Implications of sandblasting of 316 LVM stainless

steel on its ion release, in vitro corrosion behavior and

biocompatibility

J.C. Galvn
1
, M. Multigner
1
, L. Saldaa
2,3
, M. Larrea
1
, A.
Calzado-Martn
3,2
, C. Serra
4
, N. Vilaboa
3,2
, J.L. Gonzlez-
Carrasco
1,2


1
Centro Nacional de Investigaciones Metalrgicas
(CSIC), Av. Gregorio del Amo 8, 28040 Madrid (Spain).
jcgalvan@cenim.csic.es, jlg@cenim.csic.es
2
Centro de Investigacin Biomdica en Red en
Bioingeniera, Biomateriales y Nanomedicina
(CIBER-BBN), Madrid (Spain).
3
Hospital Universitario La Paz-IdiPAZ, Paseo de la
Castellana 261, 28046 Madrid (Spain).
4
CACTI - Universidade de Vigo, Campus Lagoas-
Marcosende 15, 36310 Vigo (Spain).

Introduction
The austenitic stainless steel 316 LVM combines both
good mechanic properties with a reasonable in vitro
biocompatibility and in vivo tolerance. Thus, it is used for
orthopedic applications. One method to improve the
implant osseointegration is to increase the roughness by
grit blasting. This study focus on the in vitro corrosion
behavior, ion release and biocompatibility properties of
316 LVM steel modified by sandblasting, following two
sandblasting processes of interest for the preparation of
surgical implants which are based on the use of zirconia
and alumina, respectively.

Materials and methods
Discs of 20 mm in diameter and 2 mm thick of 316LVM
steel were grit blasted by the implant manufacturer
(Surgival SL, Valencia, Spain). Blasting was performed
with two different types of particles under a pressure of
350 KPa for 2 minutes. A first set of samples has been
blasted using ZrO
2
embedded in a silica vitreous phase
microspheres sized between 125 m and 250 m. The
second set of samples has been blasted with Al
2
O
3
angular
particles of around 750 m size.
Topographic surface analysis of the as-processed
specimens was determined with an interferometer optical
profilometer. Due to the relatively high roughness of the
blasted surfaces a Vertical Scanning Interferometry mode
was used. Surface parameters were determined at 5X,
20X, and 50X magnifications, which yields fields of view
of 1.092 mm
2
, 0.068 mm
2
, and 0.011 mm
2
, respectively.
To get information of wider and more representative
fields of view (4 mm
2
), stitching of the digital images
obtained at 20X magnifications was used. Ra (nm), Rq
(nm), Rz (nm), Rt (nm), and real surface area (mm
2
) were
determined. The geometric surface area (A) was 6.41 cm
2

for all the discs. The area increase corresponds to the ratio
of measured surface area/scanned surface area and allows
determining an index area.
Microstructural characterization of surfaces and cross-
sectional views were performed by using a scanning
electron microscope (SEM) coupled with an energy
dispersive X-ray (EDX) system for chemical analysis.
The in vitro corrosion tests were carried out by soaking
the blasted and non-blasted 316 LVM stainless steel
specimens in the Ringers solution. Electrochemical
impedance spectroscopy (EIS) tests were performed in a
conventional electrochemical cell using a three electrodes
set-up. Frequency scans were carried out by applying
sinusoidal wave perturbations of 10mV in amplitude,
close to the corrosion potential. The analysed frequency
was into the 100 kHz-1 mHz range. The impedance
measurements were made after soaking the metal samples
in the Ringers solution for 5 minutes and 24 hours. The
impedance data were modelled with suitable electrical
equivalent circuits and commercial computer programs
based on complex nonlinear least-squares fitting methods.
Inductively coupled plasma optical emission spectrometry
(ICP-OES) has been applied to analyze the ion release.
With this purpose, discs of stainless steel were incubated
in 30 ml of Ringers solution in a humidified 5% CO
2

atmosphere at 37C. To prevent bacterial contamination,
this solution has been sterilized in an autoclave and
samples exposed to UV radiation. Solution released ions
were analyzed at regular intervals 24 h during 4 days. At
each time interval, 23 ml of each solution was replaced by
fresh solution aimed at similar blood that occurs in human
body renewal process. This experimental setup was
proposed by Yamamoto and Hiromoto [1], adapted to
data given by Guyton and Hall [2].
The in vitro biocompatibilitiy of the samples was studied
by using human mesenchymal stem cells from bone
marrow (hMSCs). To evaluate cell attachment and
proliferation, cells were cultured in growth medium on
polished and both rough stainless steel 316 LVM
surfaces. In order to study the ability of the surfaces to
promote osteogenic maturation, cells were switched to the
osteoblastic phenotype by incubation in osteogenic
induction media and mineralized nodule formation was
evaluated by means of Alizarin red staining.

Results and discussion
EIS show that the highest values of the impedance
correspond to the polished specimens and the lowest to the
alumina blasted specimens, thus a decrease in the corrosion
resistance of the steel following grit blasting is envisaged.
Implications of the larger area increase following blasting
are not enough to explain this different behaviour and
subsurface blasting-induced effects, playing a beneficial or
detrimental role, will be discussed. Interestingly, impedance
values increases with increasing the immersion time in
Ringers solution, despite the high concentration of Cl
-
ions
of this medium, which denotes an improvement of the
corrosion protection likely due an increase in the thickness
of the passive film in all the tested samples.
Ion release rises for the blasted conditions and it could be
partially related to the area increase. Finally,
biocompatibility tests suggest that increases in surface
roughness of stainless steel 316 LVM through blasting
processes slow down the adhesion processes cells hMSCs
to the material and the proliferation of those. That effect
is most pronounced in areas with a higher degree of
roughness. However, these topographical changes do not
interfere with the maturation ability of hMSCs cells
towards osteogenic lineage, which corroborates the
usefulness of these materials for implant surgical.

Acknowledgements
This work was supported by the Ministry of Science and
Innovation of Spain, MICINN (MAT2009-14695-C04-02,
-04 and IPT-020000-2010-0001 Projects).

References
1. A. Yamamoto, S. Hiromoto, Mater. Sci. Eng. C-
Biomimetic Supramol. Syst. 29[5] (2009) 1559-1568.
2. A.C. Guyton, J.E. Hall, Textbook of Medical
Physiology (11th edition), Elsevier Saunders,
Philadelphia, PA (2006).
Abstract #1088, 219th ECS Meeting, 2011 The Electrochemical Society
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