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Frontiers of Materials Science

ISSN 2095-025X

ISSN 2095-0268(Online)

CN 11-5985/TB

Postal Subscription Code 80-974

2018 Impact Factor: 1.701

Front. Mater. Sci.    2023, Vol. 17 Issue (2) : 230644    https://doi.org/10.1007/s11706-023-0644-x
REVIEW ARTICLE
Three-dimensional printing of biomaterials for bone tissue engineering: a review
Ahmed El-Fiqi()
Glass Research Department, Advanced Materials Technology and Mineral Resources Research Institute, National Research Centre, Cairo 12622, Egypt
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Abstract

Processing biomaterials into porous scaffolds for bone tissue engineering is a critical and a key step in defining and controlling their physicochemical, mechanical, and biological properties. Biomaterials such as polymers are commonly processed into porous scaffolds using conventional processing techniques, e.g., salt leaching. However, these traditional techniques have shown unavoidable limitations and several shortcomings. For instance, tissue-engineered porous scaffolds with a complex three-dimensional (3D) geometric architecture mimicking the complexity of the extracellular matrix of native tissues and with the ability to fit into irregular tissue defects cannot be produced using the conventional processing techniques. 3D printing has recently emerged as an advanced processing technology that enables the processing of biomaterials into 3D porous scaffolds with highly complex architectures and tunable shapes to precisely fit into irregular and complex tissue defects. 3D printing provides computer-based layer-by-layer additive manufacturing processes of highly precise and complex 3D structures with well-defined porosity and controlled mechanical properties in a highly reproducible manner. Furthermore, 3D printing technology provides an accurate patient-specific tissue defect model and enables the fabrication of a patient-specific tissue-engineered porous scaffold with pre-customized properties.

Keywords 3D printing      biomaterial ink      printability      3D printing technique      3D printed scaffold      bone tissue engineering     
Corresponding Author(s): Ahmed El-Fiqi   
Issue Date: 26 April 2023
 Cite this article:   
Ahmed El-Fiqi. Three-dimensional printing of biomaterials for bone tissue engineering: a review[J]. Front. Mater. Sci., 2023, 17(2): 230644.
 URL:  
https://academic.hep.com.cn/foms/EN/10.1007/s11706-023-0644-x
https://academic.hep.com.cn/foms/EN/Y2023/V17/I2/230644
Fig.1  (a)(b)(c) Illustrative diagrams for representative conventional techniques used in biomaterials processing into porous scaffolds. Reproduced with permission from Ref. [22] (Copyright 2021, Frontiers). (d) Schematic illustrations of electrospinning technique (this technique is limited to production of fibrous mats, i.e., 2D fibrous scaffolds, with limited thickness [23]). Reproduced with permission from Ref. [24] (Copyright 2017, John Wiley and Sons).
Manufacturing methodBenefitsPotential limitations
Solvent casting/particulate leachingRelatively simple technique that allows creation of scaffolds with regular porosity, controlled composition and pore size? Use of organic solvents precludes cells and biomolecules being included directly in scaffolds
? Difficult to control pore shape and interconnectivity
? Limited thickness of structures and mechanical properties achievable
Gas foamingEliminates use of chemical solvents? High pressures involved prohibits inclusion of cells and bioactive molecules directly into scaffolds
? Temperature labile materials may be denatured during compression moulding step
? Difficult to control pore sizes and ensure interconnectivity
Emulsification freeze-dryingDoes not require use of solid porogen? Requires use of organic solvents
? Small pore size
? Porosity often irregular
? Long processing time
Phase separation? Eliminates leaching step of porogen? Small pore sizes limit its use
? Can be combined with other techniques easily? Use of organic solvents inhibits use of bioactive molecules or cells during scaffold fabrication
Electrospinning? Creates scaffolds with large surface area for cell attachment? Organic solvents may be required, which can be harmful to cells
? Limited mechanical properties
? Simple and inexpensive technique? Difficult to incorporate precise microarchitecture into constructs
Tab.1  Limitations of conventional processing techniques (reproduced with permission from Ref. [12], Copyright 2018, Elsevier)
Fig.2  Conventional processing routes used for the production of porous ceramic scaffolds: (a) replica; (b) sacrificial template; (c) direct foaming. Reproduced with permission from Ref. [41] (Copyright 2006, John Wiley and Sons).
TechniqueAdvantagesDisadvantages
Foaming methods (general)Allows manufacturing of both closed and open-cell foams; good versatility of final part shapes, as the solution can be cast in molds without additional machiningDifficulty in achieving high interconnectivity; non-porous external surface
H2O2 foamingSimpleLow porosity control, laminar pore structure with poor 3D interconnection
Sol-gel foamingHierarchical structure can be obtained (macroporous scaffold combined with ordered mesoporous texture)Need for a high degree of control of the foaming step
In situ polymerization of organic monomer (gel-cast foaming)Highly porous ceramic; high-strength properties due to the less flawed structure and dense struts and walls producedLow pore interconnectivity
Organic phase burningout/space holderHigh mechanical strengthDifficult to obtain a homogeneous distribution of pores; poor interconnectivity
Sponge replicationReticulated pen-cell material; applicable to any ceramic material that can be dispersed into a suspension; no toxic chemicals neededMechanical properties might be poor
Tab.2  Comparison of conventional processing methods for bioceramic and bioglass porous scaffolds (reproduced with permission from Ref. [42], Copyright 2015, Frontiers)
Fig.3  Illustrative design of a biomaterial 3D printer (left) (reproduced with permission from Ref. [46], Copyright 2021, John Wiley and Sons). 3D printed porous scaffolds with well controlled porosity and pore size (right) (reproduced with permission from Ref. [47], Copyright 2022, The American Chemical Society).
Fig.4  Exemplary images of 3D printed patient-specific porous scaffolds with the same shape as the bone tissue defects: bioceramic (upper left) (reproduced with permission from Ref. [21], Copyright 2021, Elsevier); biopoymer (upper right) (reproduced with permission from Ref. [8], Copyright 2019, Elsevier); bioglass (lower panel) (reproduced with permission from Ref. [48], Copyright 2012, Elsevier).
Fig.5  3D printing techniques common for processing of biomaterials into 3D printed porous scaffolds. Reproduced with permission from Ref. [49] (Copyright 2020, John Wiley and Sons).
Fig.6  Parameters and variables affecting 3D printing techniques. Reproduced with permission from Ref. [50] (Copyright 2020, Springer Nature).
3D printing techniqueTypical materialResolutionAdvantagesShortcomings
SortMethod
Liquid-based 3D printingStereolithography (SLA)Photo-curable polymer resins50–100 μmHigh resolution, smooth surface of fabricated structuresOver-curing, which can cause overhanging parts, oxygen inhibition, brittle printed products
Digital light projection (DLP)Photo-curable polymer resins10–50 μmHigh printing speed, less affected by oxygen inhibition than SLARequiring low viscosity resins, limited mechanical properties of the printed products
Inkjet printing (IJP)Polymers, hydrogels50–300 μmRelatively high printing speed (up to 10000 drops/s), low cost, allowing printing of bioinks containing living cellsLimited materials in a narrow range of viscosity (3.5–12 mPa·s), unable to fabricate large and complex structures
PolyjetPhoto-curable polymer resins with very low viscosity and high surface tension20 μmHigh resolution, good surface quality of printed structures, relatively high printing speed, allowing fabrication of multi-lateral or multicolor objectsVery limited materials choices, expensive
Filament- or paste-based 3D printingFused deposition modelling (FDM)Polymers and their composites in the filament form100–150 μmRobust, low cost, ability to process a variety of materialsSlow printing speed, relatively low dimension precision, requiring high temperature
3D dispensingPolymers, hydrogels, ceramics, and their composites100 μm to millimetersAbility to process a variety of materials in a wide range of viscosity ((6–30)×107 mPa·s), capable of printing bioinks containing living cellsPrinting nozzle clogging, rough surface of products, relatively low printing resolution
Powder-based 3D printingSelective laser sintering (SLS)Polymer powders, ceramic powders, and composite powders50–100 μmRelatively wide range of powder materials, fabrication of complex structuresRequiring high temperature, rough surface of products, low reusability of non-sintered powders
Selective laser melting (SLM)Polymer powders, ceramic powders, metal powders, and composite powders20–100 μmAbility to process metallic materials, near net-shape fabrication, high material utilizationDifficult to control printing, balling, high residual stress, deformation issues for printed parts
Electron beam melting (EBM)Metal powders100–200 μmHigh power electron energy source, faster printing speed than SLMLower resolution and rougher surface as compared to SLM
3D powder binding (3DPB)Polymer powders, ceramic powders, and their composite power100 μmFast, low cost, allowing fabrication of multicolor objectsLow precision, rough surface, and limited mechanical strength of products
Tab.3  Comparison of different 3D printing techniques (reproduced with permission from Ref. [57], Copyright 2021, AIP Publishing)
Fig.7  (a) Rheological properties affecting printability and shape fidelity. Interplay of rheological properties in extrusion-based printing. Key aspects to assess printability in the context of extrusion- and lithography-based 3D printing technologies. Reproduced with permission from Ref. [61] (Copyright 2020, The American Chemical Society). (b) Illustration of a biomaterial ink flowability through the printing needle (i) and the formation of first printed layer of filament on the substrate (ii), and the vertical fusion of two filament layers within the 3D printed scaffold (iii). Reproduced with permission from Ref. [58] (Copyright 2020, Whioce Publishing).
Fig.8  Hierarchical structural organization of the bone tissue. Reproduced with permission from Ref. [44] (Copyright 2021, Elsevier).
Fig.9  Main approaches and different biomaterials used for treatment of bone tissue defects. Reproduced with permission from Ref. [52] (Copyright 2022, Elsevier).
Fig.10  Schematic illustration of bone tissue engineering approaches (upper panel) (reproduced with permission from Ref. [31], Copyright 2021, John Wiley and Sons). Examples of cells, growth factors, and biomaterial scaffolds used in bone tissue engineering (lower panel) (reproduced with permission from Ref. [65], Copyright 2022, Elsevier).
Scaffold characteristicDesirable features
Biocompatibility? Non-toxic breakdown products
? Non-inflammatory scaffold components, avoiding immune rejection
Biodegradability? Controlled scaffold degradation which can complement tissue ingrowth whilst maintaining sufficient support
? Degradable by host enzymatic or biological processes
? Allows invading host cells to produce their own extracellular matrix
Bioactivity? Scaffold materials that can interact with and bind to host tissue
? Osteoconductive and osteoinductive properties
? Inclusion of biological cues and growth factors to stimulate cell ingrowth, attachment and differentiation
Scaffold architecture? Interconnected pores allowing diffusion and cell migration
? Microporosity to present a large surface area for cell-scaffold interactions
? Macroporosity to allow cell migration and invasion of vasculature
? Pore size tailored to target tissue and cells
? Sufficient porosity to facilitate cell ingrowth without weakening mechanical properties
? Inbuilt vascular channels to enhance angiogenesis in vivo
Mechanical properties? Compressive elastic and fatigue strength comparable to host tissue allowing cell mechano-regulation to occur and structural integrity to remain in vivo
? Scaffold material that can be readily manipulated in the clinical environment to treat individual patient bone defects
Tab.4  General requirements for biomaterials scaffolds (reproduced with permission from Ref. [12], Copyright 2018, Elsevier)
Fig.11  Main steps involved in 3D printing process (upper panel) (reproduced with permission from Ref. [67], Copyright 2022, Elsevier). An illustrative example of 3D printing process for bone tissue engineering (lower panel) (reproduced with permission from Ref. [68], Copyright 2022, John Wiley and Sons).
Manufacturing materialBenefitsPotential limitations
Polymer? Natural polymers can be derived from extracellular matrix, ensuring high biocompatibility and low toxicity? Natural and synthetic polymers generally lack mechanical properties for load bearing
? Biodegradable? Pathological impurities such as endotoxin that may be present in natural polymers
? Often contain biofunctional molecules on their surface
? Synthetic polymers offer improved control over physical properties? Synthetic polymers are often hydrophobic and lack cell recognition sites
Ceramic? Osteoconductive and osteoinductive properties allow strong integration with host tissue? Hard and brittle when used alone
? Similar composition to host bone mineral content? May display inappropriate degradation/resorption rates, with decline in mechanical properties as a result
? Can be delivered as granules, paste or in an injectable format
Bioactive glass? Osteoconductive, osteoinductive properties? Inherent brittleness
? Adapted into clinical prosthesis already? Difficult to tune resorption rate
? Manipulation of constructs into 3D shapes to treat specific defects is challenging
? Potential for release of toxic metal ions
Metal? Biocompatible? Superior modulus can lead to stress-shielding
? Superior strength? Poor biodegradability may result in further surgery/impairment of tissue ingrowth
? Superior mechanical properties can be advantageous in situations where slow bone growth likely? Secondary release of metal ions may cause local and distal toxicity
Tab.5  Advantages and limitations of common types of biomaterials used in scaffolds fabrication for bone tissue engineering (reproduced with permission from Ref. [12], Copyright 2018, Elsevier)
Fig.12  Representative examples of common types of biomaterials used in scaffolds fabrication for bone tissue engineering. Reproduced with permission from Ref. [69]. Copyright 2019, Elsevier.
Fig.13  (a) Percentages of different 3D printing techniques investigated for bone tissue engineering. (b) Percentages of different tissue engineering applications using 3D printing techniques. (c) Comparison of uses of different 3D printing techniques over time in bone tissue engineering. Reproduced with permission from Ref. [50] (Copyright 2020, Springer-Nature).
Fig.14  Images of the stained histological sections of (a)(a?)(a??) PLGA, (b)(b?)(b??) 30%HA/PLGA, and (c)(c?)(c??) 45%HA/PLGA scaffolds, along with quantitative analyses of (d) the scaffold remains area and (e) the new bone area of the three scaffolds after implantation for 4, 12, and 24 weeks. Reproduced with permission from Ref. [123] (Copyright 2022, Elsevier).
Fig.15  (a)(a?)(a??)–(f)(f?)(f??) Magnified images of the edge and central regions of the scaffolds. M represents the scaffold material; B represents the new bone tissue; and the yellow dotted circles indicate the initial defect boundaries. 3D micro-CT reconstructed images of (g) new bone tissue, (h) quantitative analysis of the bone volume/total volume (BV/TV), (i) trabecular number (Tb.N), (j) trabecular separation (Tb.Sp), and (k) trabecular thickness (Tb.Th) of the three scaffolds after implantation for 4, 12, and 24 weeks. Reproduced with permission from Ref. [123] (Copyright 2022, Elsevier).
Fig.16  (a) Making of the defect, (b) CT imaging, (c) design of customized scaffold, and (d) 3D printing. (e)(f)(g)(h) SEM images along with photos of in-vivo implanted scaffolds. Histology results of in-vivo study: (i) red color in lower panel, (j) orange color in lower panel, and (k) blue color indicating the new formed bone. Reproduced with permission from Ref. [114] (Copyright 2019, Elsevier).
Fig.17  Illustrative diagram of PEEK scaffolds fabricated by FDM process along with in-vivo animal model for implantation (upper panel). The in-vivo bone regeneration ability in animal model as revealed by histology (lower panel). Red color refers to newly formed bone tissues. Reproduced with permission from Ref. [45] (Copyright 2020, The American Chemical Society).
Fig.18  Diagram of scaffolds preparation method (upper panel). CAD models and in-vivo study (middle and lower panels). Pink color in histology images refers to new formed bone. Yellow color in μCT reconstructed images (see the lower panel) refers to new formed bone. Reproduced with permission from Ref. [115] (Copyright 2021, Elsevier).
Fig.19  (a)(b) Pictorial diagrams of scaffold fabrication process using suspension-enclosing projection-SLA 3D printing. (c)(d)(e)(f) CAD models and μCT images. (g) Scaffold combination and in-vivo study in an animal model. Reproduced with permission from Ref. [116] (Copyright 2021, The American Chemical Society).
Fig.20  (a) CAD models and images of 3D printed HA scaffolds. (b)(c) Specific 3D printed HA scaffolds for femur and skull bone defects. (d)(e)(f)(g)(h) Mechanical behavior of the 3D printed triply periodic minimum surfaces-structured HA scaffolds. Reproduced with permission from Ref. [117] (Copyright 2022, John Wiley and Sons).
Fig.21  (a) Schematic of in-vivo femur implantation, (b) the compression test, (c) μCT images, and (d) quantitative analysis of new bone formation of the scaffolds after 4, 8, and 12 weeks of implantation. The red color refers to the new bone tissue while the white color indicates the HA scaffold material. Reproduced with permission from Ref. [117] (Copyright 2022, John Wiley and Sons).
Fig.22  Scaffold observations by μCT and SEM imaging (upper panel) along with cumulate mapping of the formed new bone (NB) (lower panel). Reproduced with permission from Ref. [118] (Copyright 2022, Elsevier).
Fig.23  Fabrication process (upper panel) and in-vivo study (middle and lower panels) of wollastonite bioceramic scaffolds. Reproduced with permission from Ref. [119] (Copyright 2022, Elsevier).
Fig.24  Illustrative schematic of (a) the scan process and generating CAD model along with (b) the DLP technique. (c)?(j) Precision testing of DLP including SEM imaging. Reproduced with permission from Ref. [120] (Copyright 2022, The American Chemical Society).
Fig.25  In-vivo animal model and in-vivo analysis of new bone formation using μCT imaging, fluorescent staining and histology. NB denotes new bone. Reproduced with permission from Ref. [120] (Copyright 2022, The American Chemical Society).
Fig.26  (a)–(h) Design and production of PEEK scaffold fabricated using FDM and titanium scaffolds (fabricated by SLM) made specifically for the patient’s maxilla. (i) Finite element models of PEEK and titanium scaffolds. (j)–(r) Design and execution of in-vivo operation. (s) Histological images of implanted PEEK and titanium scaffolds along with control. Pink color refers to new bone. Reproduced with permission from Ref. [53] (Copyright 2022, The American Chemical Society).
Fig.27  A schematic representation of (a) the 3D-printing procedures used to create scaffolds using SLS, along with (b) the macroscopic pictures, (c) design models, (d) μCT images, and (e) SEM images. Reproduced with permission from Ref. [15] (Copyright 2022, Elsevier).
Fig.28  In-vivo animal study of titanium alloy-based scaffolds with a trabecular-like structure (ITS). White: scaffold. Purple: soft tissue. Green, blue, and yellow: new bone tissue. Reproduced with permission from Ref. [15] (Copyright 2022, Elsevier).
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