Professional Documents
Culture Documents
2
Department of Metallurgical and Materials Engineering, Daelim College of Technology, Anyang 431-715, Korea
3
Department of Prosthodontics and Dentistry, Ewha Womans University, Seoul 158-710, Korea
transformation when exposed to a low-temperature range of estimated from the measurement of the diagonal dimensions
100 – 400°C for a long period of time in air or water.8,9 The of 98 N Knoop indentations after Marshall et al.12 The
phase transformation is accompanied by an abrupt decrease in toughness of the bar specimens was estimated by the inden-
strength, so that the phenomenon is called low-temperature tation method suggested by Anstis et. al.13
degradation. Aging in water vapor conditions accelerates the Hydrothermal stability of the composite specimens was
degradation. Since tetragonal zirconia abutments must be evaluated after aging for 5 h at the temperature range of
autoclave-sterilized for 20 –30 min at 120 –135°C prior to 150 –250°C in 4 MPa water vapor pressure in an autoclave.
clinical applications, the ceramic abutments should be im- The extent of hydrothermal degradation was estimated from
mune from degradation while sterilizing. The purpose of this the X-ray diffraction peak intensity ratio of the m-ZrO2 and
study is to evaluate the mechanical properties, the tetragonal t-ZrO2 phases on the aged specimen, according to Garvie and
phase stability in autoclave conditions, and biocompatibility Nicholson.14
of a newly developed (Y,Nb)-ZrO2/Al2O3 composite.
Biocompatibility Tests
The biocompatibility of the (Y,Nb)-ZrO2/Al2O3 composite
MATERIALS AND METHODS was evaluated by means of an in vitro cytotoxicity test and an
in vivo subcutaneous implant test.15,16 For the in vitro test, the
Sample Preparation cell growth and survival test and agar overlay test were
employed. The cell growth and survival test was performed
The (Y,Nb)-TZP powders were prepared by mixing 90.24
by evaluating cell growth on ten (Y,Nb)-ZrO2/Al2O3 com-
mol% ZrO2, 5.31 mol% Y2O3, and 4.45 mol% Nb2O5 using
posite disc specimens, 13 mm diameter ⫻ 1 mm thickness,
ball milling of 24 h. After drying, the mixed powders were
after 2, 4, and 6 days of mouse fibroblast (L929, ACTT) cell
calcined for 4 h at 1100°C and then attrition-milled for 3 h.
culture. Copper was used as a positive control material, and
Zirconia balls were used for the milling processes. (Y,Nb)-
no specimen was used as a negative control in culture vessels
TZP/Al2O3 composite powders were prepared by adding var-
(Nunc, Denmark). After 2, 4, and 6 days of culture, cell layers
ious amounts of Al2O3 to the (Y,Nb)-TZP powder and ball
in the dishes were trypsinized, and stained with trypan blue,
milling for 24 h. The milled slurries were dried, sieved
and cell counts were performed.
through a 100-mesh screen, die-pressed into disks and tubes,
For the agar overlay test, same-sized specimens were
and then isostatically pressed at 140 MPa. The green com-
utilized to evaluate cell viability in culture vessels (Nunc,
pacts were sintered for 2 h at 1550°C in air.
Denmark). After cell culture, 1.5% Bacto-agar (Difco, USA)
and BME solution were used for agar formation. Copper was
Property Measurements
used for the positive control material, and commercially pure
For mechanical property measurements, disk and bar speci- titanium was used for the negative control. Cells were stained
mens were employed. The disks and bars were prepared by by neutral red solution, and specimens were placed onto the
sintering of disk- and tubular-shaped composite powder com- agar layer and cultured for 24 h. The zone of discoloration
pactions, respectively. The dimension of the disk specimens and cell lysis index were evaluated according to FDI stan-
after polishing to a 1 m diamond finish was 18 mm in dards.15
diameter and 1.8 mm in thickness. A flat-on-three-ball bi- For the subcutaneous implant test, twelve 300 – 400 g
axial-fixture was used for the determination of strength of the body-weighted guinea pigs and twelve (Y,Nb)-ZrO2/Al2O3
disk specimens with a stress rate of 23 MPa 䡠 s⫺1 that was composite and twelve 3Y-TZP specimens, 10 ⫻ 7 ⫻ 1 mm,
determined from a correlation between the rate and specimen were used, respectively. Two specimens were embedded sub-
thickness in the ASTM standard.10 The bar specimens were cutaneously in the lumber area in each animal. The animals
prepared by cutting the sintered tubular-shaped composite were sacrificed after 2 and 12 weeks. After carefully remov-
specimens and the CerAdapt威 abutments radially and then ing specimens with surrounding tissue as thin as possible,
polishing to a 1 m diamond finish. The dimensions of the they were pre-fixed in 2.5% glutaraldehyde immediately, and
specimens were 1.0 ⫻ 0.7 ⫻ 9.0 mm after polishing. A the specimens were removed from the surrounding capsules.
3-point fixture was used for the determination of strength, The capsules were fixed with 1% osmium tetroxide and
where the span distance was 7 mm and the cross-head speed dehydrated in ethyl alcohol and embedded in Epon 812 for
was 0.07 mm/min. Prior to the measurements, both disks and light- and electron-microscopic observations (JEOL 1200
bar specimens were annealed for 2 h at 1200°C to remove a EX, Japan). 1 m sections were made and then stained with
residual stress from the machining procedure. A minimum of toluidine blue and 80 –100 nm sections made and then stained
ten tests was performed for each type of specimen. with uranyl acetate and lead citrate for the light- and electron-
The fracture toughness of the disk specimens was deter- microscopic observation, respectively.
mined by the indentation-strength method.11 A Vickers in-
dentation load of 98 N was placed on the center of the tensile RESULTS
face of the test specimens. A minimum of five tests was
performed for each composite specimen composition. For the Flexural strength and fracture toughness of the disk-shaped
calculation of toughness, the hardness-to-modulus ratio was (Y,Nb)-TZP/Al2O3 composites containing 0 –30 vol% of the
440 KIM ET AL.
Composite CerAdapt威
Strength, MPa 710 619
Toughness, MPa 䡠 m1/2 8.5 3.1
From the agar overlay test, the ceramic composite and pure
titanium exhibited neither discoloration nor lysis of L929
fibroblast cells. However, the positive control, copper,
showed severe cytotoxicity (Fig. 5). In vivo subcutaneous
implant tests of the composite specimens in guinea pigs
showed thick and relatively loose capsules with few inflam-
matory cell infiltration after 2 weeks, and well-defined thin
and dense fibrous capsules with elongated fibroblasts after 12
weeks under the light microscope (Fig. 6). Electron micro-
scopic observation revealed some inflammatory cells, mac-
rophage, and PMNS with slightly elongated fibroblasts after Figure 5. Agar overlay test: (a) (Y,Nb)-TZP/20 vol% Al2O3 composite
showed no cell lysis (⫻ 100); (b) pure titanium showed no cell lysis (⫻
2 weeks, and well elongated fibroblast with dense collagen 100); and (c) copper showed severe cell lysis (⫻ 100).
fibrils after 12 weeks (Fig. 7).
The in vivo subcutaneous implant test in guinea pigs was also 8. Piconi C, Maccauro G. Zirconia as a ceramic biomaterial.
performed to evaluate soft-tissue reaction. The basic screening Biomater 1999;20:1–25.
biocompatibility tests indicated that the composite material has 9. Lawson S. Environmental degradation of zirconia ceramics. J
Euro Ceram Soc 1995;15:485–502.
no cytotoxicity and presents a favorable soft-tissue response for 10. ASTM Standard F394-78. Biaxial flexure strength (modulus of
dental applications during this short-term test period. In clinical rupture) of ceramic substrate. ASTM annual book of standards,
applications, the composite abutments were placed to the sites of Vol 15.02, Sect 15. Philadelphia: Am Soc Test Mater; 1996. p.
two maxillary premolars and one maxillary first molar. Neither 446 – 450.
a failure of abutments nor an adverse soft-tissue reaction has 11. Chantikul P, Anstis GR, Lawn BR, Marshall DB. A critical
been observed for the period of 14 months since the abutments evaluation of indentation techniques for measuring fracture
toughness: II, Strength method. J Am Ceram Soc 1981;64:539 –
were connected to the implants.
543.
12. Marshall DB, Nomab T, Evans AG. A simple method for
determining elastic-modulus-to-hardness ratios using Knoop in-
CONCLUSION dentation measurements. Am Ceram Soc 1982;65:C175–C176.
13. Anstis GR, Chantikul P, Lawn BR, Marshall DB. A critical
The (Y,Nb)-TZP/Al2O3 composite showed higher strength evaluation of indentation techniques for measuring fracture
and toughness and, thus, a better flaw tolerance compared to toughness: I, direct crack measurements. J Am Ceram Soc
1981;64:533–538.
alumina-based abutments. The difference is attributed to the
14. Garvie RC, Nicholson PS. Phase analysis in zirconia systems.
t3m phase transformation of TZP and the grain bridging J Am Ceram Soc 1971;67:303–305.
whose extent was optimized with the addition of 20 vol% 15. Federation Dentaire International; Recommended standard
Al2O3. Low-temperature degradation was not observed in practices for biological evaluation of dental materials. Int Dent
(Y,Nb)-TZP/Al2O3 composites after aging at the temperature J 1980;30:140 –188.
range of 150 –250°C and 4 MPa water-vapor pressure in the 16. ISO 10993–1. Biological testing of medical and dental materials
autoclave. The hydrothermal stability of the composites was and devices — Part 1: Guidance on selection of tests. 1995.
17. Lee DY, Kim DJ, Cho DH. Low-temperature phase stability and
achieved by the inherent phase stability of (Y,Nb)-TZP due to mechanical properties of Y2O3 and Nb2O5 co-doped tetragonal
the local Y-Nb ordering in the t-ZrO2 lattice and the annihi- zirconia polycrystal ceramics. J Mater Sci Lett 1998;17:185–
lation of oxygen vacancies in Y-TZP. The in vitro and in vivo 187.
biocompatibility tests revealed neither a cytotoxicity nor a 18. Tsukuma T, Ueda TK, Shimada M. Strength and fracture tough-
significant adverse soft-tissue reaction with the composites. ness of isostatically hot-pressed composites of Al2O3 and Y2O3
The (Y,Nb)-TZP/Al2O3 composites with improved mechan- partially stabilized ZrO2 J Am Ceram Soc 1985;68:C-4 –C-5.
19. Tsukuma T, Takahata T, Shiomi M. Strength and fracture
ical properties, phase stability, and favorable biocompatibility
toughness of Y-TZP, Ce-TZP, Y-TZP/Al2O3, and Ce-TZP-
are applicable for dental implant abutment material, even Al2O3. Somiya S, Yamamoto N, Yanagida H, editors. Advances
though further research is needed to evaluate long-term bio- in ceramics, Vol 24B, science and technology of zirconia III.
logical and mechanical behavior of (Y,Nb)-TZP/Al2O3 com- Westerveille, OH: Am Ceram Soc; 1988. p 721–728.
posites for clinical use. 20. Li JF, Watanabe R. Fracture toughness of Al2O3-particle-dis-
persed Y2O3 partially stabilized zirconia. J Am Ceram Soc
1995;78:1079 –1082.
REFERENCES 21. Rice RW. Mechanism of toughening in ceramic matrix com-
posites. Ceram Eng Sci Proc 1981;2:661–701.
1. Brånemark PI, Zarb GA, Albrektsson T. Tissue-integrated pros- 22. Li P, Chen IW, Penner–Hahn JE. Effect of dopants on zirconia
theses. Chicago: Quintessense; 1985. p 11–76. stabilization — an X-ray absorption study: III, charge-compen-
2. Adell R, Lekholm U, Rockler B, Brånemark PI. A 15-year study sating dopants. J Am Ceram Soc 1994;74:1289 –1295.
of osseointegrated implants in the treatment of the edentulous 23. Guo X, Wang Z. Effect of niobia on the defect structure of
jaw. Int J Oral Surg 1981;10:387– 416. yttria-stabilized zirconia. J Euro Ceram Soc 1998;18:237–240.
3. Prestipino V, Ingber A. Esthetic high strength implant abut-
24. Kim DJ, Jung HJ, Cho DH. Phase transformation of Y2O3 and
ments. Part I. J Esthet Dent 1993;5:29 –36.
Nb2O5 doped tetragonal zirconia during low temperature ageing
4. Prestipino V, Ingber A. Esthetic high strength implant abut-
in air. Solid State Ionics 1995;80:67–73.
ments. Part II. J Esthet Dent 1993;5:63– 68.
5. Prestipino V, Ingber A. All ceramic implant abutments. Esthetic 25. Kim DJ, Jung HJ, Jang JW, Lee HL. Ionic conductivity, fracture
indications. J Esthet Dent 1996;8:255–262. toughness, and low temperature phase stability of tetragonal
6. Andersson B, Shårer P, Simon M, Bergstrom C. Ceramic im- zirconia co-doped with Y2O3 and Nb2O5. J Am Ceram Soc
plant abutments used for short-span fixed partial dentures: A 1998;81:2309 –2314.
prospective 2-year multicenter study. Int J Prosthodont 1999; 26. Schmauder S, Schubert H. Significance of internal stresses for
12:318 –324. the martensitic transformation in yttria-stabilized tetragonal zir-
7. Cales B, Stefani Y. Yttria-stabilized zirconia for improved conia polycrystals during degradation. J Am Ceram Soc 1986;
orthopedic prostheses. Wise DL, Trantolo DJ, Altobelli DE, 68:534 –540.
Yaszemski MJ, Gresser JD, Schwartz ER, editors. Encyclopedic 27. Kim DJ. Influence of aging environment on low-temperature
handbook of biomaterials and bioengineering, Part B: Applica- degradation of tetragonal zirconia alloys. J Euro Ceram Soc
tions, Vol 1. New York: Marcel Dekker; 1995. p 415– 452. 1997;17:897–-903.