The process is a combination of electrochemical reaction, acid-base reaction and precipitation reaction. 1. Water is reduced at the cathode (implant surface) 2 H2O + 2e- = H2 + 2 OH2. The hydroxide ions react with dihydrogen phosphate according OH- + H2PO4 H2O + HPO4 2-
3. Then follows the precipitation of calcium phosphates Ca2+ + H2PO42- + 2 H2O CaHPO4 x 2 H2O
5 CaHPO4 x 2 H2O + 6 OH- Ca5(PO4)3OH + 2 PO4 3- + 15 H2O The result is a composite of 2 calcium phosphate phases: Hydroxyapatite (HA) and Brushite, a physiological precursor of HA [1, 2, 3, 4, 5].
Post treatment
Test
Method
Results
Element Analysis
Lead
(EN ISO 11885-E22)
Cadmium
(EN ISO 11885-E22)
Mercury
(DIN EN 1483 E12)
7.7 110 53
Copper
(EN ISO 11885-E22)
Zinc
(EN ISO 11885-E22)
Nickel
(EN ISO 11885-E22)
0.85 ppm
A calculation by EDX analysis with a cross section of the BONIT coating shows the composite character of the coating. In the distance immediately at the surface the Ca/P ratio is 1.7 (for HA), the outer coating shows a Ca/P ratio of 1.0. (Appendix A) Infrared pattern of outer coating (deep penetration ca 2 m) with diamond ATR accessory shows the CaP phase Brushite. The HA phase isnt detectable by FTIR because the rate of HA of the whole coating is <5%. FTIR- patterns show a complete changing from the HA precursor Brushite into HA within two hours, tested in physiological buffer (Tris/HCl 0.05M, pH 7.3, 37C). (Appendix B) Data from literature confirm the rapid conversion from Brushite into HA under physiological conditions and postulate Brushite to be the temporary physiological pre-stage of the bone mineral HA [1, 2, 3, 4, 5].
Infrared Spectrometry
FTIR
Solubility
Publication: 31,4% weight loss in buffered medium Ducheyne P., (Tris(hydroxy-methyl) methylamin/HCl Biomaterials 0.05M, pH 7.3, at 37C temperature, 1990; 11(8), over a time period of 7 days). 540 The increased solubility of BONIT is responsible for more rapid bone ongrowth.
C. Biological Properties
Test
Method
Results
Cytotoxicity, L 929-Proliferation
DIN EN ISO 10993-5 BONIT coated samples do not ISO 9363-1 release substances in cytotoxic LM SOP 4-06-01 concentrations during a permanent 7 days contact of 4.5 cm2 surface area to 1 ml physiological fluid.
Adhesion after 15 min as well as proliferation after 7 days on BONIT coating was comparable high to positive control collagen I. After 30 hours in cell culture osteoblastic cells were partly covered by new precipitated fine structured crystals. (Appendix C) We start from the assumption that these new precipitations are fine HA crystals. (Abovementioned investigations by FTIR method support this assumption.) The process of BONIT certificated to be pyrogen free.
Systemically toxicity
DIN EN 10993-1
is
In several animal experiments and clinical studies the BONIT coating was proofed to be biocompatible as well as osteoinductive [6-21].
D. Physical Properties
Test
Method
Results
Morphology
REM
Thickness
15 m +/- 5 m
Tensile strength
ASTM F 1147
10 MPa
Shear strength
ASTM F 1044
13 MPa
Fatigue testing
Abrasion
resistant
Summary: BONIT is an electrochemically deposited calcium phosphate coating based upon a biomimetic process in which implants are coated in an electrolytic bath with a ~15 m thin bioactive layer of a calcium phosphate composite. The room temperature procedure provides for complete coverage of complex shapes and porous surfaces. As opposed to the monolithic crystalline structure representative of plasma-sprayed coatings, this process employs a fine crystalline structure whereby CaP crystallites are affixed to the implant surface in the shape of platelets or pins in near vertical aligment. This micro-crystalline coating accelerates bone ongrowth at the implant surface, thereby allowing faster and improved integration of the implant with controlled coating solubility. The CaP phase of BONIT is consisting of hydroxyapatite as a nucleator as well as the physiological pre-stage of the bone calcium phosphate hydroxyapatite with a Ca/P ratio of 1.0 (Brushite). Brushite is postulated as an intermediary phase in bone mineralization in vivo [1-5]. Cui et al. [22] found for the first time an amount of brushite particles in the external periostal callus of repaired femoral fractures in children. They speculated that the reason why childrens bone fractures heal faster might be related to the presence of brushite particles, that would serve as an ion reservoir for the bone matrix mineralization [22]. Villareal et al. [23] reported in vitro on a specific biologic activity for the brushite phase. They found that albumin absorption on brushite was 5x higher than on HA, they also measured for osteoblast cells in culture on pellets, a higher protein production and a higher ALP specific activity for brushite when compared to HA. Brushite has been considered as an ion reservoir of Ca and PO4 [4, 24, 25]. This high ion concentration is the reason for rapid contact osteogenesis and high mineralization; thus, the BONIT coating supports the natural healing process of a bone implant. The morphology creates an exceptional capillary effect that facilitates complete moistening of the implant surface upon the slightest contact with body fluids, specially blood. The in vivo results of BONIT can be summerized as follows: controlled resorption of BONIT coating and simultaneous substitution by new bone no inflammatory processes or foreign body reactions osseointegration without problems in shorter times.
Literature: [1] J. Xie, C. Riley, M. Kumar, K. Chittur FTIR / ATR study of protein adsorption and brushite transformation to hydroxyapatite. Biomaterials, 23 (17) (2002), 3609-16 L. Frauchinger, M. Taborelli, B.-O. Aronsson, P. Descouts Ion adsorption on titanium surfaces exposed to a physiological solution. Applied Surface Science, 143 (1) (1999), 67-77 M.W. Neuman Blood: Bone Equilibrium. Calcif Tissue Int, 34 (1982), 117-20 M.S. Johnsson, G.H. Nancollas The role of brushite and octacalcium phosphate in apatite formation. Critical Reviews in Oral Biology and Medicine, 3 (1/2) (1992), 61-82 S.V. Dorozhkin, M. Epple Die biologische und medizinische Bedeutung von Calciumphosphaten. Angew. Chemie, 114 (2002), 3260-77 S. Szmukler-Moncler, F. R. Figueiredo, P.Trisi, E.H. Rompen, R.Legrand "Immediate loading of single crowns retained by short implants. A histologic study with various surfaces in the canine mandible." Clinical Oral Implants Research, 11 (2000) 397, Abstract S. Szmukler-Moncler, F. R. Figueiredo, P.Trisi, E.H. Rompen, R.Legrand "Immediate loading of single crowns retained by short implants. A histologic study with various surfaces in the canine mandible." Clinical Oral Implants Research, 11 (2000) 397, Abstract G. Massei, P. Trisi, S. Smukler-Moncler, L. Malchiodi "Immediately loaded FBR-coated Pitt-Easy Bio-Oss implants. A histologic evaluation in 3 patients after 8-12 weeks of function." Clinical Oral Implants Research, 12 (2001) 409, Abstract L. Malchiodi, G. Massei, C. Turello, P. Masotto, M. Cassetta, G. Bortoloni, G. Cordioli, G. Del Prete, A. Della Bonna, F. Casseler, P. Masala, S. SzmuklerMoncler "Immediate loading of FBR-coated Pitt-Easy Bio-Oss implants. Preliminary results from a prospective multi-center study." Clinical Oral Implants Research, 12 (2001) 408, Abstract R. Bttcher, C. Becker, R. Semmler, T. Barth, K. Gross, P. Henriot, , W. Stermann "Shortened healing periods for FBR-coated Pitt-Easy Bio-Oss implants. Preliminary results from a prospective multi-center study in private practices." Clinical Oral Implants Research 12 (2001) 396, Abstract
[2]
[3]
[4]
[5]
[6]
[7]
[8]
[9]
[10]
Proceedings of 16th Conference of the European Society for Biomaterials, London 2001
90
mass [% ]
80
70
60
50
days in solution
Figure 1: Solubility of CaP coatings (Brushite and HA) in tris / HCl-buffer for 7 days at 37C. Composites of CaP are obtained by electrochemical deposition at 45C. The Ca/P ratio is adjustable and depends on process conditions. The solubility is changed in contrast to single phase CaP. Almost we observed a considerable rise in Ca/P ratio after immersing in tris / HCl buffer at 37C for 8 days. The Ca/P ratio is unchanged after 8 days in buffer for coating with brushite only. Conclusions The controlled and adjustable solubility of CaP composites results in a permanent local higher concentration of calcium and phosphate ions. In the first stage, due to higher soluble CaP phases the local increase in ion concentrations provides favourable conditions for proliferation and differentiation of osteoblast like cells in bone. CaP phases with less solubility represent an ion source over a longer period during bony anchorage. The morphology of these CaP-composite coatings in the shape of platelets or needles in nearly vertical alignment causes an exceptional capillary effect which enables a complete moistening of the surface and represents therefore an ideal temporary matrix for bone regeneration. References 1. S. H. Maxian et al. (1993) J. Biomed. Mat. Res. 27, 617 2. S. Szmukler-Moncler et al. (2000) Bioceramics, 13, 395-8 3. J. M. Zhang et al. (1998) J. Mater. Sci. Lett., 17, 1077- 9
HISTOMORPHOMETRICAL AND MECHANICAL EVALUATION OF VARIOUS SURFACES ON TITANIUM TESTBODIES PLACED INTO FEMORA OF THE GTTINGER MINIPIG CAN A RESORBABLE CA-P COATING INCREASE THE OSTEOINTEGRATION ?
+*Schwarz, M.L.; Kowarsch, M.; Rose, S.B.; Jani, L. +University Hospital Mannheim, Mannheim, Germany
Introduction: The quality of the surface of implants affects their osteointegration. Under this premise we can distinguish between a surface-altering and an applying procedure. The last one is used to design porous surfaces which enables the bony tissue to grow into the pores, creating a stable bond due to a form-closed, bone-implant contact. Osteointegration can be further improved by applying an HA coating. Using this procedure a further interface is created thus possibly making the fate of the HA coating uncertain. The use of soluble Ca-P promises the same advantages as the stable HA, without having to create another interface, and making the implant surface directly accessible to the bone tissue due to the early dissolution of the coating. The objective of this work was to compare four (A, B, C, D) different surfaces with each other and to investigate in which way they differentiate in respect to their osteointegration and their mechanical fixation. The target criteria were the histological integration and the maximal sheare-strength. A further test was to ascertain whether the site of the implant (intertrochanteric or intercondylar) had any effect on the findings. Material : Four cylindrical test specimens with rated dimensions of 8 mm x 30 mm were implanted transcortically into the femora of 21 adult Gttinger minipigs using an exact press-fit technique: two intertrochanteric and two intercondylar respectively. One animal had to be replaced as a result of a hematogenous infection. The surfaces of A were glass-blasted (Ra: 0.8 m), those of B sandblasted (Ra: 4.3 m), C and D were given a porous TPS (titan plasma spray) coating (Fa. XXX, Rostock, Germany) (Ra: 28.4 m), D in addition was treated electrochemically with a layer of Bonit (Fa. XXX, Rostock, Germany) (Ra: 28.4 m). Roughness gauging was performed with a Perthometer Concept (Fa. Mahr GmbH, Gttingen, Germany). 20 test specimens of each type of surface were implanted. 10 were tested histologically, the other 10 mechanically using a pull-out test. In a cross-over design the respectively left and right facing implants were evaluated mechanically and histologically. The fresh bone was subjected to mechanical testing immediately following the scarifying of the animals. The histological evaluation was performed on a calibrated image assessment program following the dyeing according to Masson Goldner. Three comparable sections per implant were taken to measure the direct osseous growth on the implant and to analyse the periprothetic tissue. The implants, together with the surrounding bone structure, were cut along their axis. The direction of the cut ran parallel to the direction of load. The animals were sacrificed approx. 12 weeks following the operative treatment (Range: 84, 97, Median 88 d). The experiment was granted permission by the Regierungsprsidium Karlsruhe on 29.11.1999, Ref. No. 359185.81/128/99. Statistical evaluation was performed with the aid of SAS and EXCEL programs. Results : The mechanical tests showed definite differences between individual groups (Table 1). The additional Bonit coating brought about an approximately 13% greater strength. No significant differences were found in the rigidity of the bone/implant bonding. The histomorphometric investigation showed definite differences. All surfaces showed significantly varying results (Fig. 1). The measurements were: Type A 1.9% (1.1), Type B 10.5% (3.6), Type C 22.4% (4.5), and Type D 48.8% (4.5). Neither the mechanical nor the histomorphometric examinations showed significant differences between the intertrochanteric and the intercondylar types of implants. Histologically, it was possible to see that in the case of Type A and B bone trabeculae were developing parallel to the surface of the implant, whereas in the case of Type C and D the bone trabeculae went directly onto the surface of the implant. In all the types the bone trabeculae were stronger developed close to the implant rather than further away from the implant. This was particularly obvious in the case of the Type A surface.
In the case of Type A and B we found more branch points with 3 or 4 arms as well as hole number (lacunae) than in Types C and D. The number of the trabeculae and the trabecular termini provided similar values. shear-strength stdev. stiffness stdev. [N/mm] [N/mm] A 0.7* 0.3 11.6 7.8 B 3.2* 0.6 13.3 2.1 C 6.5* 1.5 10.5 4.6 D 7.3 1.9 11.1 3.2 Table 1: Results of the pull - out tests and the stiffness analysis. Statistical significant (*) differences were seen with the shear - strength between all groups, except for group C and D. Regarding the stiffness no statistical significant differences were seen.
bony ongrowth
100 80 % 60 40 20 0 Glasperl Korund TPS BONIT
group
Fig.1: Types A (glass-blasted) B (sandblasted, Korund), C (TPS) and D (TPS +Bonit) were significantly different from each other. Discussion: We were able to confirm that an increased roughness results in an improved osteointegration, accompanied by a higher mechanical strength. The results show a definite upward movement when a porous surface (TPS) is used. In this case the spatial recesses provide not only a better mechanical strength, but the TPS coating used improves bone ongrowth. Following pull-out tests the implants having Type C and D surfaces still had fragments of bone adhering to them, which proves that the breaking takes place not only at the bone-implant interface, but also within the periprothetic bone. The additional coating with a soluble CaP (Bonit) brought about a definite increase in osteointegration whose significance in the histological finding was reflected in an increased, but not significantly higher mechanical strength. It was not possible to establish a clear indication that the soluble CaP has a positive effect on the periprothetic tissue compared to the usual histomorphometric parameters. Surfaces A and B, rather, showed indications of a more stable bone structure, but had developed a kind of periprothetic bone lamella. The present findings show that the site of the implant (intertrochanteric or intercondylar) does not necessarily produce different results. Thus, we consider the present transcortical model of the Gttinger Minipig as a reproducible approach.