Table of Contents
Table of Contents ........................................................................................................................................ ii
List of Abbreviations .................................................................................................................................. iii
Introduction ................................................................................................................................................. 1
Significance of Research ............................................................................................................................. 2
Hypothesis ................................................................................................................................................... 3
Objectives .................................................................................................................................................... 3
Proposed methods and tasks ........................................................................................................................ 4
1. Successful fabrication of Zn-based alloy cubes of minimal defects ................................................4
2. Design and fabrication of cubic scaffolds fabricated by SLM .........................................................4
3. Design and fabrication of customized bone scaffolds ......................................................................5
4. Physical and mechanical characterization of customized bone scaffold .........................................5
5. In vitro study 1 and 2 ..........................................................................................................................5
6. In vivo study: evaluation of custom-made scaffold in a critical-sized animal defect model
(optional via collaboration) ................................................................................................................................6
Statistical Analysis ...................................................................................................................................... 6
Research Environment ................................................................................................................................ 6
Research Timeline ....................................................................................................................................... 6
Expected Conclusions ................................................................................................................................. 7
References ................................................................................................................................................... 8
ii
List of Abbreviations
3D Three-dimensional
AM Additive manufacturing
CT Computer tomography
EBM Electron beam machining
ECM Extracellular matrix
Fe Iron
FTIR Fourier transform infrared spectroscopy
Mg Magnesium
MRI Magnetic resonance imaging
SBF Simulated body fluid
SLM Selective laser melting
SEM Scanning electron microscopy
Zn Zinc
iii
Introduction
Bone remodeling is a dynamic process which repeats for thousands of times during a human
lifetime in order to maintain the construction of bone. Briefly, it starts with bone resorption state
and finishes with the formation of new bone [1]. Although this process occurs naturally, it is
different when patients experience large bone defects, for instance, due to injuries or cancers. In
these cases, bone cannot remodel naturally, and so other methods, like bone grafting, are utilized.
It is a procedure, which uses from a bone extracted from elsewhere in order to promote bone
formation. In spite of considering such a method as a gold standard, it has some drawbacks like
donor site morbidity and infection. In this regard, alternative methods have guided researchers
towards tissue engineering [2,3].
Tissue engineering is a multidisciplinary field including the combination of engineering and
life science in order to provide a suitable structural framework to restore, protect, or augment the
tissue functions [4]. From tissue engineering point of view, three-dimensional (3D)
biocompatible scaffolds needed for bone regeneration have to mimic both biological and
mechanical properties of the extracellular matrix (ECM) in order to control the cell function,
diffusive capability and induce bone regeneration [5,6]. Furthermore, high diffusivity is
necessary for 3D scaffolds in order to facilitate the mass transport of oxygen and nutrients [7].
To access this approach, the average pore size of scaffolds should be in the range of 300-400
micron to allow osteons to grow into the scaffold [8]. In this regard, a complete library for tissue
engineering scaffold structure and its features was reported by Cheah et al. [9,10].
There are several techniques applied to fabricate 3D metal scaffolds such as liquid state
processing, solid state processing, electro-deposition and vapor deposition. While the shape and
size of the pores are tunable in these techniques, the mentioned conventional methods can
fabricate randomly organized porous structure [11]. Other modern techniques developed for the
fabrication of 3D scaffolds are additive manufacturing (AM) methods. These techniques are
based on joining materials layer by layer. They have undoubtedly revolutionized the fabrication
of scaffolds by improving the ability to control pore size and interconnectivity [12].
According to working principal, AM processes are divided into three sub-groups of laser-
based (e.g. selective laser melting [13]), printing-based (e.g. 3-D printing [14]) and extrusion-
based (e.g. fused deposition modeling [15]) techniques. The selective laser melting (SLM)
process is able to produce fully dense metallic parts. This technique has revolutionized the
fabrication of novel implants, fabricated by metal powders, which can be utilized in load-bearing
applications [16]. In this project, SLM technique is used in order to fabricate customized bone
scaffolds including novel biodegradable Zn-based alloys. It was reported that Zinc (Zn) is
considered as an appropriate material for bone implants due to its properties. It stimulates bone
fracture healing, reduces bone loss (postmenopausal patients), improves bone mineralization and
skeletal strength [17,18]. Moreover, it was reported that magnesium (Mg) implants degrade fast.
On the other hand, iron (Fe) implants degrade slowly and in this way, Zn is very promising as an
1
alternative solution for biodegradable implants [19].
Significance of Research
As it is mentioned, bone grafting methods have their problems like infection, acute pain,
disease transmission and so on, which have numerous post-surgery complications. From the
clinical point of view, bioactive scaffolds can tackle these issues. For instance, in spite of
autograft method, bone healing does not need patient's tissue donation in this method.
Additionally, these scaffolds can be sterilized in order to reduce the risk of infections, which is
predictable in allograft method. Bioactive scaffolds are more effective economically rather than
using growth factors like bone morphogenetic protein-2 [20]. In addition, in some clinical cases,
a huge amount of bone removal is needed, and so methods like autograft are not possible. In this
regard, AM techniques have gained increasing attention and, they are already successfully
reshaping current supply chains for customized orthopedic implants, minimizing high-costs and
long lead times.
From the material point of view, polymers, ceramics, and metals are utilized in bone tissue
engineering applications like bone defect repair. The strength and toughness required for load-
bearing applications play a decisive role in material selection so that, for instance, polymers are
not good candidates for load-bearing applications [21,22]. Taking ceramics as an outstanding
example, in spite of their suitable biological properties, they are brittle and cannot be dissolved
in the body [23]. To tackle the mentioned issues, researchers have focused on bulk metallic
implants for load-bearing applications. Although titanium scaffolds have been broadly used for
load-bearing applications, once implanted they remain in the body permanently until failure.
While failed, revision surgeries are often needed but with higher risk of failure. Recently, more
efforts have devoted to the fabrication of biodegradable metallic scaffolds made by
biocompatible materials like Zn [24]. Zn is considered vital for biological functions in the body
due to its effects on cellular metabolism [25]. According to the literature review, there are
limited numbers of research studies conducted on Zn alloys as biodegradable implants
[22,26,27]. In order to enhance the strength and plasticity of Zn-based implants, adding alloying
elements is proposed [28]. However, Zn-based alloys are considered as multi-phase metals with
localized corrosion [29]. Hence, the level of added materials to the Zn-based implant should be
investigated carefully. Zn-based biodegradable implants can be categorized in various groups
like Zn-Mg, Zn-Sr, Zn-Fe alloys and so on [28].
Regarding the limitations of gold standard methods like autograft as well as applicable
materials for load-bearing applications, researchers are going toward gaining benefit from AM
techniques to design and fabricate custom-made biodegradable bone scaffolds. These kinds of
scaffolds, fabricated according to magnetic resonance imaging (MRI) or computer tomography
(CT) scan data, are custom-made so that they can be fabricated according to scan data of patients
and used as final products. It can enhance the accuracy and reduce surgery lead-time in
comparison with conventional methods. In this way, from patient customized bone scaffolds to
patient specific implants can be manufactured via this method [30]. Surgeons defined the size of
2
a bone graft based on their experience and shaped it during the surgery, which led to an
inaccurate and time-consuming approach [31–34]. Alberty was in the group of first researchers
who fabricated models via CT scan data in 1979 [35]. Recently, the fabrication of customized
scaffold has a significant importance. As an example, this kind of scaffold was used for
augmentation of an atropic mandibular ridge reported by Mangano et al., recently [36].
Overall, the production of customized scaffolds can potentially provide an alternative to the
current bone grafting methods for bone regeneration with a lower risk of infections. This can
improve the quality of life for patients, and is the ultimate goal of this PhD project. In addition, it
can lead to lower surgery time due to lack of any trial and error during the surgery. It is worth
mentioning that the AM global market is estimated to have a 13.5% growth by 2017 [30]. This
PhD project will thus concentrate on the fabrication of Zn-based alloy scaffolds for customized
load-bearing orthopedic applications.
Hypothesis
It is hypothesized that AM technology together with image processing can be implemented to
fabricate customized bone scaffolds with a novel concept of Zn-based material. As it was
mentioned, Zn-based scaffolds were reported with less than ten publications as a hot topic in the
group of biodegradable metals [28]. In addition, regarding low strength and plasticity of pure Zn,
adding alloying elements can be a good solution. To do so, a design approach is developed to
fabricate customized bone scaffolds, via CT scan data and 3D printing, which can heal bone
injuries as a safe and available reconstruction method rather than bone grafting techniques.
Following objectives indicate the way utilized in order to achieve the main goal of this research.
Objectives
In this PhD project, the following interrelated researches are conducted to confirm the
biocompatibility and biodegradability of scaffolds.
Successful fabrication of Zn-based alloy cubes of minimal defects
Design and fabrication of cubic scaffolds by SLM technique
o Optimization of fabricated scaffolds from geometrical point of view based on
literature review
Design and fabrication of customized bone scaffolds
o Data acquisition and CT scan data processing
o Design and fabrication of unit cell and porous structure
Physical and mechanical characterization of customized bone scaffolds
3
In vitro studies 1 and 2
o Evaluation of the bioactivity of scaffolds in simulated body fluid (SBF)
o Evaluation of cellular responses of cells to customized scaffolds
In vivo study: evaluation of custom-made scaffolds in a critical-sized animal defect
model
Proposed methods and tasks
1. Successful fabrication of Zn-based alloy cubes of minimal defects
An important task is to fabricate Zn alloys with minimal defects. It is obvious that Zn has low
temperature of evaporation, and so its fabrication by SLM or electron beam machining (EBM) is
very challenging. Hence, optimization of SLM processing for Zn alloys is one of the major
components of this PhD project.
In this step, with respect to the literature review, some cubes are developed using SLM. One
of the Zn-based alloys like Zn-Mg, containing up to 10.0 wt. % Mg can be selected and
evaluated in comparison with pure Zn. Mg can enhance mechanical properties of pure Zn
implants regarding the available literature review. In this way, before the fabrication of scaffolds,
optimized cubes can be fabricated by defining appropriate laser power, scan speed, scan strategy,
process atmosphere, process pressure and, other relevant factors. It is worth mentioning that the
optimum machine parameters are defined in this step with the assistance of an optimization
procedure.
2. Design and fabrication of cubic scaffolds fabricated by SLM
In the previous objective, the main goal was the fabrication of simple cubes fabricated by Zn
alloys. It shows its importance due to low temperature of evaporation of Zn. In this step, with the
assistance of the obtained optimized parameters (first objective) for Zn alloys processing,
scaffolds are fabricated. As it was mentioned, Zn is considered as an appropriate biocompatible
metal with good biodegradation rate. This biocompatible metal can be applied for the fabrication
of porous scaffolds for bone tissue engineering applications. In this project, in order to improve
biological properties, surface treatment can also be applied.
In order to evaluate the effects of pore size and shape on the characteristics of scaffolds, some
different groups are fabricated regarding literature review. As an outstanding example, it was
reported that the proper pore size for osteons is 300 to 400 micron [8]. Therefore, in this step, by
considering the required biological requirements, the optimized parameters are determined with
respect to the desired application. For instance, parameters, like spacing between fibers, are
defined so that scaffolds can be fabricated with appropriate biological properties.
4
3. Design and fabrication of customized bone scaffolds
In this step, raw CT scan data is obtained from CT scan radiography of an animal with a
critical-sized bone defect, which is not healed normally. CT scan data acquisition is then
completed by saving in DICOM format and importing into medical software like Mimics. The
CT scan data is then processed to reduce probable noises. The unnecessary parts are removed
and the desired section is saved in STL format as a 3D model. In order to evaluate the accuracy
of the generated STL file, software like CloudCompare can be used to compare STL file with
raw CT scan data. Once the accuracy is confirmed, a unit cell with required porosity and
structure is designed. In the next step, a customized 3D model, which can be fitted into the defect
site, is designed and then, the prepared unit cell is projected onto this 3D model. In this way,
customized scaffold with appropriate porous structure is designed. The unit cell is designed so
that 3D porous scaffold can be fabricated without any support during the printing process. In the
final step, the fabricated scaffold can be treated by a well-chosen heat treatment procedure [11].
It is worth mentioning that the porosity, pore size and, manufacturing constraints of customized
bone scaffold are determined so that it can match bone stiffness.
4. Physical and mechanical characterization of customized bone scaffold
In this step, all of cubic scaffolds as well as customized scaffolds are evaluated. The particle
size distribution of the utilized metallic powders during the SLM printing is determined via
transmission electron microscopy (TEM). The porosity of scaffolds is evaluated using
Archimedes' principle. In addition, the morphology of the scaffolds are evaluated using scanning
electron microscopy (SEM) and SEM images can be imported into open source software like
ImageJ for measurement. In detail, structures of the materials are observed using an optical
metallographic microscope and scanning electron microscope with EDS analyzer. The chemical
composition of scaffold can be analyzed by X-ray fluorescence spectroscopy. The mechanical
properties of scaffolds are determined using compression as well as tensile test. Moreover,
corrosion behavior of should be evaluated by electrochemical tests and Vickers hardness is
determined by a hardness tester.
5. In vitro study 1 and 2
The ability of scaffold in the formation of calcium phosphate-like precipitates on the surfaces
of the fabricated scaffold is evaluated by soaking in SBF solution prepared according to Kokubo
protocol [37]. The formation of calcium phosphate-like precipitates can be confirmed by SEM
images of scaffolds after immersion in SBF for 28 days. In addition, in another study, the
evaluation of cellular responses of cells to customized scaffolds can be done by another in vitro
study.
5
6. In vivo study: evaluation of custom-made scaffold in a critical-sized animal
defect model (optional via collaboration)
In this step, the fabricated customized scaffold can be evaluated by an in vivo study. The
animal with a critical-sized defect, selected for gaining CT scan data, is used for this study. The
customized scaffold designed and fabricated according to the defect site is implanted in the
defect site. After a while, histological analysis can confirm the effect of scaffold on the defect
site. The animal study for the proposed research should be approved by the Animal Research
Ethics Board.
Statistical Analysis
All data are represented as mean ± SD and subjected to one-way analyses of variance (one-
way ANOVA). Significance levels are set at p < 0.05. In all statistical analysis, normality and
equal variance are assumed.
Research Environment
The recently installed SLM 125HL 3D metal printer (SLM Solutions) at Macquarie
University is available to the proposed research. Through the connection of my prospective
supervisor, Dr Wei Xu (holding honorary appointments at the University of Melbourne and
RMIT University), I have the full access to two 3D metal printers (SLM Solutions, SLM250 and
SLM 125) at RMIT and one 3D meter printer (Renishaw AM250) at the University of
Melbourne. Advanced characterization techniques needed for the proposed research are also
available at both Macquarie University and RMIT with full access.
Research Timeline
Step 1: Lab induction and training, literature review, pre-preparation work
Step 2: Design and fabrication of simple cubes
Step 3: Design and fabrication of cubic scaffolds
Step 4: Design and fabrication of customized scaffolds
Step 5: Characterization
Step 6: In vitro study
Step 7: In vivo study
Step 8: Investigation of the obtained results and preparation of draft of papers
Step 9: Preparation of the PhD thesis
6
Gantt chart
Year 1 Year 2 Year 3
Tasks
Q1 Q2 Q3 Q4 Q1 Q2 Q3 Q4 Q1 Q2 Q3 Q4
Lab induction and training,
literature review, pre-preparation
work
Design and fabrication of simple
cubes
Design and fabrication of cubic
scaffolds
Design and fabrication of
customized scaffolds
Characterization
In vitro Study
In vivo study
Investigation of the obtained results
and preparation of draft of papers
Preparation of the PhD thesis
Expected Conclusions
The main aim of this project is to characterize a novel customized scaffolds of Zn-based
alloys developed using a SLM machine. It is expected that these customized scaffolds have an
increase in elastic modulus rather than pure one owing to the presence of materials like Mg,
which can enhance mechanical properties of Zn implants regarding the literature review. In
addition, immersing the customized scaffold in SBF may indicate the significant formation of
hydroxyapatite crystals on the surface of the scaffold owing to the presence of selected materials.
According to expected results, the developed scaffold may be applied as a bioactive bone
substitute. However, evaluating of this scaffold should be explored by performing some in vitro
and in vivo studies as it was discussed.
7
References
[1] J. Ren, K.A. Blackwood, A. Doustgani, P.P. Poh, R. Steck, M.M. Stevens, et al., Melt-
electrospun polycaprolactone strontium-substituted bioactive glass scaffolds for bone
regeneration, J Biomed Mater Res Part A. 102 (2014) 3140–3153.
[2] E. Farré-Guasch, J. Wolff, M.N. Helder, E.A.J.M. Schulten, T. Forouzanfar, J. Klein-
Nulend, Application of Additive Manufacturing in Oral and Maxillofacial Surgery, J Oral
Maxillofac Surg. (2015) 1–11.
[3] C.-Y. Lin, R.M. Schek, A.S. Mistry, X. Shi, A.G. Mikos, P.H. Krebsbach, et al.,
Functional bone engineering using ex vivo gene therapy and topology-optimized,
biodegradable polymer composite scaffolds, Tissue Eng. 11 (2005) 1589–1598.
[4] R. Langer, Perspectives and challenges in tissue engineering and regenerative medicine,
Adv Mater. 21 (2009) 3235–3236.
[5] E. Boccardi, I. V Belova, G.E. Murch, A.R. Boccaccini, T. Fiedler, Oxygen diffusion in
marine-derived tissue engineering scaffolds, J Mater Sci Mater Med. 26 (2015) 1–9.
[6] W. Teo, W. He, S. Ramakrishna, Electrospun scaffold tailored for tissue specific
extracellular matrix, Biotechnol J. 1 (2006) 918–929.
[7] E. Sachlos, J.T. Czernuszka, Making tissue engineering scaffolds work. Review: the
application of solid freeform fabrication technology to the production of tissue
engineering scaffolds, Eur Cell Mater. 5 (2003) 39–40.
[8] M.G. Yeo, G.H. Kim, Preparation and characterization of 3D composite scaffolds based
on rapid-prototyped PCL/β-TCP struts and electrospun PCL coated with collagen and HA
for bone regeneration, Chem Mater. 24 (2011) 903–913.
[9] C.M. Cheah, C.K. Chua, K.F. Leong, S.W. Chua, Development of a tissue engineering
scaffold structure library for rapid prototyping. Part 2: parametric library and assembly
program, Int J Adv Manuf Technol. 21 (2003) 302–312.
[10] C.M. Cheah, C.K. Chua, K.F. Leong, S.W. Chua, Development of a tissue engineering
scaffold structure library for rapid prototyping. Part 1: investigation and classification, Int
J Adv Manuf Technol. 21 (2003) 291–301.
[11] X. Wang, S. Xu, S. Zhou, W. Xu, M. Leary, P. Choong, et al., Topological design and
additive manufacturing of porous metals for bone scaffolds and orthopaedic implants: A
review, Biomaterials. 83 (2016) 127–141.
[12] S.M. Giannitelli, P. Mozetic, M. Trombetta, A. Rainer, Combined additive manufacturing
approaches in tissue engineering, Acta Biomater. 24 (2015) 1–11.
doi:10.1016/[Link].2015.06.032.
[13] S. Bremen, W. Meiners, A. Diatlov, Selective laser melting, Laser Tech J. 9 (2012) 33–38.
8
[14] N.E. Fedorovich, J.R. De Wijn, A.J. Verbout, J. Alblas, W.J.A. Dhert, Three-dimensional
fiber deposition of cell-laden, viable, patterned constructs for bone tissue printing, Tissue
Eng Part A. 14 (2008) 127–133.
[15] J.-T. Schantz, A. Brandwood, D.W. Hutmacher, H.L. Khor, K. Bittner, Osteogenic
differentiation of mesenchymal progenitor cells in computer designed fibrin-polymer-
ceramic scaffolds manufactured by fused deposition modeling, J Mater Sci Mater Med. 16
(2005) 807–819.
[16] M. Gieseke, C. Noelke, S. Kaierle, V. Wesling, H. Haferkamp, Selective laser melting of
magnesium and magnesium alloys, 2013.
[17] A.C. Hänzi, I. Gerber, M. Schinhammer, J.F. Löffler, P.J. Uggowitzer, On the in vitro and
in vivo degradation performance and biological response of new biodegradable Mg–Y–Zn
alloys, Acta Biomater. 6 (2010) 1824–1833.
[18] M.R. Towler, D. Boyd, C. Freeman, I.M. Brook, P. Farthing, Comparison of in vitro and
in vivo Bioactivity of SrO—CaO—ZnO—SiO2 Glass Grafts, J Biomater Appl. 23 (2009)
561–572.
[19] E. Mostaed, S. Lofferdo, D. Mantovani, M. Vedani, Fabrication of zinc alloy minitubes
for biodegradable stent applications, Eur Cells Mater. 30 (2015) 6.
[20] K.S. Cahill, J.H. Chi, A. Day, E.B. Claus, Prevalence, complications, and hospital charges
associated with use of bone-morphogenetic proteins in spinal fusion procedures, J Am
Med Assoc. 302 (2009) 58–66.
[21] Y.B. Wang, X.H. Xie, H.F. Li, X.L. Wang, M.Z. Zhao, E.W. Zhang, et al., Biodegradable
CaMgZn bulk metallic glass for potential skeletal application, Acta Biomater. 7 (2011)
3196–3208.
[22] D. Vojtech, J. Kubasek, J. Serak, P. Novak, Mechanical and corrosion properties of newly
developed biodegradable Zn-based alloys for bone fixation, Acta Biomater. 7 (2011)
3515–3522. doi:10.1016/[Link].2011.05.008.
[23] M. Vogel, C. Voigt, U.M. Gross, C.M. Müller-Mai, In vivo comparison of bioactive glass
particles in rabbits, Biomaterials. 22 (2001) 357–362.
[24] M.T. Andani, N.S. Moghaddam, C. Haberland, D. Dean, M.J. Miller, M. Elahinia, Metals
for bone implants. Part 1. Powder metallurgy and implant rendering, Acta Biomater. 10
(2014) 4058–4070.
[25] S. Zhang, X. Zhang, C. Zhao, J. Li, Y. Song, C. Xie, et al., Research on an Mg–Zn alloy
as a degradable biomaterial, Acta Biomater. 6 (2010) 626–640.
[26] H.T. Yang, Z.H. Wang, H.F. Li, Y.F. Zheng, Y.W. Li, R. Li, Hemocompatibility of newly
designed bioabsorbable Zn / ZnO composites, Eur Cells Mater. 30 (2015) 72.
[27] M. Montani, A.G. Demir, E. Mostaed, M. Vedani, B. Previtali, Selective laser melting of
pure Fe and pure Zn for biodegradable implants, Eur Cells Mater. 30 (2015) 2.
9
[28] H. Li, Y. Zheng, L. Qin, Progress of biodegradable metals, Prog Nat Sci Mater Int. 24
(2014) 414–422.
[29] X. Liu, J. Sun, K. Qiu, Y. Yang, Z. Pu, L. Li, et al., Effects of alloying elements (Ca and
Sr) on microstructure, mechanical property and in vitro corrosion behavior of
biodegradable Zn–1.5 Mg alloy, J Alloys Compd. 664 (2016) 444–452.
[30] A. Al-Ahmari, E.A. Nasr, K. Moiduddin, M. Alkindi, A. Kamrani, Patient specific
mandibular implant for maxillofacial surgery using additive manufacturing, in: Ind Eng
Oper Manag (IEOM), 2015 Int Conf, IEEE, 2015: pp. 1–7.
[31] W.L. Grayson, M. Fröhlich, K. Yeager, S. Bhumiratana, M.E. Chan, C. Cannizzaro, et al.,
Engineering anatomically shaped human bone grafts, Proc Natl Acad Sci. 107 (2010)
3299–3304.
[32] J. Li, L. Zhang, S. Lv, S. Li, N. Wang, Z. Zhang, Fabrication of individual scaffolds based
on a patient-specific alveolar bone defect model, J Biotechnol. 151 (2011) 87–93.
[33] J. Rustemeyer, A. Busch, A. Sari-Rieger, Application of computer-aided
designed/computer-aided manufactured techniques in reconstructing maxillofacial bony
structures, Oral Maxillofac Surg. 18 (2014) 471–476.
[34] F. Mangano, P. Zecca, S. Pozzi Taubert, A. Macchi, M. Ricci, G. Luongo, et al.,
Maxillary sinus augmentation using computer-aided design/computer-aided
manufacturing (CAD/CAM) technology, Int J Med Robot Comput Assist Surg. 9 (2013)
331–338.
[35] N. Šimunić, D. Vidović, D. Bursać, I. Matković, Application of 3D printed drill guides in
implant dentistry, in: 6th Eur Conf Int Fed Med Biol Eng, Springer, 2015: pp. 383–386.
[36] F.G. Mangano, P.A. Zecca, R. Van Noort, S. Apresyan, G. Iezzi, A. Piattelli, et al.,
Custom-Made Computer-Aided-Design/Computer-Aided-Manufacturing Biphasic
Calcium-Phosphate Scaffold for Augmentation of an Atrophic Mandibular Anterior
Ridge, Case Rep Dent. 2015 (2015) 1–11.
[37] T. Kokubo, H. Takadama, How useful is SBF in predicting in vivo bone bioactivity?,
Biomaterials. 27 (2006) 2907–2915.
10
