic Rats by Human Umbilical Cord-Derived Mesenchymal Stem Cells. Stem Cells Int, 2020:7430968. https://doi.org/10.1155/2020/7430968
Cook GW, Benton MG, Akerley W, et al., 2020, Structural Variation and its Potential Impact on Genome Instability: Novel Discoveries in the EGFR Landscape by Long-Read Sequencing. PLoS One, 15:e0226340. https://doi.org/10.1371/journal.pone.0226340
Deng W, Chen H, Su H, et al., 2020, IL6 Receptor Facilitates Adipogenesis Differentiation of Human Mesenchymal Stem Cells Through Activating P38 Pathway. Int J Stem Cells, 13:142–50. https://doi.org/10.15283/ijsc19073
Anderson JM, Rodriguez A, Chang DT, 2008, Foreign Body Reaction to Biomaterials. Semin Immunol, 20:86–100. https://doi.org/10.1016/j.smim.2007.11.004
Henrich D, Seebach C, Nau C, et al., 2016, Establishment and Characterization of the Masquelet Induced Membrane Technique in a Rat Femur Critical-Sized Defect Model. J Tissue Eng Regen Med, 10:E382–96. https://doi.org/10.1002/term.1826
Al-Maawi S, Wang X, Sader R, et al., 2021, Multinucleated Giant Cells Induced by a Silk Fibroin Construct Express Proinflammatory Agents: An Immunohistological Study. Materials (Basel), 14:1–19. https://doi.org/10.3390/ma14144038
Söhling N, Ondreka M, Kontradowitz K, et al., 2022, Early Immune Response in Foreign Body Reaction is Im-Plant/Material Specific. Materials (Basel), 15:2195. https://doi.org/10.3390/ma15062195
Persson T, Monsef N, Andersson P, et al., 2003, Expression of the Neutrophil-Activating CXC Chemokine ENA-78/CXCL5 by Human Eosinophils. Clin Exp Allergy, 33:531–7. https://doi.org/10.1046/j.1365-2222.2003.01609.x
Lorenzo J, 2016, The Effects of Immune Cell Products (Cytokines and Hematopoietic Cell Growth Factors) on Bone Cells. In: Osteoimmunology. Cambridge, Massachusetts: Academic Press. p143–67. https://doi.org/10.1016/B978-0-12-800571-2.00009-8
Harada A, Sekido N, Akahoshi T, et al., 1994, Essential Involvement of Interleukin-8 (IL-8) in Acute Inflammation. J Leukoc Biol, 56:559–64. https://doi.org/10.1002/jlb.56.5.559
Hehlgans T, Pfeffer K, 2005, The Intriguing Biology of the Tumour Necrosis Factor/Tumour Necrosis Factor Receptor Superfamily: Players, Rules and the Games. Immunology, 115:1–20. https://doi.org/10.1111/j.1365-2567.2005.02143.x
Rossi DL, Vicari AP, Franz-Bacon K, et al., 1997, Identification Through Bioinformatics of Two New Macrophage Proinflammatory Human Chemokines: MIP-3alpha and MIP-3beta. J Immunol, 158:1033–6.
Downloads
Published
Issue
Section
License
Copyright (c) 2022 Nicolas Söhling, Shahed Al Zoghool, Eva Schätzlein, Jonas Neijhoft, Karla Mychellyne Costa Oliveira, Liudmila Leppik, Ulrike Ritz, Edgar Dörsam, Johannes Frank, Ingo Marzi, Andreas Blaeser, Dirk Henrich
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
