Optimisation of the mechanical and handling properties of an injectable calcium phosphate cement

Calcium phosphate cements have the potential to be successful in minimally invasive surgical techniques, like that of vertebroplasty, due to their ability to be injected into a specific bone cavity. These bone cements set to produce a material similar to that of the natural mineral component in bone. Due to the ceramic nature of these materials they are highly brittle and it has been found that they are difficult to inject. This study was carried out to determine the factors that have the greatest effect on the mechanical and handling properties of an apatitic calcium phosphate cement with the use of a Design of Experiments (DoE) approach. The properties of the cement were predominantly influenced by the liquid:powder ratio and weight percent of di-sodium hydrogen phosphate within the liquid phase. An optimum cement composition was hypothesised and tested. The mechanical properties of the optimised cement were within the clinical range for vertebroplasty, however, the handling properties still require improvement.     

Novel calcium phosphate composite bone cement: strength and bonding properties

A new high-strength cement prepared from calcium phosphate and calcium aluminate has been developed and was evaluated for potential use in bone and joint repair applications. Cement specimens were aged under simulated physiological conditions. The compressive strength of the cement was determined at time intervals 1 h after setting up to 52 weeks. A compressive strength of 111.6±12.9 MPa was measured at 4 weeks, with the cement attaining 64% of this maximum strength within 4 h of preparation. Compressive strength greater than 90 MPa was maintained up to 52 weeks. The strength of the cement–prosthesis interface was studied using a pull-out test. Polished, 316L stainless steel rods were implanted in canine cadaver femurs to simulate a cemented hip prosthesis. At 4, 24 h, and 60 days post implantation, the force required to displace the rod was measured. Mean interfacial shear strengths of 1.17±0.25, 1.11±0.21, and 1.11±0.32 MPa were observed at respective time-periods.     

On the development of an apatitic calcium phosphate bone cement

Development of an apatitic calcium phosphate bone cement is reported. 100 μ Particles of tetracalcium phosphate (TTCP) and dicalcium phosphate dihydrate (DCPD) were mixed in equimolar ratio to form the cement powder. The wetting medium used was distilled water with Na₂HPO₄ as accelerator to manipulate the setting time. The cement powder, on wetting with the medium, formed a workable putty. The setting times of the putty were measured using a Vicat type apparatus and the compressive strength was determined with a Universal Testing Machine. The nature of the precipitated cement was analyzed through X-ray diffraction (XRD), fourier transform infrared spectrometry (FTIR) and energy dispersive electron microprobe (EDAX). The results showed the phase to be apatitic with a calcium-to-phosphorous ratio close to that of hydroxyapatite. The microstructure analysis using scanning electron microscopy (SEM) showed hydroxyapatite nano-crystallite growth over particulate matrix surface. The structure has an apparent porosity of ∼ 52%. There were no appreciable dimensional or thermal changes during setting. The cement passed the in vitro toxicological screening (cytotoxicity and haemolysis) tests. Optimization of the cement was done by manipulating the accelerator concentration so that the setting time, hardening time and the compressive strength had clinically relevant values.     

一种可注射可降解磷酸钙骨水泥的结构与性能

通过采用部分结晶磷酸钙和磷酸氢钙制备了新型可注射可降解磷酸钙骨水泥.研究表明:该材料具备优良的可注射性能,并通过添加变性淀粉,显著改善了材料的抗溃散性能,骨水泥的水化产物是直径和长度在分别约为100和1000nm左右的棒状类骨羟基磷灰石.所研制的骨水泥在体温(37℃)条件下凝结较快,而在室温(25℃)和冷藏温度(5℃)可在较长时间保持不固化,这就为骨水泥的临床应用提供了很有利的条件.体外溶血试验、体外细胞毒试验、热原性试验、小鼠的急性毒性试验、微核试验、豚鼠的致敏性试验、小鼠的肌内埋植试验及兔的骨内埋植试验等一系列毒性及生物相容性试验表明该材料无毒副作用,具有良好的生物相容性.复合rhBMP-2的可注射磷酸钙骨水泥植入猕猴椎体后的近远期影像学和组织学观察表明,骨水泥可降解且降解和新骨长入基本同步.     

磷酸钙骨水泥化学活性及机械性能研究

在研究磷酸四钙制备方法的基础上,讨论其化学活性,发现该物质在高温时热稳定性不好,易于向能量更低的羟基磷灰石转化;在室温下又容易吸附空气中的水分子,发生缓慢水解.因此最好真空保存.     

The optimum zinc content in set calcium phosphate cement for promoting bone formation in vivo

The final aim of our study is to develop a novel calcium phosphate cement based on zinc-containing α-tricalcium phosphate (αZnTCP) and evaluate its potential as bonegraft material in vivo. In the present study, in vivo efficacy of zinc in hardened bodies of αZnTCP was explored. The hardened bodies prepared from αZnTCP with zinc content of 0.00, 0.04, 0.08, 0.11 and 0.19 wt % were prepared by mixing pure αTCP or αZnTCP powder with 12 wt% sodium succinate solution at a solid-to-liquid ratio of 2.0. Due to the release of zinc ions into the physiological salt solution during curing, the zinc content in the hardened bodies was calculated to be 0.00, 0.03, 0.06, 0.10 and 0.18 wt%, respectively. The hardened bodies were implanted in the femora and tibia of white rabbits for 4 weeks. Histological and histomorphometric evaluation showed that the hardened body containing 0.03 wt% zinc, significantly promoted more new bone formation without evoking adverse tissue reactions than that without zinc. The hardened bodies containing 0.06 and 0.10 wt% zinc also resulted in the increase in numbers of active osteoblasts surrounding the new bone but caused inflammation at the implant sites. Results of this study indicate that the hardened body prepared with αZnTCP is superior to that prepared with αTCP in promoting new bone formation due to the release of zinc ions. This study also indicates that the optimum amount of zinc in the hardened body is about 0.03 wt % to avoid inflammatory reaction.     

In situ study on the curing process of calcium phosphate bone cement

The aim of this study was to follow the entire curing process of modified alpha-TCP cement, and to explore how the liquid phase affects the curing reaction. Two calcium phosphate bone cements (CPCs) with a variety of aqueous solution were studied for comparison. In situ X-ray diffraction analysis and pH testing were employed to follow the chemical reaction, while quantitative ultrasonic measurement (QUS) was carried out to monitor the physical change. Results showed that CPC powders were completely consumed after 72 h. Two steps were presented in apatite formation. The first step was the precipitation of carbonated hydroxyapatite (CHA), and in the second step, conversion of calcium deficient hydroxyapatite (CDHA) was the dominant reaction. Finally, CPCs were fully converted to apatite except the cement with NaH2PO4 as liquid phase, because acidic environment inhibited the conversion of apatite. The pH increased linearly after mixing, when supersaturation was reached, it decreased to pH approximately 6.0 gradually. Ultrasound measurement indicated that the variation of speed of sound (SOS) was related to both apatite formation and microstructural evolution. Ultrasonic attenuation coefficient (UAC) was able to quantitatively describe the curing process from viscous paste to elastic solid as a function of curing time. Moreover, the curing reaction conformed to classical dissolution-precipitation mechanism.     

The mechanical properties of recovered PMMA bone cement: A preliminary study

Samples of polymethylmethacrylate (PMMA) bone cement, used in the fixation of hip prostheses, have been recovered from 11 patients after in service life spans of between 15 and 24 years. Eighteen samples in total have been recovered from the acetabular and/or femoral cement. Samples were subjected to three point bending tests, their density, porosity and microhardness determined and all specimens were examined using EDX and X-ray techniques. Since the porosity of many of the samples is very high, the continuous matrix properties are inferred from the performance of individual specimens. No evidence has been found to suggest that the PMMA has deteriorated whilst in-vivo and the mechanical properties of the cement matrices appear to be comparable to freshly made PMMA.     

磷酸钙/纤维蛋白胶复合支架材料的结构及力学性能分析

用可吸收磷酸钙骨水泥和纤维蛋白胶按一定比例体外构建复合支架材料,通过XRD、SEM、抗压实验和空隙率测试等方法对其结构及力学性能进行分析.结果发现:由于加入纤维蛋白胶,复合支架材料在一定程度上延长了磷酸钙骨水泥的初凝时间,但并不影响磷酸钙骨水泥的终凝时间;同时,加入纤维蛋白胶改变了骨水泥固化体的微观结构,提高了骨水泥的抗压强度,其最大抗压强度达到14MPa,弹性模量在96.64～269.39MPa之间,空隙率为38.8%.与在同样条件下制备的磷酸钙骨水泥比较,复合支架材料的抗压强度增强了55.6%,而空隙率仅仅下降了6.9%;XRD分析显示,复合支架材料并不影响磷酸钙骨水泥的最终的转化,其结晶结构仍是羟基磷灰石结构,是更好的骨组织工程支架材料.     

可注射磷酸钙骨水泥及其应用

可注射磷酸钙骨水泥作为一种新型人工骨替代材料,以其良好的生物相容性和骨传导性被广泛应用于临床骨缺损和牙缺损的修复.本文介绍了可注射磷酸钙骨水泥的种类和特性,指出了存在的问题和应用前景.     

In vivo behaviour of three calcium phosphate cements and a magnesium phosphate cement

Three types of calcium phosphate cements and one magnesium phosphate cement were implanted subcutaneously in rats under exclusion of direct cellular contact. Retrieval times were either 1, 2, 4 or 8 weeks. Before and after retrieval the compressive strength, the diametral tensile strength, the quantitative chemical composition, the qualitative phase composition, the FTIR spectrum and the microstructure were determined. The three calcium phosphate cements maintained their strength during implantation. The phase DCPD was completely transformed into a Na- and CO₃-containing apatite, the phases DCP and CDHA only partially. It could not be ruled out that OCP is also transformed into a bone-mineral-like apatite to a certain extent. That this latter process occurs much faster during the turn-over of living bone, is probably due to the very small crystal size of the OCP particles in bone.     

Characterization of a novel calcium phosphate composite bone cement: Flow, setting, and aging properties

The flow, setting, and aging characteristics of a newly developed calcium phosphate/calcium aluminate composite orthopaedic cement were studied. The effect of vibration on the flow of the cement paste was studied and found to greatly enhance placement. The setting times of this cement were dependent on temperature and decreased with increasing temperatures. At 37^∘C, the working and setting times were 6.3 ± 0.3 and 12.8 ± 0.4 minutes, respectively. Hydration and conversion of the cement phases continued while specimens were stored under simulated, physiological conditions. A cumulative increase in mass of 8.23 ± 0.65% was observed over a 14 month test period. During this time, the cement was found to expand slightly, 0.71 ± 0.39%. X-ray diffraction was used to characterize the crystalline phases present during hydration and conversion. The calcium aluminate in the cement hydrated and formed calcium aluminate chloride hydrates, while no changes were observed in the β-tricalcium phosphate during the testing period.     

PVA复合磷酸钙骨水泥的制备和性能研究

将含有不同质量分数聚乙烯醇(PVA)的PVA-KH2PO4-Na2HPO4体系缓冲溶液作为骨水泥的调和液,将其与磷酸钙骨水泥粉末混合后成型。将试样在接近生理条件(相对湿度100%,温度(37±1)℃)下养护24h,发现PVA掺入量为1%时的抗压强度达到31.71MPa,比未掺入的提高了将近70%。     

Fabrication and mechanical properties of calcium phosphate cements (CPC) for bone substitution

Calcium phosphate cements have been used in medical and dental applications for many years. However, their low strength and their high brittleness prohibit their use in many stress-bearing locations, which would require an improvement in mechanical properties. The influence of microstructural parameters on the latter has nevertheless barely been investigated in a systematic manner. To this aim, apatite cements have been fabricated through α-TCP (α-tricalcium phosphate) hydrolysis, and their mechanical properties have been measured (Young's modulus, fracture toughness and compressive strength), as a function of various parameters (particle size, liquid-to-powder ratio, amount and morphology of porosity, including macropores created by mannitol particles used as porogen). Five days following the mixing of phases, identification and microstructural observation indicated the presence of unreacted α-TCP particles, exhibiting very weak links with the apatite matrix and often surrounded by microcracks. The latter provoke a decrease in Young's modulus. The coarser the microstructure, the larger the critical flaw size causing fracture. In the case of macroporous materials, the critical flaw size increases with macroporosity. The knowledge gained should allow, in the end, to improve mechanical properties by controlling the microstructure, and to find a better compromise between strength and biological behaviour.     

Alginate combined calcium phosphate cements: mechanical properties and in vitro rat bone marrow stromal cell responses

Here, we prepared self-setting calcium phosphate cements (CPCs) based on α-tricalcium phosphate with the incorporation of sodium alginate, and their mechanical properties and in vitro cellular responses were investigated. The addition of alginate enhanced the hardening reaction of CPCs showing shorter setting times within a range of powder-to-liquid ratios. When immersed in a body simulating fluid the alginate-CPCs fully induced a formation of an apatite crystalline phase similar to that of bare CPCs. The compressive and tensile strengths of the CPCs were found to greatly improve during immersion in the fluid, and this improvement was more pronounced in the alginate-CPCs. As a result, the alginate-CPCs retained significantly higher strength values than the bare CPCs after 3–7 days of immersion. The rat bone marrow derived stromal cells (rBMSCs) cultured on the alginate-CPCs initially adhered to and then spread well on the cements surface, showed an on-going increase in the population with culture time, and differentiated into osteoblasts expressing bone-associated genes (collagen type I, osteopontin and bone sialoprotein) and synthesizing alkaline phosphatase. However, the stimulated level of osteogenic differentiation was not confirmative with the incorporation of alginate into the CPC composition based on the results. One merit of the use of alginate was its usefulness in forming CPCs into a variety of scaffold shapes including microspheres and fibers, which is associated with the cross-link of alginate under the calcium-containing solution.     

Controlled release of gentamicin from calcium phosphate/alginate bone cement

Calcium phosphate cement (CPC) is a good biomaterial for bone defect repair and as a delivery system for active agents. The aim of this study was to explore the physicochemical properties and in vitro soaking and release behaviors of gentamicin-loaded CPC with and without alginate; in particular, for biocompatibility. MTT colorimetric assay and RT-PCR were used to detect U2OS cell viability and level of cyclooxygenase-2 (COX-2), respectively. As a result, the setting time increased after the addition of 0.5% alginate and 5% gentamicin, reaching 19 min—significantly higher than the 8 min taken by the CPC control, demonstrating the adverse effect of alginate and gentamicin on the setting reaction of CPC. Gentamicin might reduce the diametral tensile strength, while alginate did not affect the strength. The rate of gentamicin release from CPC can be extended by the presence of alginate. The addition of gentamicin did not show signs of impaired cell viability, but alginate enhanced the cell viability. COX-2 expression of U2OSs cultured in the alginate-containing cement extract was about one-third level of the cement extract without alginate. Alginate-containing CPC is not only useful as a reservoir for antibiotic delivery but it also helps stimulate bone regeneration.     

Relationship between apatite-forming ability and mechanical properties of bioactive PMMA-based bone cement modified with calcium salts and alkoxysilane

Polymethylmethacrylate (PMMA)-based bone cement is used for the fixation of artificial joints in orthopaedics. However, the fixation is liable to loosen in the body, because the cement does not bond to living bone. So-called bioactive ceramics bond directly to living bone through the apatite layer formed on their surfaces in the body. We previously revealed that modification using γ-methacryloxypropyltrimethoxysilane (MPS) and water-soluble calcium salts such as calcium acetate and calcium hydroxide was effective for providing the PMMA-based bone cement with apatite-forming ability in a simulated body fluid (SBF, Kokubo solution) that closely reproduces the body environment. However, the effect of the chemical reaction forming the apatite on the mechanical properties of the cements has not been clarified. The present work aimed to investigate this issue from the viewpoint of the interface structure between the apatite and the cement. The surface of the cement modified with calcium acetate and MPS was fully covered with newly formed apatite after soaking in Kokubo solution within 7 days, while half of the surface area of the cement modified with calcium hydroxide and MPS was covered with the apatite. The bending strength of the modified cements decreased after soaking in Kokubo solution. Porous structure was observed in the region about 50–100 μm in depth from the top surface because of release of the Ca²⁺ ions by both modified cements after soaking in Kokubo solution. The decrease in bending strength of the modified cements could be attributed to the formation of the pores. In addition, the pores on the top surfaces of the cements were filled with the newly formed apatite. The apatite formation would be effective not only for bioactivity but also for decreasing the reduction of mechanical strength.     

Morphology and mechanical behavior of TTCP-derived calcium phosphate cement subcutaneously implanted in rats

A pre-hardened, TTCP-derived CPC was immersed in Hanks’ solution as well as subcutaneously implanted into abdomen of rats. The implant-soft tissue interfacial morphology was examined and properties of the CPC were evaluated and compared under in vitro and in vivo conditions. The results indicate that the surface of immersed samples appeared rougher and more porous than that of implanted samples and was covered with a layer of fine apatite crystals. The CPC samples implanted for 4 weeks or longer were surrounded by a layer of fibrous tissue, which was further surrounded by a soft tissue capsule comprising numerous fat cells. The soft tissue capsule had a non-uniform distribution in thickness, which increased most significantly between 4 weeks and 12 weeks after implantation. None of polymorphic cells, osteoblast cells or bone cells adjacent to the implant were observed. The majority of original TTCP powder was transformed into apatite after 1 day of either immersion in Hanks’ solution or implantation. The average porosity values of samples immersed in Hanks’ solution for 4 weeks or longer were significantly larger than those immersed for 1 day or 1 week. The porosity values of samples implanted for different times were not significantly different. The DTS values of Hanks’ solution-immersed samples largely decreased after a few weeks of immersion. The implanted samples maintained their strengths throughout the study.     

In vivo release of rhBMP-2 loaded porous calcium phosphate cement pretreated with albumin

In this study, the release of rhBMP-2 loaded porous Ca-P cement was studied in vitro and in vivo. We hypothesized that adsorption sites of Ca-P ceramic with high affinity for rhBMP-2 can be blocked by pretreatment of the ceramic with albumin prior to rhBMP-2 loading, which would result in weaker rhBMP-2 binding and enhanced release of rhBMP-2. Preset porous Ca-P cement discs with a diameter of 6.35 mm (volume: 75 mm³) were pretreated by incubation in a solution of 10% rat serum albumin for 24 h or in ddH₂O (control group) prior to administration of 5 μg radiolabeled ¹³¹I-rhBMP-2. Release was assessed in vitro in phosphate buffered saline (PBS) and fetal calf serum and in vivo by longitudinal scintigraphic imaging of radiolabeled ¹³¹I-rhBMP-2 and gamma counting of dissected implants. In vitro release from pretreated discs was higher during the first day. For both formulations, release in PBS was limited compared to release in serum. In vivo release considerably exceeded in vitro release. In vivo release kinetics showed no significant difference of half-lives between pretreated and control discs. Both formulations showed sustained release during at least 4 weeks. Ex vivo gamma counting of retrieved samples confirmed scintigraphic results and showed that the capsule and surrounding tissues only contained a minor fraction rhBMP-2. We conclude that 1. scintigraphy of ¹³¹I-labeled rhBMP-2 provides a reliable method for longitudinal measurement of rhBMP-2 release kinetics in vivo. 2. albumin pretreatment of porous Ca-P cement does not results in relevant increase of initial release of rhBMP-2 in vivo, and 3. preset rhBMP-2 loaded porous Ca-P cement discs exhibit one phase exponential release kinetics in the rat ectopic model, characterized by a retention of 20–30% after 4 weeks.     

Effect of various additives and temperature on some properties of an apatitic calcium phosphate cement

The effect of additives and temperature on setting time, swelling time and compressive strength of a previously developed apatitic calcium phosphate cement was investigated. Setting was faster at body temperature than at room temperature. Early contact with aqueous solutions resembling blood and other body fluids had no effect. Deliberate additions of soluble carbonates, pyrophosphate or magnesium salts to the cement powder retarded or even inhibited setting. However, additions of calcium pyrophosphate, -tertiary calcium phosphate or sintered hydroxyapatite to the cement powder in amounts up to 10% had no effect on the cement properties. Several organic substances were used as additives. They all retarded the setting and decreased the strength of the cement considerably.