Calcium phosphate cements for bone engineering and their biological properties

Calcium phosphate cements （CPCs） are frequently used to repair bone defects. Since their discovery in the 1980s, extensive research has been conducted to improve their properties, and emerging evidence supports their increased application in bone tissue engineering. Much effort has been made to enhance the biological performance of CPCs, including their biocompatibility, osteoconductivity, osteoinductivity, biodegradability, bioactivity, and interactions with cells. This review article focuses on the major recent developments in CPCs, including 3D printing, injectability, stem cell delivery, growth factor and drug delivery, and pre- vascularization of CPC scaffolds via co-culture and tri-culture techniques to enhance angiogenesis and osteogenesis.

REINFORCEMENT OF CALCIUM PHOSPHATE CEMENTS WITH PHOSPHORYLATED CHITIN

Phosphorylated chitins (P-chitins) as the additives of calcium phosphate cements (CPCs) were prepared by the phosphorylation of chitin with phosphorus pentoxide in methanesulfonic acid. Their physical properties and effects on CPCs from monocalcium phosphate monohydrate (MCPM) and calcium oxide (CaO) or dicalcium phosphate dihydrate (DCPD) and calcium hydroxide [Ca(OH)(2)] were investigated. Addition of P-chitin (M-w = 2.60 x 10(4); degree of substitution, DS = 0.68) to the liquid phase in amounts up to 3 wt.% for MCPM and CaO cements or 1.5 wt.% for DCPD and Ca(OH)(2) cements could enhance the mechanical strength considerably, while little influence on the setting time was observed. However, further addition of P-chitin will cause no setting.

Effect of Additives on the Morphology of the Hydrated Product and Physical Properties of a Calcium Phosphate Cement

The morphology of a hydrated calcium phosphate cement （CPC） doped with several normally used additives was investigated by scanning electron microscopy （SEM） and the compressive strength of the cement was determined in this study. The hydrated products of CPC without additives was rod-like hydroxyapatite （HA） grains with around 2-5 μm in length and 100 nm in width. The addition of Sr obviously decreased the crystal size of the rod-like grains. CPCs containing carbonate, collagen and gelatin showed flake-like crystal morphology. Crylic acid-containing CPC presented flocculus-like structure. And malic acid-containing CPC exhibited oriented flake-like structure. The X-ray diffraction （XRD） analysis showed that the additives used in this study did not alter the hydration products of the cement. The compressive strength tests indicated that the compressive strength of the cement with rod-like morphology HA crystals was much higher than that of the cement with flake-like morphology HA crystals, and the cement with oriented flake-like morphology HA crystals exhibited the poorest compressive strength.

Development and characterization of α-tricalcium phosphate/monocalcium aluminate composite bone cement

Calcium phosphate cements have received much attention in recent decades owing to their biocompatibility, in situ handling, and shaping abilities. However, their low initial mechanical strength is still a major limitation. On the other hand, calcium aluminate cements (CACs) set fast and have a high initial strength and good corrosion resistance in contact with body fluids, making them excellent dental restorative materials. Therefore, the chemical, mechanical and biological properties of new-TCP/CA cement after aging in simulated body fluid (SBF) were investigated. The results indicated that the composites have setting times not appropriated for immediate applications and have degradation rates higher than those of the traditional CPCs. Moreover, the compressive strength of composite was lower than 5MPa and did not increase with SBF immersion. However, the α-TCP/CA composites showed a higher bioactivity at early stages and were not only more biocompatible but also more noncytotoxic.

Effect of carbon fiber on calcium phosphate bone cement

The calcium phosphate cement (α-TCP/TTCP) was reinforced with oxidation-t reated carbon fibers. The effect of aspect ratio and content of carbon fiber on the compression strength and bending strength of the hardened body was discussed . The results show that the reinforcing effect is optimal as the aspect ratio is 375 and the additive amount is 0.3%(mass fraction). Under this condition,the c ompressive strength is increased by 55% (maximum 63.46 MPa),and the bending st rength is nearly increased by 100% (maximum 11.95 MPa),respectively. However,if the additive quantity and aspect ratio are too high,the effect of the carbon fibers is limited because it can not be dispersed uniformly in the hardened bod y. The biological evaluation indicates that the calcium phosphate cement reinfor ced by carbon fibers has good biocompatibility.

The Effect of Premixed Schedule on the Crystal Formation of Calcium Phosphate Cement-chitosan Composite with Added Tetracycline

In this study, calcium phosphate cements （CPC） were prepared by mixing cement powders of tetracalcium phosphate （TTCP） with a cement liquid of phosphate acid saline solution. Tetracycline （TTC）-CPC, chitosan-CPC and chitosan-TTC-CPC were investigated with different premixed schedule. It was demonstrate that both TTC and chitosan worked on the phase transition and crystal characteristics. TTCP mixed with phosphate acid saline solution had similar features of Fourier transform-infrared spectrometry （FT-IR） no matter it was mixed with chitosan or TTC or both. TTC premixed with cement liquid or powder had significant different features of FT-IR and 876 cm-1 seemed to be a special peak for TTC when TTC was premixed with cement liquid. This was also supported by XRD analysis, which showed that TTC premixed with cement liquid improved phase transition of TTCP to OCP. Chitosan, as organic additive, regulates the regular crystal formation and inhibits the phase transition of TTCP to OCP, except when it is mingled with cement liquid premixed with TTC in field scanning electron microscope. It was concluded that the premixed schedule influences the crystal formation and phase transition, which may be associated with its biocompatibility and bioactivities in vivo.

The In-situ Reinforcement of Calcium Phosphate Cement and Its Micro-structural Analysis

Carbon nanotubes （ CNTs ） and polyacrylic acid were employed to modify the setting process and hydration products of β-TCP/TTCP calcium phosphate cement. The micro-structure of hydration product and the fashion of how additives and hydration particles interconnected were investigated. With the modification effect of CNTs , the setting particles and CNTs got winded and interconnected and thus made the composite more compact and denser.

Effect of Substitutional Sr Ion on Mechanical Properties of Calcium Phosphate Bone Cement

A novel calcium phosphate cement was developed by adding different amount of SrO to tetracalcium phosphate during the fabrication process. The experimental results show that compressive strength of cements based on tetracalcium phosphate doped with SrO significantly increases with the increase of SrO content, approximately 60 MPa, whilst the mechanical behavior of CPC slightly decreases if 0.7wt% SrO is added. X-ray diffraction measurement confirms the setting reaction of doped cements is similar to that of pure calcium phosphate cement （CPC）. Low crystalline hydroxyapatite （HA） is found to be the main constituent of set cement. A mechanical reinforcement effect is resulted from the substitution of Sr ion to Ca2＋ in tetracalcium phosphate （TTCP）, accelerating HA crystal formation and a more cross-linked cement structure. In vitro bioactivity tests showed that CPC added with 0.5wt% SrO had a more rapid degradation compared with pure CPC.

Augmentation of Pedicle Screw Fixation with Calcium Phosphate Cement

To determine whether a biodegradable calcium phosphate cement(CPC) provides significant augmentation of pedicle screw fixation or not,an in vitro biomechanical study was carried out to evaluate the biomechanical effect of CPC in the restoration and augmentation of pedicle screw fixation.Axial pullout test and cyclic bending resistance test were employed in the experiment,and polymethylmethacrylate (PMMA) was chosen as control.The results demonstrate that the pullout strengths following CPC restoration and augmentation are 74% greater on an average than those of the control group,but less than those of PMMA restoration group and augmentation group respectively (increased by 126% versus control).In cyclic bending resistance test,the CPC augmented screws are found to withstand a greater number of cycles or greater loading with less displacement before loosening,but the augmentation effect of PMMA is greater than that of CPC.

Microstructure and Mechanical Properties of Calcium Phosphate Cement/Gelatine Composite Scaffold with Oriented Pore Structure for Bone Tissue Engineering

The macroporous calcium phosphate（CPC） cement with oriented pore structure was prepared by freeze casting. SEM observation showed that the macropores in the porous calcium phosphate cement were interconnected aligned along the ice growth direction. The porosity of the as-prepared porous CPC was measured to be 87.6% by Archimede＇s principle. XRD patterns of specimens showed that poorly crystallized hydroxyapatite was the main phase present in the hydrated porous calcium phosphate cement. To improve the mechanical properties of the CPC scaffold, the 15% gelatine solution was infiltrated into the pores under vacuum and then the samples were freeze dried to form the CPC/gelatine composite scaffolds. After reinforced with gelatine, the compressive strength of CPC/gelatine composite increased to 5.12 MPa, around fifty times greater than that of the unreinforced macroporous CPC scaffold, which was only 0.1 MPa. And the toughness of the scaffold has been greatly improved via the gelatine reinforcement with a much greater fracture strain. SEM examination of the specimens indicated good bonding between the cement and gelatine. Participating the external load by the deformable gelatine, patching the defects of the CPC pores wall, and crack deflection were supposed to be the reinforcement mechanisms. In conclusion, the calcium phosphate cement/gelatine composite with oriented rmre structure nrenared in this work might be a potential scaffold for bone tissue engineerinm

Evaluation of Calcium Phosphate Cement As a Root Canal Sealer Filling Material

Calcium phosphate cement for root end sealing was obtained by in mixing alpha-tricalcium phosphate and additives with an aqueous solation of citric. Powder and liquid were mixed at a ratio of 1.25g/mL. The biocompatibility of this material was investigated primarily by subcutaneous implantation tests. Then calcium phosphate cement was used to fill three adult dogs' root canal, both calcium hydroxide paste and hydroxyapatite paste as control. The animals were killed at 4, 12, 20 weeks postoperatively respectively. The effects of different materials on the apical closure, restoration of periapical tissues and adaptability to the dentinal surface were examined by optical and electronic microscope. The observation at 20 weeks shows that the calcium phosphate cement has the potentialities of being a root canal sealer filling material available for pulpless teeth with open-apex and destructive periapical tissue.

Preparation of a Novel Calcium Phosphate Cement Using N-methylene Phosphonic Chitosan as a Gelling Agent

A modified chitosan （ N-methylene phosphonic Chitosan, NMPC） was synthesized to improve solubility and ability to bind calcium ion. The properties of the raw material chitosan and its derivative NMPC were characterised using FTIR , ^1H- NMR . The aim of this study was to enhance the compressive CPC by reinforcing with NMPC. A formulation consisting of CPC powder , buffer solution and gelling agent was used for preparation of the CPC. CPC powder coasisted of tetracalcium phosphate（ TTCP ） and dicalcium phosphate anhydrous （ DCPA ）. NMPC which acted as the gelling ageut was dissohed into KH2PO4-Na2 HPO4 buffer solution. Each specimen in the mold was sandciched between two fritted glass sides and kept for 24 hours. Compressive strengths were determined, the setting product was identified using X-ray diffraction and scanning electron microscopy was used to investigate the hydroxyapatite particles size and porosity. The experimental results showed that the dominating influence on the compressive strengths of CPC-AMPC was the HA panicle size, its uniformity and appropriate porosity.

Preparation and Compressive Strength of Calcium Phosphate Bone Cement Containing N,O-carboxymethyl Chitosan

N, O-carboxymethyl chitosan （ CMCTS ）, a kind of biodegradable organic substance, was added to calcium phosphate bone cement （CPC） to prodnce a composite more similar in composition to human bone. The compressive strength of the new material was inereased by 10 times compared with conventional CPC.

Microstructure and Mechanical Properties of Calcium Phosphate Cement/gelatine Composite Scaffold with Oriented Pore Structure for Bone Tissue Engineering

In this study,the macroporous calcium phosphate cement with oriented pore structure was prepared by freeze casting.SEM observation showed that the macropores in the porous calcium phosphate cement were interconnected aligned along the ice growth direction.The porosity of the as-prepared porous CPC was measured to be 87.6% by Archimede's principle.XRD patterns of specimens showed that poorly crystallized hydroxyapatite was the main phase present in the hydrated porous calcium phosphate cement.To improve the mechanical properties of the CPC scaffold,the 15% gelatine solution was infiltrated into the pores under vacuum and then the samples were freeze dried to form the CPC/gelatine composite scaffolds.After reinforced with gelatine,the compressive strength of CPC/gelatin composite increased to 5.12 MPa,around 50 times greater than that of the unreinforced macroporous CPC scaffold,which was only 0.1 MPa.And the toughness of the scaffold has been greatly improved via the gelatine reinforcement with a much greater fracture strain.SEM examination of the specimens indicated good bonding between the cement and gelatine.In conclusion,the calcium phosphate cement/gelatine composite with oriented pore structure prepared in this study might be a potential scaffold for bone tissue engineering.

Bone tissue engineering via nanostructured calcium phosphate biomaterials and stem cells

Tissue engineering is promising to meet the increasing need for bone regeneration. Nanostructured calcium phosphate （CAP） biomaterials/scaffolds are of special interest as they share chemical/crystallographic similarities to inorganic components of bone. Three applications of nano-CaP are discussed in this review： nanostructured calcium phosphate cement （CPC）; nano-CaP composites; and nano-CaP coatings. The interactions between stem cells and nano-CaP are highlighted, including cell attachment, orientation/ morphology, differentiation and in vivo bone regeneration. Several trends can be seen： （i） nano-CaP biomaterials support stem cell attachment/proliferation and induce osteogenic differentiation, in some cases even without osteogenic supplements; （ii） the influence of nano-CaP surface patterns on cell alignment is not prominent due to non-uniform distribution of nano-crystals; （iii） nano-CaP can achieve better bone regeneration than conventional CaP biomaterials; （iv） combining stem cells with nano-CaP accelerates bone regeneration, the effect of which can be further enhanced by growth factors; and （v） cell microencapsulation in nano-CaP scaffolds is promising for bone tissue engineering. These understandings would help researchers to further uncover the underlying mechanisms and interactions in nano-CaP stem cell constructs in vitro and in vivo, tailor nano-CaP composite construct design and stem cell type selection to enhance cell function and bone regeneration, and translate laboratory findings to clinical treatments.

Calcium Phosphate Biomaterials: An Update

Current calcium phosphate （ CaP ） biomaterials for bone repair, substitution, augmentation and regeneration include hydroxyapatite （ HA ） from synthetic or biologic origin, beta-tricaicium phosphate （ β- TCP ）, biphasic calcium phosphate （BCP）, and are available as granules, porous blocks, components of compashes （CaP/pollymer） cements, and as coatings on orthopedic and dental implants. Experimental calcium phosphate biomaterials include CO3^- and F-substituted apatites, Mg-and Zn-substituted β-TCP, calcium phosphate glasses, This paper is a brief review of the different types of CaP biomaterials and their properties such as bioactivity , osteoconductivity , osteoinductivity.

CPC/PMMA双向骨水泥性能及生物相容性的初步研究

目的将聚甲基丙烯酸甲酯骨水泥(PMMA)与磷酸钙骨水泥(CPC)制成一种注射型混合骨水泥,利用各自的特性,优势互补,从而使该混合骨水泥成为理想的椎体成形术及椎体后凸成形术填充材料。方法实验材料为注射型PMMA骨水泥和注射型CPC骨水泥。首先将PMMA与CPC按质量比1∶2、1∶1、2∶1均匀混合,制成不同比例的混合骨水泥,然后将PMMA、CPC、混合骨水泥(1∶2、1∶1、2∶1)制成标准试件(直径6mm、高12mm),最后对试件进行力学性能、凝固温度、凝固时间、电镜扫描、X线衍射、动物肌肉内植入实验,初步评估混合骨水泥的性能及生物相容性。结果 PMMA与CPC在1∶1比例混合时理化性能满意,其抗压强度较CPC提高约120%,凝固过程中的最高温度低于45℃,凝固时间约12～14 min,微观结构与CPC相似,孔径在100～300μm之间分布最多(约70%),大部分孔与孔之间有10～20μm小孔贯通。动物肌肉内植入实验显示混合骨水泥(1∶1)的组织相容性满意,但降解速度缓慢。结论 PMMA与CPC在1∶1比例混合能有效克服单一材料缺点,优势互补,从而使该混合骨水泥有望成为理想的椎体成形术及椎体后凸成形术填充材料。

硅酸盐生物活性玻璃纤维对磷酸钙骨水泥性能的影响

背景:磷酸钙骨水泥的力学性能差、成骨能力弱等问题阻碍了其进一步发展。硅酸盐生物活性玻璃因具有良好的生物活性及成骨能力而备受青睐,同时其纤维结构能够加强材料的力学强度。目的:探究硅酸盐生物活性玻璃纤维复合磷酸钙骨水泥的力学性能、生物相容性及成骨效果。方法:将不同质量百分比(0%,10%,20%)的硅酸盐生物活性玻璃纤维分别加入磷酸钙骨水泥固相中,与液相混合后固化48 h,得到磷酸钙骨水泥与硅酸盐生物活性玻璃纤维复合磷酸钙骨水泥,表征骨水泥的力学性能、凝固时间及离子析出情况。将3组骨水泥浸提液分别与MC3T3-E1细胞共培养,通过CCK-8检测、活死染色、鬼笔环肽染色评估材料的细胞相容性;成骨诱导后,通过碱性磷酸酶染色、茜素红染色、RUNX2免疫荧光染色及RT-PCR检测评估材料的成骨诱导能力。结果与结论:①随着硅酸盐生物活性玻璃纤维含量的增加,骨水泥的抗压强度与抗弯强度均升高,凝固时间延长;将骨水泥浸泡于模拟体液中均可检测到硅离子、钙离子、磷离子析出,并且随着硅酸盐生物活性玻璃纤维含量的增加,骨水泥释放的硅离子、磷离子质量浓度升高,钙离子质量浓度降低;②活死染色、鬼笔环肽染色结果显示,硅酸盐生物活性玻璃纤维复合磷酸钙骨水泥对MC3T3-E1细胞无毒性作用;CCK-8检测结果显示,硅酸盐生物活性玻璃纤维复合磷酸钙骨水泥可促进MC3T3-E1细胞的增殖;③随着骨水泥中硅酸盐生物活性玻璃纤维含量的增加,MC3T3-E1细胞的碱性磷酸酶活性及细胞外钙沉积均增加,RUNX2蛋白表达增加,碱性磷酸酶、骨钙素、骨桥蛋白、RUNX2 mRNA表达增加;④结果表明,硅酸盐生物活性玻璃纤维可增强磷酸钙骨水泥的力学性能与成骨诱导能力,其中以20%硅酸盐生物活性玻璃纤维的效果更明显。

CPC/CaO_2氧缓释复合材料的制备及其释氧性能探讨

利用过氧化钙(CaO2)遇水释放氧气的特性,采用混合法将CaO2载入到磷酸钙骨水泥(CPC)中,并通过挤出-滚圆造粒方法制备出CPC/CaO2氧缓释复合材料,旨在为污染地下水的好氧生物修复技术提供一种长期、高效的供氧源。由X射线衍射(XRD)分析可知,通过固相反应制取的CPC主要成分为Ca5(PO4)3(OH)和Ca10(PO4)6(OH)2,具有较快的固化时间和固化效果;CPC/CaO2氧缓释复合材料分3个阶段进行释氧,其中2、3阶段分别符合一级和零级释氧动力学特征,且其释氧速率和释氧周期得到明显改善。本研究成果可用于实际地下水修复工程,解决污染地下水原位生物修复过程中溶解氧(DO)的输送问题。