Conclusions

To investigate the efficacy of the CO2 laser as a means for improving the
bioactivity and biointegration of bone implants materials, work was con-
ducted to alter the surface properties of two widely used bioinert ceramics,
magnesia–partially stabilised zirconia (MgO–PSZ) and yttria–partially sta-
bilised zirconia (Y–PSZ), and two established biograde metals, Ti–6Al–4V
alloy and 316 LS stainless steel. More specifically, the ability of the CO2
laser to modify the wettability characteristics of the materials and induce
functional groups, thereby allowing bone-like apatite formation, protein
adsorption and cells to be manipulated, was studied. Valuable inroads
have been made as a result of this work for establishing the CO2 laser as a
novel and viable technique for improving the biocompatibility of implant
materials.
The CO2 laser surface treatment of the MgO–PSZ brought about a
reduction in the contact angle, y, formed between the MgO–PSZ and the
control test liquids, providing a clear indication that the wettability char-
acteristics of the material were modified. Moreover, the extent of this
wettability characteristics modification was demonstrated to be variable
and controllable by means of manipulation of the CO2 laser operating
parameters. Changes in the wettability characteristics of the MgO–PSZ
were attributed to the following factors: (a) an increase in surface roughness,
(b) incorporation of oxygen at the MgO–PSZ surface resulting from CO2
p
laser treatment and (c) the increase in the polar component, gsv , of the
surface energy resulting from the melting and re-solidification of the MgO–
p
PSZ surface. In addition, gsv for the MgO–PSZ was seen to increase as the
crystal size and the presence of the tetragonal phase increased after the CO2
p
laser treatment. Further analysis revealed that gsv , by way of the re-solidified
microstructure, was the primary influential factor governing changes in y
and hence the wettability characteristics of the MgO–PSZ. Incorporation of

Laser Surface Treatment of Bio-Implant Materials L. Hao and J. Lawrence

# 2005 John Wiley & Sons, Ltd
180 Conclusions

oxygen at the surface was also shown to influence, to a lesser extent, changes
in the wettability characteristics, while surface roughness was found to play
a very minor role in inducing changes in the wettability characteristics of the
MgO–PSZ.
The bioactivity of the untreated and CO2 laser modified MgO–PSZ was
investigated in SBF, while protein adsorption and hFOB cells were used to
examine the in vitro biological response. It was demonstrated that the CO2
laser treatment could improve the bioactivity of the MgO–PSZ surface by
generating functional groups to facilitate the formation of bone-like apatites.
The apatite formed readily on the CO2 laser treated MgO–PSZ samples, with
relatively high amounts of hydroxyl groups being generated. In contrast,
no apatite formation was observed on the untreated MgO–PSZ samples
and consequently few hydroxyl groups were generated. Further analysis
revealed that the Zr–OH groups on the surface of the CO2 laser treated
MgO–PSZ samples were the functional groups facilitating the apatite
formation; the surface melting on the MgO–PSZ induced by the CO2 laser
processing provided the Zr4 þ ion and OH ion and in turn created the Zr–
OH group. Compared with the untreated MgO–PSZ, the CO2 laser treatment
brought about a thinner adsorbed albumin layer and a thicker adsorbed
fibronectin layer on the MgO–PSZ surface. Whereas the albumin adsorption
decreased, the fibronectin increased with increased wettability, indicating
that the wettability of the MgO–PSZ was the predominant factor governing
p
protein adsorption. Further, the observed effect of gsv on protein adsorption
implied that protein adsorption on the MgO–PSZ surface was probably due
to polar and chemical interactions. Better hFOB osteoblast cell responses
were witnessed on the CO2 laser treated MgO–PSZ samples in comparison
with untreated samples. Generally, the cell cover density increased with
increasing CO2 laser power density. The change in topography induced by
the CO2 laser treatment is certain to be one of the factors influencing the
hFOB osteoblast, but its role will be minor. The improved wettability
characteristics of the MgO–PSZ due to enhanced surface energy brought
p
by the CO2 laser treatment, especially gsv , played a significant role in
precipitating initial cell attachment and spreading in high numbers, conse-
quently enhancing long-term cell adhesion and growth.
The CO2 laser surface treatment of the Ti–6Al–4V alloy brought about a
reduction in the y formed between the Ti–6Al–4V alloy and the simulated
physiological liquids, signifying that the wettability characteristics of the
material were modified. It was found that modification of the surface
roughness, surface oxygen content and surface energy of the Ti–6Al–4V
alloy following the CO2 laser treatment were the factors influencing the
wettability characteristics. It was found that the wettability characteristics of
the Ti–6Al–4V alloy were, after the surface roughness, influenced by the
surface oxygen content and, to some extent, by the microstructure. The
Conclusions 181

reductions in y occasioned by the CO2 laser treatment were found to

contribute to an augmentation in the work adhesion of selected physiologi-
cal liquids (SBF and SBF þ BSA) on the Ti–6Al–4V alloy. As both SBF and
SBF þ BSA have close chemical compositions to human body fluids, the
increase in the work adhesion of the Ti–6Al–4V alloy surface towards these
fluids would mean better suitability of the Ti–6Al–4V alloy for use as a
biomaterial after the CO2 laser treatment.
Apatite formation on the untreated and CO2 laser treated Ti–6Al–4V alloy
after soaking in SBF was used to investigate bioactivity. In addition, protein
adsorption and the hFOB cell response were used to examine the in vitro
biological response on the untreated and CO2 laser treated Ti–6Al–4V alloy.
The fact that apatite nuclei were observed only on the CO2 laser modified
Ti–6Al–4V alloy samples demonstrated that the CO2 laser treatment was
capable of improving the bioactivity of the Ti–6Al–4V alloy; no apatite nuclei
appeared on the untreated samples. It is believed that the CO laser-induced
oxidised surface layer on the Ti–6Al–4V alloy generated the hydroxide ions
in the water and resulted in the nucleation of the apatite. The CO2 laser
treatment brought about a thinner adsorbed albumin layer and a thicker
adsorbed fibronectin layer on the Ti–6Al–4V alloy compared with the
untreated samples. Moreover, the albumin adsorption was seen to decrease,
while the fibronectin increased, with increasing wettability of the Ti–6Al–4V
alloy. This would suggest that the wettability characteristics of the Ti–6Al–
4V alloy are the chief driver for protein adsorption. Further, the observed
p
effect of gsv on protein adsorption implied that the protein adsorption on the
Ti–6Al–4V alloy surface was most likely due to the polar and chemical
interactions. One-day cell adhesion tests showed that cells not only adhered
and spread better but also grew faster on the CO2 laser treated Ti–6Al–4V
alloy sample than on either the untreated or mechanically roughened (the
traditional method of improving cell adhesion) samples. Additionally,
compared with the untreated sample, MTT cell proliferation analysis
revealed that mechanical roughening of the surface of the Ti–6Al–4V alloy
resulted in only a slight enhancement, while the CO2 laser treatment brought
about a considerable increase. Although surface roughness is surely one of
the factors influencing cell adhesion and proliferation, certain other aspects
of wettability characteristics – surface oxygen content and – were found to
play an important role in promoting cell proliferation. Indeed, it was evident
that the better wettability characteristics of the CO2 laser treated Ti–6Al–4V
alloy were responsible for the improved MTT value. Thus it would be
reasonable to maintain that a correlation exists between the CO2 laser
induced wettability characteristics of the Ti–6Al–4V and the hFOB osteoblast
cell bioactivity. Moreover, it is apparent from the results that the CO2 laser
treatment could be a more effective way to improve osteoblast cell adhesion
than the traditional methods currently available.
182 Conclusions

To determine the presence of generic features of the CO2 laser surface

processing technique for improving the biocompatibility of ceramic and
metal biomaterials, surface modifications of the Y–PSZ and the 316 LS
stainless steel were conducted. Changes in the surface properties of the
Y–PSZ and the 316 LS stainless steel following the CO2 laser irradiation were
analysed and the in vitro biological responses of the untreated and the laser
modified materials were evaluated.
The CO2 laser surface treatment of both the MgO–PSZ and the Y–PSZ
bioinert ceramics caused a general reduction in y, suggesting that CO2 laser
induced changes to the wettability characteristics of these bioinert ceramic
materials are generic. It is possible to claim this as increases in the surface
p
roughness, surface oxygen content and gsv of both the MgO–PSZ and the Y–
PSZ following the CO2 laser treatment altered in relation to one another, an
occurrence that could only take place if the changes were generic. The
predominant factor governing modification of the wettability characteristics
p
of both the MgO–PSZ and the Y–PSZ was identified as the increase in gsv ,
further reinforcing the proposition that the changes are generic. Improve-
ments in the cell response of the MgO–PSZ and the Y–PSZ after the CO2
laser surface treatment were correlated directly to CO2 laser induced
enhancement of the wettability characteristics. On account of the generic
effects of the CO2 laser surface treatment on the surface roughness and
wettability, improvements in the cell response of the bioinert ceramics after
the CO2 laser surface treatment appear to be generic.
Observed changes in the wettability characteristics of the Ti–6Al–4V alloy
and the 316 LS stainless steel when surface treated with the CO2 laser were
deemed to be generic as increases in the surface roughness, surface oxygen
p
content and gsv of both biometals appear to have altered in relation to one
another. Additional verification of the generic nature of the wettability
characteristics modification of both biometals comes from the fact that the
leading aspect governing the modification of the wettability characteristics
of both biometals was the increase in surface roughness. Because surface
roughness is the predominant mechanism active in influencing the wett-
ability characteristics of other laser treated metallic materials, it seems
highly likely that the laser induced changes to the wettability characteristics
of most metals will actually be generic. Favourable protein adsorption
and improvements in the cell response of the Ti–6Al–4V alloy and the
316 LS stainless steel after the CO2 laser surface treatment were correlated
directly to CO2 laser induced enhancement of the surface roughness and
wettability characteristics of these biometals. Because of the generic effects
of the laser surface treatment on the surface roughness and wettability
characteristics of other metals, it seems valid to say that improvements in the
cell response of these biometals after the CO2 laser surface treatment are
generic.
Conclusions 183

The major focus of this work was the employment of a CO2 laser for
the surface processing of bioinert ceramics and biograde metals that are
widely used as load-bearing bone implants. This contemporary research, in
conjunction with the findings of others, firmly establishes the potential of
the CO2 laser for improving the biocompatibility of a variety of other
biomaterials.