Oshida Y. Bioscience and Bioengineering of Titanium Materials
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revealed surface of implants, and the surrounding environments, and the selective
dissolution of the alloy constituents into the surrounding tissues. Besides, it is
generally believed that the solid powder such as wear/friction products particles
have allergic reaction to the living tissue, so that the biological effects of wear
products (debris) should be considered separately from the biocompatibility of
the implants. Particulate wear debris is detected in histocytes/macrophages of
granulomatous tissues adjacent to loose joint prostheses. Such cell–particle inter-
actions have been simulated in vitro by challenging macrophages with particles
doses according to weight percent, volume percent, and number of particles.
Macrophages stimulated by wear particles are expected to synthesize numerous
factors affecting events in the bone–implant interface. Wear debris from orthope-
dic joint implants initiates a cascade of complex cellular events that can result in
aseptic loosening of the prosthesis [5-67]. Macrophages are cells, which are
mainly involved in phagocytosis and signaling and maintaining inflammation that
leads to cell damage in soft tissues and bone resorption [5-68, 5-69]. The pres-
ence of large particles provides the major stimulus for cell recruitment and gran-
uloma formation, whereas the small particles are likely to be the main stimulus
for the activation of cells which release pro-inflammatory products. Apoptosis
can be morphologically recognized by a number of features, such as loss of spe-
cialized membrane structures, blebbing, condensation of the cytoplasm, conden-
sation of nuclear chromatin, and splitting of the cell into a cluster of
membrane-bound bodies [5-68, 5-69]. Recent advances in the understanding of
the mechanisms of osteoclastogenesis and osteoclast activation at the cellular and
molecular levels have indicated that bone marrow-derived macrophages may play
a dual role in osteolysis associated with the total joint replacement: (1) the major
cell in host defense responds to UHMWPE particles via the production of
cytokines, and (2) the precursors for the osteoclasts responsible for the ensuing
bone resorption [5-70].
To avoid such wear debris toxicity, the wear resistance needs to be improved.
Güleryüz et al. [5-71] performed a comparative investigation of thermal oxidation
treatment for Ti-6Al-4V to determine the optimum oxidation conditions for eval-
uation of corrosion-wear properties. Characterization of modified surface layers
was made by means of microscopic examinations, hardness measurements and X-
ray diffraction analysis. Optimum oxidation condition was determined according
to the results of accelerated corrosion tests made in 5 M HCl solution It was
reported that (i) Ti-6Al-4V alloy exhibited excellent resistance to corrosion after
oxidation at 600°C for 60 h, and (ii) this oxidation condition achieved 25 times
higher wear resistance than the untreated alloy during a reciprocating wear test
conducted in a 0.9% NaCl solution [5-71].
Mechanical and Tribological Behaviors 119
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Chapter 6
Biological Reaction
6.1. Toxicity 127
6.2. Cytocompatibility (Toxicity to Cells) 130
6.3. Allergic Reaction 133
6.4. Metabolism 136
6.5. Biocompatibility 138
6.6. Biomechanical Compatibility 143
References 148
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Chapter 6
Biological Reaction
Titanium materials are exposed to biological environments, and their behaviors in
such environments need to be known. This chapter will discuss biological reac-
tions in general.
6.1. TOXICITY
Toxicity can be measured by the effects on the target (organism, organ, or tissue).
Because individuals typically have different levels of response to the same dose of a
toxin, a population-level measure of toxicity is often used which relates the proba-
bility of an outcome for a given individual in a population. One such measure is the
LD50 (LD stands for lethal dose), which is a concentration measure for a toxin at
which 50% of the members of an exposed population die from exposure [6-1]. When
such data does not exist, estimates are made by comparison to known similar toxic
substances, or to similar exposures in similar organisms. Then the safety factors
must be built in to protect against the uncertainties of such comparisons, in order to
improve protection against these unknowns. Toxicity can refer to the effect on a
whole organism (such as a human, a bacterium, or a plant), or to a substructure (such
as the liver). In the science of toxicology, the subject of such study is the effect of an
external substance or condition and its deleterious effects on living things: organ-
isms, organ systems, individual organs, tissues, cells, and subcellular units. Toxicity
of a substance can be affected by many different factors, such as the route of admin-
istration, the time of exposure, the number of exposures, the physical form of the
toxin (solid, liquid, gaseous), the genetic makeup of an individual, an individual’s
overall health, and many others.
There are generally three types of toxic entities: chemical, biological, and phys-
ical.
(1) Chemicals include both inorganic substances, such as lead, hydrofluoric
acid, and chlorine gas, as well as organic compounds such as ethyl alcohol, most
medications, and poisons from living things.
Chemicals (which can be toxic) should include corrosion product such as
oxides, chlorides, or sulfides as well as dissolved metallic ions through anodic
reaction. The toxicity is related to primary biodegradation products (simple and
complex cations and anions), particularly those of higher atomic weight metals.
Factors to be considered include (1) the amount dissolved by biodegradation per
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time unit, (2) the amount of material removed by metabolic activity in the same
time unit, and (3) quantities of solid particles and ions deposited in the tissue and
any associated transfers to the systemic system. Hence, the extent of toxicity is
indirectly related to corrosion rate [6-2–6-5]. An in vivo corrosion experiment can
always be followed up by a tissue examination. This type of tissue reaction is
observed for both corrosion-resistant materials (Mo-containing stainless steel and
Co-Cr-Mo alloys) and for the strongly corroding metals Fe, Mo, and Al. It must
be assumed that the unwanted corrosion product itself is the determining factor.
Fe, Mo, and Al, as non-toxic metals, do not trigger an extreme tissue reaction
despite high corrosion rates (about 300 times that of stainless steel). On the other
hand, the very low corrosion rates of stainless steel and Co-Cr alloys, comparable
to that of Ti, Nb, and Zr, release sufficient quantities of the highly toxic elements
nickel and cobalt, to trigger a noticeable tissue reaction. It should be mentioned at
this point that no limiting concentration is commonly admitted for an allergic reac-
tion. Gold and silver are also to be found in this middle group, as these noble met-
als can also corrode in living tissue. As demonstrated above, a low corrosion rate
alone is not sufficient to guarantee compatibility, nor is the elemental dose the
only determining factor. It would appear that the intrinsic toxicity of the element
and its ability to bind to macromolecules are equally important. Chemical data for
Ti, Zr, Nb, and Ta show that organic complexes of this type are unlikely, and it is
also known that organo-metallic compounds of these elements, as far as they exist,
are unstable [6-6].
There are certain types of elements which are essential for living organisms and
types of elements are different between kinds of living substances. For plants,
there are macronutrients inlcuding N, P, S, Ca, Mg, and Fe, and micronutrients
such as Mn, Zn, Cu, Mo, B, and Cl [6-7]. On the other hand, essential trace ele-
ments for living organisms (particularly, the higher animals) include Fe, F, Si, Zn,
Sr, Pb, Mn, Cu, Sn, Se, I, Mo, Ni, B, Cr, As, Co, and V. Among these elements, the
following nine elements are considered as the most important bioelements, since
they are able to exist as a constituting element for metalloproteins, metalloen-
zymes, or even vitamins: Fe, Zn, Mn, Cu, Se, Mo, Ni, Co, and V [6-8, 6-9].
Despite reports associating tissue necrosis with implant failure, the degree to
which processes, such as metal toxicity, negatively impact implant performance is
unknown. Hallab et al. [6-10] evaluated representative human peri-implant cells
(i.e., osteoblasts, fibroblasts, and lymphocytes) when challenged by Al
3⫹
, Co
2⫹
,
Cr
3⫹
, Fe
3⫹
, Mo
5⫹
, Ni
2⫹
, and V
3⫹
chloride solutions (and Na
2⫹
as a control) over a
wide range of concentrations (0.01–10.0 mM). It was reported that (i) differential
effects were found to be less a function of the cell type than of the composition
and concentration of metal challenge, (ii) no preferential immuno-suppression was
demonstrated, and (iii) soluble Co released from Co-Cr alloy and V released from
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