Bioactive Carbonate Apatite Cement with Enhanced Compressive Strength via Incorporation of Silica Calcium Phosphate Composites and Calcium Hydroxide.

Carbonate apatite (CO3Ap) is a bioceramic material with excellent properties for bone and dentin regeneration. To enhance its mechanical strength and bioactivity, silica calcium phosphate composites (Si-CaP) and calcium hydroxide (Ca(OH)2) were added to CO3Ap cement. The aim of this study was to investigate the effect of Si-CaP and Ca(OH)2 on the mechanical properties in terms of the compressive strength and biological characteristics of CO3Ap cement, specifically the formation of an apatite layer and the exchange of Ca, P, and Si elements. Five groups were prepared by mixing CO3Ap powder consisting of dicalcium phosphate anhydrous and vaterite powder added by varying ratios of Si-CaP and Ca(OH)2 and 0.2 mol/L Na2HPO4 as a liquid. All groups underwent compressive strength testing, and the group with the highest strength was evaluated for bioactivity by soaking it in simulated body fluid (SBF) for one, seven, 14, and 21 days. The group that added 3% Si-CaP and 7% Ca(OH)2 had the highest compressive strength among the groups. SEM analysis revealed the formation of needle-like apatite crystals from the first day of SBF soaking, and EDS analysis indicated an increase in Ca, P, and Si elements. XRD and FTIR analyses confirmed the presence of apatite. This combination of additives improved the compressive strength and showed the good bioactivity performance of CO3Ap cement, making it a potential biomaterial for bone and dental engineering applications.

Inhibition of osteoclast activities by SCPC bioceramic promotes osteoblast-mediated graft resorption and osteogenic differentiation.

Maximizing vital bone in a grafted site is dependent on a number of factors. These include resorption or turnover of the graft material, stimulation of bone formation pathway without a need for biological molecules added to the site and inhibition of cellular activities that compromise the mineralization of new bone matrix. In the present study, the dissolution profile of silica-calcium phosphate composite (SCPC) in physiological solution was measured and the data were fed to (ANN-NARX) prediction model to predict the time required for complete dissolution. The inductively coupled plasma-optical emission spectrometer ionic composition analysis of the culture medium incubated for 3 days with SCPC showed 57% decrease in Ca concentration and a significant increase in the concentration of Si (13.5 ± 1.8 µg/ml), P (249.4 ± 22 µg/ml), and Na (9.3 ± 0.52 µg/ml). In conjunction with the release of Si, P, and Na ions, the bone resorptive activity of osteoclasts was inhibited as indicated by the significant decrease in multinucleated tartrate resistant acidic phosphate stained cells and the volume of resorption pits on bone slices. In contrast, addition of SCPC to hBMSC cultured in conventional medium promoted higher Runt-related transcription factor 2 (p < .05), osteocalcin (p < .01), and bone sialo protein (p < .01) than that expressed by control cells grown in the absence of SCPC. The predicted dissolution time of 200 mg of porous SCPC particles in 10 ml phosphate buffered saline is 6.9 months. An important byproduct of the dissolution is inhibition of osteoclastic activity and promotion of osteoblastic differentiation and hence bone formation.

Synthesis and characterization of porous bioactive SiC tissue engineering scaffold.

Silicon carbide (SiC) is an inert material with excellent biocompatibility properties. A major issue that limits its use as a medical device is the difficult processing technique that requires hot pressing at a temperature (>2,000o C) and pressure (1,000-2,000 atm). In the present study, we developed a protocol to synthesize a porous SiC scaffold by pressing the powder at 50 MPa and heating at 900o C/2 hr. The surface of SiC was chemically modified by NaOH to facilitate sintering and induce bioactivity. Porous discs with 51.51 ± 3.17% porosity and interconnected pores in the size range from 1 to 1,000 µm were prepared using 40% PEG. The average compressive strength and Young's modulus of the scaffolds were 1.94 ± 0.70 and 169.2 ± 0.08 MPa, respectively. FTIR analysis confirmed the formation of biomimetic hydroxyapatite layer after 2 hr of immersion in simulated body fluid. The Ca/P ratio was dependent on the concentration of the silanol groups created on the material surface. Increasing the atomic % of silicon on the SiC surface from 33.27 ± 9.53% to 45.13 ± 4.74% resulted in a 76% increase in the osteocalcin expression by MC3T3-E1 cells seeded on the material after 7 days. The cells colonized the entire thickness of the template and filled the pores with mineralized extracellular matrix after 14 days. Taken all together, the porous SiC scaffolds can serve as a bone graft for tissue reconstruction and cell delivery in trauma surgery.

Tissue Engineering: What is New?

Soft and hard tissue engineering has expanded the frontiers of oral/maxillofacial augmentation. Soft tissue grafting enhancements include improving flap prevascularization and using stem cells and other cells to create not only the graft, but also the vascularization and soft tissue scaffolding for the graft. Hard tissue grafts have been enhanced by osteoinductive factors, such as bone morphogenic proteins, that have allowed the elimination of harvesting autogenous bone and thus decrease the need for other surgical sites. Advancements in bone graft scaffolds have developed via seeding with stem cells and improvement of the silica/calcium/phosphate composite to improve graft characteristics and healing.

Calcium release and physical properties of modified carbonate apatite cement as pulp capping agent in dental application.

BACKGROUND: Carbonate apatite (CO3Ap) and silica-calcium phosphate composite (SCPC) are bone substitutes with good prospect for dental application. SCPC creates a hydroxyapatite surface layer and stimulate bone cell function while, CO3Ap induce apatite crystal formation with good adaptation providing good seal between cement and the bone. Together, these materials will add favorable properties as a pulp capping material to stimulate mineral barrier and maintain pulp vitality. The aim of this study is to investigate modification of CO3Ap cement combined with SCPC, later term as CO3Ap-SCPC cement (CAS) in means of its chemical (Calcium release) and physical properties (setting time, DTS and pH value). METHODS: The study consist of three groups; group 1 (100% calcium hydroxide, group 2 CO3Ap (60% DCPA: 40% vaterite, and group 3 CAS (60% DCPA: 20% vaterite: 20% SCPC. Distilled water was employed as a solution for group 1, and 0.2 mol/L Na3PO4 used for group 2 and group 3.Samples were evaluated with respect to important properties for pulp capping application such as pH, setting time, mechanical strength and calcium release evaluation. RESULTS: The fastest setting time was in CO3Ap cement group without SCPC, while the addition of 20% SCPC slightly increase the pH value but did not improved the cement mechanical strength, however, the mechanical strength of both CO3Ap groups were significantly higher than calcium hydroxide. All three groups released calcium ions and had alkaline pH. Highest pH level, as well as calcium released level, was in the control group. CONCLUSION: The CAS cement had good mechanical and acceptable chemical properties for pulp capping application compared to calcium hydroxide as a gold standard. However, improvements and in vivo studies are to be carried out with the further development of this material.

Novel Bioceramic Urethral Bulking Agents Elicit Improved Host Tissue Responses in a Rat Model.

Objectives. To test the physical properties and host response to the bioceramic particles, silica-calcium phosphate (SCPC10) and Cristobalite, in a rat animal model and compare their biocompatibility to the current clinically utilized urethral bulking materials. Material and Methods. The novel bulking materials, SCPC10 and Cristobalite, were suspended in hyaluronic acid sodium salt and injected into the mid urethra of a rat. Additional animals were injected with bulking materials currently in clinical use. Physiological response was assessed using voiding trials, and host tissue response was evaluated using hard tissue histology and immunohistochemical analysis. Distant organs were evaluated for the presence of particles or their components. Results. Histological analysis of the urethral tissue five months after injection showed that both SCPC10 and Cristobalite induced a more robust fibroblastic and histiocytic reaction, promoting integration and encapsulation of the particle aggregates, leading to a larger bulking effect. Concentrations of Ca, Na, Si, and P ions in the experimental groups were comparable to control animals. Conclusions. This side-by-side examination of urethral bulking agents using a rat animal model and hard tissue histology techniques compared two newly developed bioactive ceramic particles to three of the currently used bulking agents. The local host tissue response and bulking effects of bioceramic particles were superior while also possessing a comparable safety profile.

Acceleration of Alveolar Ridge Augmentation Using a Low Dose of Recombinant Human Bone Morphogenetic Protein-2 Loaded on a Resorbable Bioactive Ceramic.

PURPOSE: The aim of the present study was to evaluate the effect of a porous silica-calcium phosphate composite (SCPC50) loaded with and without recombinant human bone morphogenetic protein-2 (rhBMP-2) on alveolar ridge augmentation in saddle-type defects. MATERIALS AND METHODS: Micro-granules of SCPC50 resorbable bioactive ceramic were coated with rhBMP-2 10 mg and then implanted into a saddle-type defect (12 × 7 mm) in a dog mandible and covered with a collagen membrane. Control groups included defects grafted with SCPC50 granules without rhBMP-2 and un-grafted defects. Bone healing was evaluated at 8 and 16 weeks using histologic and histomorphometric techniques. The increase in bone height and total defect fill were assessed for each specimen using the ImageJ 1.46 program. The release kinetics of rhBMP-2 was determined in vitro. The height of the bone in the grafted defects and the total defect fill were statistically analyzed. RESULTS: SCPC50 enhanced alveolar ridge augmentation as indicated by the increased vertical bone height, bone surface area, and bone volume after 16 weeks. SCPC50-rhBMP-2 provided a sustained release profile of a low effective dose (BMP-2 4.6 ± 1.34 pg/mL per hour) during the 1- to 21-day period. The slow rate of release of rhBMP-2 from SCPC50 accelerated synchronized complete bone regeneration and graft material resorption in 8 weeks. Successful rapid reconstruction of the alveolar ridge by SCPC50 and SCPC50-rhBMP-2 occurred without any adverse excessive bone formation, inflammation, or fluid-filled voids. CONCLUSIONS: Results of this study suggest that SCPC50 is an effective graft material to preserve the alveolar ridge after tooth extraction. Coating SCPC50-rhBMP-2 further accelerated bone regeneration and a considerable increase in vertical bone height. These findings make SCPC50 the primary choice as a carrier for rhBMP-2. SCPC50-rhBMP-2 can serve as an alternative to autologous bone grafting.

Rat animal model for preclinical testing of microparticle urethral bulking agents.

OBJECTIVES: To develop an economic, practical and readily available animal model for preclinical testing of urethral bulking therapies, as well as to establish feasible experimental methods that allow for complete analysis of hard microparticle bulking agents. METHODS: Alumina ceramic beads suspended in hyaluronic acid were injected into the proximal urethra of 15 female rats under an operating microscope. We assessed overall lower urinary tract function, bulking material intraurethral integrity and local host tissue response over time. Microphotographs were taken during injection and again 6 months postoperatively, before urethral harvest. Urinary flow rate and voiding frequency were assessed before and after injection. At 6 months, the urethra was removed and embedded in resin. Hard tissue sections were cut using a sawing microtome, and processed for histological analysis using scanning electron microscopy, light microscopy and immunohistochemistry. RESULTS: Microphotographs of the urethra showed complete volume retention of the bulking agent at 6 months. There was no significant difference between average urinary frequency and mean urinary flow rate at 1 and 3 months postinjection as compared with baseline. Scanning electron microscopy proved suitable for evaluation of microparticle size and integrity, as well as local tissue remodeling. Light microscopy and immunohistochemistry allowed for evaluation of an inflammatory host tissue reaction to the bulking agent. CONCLUSIONS: The microsurgical injection technique, in vivo physiology and novel hard tissue processing for histology, described in the present study, will allow for future comprehensive preclinical testing of urethral bulking therapy agents containing microparticles made of a hard material.

Early osteoblast responses to orthopedic implants: Synergy of surface roughness and chemistry of bioactive ceramic coating.

Pro-osteogenic stimulation of bone cells by bioactive ceramic-coated orthopedic implants is influenced by both surface roughness and material chemistry; however, their concomitant impact on osteoblast behavior is not well understood. The aim of this study is to investigate the effects of nano-scale roughness and chemistry of bioactive silica-calcium phosphate nanocomposite (SCPC50) coated Ti-6Al-4V on modulating early bone cell responses. Cell attachment was higher on SCPC50-coated substrates compared to the uncoated controls; however, cells on the uncoated substrate exhibited greater spreading and superior quality of F-actin filaments than cells on the SCPC50-coated substrates. The poor F-actin filament organization on SCPC50-coated substrates is thought to be due to the enhanced calcium uptake by the ceramic surface. Dissolution analyses showed that an increase in surface roughness was accompanied by increased calcium uptake, and increased phosphorous and silicon release, all of which appear to interfere with F-actin assembly and osteoblast morphology. Moreover, cell attachment onto the SCPC50-coated substrates correlated with the known adsorption of fibronectin, and was independent of surface roughness. High-throughput genome sequencing showed enhanced expression of extracellular matrix and cell differentiation related genes. These results demonstrate a synergistic relationship between bioactive ceramic coating roughness and material chemistry resulting in a phenotype that leads to early osteoblast differentiation.

Tissue engineering scaffold for sequential release of vancomycin and rhBMP2 to treat bone infections.

The ability of silica calcium phosphate nanocomposite (SCPC75) for the controlled sequential delivery of vancomycin (Vanc) and rhBMP2 was evaluated. Fourier transform infrared spectroscopy analyses of the SCPC75 showed an increase in the bond energy of the PO4 (-3) due to the interactions with negatively charged moieties of Vanc. Furthermore, a decrease in the bond energy of the Si-O-Si functional groups was observed after rhBMP2 adsorption. In conjunction with the differences in bond site and bond energy at the ceramic/drug interface, significant differences in drug release kinetics and bioceramic dissolution rate were found. UV-vis spectrometry showed a burst release of Vanc in the first 8 h followed by a sustained release stage for up to 28 days. ELISA showed first-order release kinetics of rhBMP2 without burst release. The rhBMP2 release from SCPC75 was associated with a significantly lower rate of Ca and a higher rate of Si dissolutions when compared with Vanc release over identical time periods. Differences in the release kinetic profiles of Vanc and rhBMP2 from the SCPC75-Vanc/SCPC75-rhBMP2 scaffolds at 70/30, 50/50, or 20/80 ratios allowed for sequential drug release profiles that could be exploited to customize doses and release duration of each drug. The released rhBMP2 significantly upregulated MC3T3-E1 expression of collagen type I, osteopontin, and osteocalcin mRNA by 12.6-, 3.3-, and 2.4-fold, respectively. The released Vanc demonstrated bactericidal effects on Staphylococcus aureus in vitro. These results suggest the potential of SCPC75-Vanc-rhBMP2 scaffolds in the treatment of damaged and/or infected bone.

Alveolar ridge preservation using resorbable bioactive ceramic composite: a histological study.

PURPOSE: The purpose of this study was to histologically evaluate newly generated vital bone using porous granules of bioactive and resorbable silica-calcium phosphate nanocomposite (SCPC) in extraction sockets. MATERIAL AND METHODS: Six patients with a non-restorable maxillary central incisor requiring extraction followed by implant placement participated in the study. Extraction sockets were grafted with granules of SCPC. After 6 months, a bone core sample was retrieved from the center of the healed socket for histologic analysis, and dental implants were placed. Alveolar bone width was clinically assessed immediately after tooth extraction and 6 months after bone grafting, at the time of implant placement. Alveolar bone height was radiographically assessed immediately after tooth extraction and 6 months after extraction. RESULTS: Histomorphometric analyses of sockets grafted with SCPC for 6 months revealed 46.8% +/- 14% new vital bone and 2.5% +/- 1.5% graft material remnants. In these sockets, the mean bone height resorption over the 6-month period of healing was 1.6 mm +/- 1.5 mm. The mean bone width resorption of 2 mm +/- 0.7 mm was found at the bone crest. CONCLUSION: The results of this study suggest that SCPC graft material reduces the amount of change in alveolar ridge dimensions after tooth extraction and facilitates the regeneration of new vital bone.

Mechanical properties and cytotoxicity of a resorbable bioactive implant prepared by rapid prototyping technique.

Bioceramic processing using rapid prototyping technique (RPT) results in a fragile device that requires thermal treatment to improve the mechanical properties. This investigation evaluates the effect of thermal treatment on the mechanical, porosity, and bioactivity properties as well as the cytotoxicity of a porous silica-calcium phosphate nanocomposite (SCPC) implant prepared by RPT. Porous SCPC implant was subject to 3-h treatment at 800°C, 850°C, or 900°C. The compressive strength (s) and modulus of elasticity (E) were doubled when the sintering temperature is raised from 850 to 900°C measuring (s = 15.326 ± 2.95 MPa and E = 1095 ± 164 MPa) after the later treatment. The significant increase in mechanical properties takes place with minimal changes in the surface area and the percentage of pores in the range 1-356 µm. The SCPC implant prepared at 900°C was loaded with rh-BMP-2 and grafted into a segmental defect in the rabbit ulna. Histology analyses showed highly vascularized bone formation inside the defect. Histopathological analyses of the liver, spleen, kidney, heart, and the lung of rabbits grafted with and without SCPC demonstrated healthy tissues with no signs of toxicity or morphology alterations. Results of the study suggest that it is possible to engineering the mechanical properties of the SCPC implant without compromising its bioactivity. The enhanced bone formation inside the porous SCPC facilitated cell-mediated graft resorption and prohibited any accumulation of the material in the body organs.

Machining of a bioactive nanocomposite orthopedic fixation device.

Bioactive ceramics bond to bone and enhance bone formation. However, they have poor mechanical properties which restrict their machinability as well as their application as load bearing implants. The goal of this study was to machine bioactive fixation screws using a silica-calcium phosphate nanocomposite (SCPC50). The effect of compact pressure, holding time, and thermal treatment on the microstructure, machinability, and mechanical properties of SCPC50 cylinders were investigated. Samples prepared by powder metallurgy technique at compact pressure range of 100-300 MPa and treated at 900°C/1 h scored a poor machinability rating of (1/5) due to the significant formation of amorphous silicate phase at the grain boundaries. On the other hand, lowering of compact pressure and sintering temperature to 30 MPa/3 h and 700°C/2 h, respectively, minimized the formation of the amorphous phase and raised the machinability rating to (5/5). The modulus of elasticity and ultimate strength of machinable SCPC50 were 10.8 ± 2.0 GPa and 72.8 ± 22.8 MPa, respectively, which are comparable to the corresponding values for adult human cortical bone. qRT-PCR analyses showed that bone cells attached to SCPC50 significantly upregulated osteocalcin mRNA expression as compared to the cells on Ti-6Al-4V. Moreover, cells attached to SCPC50 produced mineralized bone-like tissue within 8 days. On the other hand, cells attached to Ti-6Al-4V failed to produce bone mineral under the same experimental conditions. Results of the study suggest that machinable SCPC50 has the potential to serve as an attractive new material for orthopedic fixation devices.

Promotion of pro-osteogenic responses by a bioactive ceramic coating.

The objective of this study was to analyze the responses of bone-forming osteoblasts to Ti-6Al-4V implant material coated with silica-calcium phosphate nanocomposite (SCPC50). Osteoblast differentiation at the interface with SCPC50-coated Ti-6Al-4V was correlated to the adsorption of high amount of serum proteins, high surface affinity to fibronectin, Ca uptake from and P and Si release into the medium. SCPC50-coated Ti-6Al-4V adsorbed significantly more serum protein (p < 0.05) than control uncoated substrates. Moreover, Western blot analysis showed that the SCPC50 coating had 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 substrates than that on the surface of the control uncoated substrates. 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-6A1-4V substrates. 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 substrates. These results suggest that SCPC50 coating could enhance bone integration with orthopedic and maxillofacial implants while minimizing the induction of inflammatory bone cell responses.

Evaluation of a bioresorbable drug delivery system for the treatment of hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) represents a major global health burden. Typically HCC responds poorly to chemotherapy, and such approaches to treat HCC are commonly associated with severe hepatic and/or systemic toxicity. The aim of this study was to evaluate a porous resorbable silica-calcium phosphate nanocomposite (SCPC) as a controlled release vehicle for cisplatin. Particles of two different formulations--SCPC50 and SCPC75, containing 19.49 and 32.9 mol % silica, respectively--were loaded with cisplatin by immersion treatment and pressed into discs. In vitro release kinetics studies of cisplatin from SCPC50 and SCPC75 demonstrated an initial burst release of 0.39 ± 0.04 mg (of the 1.49 mg total loaded) and 0.87 ± 0.07 mg (of the 2.34 mg total loaded), respectively. Over the following 44-day period. SCPC75-cisplatin hybrid produced a significantly higher sustained cisplatin release than that released from SCPC50. Cisplatin release correlated well with the surface area, and silica dissolution kinetics of the SCPC carrier. Treatment of rat HCC cells (H4IIE) with cisplatin released from SCPC-cisplatin hybrids induced apoptotic cell death in H4IIE cells in vitro. Results of this study suggest that SCPC composites may be of potential use for the treatment of HCC in vivo.

Electrophoretic deposition of bioactive silica-calcium phosphate nanocomposite on Ti-6Al-4V orthopedic implant.

Bioactive silica-calcium phosphate nanocomposite (SCPC) has been coated on Ti-6Al-4V implant employing an electrophoretic deposition (EPD) technique. The effects of composition and pH of the suspending medium 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. Discs of Ti-6Al-4V were passivated, coated with SCPC50 (200 nm-10 µm) and thermally treated at 600-800°C 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°C, 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. SEM-EDX analyses of SCPC50-coated Ti-6Al-4V preimmersed in PBS for 7 days showed the formation of a Ca-deficient hydroxyapatite surface layer. ICP-OES analyses of the immersing solution (n = 6) showed an increase in the ionic concentration of Si from 3.3 ± 0.9 to 5.0 ± 1.2 ppm between days 1 and 4; after which no significant change in the Si concentration was measured. Bone marrow mesenchymal stem 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. Results of the study suggest that bioactive SCPC50 can efficiently be coated on Ti-6Al-4V using EPD. The SCPC50 coating has the potential to enhance bone integration with the orthopedic implant.

Effects of exogenous phosphorus and silicon on osteoblast differentiation at the interface with bioactive ceramics.

In this study, we have investigated the effects of dissolved phosphorus and silicon on osteoblast differentiation in vitro. Neonatal rat calvarial osteoblasts were seeded on silica-calcium phosphate composites (SCPCS), hydroxyapatite (HA-200), and tissue culture polystyrene (TCPS) and incubated over 4 days in media containing 0 {minimal essential medium [MEM] (-)} or 3 mM ß-glycerophosphate [MEM (+)]. Inductively coupled plasma analysis showed that P-content in original MEM (+) was 225% higher than that in MEM (-). Moreover, P-content in MEM (+) significantly increased to 3.4-4.4 mM and 3.6-4.7 mM after 2 and 4 days incubation with SCPC, respectively, owing to material dissolution and exogenous phosphate supplementation. In contrast, P-content in MEM (+) showed no change upon incubation with HA or TCPS. The P-content in MEM (-) incubated with SCPC was considerably lower than that in MEM (+). SCPC exhibited controlled Si-release in cell culture media [MEM (-) or MEM (+)], with Si-rich SCPC showing a significantly greater dissolution than Si-poor SCPC. Moreover, SCPC, unlike HA, demonstrated a cell- and solution-mediated dissolution over 4 days. Quantitative real-time PCR showed that in MEM (-), osteocalcin and osteopontin mRNA expression on Si-rich SCPC was significantly greater than that on HA, suggesting that Si plays an important role in enhancing bone-cell differentiation. However, osteoblast phenotypic expression on SCPC was significantly decreased after 4 days incubation in MEM (+), indicating that sustained exposure to elevated P-levels in the media can downregulate osteoblast function. Our results demonstrate that the controlled dissolution of SCPC provides a natural stimulus for bone-cell differentiation in vitro and could obviate the need of exogenous phosphate supplementation.

A ceramic-based anticancer drug delivery system to treat breast cancer.

Drug delivery systems offer the advantage of sustained targeted release with minimal side effect. In the present study, the therapeutic efficacy of a porous silica-calcium phosphate nanocomposite (SCPC) as a new delivery system for 5-Fluorouracil (5-FU) was evaluated in vitro and in vivo. In vitro studies showed that two formulations; SCPC50/5-FU and SCPC75/5-FU hybrids were very cytotoxic for 4T1 mammary tumor cells. In contrast, control SCPCs without drug did not show any measurable toxic effect. Release kinetics studies showed that SCPC75/5-FU hybrid provided a burst release of 5-FU in the first 24 h followed by a sustained release of a therapeutic dose (30.7 microg/day) of the drug for up to 32 days. Moreover, subcutaneous implantation of SCPC75/5-FU hybrid disk in an immunocompetent murine model of breast cancer stopped 4T1 tumor growth. Blood analyses showed comparable concentrations of Ca, P and Si in animals implanted with or without SCPC75 disks. These results strongly suggest that SCPC/5-FU hybrids can provide an effective treatment for solid tumors with minimal side effects.

Resorbable bioactive ceramic for treatment of bone infection.

Polymethylmethacrylate (PMMA) beads have been widely used in the treatment of bone infection over the last three decades. Although PMMA does offer a mechanism to quickly and effectively administer a localized dose of antibiotic to the site of infection, its efficacy is limited by its nonresorbability and nonbioactivity. Resorbable bioactive silica-calcium phosphate nanocomposite (SCPC75) was investigated as a novel controlled release carrier of vancomycin for the treatment of osteomyelitis. SCPC75 particles adsorbed significantly higher amount of vancomycin compared with PMMA. Moreover, SCPC75 provided a sustained release kinetics of therapeutic dose of vancomycin up to 35 days. The novel resorbable ceramic was able to release 95.5% of the adsorbed drug in an average dose of 12 microg/mL/day over 480 h (35 days). In conjunction with the sustained drug release, a controlled dissolution rate that led to 40% mass loss of SCPC75 was observed. On the other hand, PMMA provided a sustained release of a therapeutic dose of vancomycin for 14 days after which minimal concentration of the drug was detected. Moreover, PMMA retained 32% of the drug adsorbed onto its surface. The SCPC-vancomycin implant can serve a dual function: provide a sustained therapeutic dose of antibiotic to eradicate infection and stimulate bone cell differentiation and new bone formation. (c) 2010 Wiley Periodicals, Inc. J Biomed Mater Res, 2010.

Effect of bioactive glass crystallization on the conformation and bioactivity of adsorbed proteins.

Protein adsorption onto the surface of a biomaterial mediates cell adhesion and enhances tissue-implant integration. In a previous study, we demonstrated that crystallization of bioactive glass (BG) significantly increased the negative zeta potential and decreased serum protein adsorption onto the material surface. In this study, the conformation of protein adsorbed onto the surface of amorphous bioactive glass (ABG) and crystallized bioactive glass (CBG) was analyzed and correlated to bone marrow mesenchymal stem cell adhesion and spreading. ABG and CBG were immersed in three different protein solutions containing 10% fetal bovine serum, bovine serum albumin (BSA), and fibronectin (FN) for 4 h at 37 degrees C. Grazing angle Fourier transform infrared spectroscopy (GA-FTIR) demonstrated that the ratio of (amide I)/(amide II) functional groups of all proteins adsorbed onto ABG was greater than that for proteins adsorbed onto CBG. The Gaussian curve fitting analysis suggests that the significant expression of amide I, rich in charged and flexible unordered secondary structure of adsorbed FN, stimulated bone cell adhesion and spreading on the surface of ABG. CBG enforces protein conformation that exposes amide II, rich in neutral and stable beta-sheet structure and alpha-helix, which limited cell adhesion and spreading. Although ABG adsorbed significantly higher quantity of BSA than FN, GA-FTIR analyses showed that the ratio of amide I/amide II was significantly higher for adsorbed FN. Therefore, the intensity of amide I or amide II bands cannot be taken as a measure of the quantity of adsorbed protein.