Bioactive ceramic coating on orthopedic implants for enhanced bone tissue integration
Analytics
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Abstract
Tissue integration between bone and orthopedic implant is essential for implant fixation and longevity. An immunological response leads to fibrous encapsulation of metallic implants leading to implant instability and failure. Bioactive ceramics have the ability to directly bond to bone; however, they have limited mechanical strength for load bearing applications. Coating bioactive ceramics on metallic implant offers the exciting opportunity to enhance bone formation without compromising the mechanical strength of the implant. In the present study, we have developed a novel bioactive silica-calcium phosphate nanocomposite (SCPC) coating on medical grade Ti-6Al-4V orthopedic implant using electrophoretic deposition (EPD) and evaluated bone tissue response to the coated implant at the cellular level. The effect of SCPC composition and suspending medium pH on the zeta potential of three different SCPC formulations; SCPC25, SCPC50 and SCPC75 were analyzed. The average zeta potential of SCPC50 in pure ethanol was more negative than that of SCPC25 or SCPC75; however the difference was not statistically significant. Ti-6Al-4V discs were passivated, coated with SCPC50 (200 nm - 10 μm) and thermally treated at 600 - 800 ºC to produce a coating thickness in the range of 43.1 ± 5.7 to 30.1 ± 4.6 μm. After treatment at 600, 700 and 800 ºC, the adhesion strength at the SCPC50/Ti-6Al-4V interface was 42.6 ± 3.6, 44.7 ± 8.7 and 47.2 ± 4.3 MPa, respectively. XRD analyses of SCPC50 before and after EPD coating indicated no change in the crystallinity of the material. Fracture surface analyses showed that failure occurred within the ceramic layer or at the ceramic/polymer interface; however, the ceramic/metal interface was intact in all samples. The adhesion strength of SCPC50-coated substrates after immersion in PBS for 2 days (11.7 ± 3.9 MPa) was higher than that measured on commercially available hydroxyapatite (HA) coated substrates (5.5 ± 2.7 MPa), although the difference was not statistically significant. SEM - EDX analyses of SCPC50-coated Ti-6Al-4V pre-immersed in PBS for 7 days showed the formation of a Ca-deficient HA surface layer. Bone cells attached to the SCPC50-coated implants expressed significantly higher (p < 0.05) alkaline phosphatase activity (82.4 ± 25.6 nmoles p-NP/mg protein/min) than that expressed by cells attached to HA-coated or uncoated implants. Protein adsorption analyses showed that SCPC50-coated substrates adsorbed significantly more (p < 0.05) serum protein (14.9 ± 1.2 µg) than control uncoated substrates (8.9 ± 0.7 µg). Moreover, Western blot analysis showed that the SCPC50 coating has a high affinity for serum fibronectin. Protein conformation analyses by FTIR showed that the ratio of the area under the peak for amide I/amide II bands was significantly higher (p < 0.05) on the surface of SCPC50-coated substrate (5.0 ± 0.6) than that on the surface of the control uncoated substrates (2.2 ± 0.3). Moreover, ICP-OES analyses indicated that SCPC50-coated substrates withdrew Ca ions from, and released P and Si ions into, the tissue culture medium, respectively. In conjunction with the favorable protein adsorption and modifications in medium composition, MC3T3-E1 osteoblast-like cells attached to SCPC50-coated substrates expressed 10-fold higher level of mRNA encoding osteocalcin and had significantly higher production of osteopontin and osteocalcin proteins than cells attached to the uncoated Ti-6Al-4V substrate. In addition, osteoblast-like cells attached to the SCPC50-coated substrates produced significantly lower levels of the inflammatory and osteoclastogenic cytokines, IL-6, IL-12p40 and RANKL than those attached to uncoated Ti-6Al-4V. Surface topography analyses using AFM suggested that the SCPC50 particles deposit onto the metal surface in a manner that preferentially fills the grooves on the substrate created during substrate preparation. An increase in the surface roughness of the SCPC50-coated substrate from 217.8 ± 54.6 nm to 284.3 ± 37.3 nm was accompanied by enhanced material dissolution, reduced cell proliferation and poor actin cytoskeleton organization, which are characteristics typical of differentiating bone cells on bioactive ceramic surfaces. Results of the study demonstrate that bioactive SCPC50 can efficiently be coated on Ti-6Al-4V using EPD. Moreover, the in vitro bone cell response suggests that SCPC50-coating has the potential to enhance bone integration with orthopedic and maxillofacial implants while minimizing the induction of inflammatory bone cell responses.