Materials Letters 274 (2020) 128057

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Materials Letters
journal homepage: www.elsevier.com/locate/mlblue

Composite dip coating improves biocompatibility of porous metallic

scaffolds
Joseph Deering a, Amanda Clifford a, Andrew D’Elia a, Igor Zhitomirsky a,b, Kathryn Grandfield a,b,⇑
a
Department of Materials Science and Engineering, McMaster University, Hamilton, ON, Canada
b
School of Biomedical Engineering, McMaster University, Hamilton, ON, Canada

a r t i c l e i n f o a b s t r a c t

Article history: Porous materials are becoming more common for bone implants, and it is increasingly important to find
Received 17 April 2020 surface modification strategies that affect both the implant exterior and porous interior. In this study,
Received in revised form 26 May 2020 selective laser melting (SLM) was used to create porous stainless steel implants 8 mm in diameter, which
Accepted 26 May 2020
were subsequently dip coated with a composite polymethylmethacrylate-alumina (PMMA-Al2O3) film.
Available online 27 May 2020
Imaging with electron microscopy found evidence of the films at a depth of 2.2 mm into the porous
implants, with dual-scale topography created by the native SLM stainless steel substrate and alumina
Keywords:
nanoparticles. Energy dispersive X-ray spectroscopy confirmed the presence of the coating along the
Dip coating
Polymethylmethacrylate
periphery of interior pores. In vitro tests with osteoblast-like cells showed greater cell metabolism on
Additive manufacturing composite-coated samples compared to uncoated dense samples after seven days of culture.
Porous materials Ó 2020 Elsevier B.V. All rights reserved.
Osseointegration
Sol–gel preparation

1. Introduction effective for depositing bioactive and organic coating materials on

the interior of porous structures. Where alumina-based ceramic
Porous metallic implant materials are becoming increasingly materials have been traditionally used for bulk implant compo-
common for usage in bone and joint replacement due to their abil- nents due to their excellent biocompatibility in bone applications
ity to mitigate stress-shielding effects [1]. The use of additive man- [13], their integration in nanoparticle form on the interior of por-
ufacturing enables effective scaffold design in bone implants by ous constructs has not been evaluated. Polymethylmethacrylate
creating tunable mechanical performance [2] and channels for (PMMA), commonly known as a primary constituent in some bone
mass transport [3] by implementing porosity. cements, has recently been shown to have favourable osteogenic
To encourage bone ingrowth, implants have been coated with effects as an implant coating [14]. The potential for PMMA-Al2O3
composite materials, which can contain bioceramic particles such organic composite material to coat the interior of porous metallic
as calcium phosphate [4], rutile [5], and alumina [6]. One key lim- scaffolds remains unexplored.
itation of conventional coating techniques, such as plasma spray- The objective of this work was the development of a facile dip
ing, is their line-of-sight processing [7], which prevents uniform coating method to deposit a composite PMMA-Al2O3 coating on
coating deposition in the interior of porous structures, and their the interior of an additively manufactured porous scaffold. Scan-
high-temperature processing. Changing the surface composition ning electron microscopy (SEM) was used to confirm deposition,
and topography in the interior of porous specimens is possible by and the potential for improving osseointegration was explored
electrochemical methods, such as micro-arc oxidation [8,9], where with an in vitro cell metabolism assay.
oxides of the base metal, calcium, or phosphorus are added to the
surface. Composite materials have also been developed as a feed-
stock for additive manufacturing processes as a means of modify- 2. Methods
ing the interior of porous structures compared to structures
produced by traditional titanium feedstocks [10]. Recently, immer- 2.1. Scaffold production
sion techniques [11] and electrophoretic deposition [12] have been
Cylinders (h = 8 mm, ø = 8 mm) were designed in Autodesk Net-
fabb and hollowed to create body-centred cubic lattice struts
⇑ Corresponding author. rotated 45° about the X and Y axes. Fully dense implants as well
E-mail address: [email protected] (K. Grandfield). as porous implants with strut diameters of 450 mm with a unit cell

https://doi.org/10.1016/j.matlet.2020.128057
0167-577X/Ó 2020 Elsevier B.V. All rights reserved.
2 J. Deering et al. / Materials Letters 274 (2020) 128057

spacing of 1.2 mm, and therefore pores with an approximate throat analyzed by SEM imaging (FEI Magellan 400) and energy dispersive
diameter of 275 mm were formed. Scaffolds were built using 304L X-ray spectroscopy (EDS) using an accelerating voltage of 10 kV.
stainless-steel powder (<45 mm, Carpenter Additive) using the
selective laser melting (SLM) process (EOSINT M280, Germany). 2.3. Cell viability
Laser power was set to 200 W, scan speed 800 mm/s, hatch spacing
80 mm, and layer thickness 40 mm during fabrication. Metal scaf- Saos-2 osteosarcoma cells (ATCC) were seeded on additively
folds were cleaned with ethanol and deionized water in an ultra- manufactured solid implants, porous implants, and PMMA-Al2O3
sonic bath for 20 min. Scaffolds were either kept whole or coated porous implants at a density of 10,000 cells/cm2 in McCoy’s
sectioned longitudinally for subsequent coating. Modified 5A media. Cells were cultured at 37 °C in an atmosphere
of 5% CO2 for seven days. After one, three, and seven days, a solu-
2.2. Coating deposition and characterization tion of 5% alamarBlue reagent in media (Life Technologies Inc.) was
added to wells for 60 min. The dye was pipetted into a separate
Polymethylmethacrylate (MW ~ 120,000, Millipore Sigma) and plate and fluorescence was measured at an excitation-emission
alumina (0.13 mm, Al2O3, Baikowski) particles were obtained. wavelength of 540–580 nm. Statistical significance was deter-
Under continuous stirring, 10 g L 1 PMMA was added to a mixed mined using a two-way ANOVA in R with Tukey’s HSD test and
solvent containing 20% deionized water and 80% isopropanol (Mil- p < 0.05.
lipore Sigma) and heated to 55 °C, at which point the PMMA fully
dissolved, only marginally increasing the viscosity. The PMMA 3. Results and discussion
solution was cooled back to room temperature, and Al2O3 particles
were added to a concentration of 10 g L-1. Metallic scaffolds were 3.1. Coating deposition
coated whole to produce specimens for cell viability assays or in
halves for easier investigation of the midplane with electron Dip coating with the PMMA-Al2O3 composite produced a coat-
microscopy. Scaffolds were attached to copper tape (see Fig. S1), ing on both the exterior and interior of the scaffolds. Where PMMA
immersed into the PMMA-Al2O3 suspension under sonication for bonds to the substrate may be governed by bidentate ligands, the
one minute, removed, and air dried at room temperature for 24 h mechanical strength of the composite coating on the stainless steel
to evaporate solvent. substrate is possibly increased compared to other coatings with
The mid-plane of the scaffold was sputter coated with carbon monodentate bonding coordination. Higher magnification images
for conductivity, mounted to a SEM stub with silver paint and of a representative pore (Fig. 1A) illustrate the presence of the

Fig. 1. (A) Image of a representative pore on the interior of the scaffold. Thickness of PMMA-Al2O3 around the pore periphery is not constant. (B) Surface topography of
PMMA-Al2O3 composite coating at the scaffold interior. (C) Surface topography of the PMMA-Al2O3 composite coating at the scaffold exterior. The Al2O3 distribution is
comparable at scaffold exterior and interior sites.

Fig. 2. (A) Cross-sectional SEM image with bright regions representative of the coating. Sampling sites at various distances to exterior correspond to EDS maps at depths of
0.4 mm (Row B), 1.2 mm (Row C) and 2.2 mm (row D). Increased intensity of aluminum and carbon at the pore periphery indicates complete penetration of the coating into
the interior pores.
 J. Deering et al. / Materials Letters 274 (2020) 128057 3

penetrated at least 2.2 mm into the scaffold interior, indicating

that this immersion technique is suitable for coating the complete
interior of porous metallic implants with a biomedical composite
material. The alumina nanoparticles also contributed to a nanos-
cale topography around the pore periphery. In vitro characteriza-
tion of the coated scaffolds with Saos-2 cells showed statistically
higher rates of cell metabolism when compared to fully dense
structures with the same geometry. Therefore, the dip coating
method is a promising approach for creating composite coatings
on porous implants to improve the potential osteoconductivity of
their interior pores. The successful demonstration of Al2O3
nanoparticles, PMMA, and stainless steel scaffolds as pilot materi-
als should lead to investigation of other additives and substrates,
such as titanium alloys, for polymer-composite coatings on porous
Fig. 3. Saos-2 cell viability on solid, porous, and coated porous AM parts. Porous SLM implants. Future work should also evaluate the limits of
implants coated with the PMMA-Al2O3 composite coating showed significantly
PMMA-Al2O3 integration in larger-sized implants.
higher cell proliferation after seven days compared to solid components, while all
porous implants showed significantly more cell proliferation after seven days.

CRediT authorship contribution statement

composite coating at an interior site, where non-uniform thickness
is observed around the periphery of the pore. There is also evidence
Joseph Deering: Conceptualization, Investigation, Formal anal-
of sintered stainless steel particles on the lattice struts within the
ysis, Writing - original draft. Amanda Clifford: Investigation,
pores, adding elements of microscale topography on both interior
Methodology, Writing - review & editing. Andrew D’Elia:
and exterior lattice sites. Nanoparticles of Al2O3 were observed to
Investigation, Methodology, Writing - review & editing. Igor
be uniformly distributed through the PMMA matrix without aggre-
Zhitomirsky: Conceptualization, Supervision, Funding acquisition,
gation (Fig. 1B) at interior sites of the scaffolds. These nanoparti-
Writing - review & editing. Kathryn Grandfield: Conceptualiza-
cles, in conjunction with inherent striations in the PMMA matrix,
tion, Supervision, Funding acquisition, Writing - review & editing.
add elements of nanoscale topography to interior and exterior sites
in the scaffold, which can be favourable for osseointegration [15].
Coating deposition on the interior appears consistent with the Declaration of Competing Interest
exterior (Fig. 1C), where Al2O3 is evenly distributed through the
PMMA. The authors declare that they have no known competing finan-
EDS maps at interior scaffold sites (Fig. 2A) on the cross-section cial interests or personal relationships that could have appeared
are shown in Fig. 2B, 2C, and 2D. Each site has a different distance to influence the work reported in this paper.
to the scaffold exterior. Elemental signals characteristic to the
uncoated stainless-steel (Fe, Ni) were uniform on the cross- Acknowledgements
sectional surface, while signals characteristic to the composite
coating (C, Al) displayed higher intensity directly around the pore The authors acknowledge the support of the Natural Sciences
periphery at depths of 0.4 mm, 1.2 mm, and 2.2 mm. and Engineering Research Council of Canada (NSERC) (RGPIN-
2014-06053 and RGPIN-2018-04014). JD was supported by an
3.2. Cell viability NSERC PGS-D. Electron microscopy was conducted at the Canadian
Centre for Electron Microscopy, and cell culture in the Biointer-
The results of cell viability assays on the solid implants, porous faces Institute at McMaster University.
implants, and PMMA-Al2O3 coated porous implants are shown in
Fig. 3. After seven days of culture, the PMMA-Al2O3 coated scaf- Appendix A. Supplementary data
folds significantly outperformed dense stainless-steel samples,
confirming what has been shown previously on two-dimensional Supplementary data to this article can be found online at
substrates [14]. Statistically higher cell metabolism was also https://doi.org/10.1016/j.matlet.2020.128057.
observed from day one to day seven on both the porous implants
without coating and PMMA-Al2O3 coated porous implants. These
References
results suggest that the addition of an interconnected porosity net-
work with a diameter of 275 mm allows for rapid cellular prolifer- [1] S. Arabnejad, B. Johnston, M. Tanzer, D. Pasini, Fully porous 3D printed
ation through the porous interior of the scaffold relative to the titanium femoral stem to reduce stress-shielding following total hip
control. The addition of PMMA-Al2O3 composite coatings on por- arthroplasty, J. Orthopaed. Res. 35 (8) (2017) 1774–1783.
[2] X. Wang, S. Xu, S. Zhou, W. Xu, M. Leary, P. Choong, et al., Topological design
ous scaffolds further improves osseointegration potential relative and additive manufacturing of porous metals for bone scaffolds and
to uncoated implants. By coating the SLM implant surface, surface orthopaedic implants: a review, Biomaterials 83 (2016) 127–141.
topography, chemistry, and wettability are modified, and it is prob- [3] A.A. Zadpoor, Additively manufactured porous metallic biomaterials, J. Mater.
Chem. B 7 (26) (2019) 4088–4117.
able that metallic particle release is impeded. All of these factors [4] N. Eliaz, N. Metoki, Calcium phosphate bioceramics: a review of their history,
can change the dynamics of cell-surface interaction, contributing structure, properties, coating technologies and biomedical applications,
to the resulting increase in cell viability seen here. Materials 10 (4) (2017) 334.
[5] A. Clifford, B.E.J. Lee, K. Grandfield, Zhitomirsky I. Biomimetic modification of
poly-l-lysine and electrodeposition of nanocomposite coatings for orthopaedic
applications, Colloids Surfaces B Biointerfaces 176 (2018) (Polym. (United
4. Conclusions Kingdom). 121 2017):115–21.
[6] A.J. McManus, R.H. Doremus, R.W. Siegel, R. Bizios, Evaluation of
PMMA-Al2O3 composite films were successfully deposited on cytocompatibility and bending modulus of nanoceramic/polymer
composites, J. Biomed. Mater. Res. A 72A (1) (2005) 98–106.
the porous interior of SLM stainless steel implants using a facile [7] E. Munting, The contributions and limitations of hydroxyapatite coatings to
dip coating method. SEM and EDS maps confirmed that the coating implant fixation, Int. Orthop. 20 (1) (1996) 1–6.
4 J. Deering et al. / Materials Letters 274 (2020) 128057

[8] P. Xiu, Z. Jia, J. Lv, C. Yin, Y. Cheng, K. Zhang, et al., Tailored surface treatment of [12] V.O. Kollath, Q. Chen, S. Mullens, J. Luyten, K. Traina, A.R. Boccaccini, et al.,
3D printed porous Ti6Al4V by microarc oxidation for enhanced Electrophoretic deposition of hydroxyapatite and hydroxyapatite–alginate on
osseointegration via optimized bone in-growth patterns and interlocked rapid prototyped 3D Ti6Al4V scaffolds, J. Mater. Sci. 51 (5) (2016) 2338–2346.
bone/implant interface, Acs Appl. Mater Inter. 8 (28) (2016) 17964–17975. [13] P. Zeng, Biocompatible alumina ceramic for total hip replacements, Mater. Sci.
[9] Z. Chen, X. Yan, Y. Chang, S. Xie, W. Ma, G. Zhao, et al., Effect of polarization Tech. Ser. 24 (5) (2013) 505–516.
voltage on the surface componentization and biocompatibility of micro-arc [14] A. D’Elia, J. Deering, A. Clifford, B.E.J. Lee, K. Grandfield, I. Zhitomirsky,
oxidation modified selective laser melted Ti6Al4V, Mater. Res. Express 6 (8) Electrophoretic deposition of polymethylmethacrylate and composites for
(2019) 086425. biomedical applications, Colloids Surf. B Biointerfaces 188 (2019) 110763.
[10] H. Attar, S. Ehtemam-Haghighi, N. Soro, D. Kent, M.S. Dargusch, Additive [15] O. Omar, D. Karazisis, A. Ballo, S. Petronis, H. Agheli, L. Emanuelsson, et al., The
manufacturing of low-cost porous titanium-based composites for biomedical role of well-defined nanotopography of titanium implants on
applications: advantages, challenges and opinion for future development, J. osseointegration: cellular and molecular events in vivo, Int. J. Nanomed. 11
Alloy. Compd. 827 (2020) 154263. (2016) 1367–1382.
[11] C. Domínguez-Trujillo, E. Peón, E. Chicardi, H. Pérez, J.A. Rodríguez-Ortiz, J.J.
Pavón, et al., Sol-gel deposition of hydroxyapatite coatings on porous titanium
for biomedical applications, Surf Coatings Technol. 333 (2018) 158–162.