Synthetic amorphous calcium phosphates (ACPs): preparation, structure, properties, and biomedical applications.

Amorphous calcium phosphates (ACPs) represent a metastable amorphous state of other calcium orthophosphates (abbreviated as CaPO4) possessing variable compositional but rather identical glass-like physical properties, in which there are neither translational nor orientational long-range orders of the atomic positions. In nature, ACPs of a biological origin are found in the calcified tissues of mammals, some parts of primitive organisms, as well as in the mammalian milk. Manmade ACPs can be synthesized in a laboratory by various methods including wet-chemical precipitation, in which they are the first solid phases, precipitated after a rapid mixing of aqueous solutions containing dissolved ions of Ca2+ and PO43- in sufficient amounts. Due to the amorphous nature, all types of synthetic ACPs appear to be thermodynamically unstable and, unless stored in dry conditions or doped by stabilizers, they tend to transform spontaneously to crystalline CaPO4, mainly to ones with an apatitic structure. This intrinsic metastability of the ACPs is of a great biological relevance. In particular, the initiating role that metastable ACPs play in matrix vesicle biomineralization raises their importance from a mere laboratory curiosity to that of a reasonable key intermediate in skeletal calcifications. In addition, synthetic ACPs appear to be very promising biomaterials both for manufacturing artificial bone grafts and for dental applications. In this review, the current knowledge on the occurrence, structural design, chemical composition, preparation, properties, and biomedical applications of the synthetic ACPs have been summarized.

Functionalized calcium orthophosphates (CaPO4) and their biomedical applications.

Due to the chemical similarity to natural calcified tissues (bones and teeth) of mammals, calcium orthophosphates (abbreviated as CaPO4) appear to be good biomaterials for creation of artificial bone grafts. However, CaPO4 alone have some restrictions, which limit their biomedical applications. Various ways have been developed to improve the properties of CaPO4 and their functionalization is one of them. Namely, since surfaces always form the interfaces between implanted grafts and surrounding tissues, the state of CaPO4 surfaces plays a crucial role in the survival of bone grafts. Although the biomedically relevant CaPO4 possess the required biocompatible properties, some of their properties could be better. For example, functionalization of CaPO4 to enhance cell attachment and cell material interactions has been developed. In addition, to prepare stable formulations from nanodimensional CaPO4 particles and prevent them from agglomerating, the surfaces of CaPO4 particles are often functionalized by sorption of special chemicals. Furthermore, there are functionalizations in which CaPO4 are exposed to various types of physical treatments. This review summarizes the available knowledge on CaPO4 functionalizations and their biomedical applications.

Biphasic calcium phosphates bioceramics (HA/TCP): Concept, physicochemical properties and the impact of standardization of study protocols in biomaterials research.

Biphasic calcium phosphates (BCP) bioceramics have become the materials of choice in various orthopedic and maxillofacial bone repair procedures. One of their main advantages is their biodegradation rate that can be modified by changing the proportional ratio of the composition phases. For enhanced bone tissue regeneration, the bioactivity of BCP should be increased by optimizing their physicochemical properties. To date, the ideal physicochemical properties of BCP for bone applications have not been defined. This is mostly related to lack of standard study protocols in biomaterial science especially with regards to their characterizations and clinical applications. In this paper we provided a review on BCP and their physicochemical properties relevant to clinical applications. In addition, we summarized the available literature on their use in animal models and evaluated the influences of different composition ratios on bone healing. Controversies in literature with regards to ideal composition ratio of BCP have also been discussed in detail. We illustrated the discrepancies in study protocols among researchers in animal studies and emphasized the need to develop and follow a set of generally accepted standardized guidelines. Finally; we provided general recommendations for future pre-clinical studies that allow better standardization of study protocols. This will allow better comparison and contrast of newly developed bone substitute biomaterials that help further progress in the field of biomaterial science.

The effect of simulating body fluid on the structural properties of hydroxyapatite synthesized in the presence of citric acid.

In present work, the effect of citric acid (CA) addition in different amounts (0, 1, 5 and 10 ml) on the structure of hydroxyapatite (HAp) was investigated using X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM) and energy dispersive X-ray (EDX) spectroscopy techniques. The crystallite dimensions, lattice parameters, unit cell volume, crystallinity percentage and Ca/P molar ratio were found to be affected by the CA content. To investigate the influence of CA on the bioactive properties of the HAp samples and to determine the optimum amount of CA, in vitro soaking tests in simulated body fluid (SBF) were performed. Although the samples' morphology was found to be affected by neither the amount of CA nor the soaking time in SBF, the soaking results revealed that the maximum changes in the Ca/P ratio were found for the HAp samples prepared in the presence of the highest amounts of CA, which pointed out to the highest bioactivity of these samples.

Calcium orthophosphates (CaPO4): occurrence and properties.

The present overview is intended to point the readers' attention to the important subject of calcium orthophosphates (CaPO4). This type of materials is of the special significance for the human beings because they represent the inorganic part of major normal (bones, teeth and antlers) and pathological (i.e., those appearing due to various diseases) calcified tissues of mammals. For example, atherosclerosis results in blood vessel blockage caused by a solid composite of cholesterol with CaPO4, while dental caries and osteoporosis mean a partial decalcification of teeth and bones, respectively, that results in replacement of a less soluble and harder biological apatite by more soluble and softer calcium hydrogenorthophosphates. Therefore, the processes of both normal and pathological calcifications are just an in vivo crystallization of CaPO4. Similarly, dental caries and osteoporosis might be considered as in vivo dissolution of CaPO4. In addition, natural CaPO4 are the major source of phosphorus, which is used to produce agricultural fertilizers, detergents and various phosphorus-containing chemicals. Thus, there is a great significance of CaPO4 for the humankind and, in this paper, an overview on the current knowledge on this subject is provided.

Calcium Orthophosphate-Containing Biocomposites and Hybrid Biomaterials for Biomedical Applications.

The state-of-the-art on calcium orthophosphate (CaPO4)-containing biocomposites and hybrid biomaterials suitable for biomedical applications is presented. Since these types of biomaterials offer many significant and exciting possibilities for hard tissue regeneration, this subject belongs to a rapidly expanding area of biomedical research. Through the successful combinations of the desired properties of matrix materials with those of fillers (in such systems, CaPO4 might play either role), innovative bone graft biomaterials can be designed. Various types of CaPO4-based biocomposites and hybrid biomaterials those are either already in use or being investigated for biomedical applications are extensively discussed. Many different formulations in terms of the material constituents, fabrication technologies, structural and bioactive properties, as well as both in vitro and in vivo characteristics have been already proposed. Among the others, the nano-structurally controlled biocomposites, those containing nanodimensional compounds, biomimetically fabricated formulations with collagen, chitin and/or gelatin, as well as various functionally graded structures seem to be the most promising candidates for clinical applications. The specific advantages of using CaPO4-based biocomposites and hybrid biomaterials in the selected applications are highlighted. As the way from a laboratory to a hospital is a long one and the prospective biomedical candidates have to meet many different necessities, the critical issues and scientific challenges that require further research and development are also examined.

Calcium orthophosphate deposits: Preparation, properties and biomedical applications.

Since various interactions among cells, surrounding tissues and implanted biomaterials always occur at their interfaces, the surface properties of potential implants appear to be of paramount importance for the clinical success. In view of the fact that a limited amount of materials appear to be tolerated by living organisms, a special discipline called surface engineering was developed to initiate the desirable changes to the exterior properties of various materials but still maintaining their useful bulk performances. In 1975, this approach resulted in the introduction of a special class of artificial bone grafts, composed of various mechanically stable (consequently, suitable for load bearing applications) implantable biomaterials and/or bio-devices covered by calcium orthophosphates (CaPO4) to both improve biocompatibility and provide an adequate bonding to the adjacent bones. Over 5000 publications on this topic were published since then. Therefore, a thorough analysis of the available literature has been performed and about 50 (this number is doubled, if all possible modifications are counted) deposition techniques of CaPO4 have been revealed, systematized and described. These CaPO4 deposits (coatings, films and layers) used to improve the surface properties of various types of artificial implants are the topic of this review.

Strontium substituted hydroxyapatites: Synthesis and determination of their structural properties, in vitro and in vivo performance.

The objective of this study is to present a detailed report related to the synthesis and characterization of strontium substituted hydroxyapatites. Based on this purpose, hydroxyapatite (HAp) bioceramics with different amounts of strontium (e.g., 0, 0.45, 0.90, 1.35, 1.80 and 2.25 at.%) were prepared using a sol-gel method. The effects of Sr substitution on the structural properties and biocompatibility of the samples were studied by X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy and scanning electron microscopy (SEM) techniques, in vitro and in vivo tests. All the samples composed of the nanoparticles ranging from 21 to 27 nm. The presence of Sr at low levels influenced the crystal size, crystallinity degree, lattice parameters and volume of the unit cell of the HAp. Both in vitro conditions and soaking period in simulated body fluid (SBF) significantly affected these properties. Especially, the (Ca+Sr)/P molar ratio gradually decreases with increasing soaking period in SBF. Animal experiments revealed the bone formation and osseointegration for all samples, and as compared with other groups, more reasonable, were observed for the sample with the lowest Sr content.

Structural and dielectric properties of yttrium-substituted hydroxyapatites.

Hydroxyapatite (HAp) samples doped with 0, 2 and 4 at.% of yttrium (Y) were characterized using X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy attached with energy dispersive X-ray (EDX) spectroscopy, antimicrobial activity tests and dielectric studies. The hydroxyl groups observed in FTIR spectra confirmed the formation of HAp phase in the studied samples. The crystallite size, crystallinity degree and lattice parameters of the samples were changed with Y content. The volume of the unit cell was gradually decreased with the addition of Y. Undoped and Y-containing HAp samples were screened to determine their in vitro antimicrobial activities against the standard strains. It was found that no samples have any antimicrobial effect. The relative dielectric permittivity and dielectric loss are affected by Y content. While the alternating current conductivity increases with increasing frequency, it decreases with increasing Y content.

Synthesis and characterization of Ce-substituted hydroxyapatite by sol-gel method.

Both undoped hydroxyapatite (HAp) and three Ce-substituted HAp samples with variable amounts (from 0.5 to 2 at.%) of Ce were synthesized by sol-gel method. The samples were studied by X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM) and energy dispersive X-ray (EDX) spectroscopy to determine the crystallite size, crystallinity degree, phases, functional groups, morphology and elemental composition. In all samples, the amount of HAp exceeded 92%, while the amount of admixture ß-TCP was always below 8% and no changes were observed by addition of Ce. The crystallinity degree of the samples was always within 84-89%, while the calculated dimensions of crystallites appeared to be within 26-35nm. The microstructure and elemental composition of all the samples were found to be affected by the addition of Ce.

Calcium orthophosphate coatings on magnesium and its biodegradable alloys.

Biodegradable metals have been suggested as revolutionary biomaterials for bone-grafting therapies. Of these metals, magnesium (Mg) and its biodegradable alloys appear to be particularly attractive candidates due to their non-toxicity and as their mechanical properties match those of bones better than other metals do. Being light, biocompatible and biodegradable, Mg-based metallic implants have several advantages over other implantable metals currently in use, such as eliminating both the effects of stress shielding and the requirement of a second surgery for implant removal. Unfortunately, the fast degradation rates of Mg and its biodegradable alloys in the aggressive physiological environment impose limitations on their clinical applications. This necessitates development of implants with controlled degradation rates to match the kinetics of bone healing. Application of protective but biocompatible and biodegradable coatings able to delay the onset of Mg corrosion appears to be a reasonable solution. Since calcium orthophosphates are well tolerated by living organisms, they appear to be the excellent candidates for such coatings. Nevertheless, both the high chemical reactivity and the low melting point of Mg require specific parameters for successful deposition of calcium orthophosphate coatings. This review provides an overview of current coating techniques used for deposition of calcium orthophosphates on Mg and its biodegradable alloys. The literature analysis revealed that in all cases the calcium orthophosphate protective coatings both increased the corrosion resistance of Mg-based metallic biomaterials and improved their surface biocompatibility.

A detailed history of calcium orthophosphates from 1770s till 1950.

Due to the chemical similarity with human bones and teeth, calcium orthophosphates are the inorganic substances of a special importance for the human being: they appear to be the excellent compounds to construct artificial bone grafts. In addition, calcium orthophosphates are necessary for both animals and plants as the source of important chemical elements. Obviously, these facts have not become apparent immediately; thus, providing the detailed annals of the knowledge development on the subject is the purpose of this review. The chosen time scale started with the earliest available studies of 1770s (to the best of my findings, calcium orthophosphates had been unknown before), passed through the entire 19th century and finished in 1950, because since then the amount of publications rapidly increased and the subject became too broad. In addition, since publications of the second half of the 20th century are easily accessible, other scientists have already reviewed the substantial amount of them. Many forgotten and poorly known historical facts, names, approaches, concepts and misconceptions have been extracted from the old publications. To maximize objectivity, an extensive quotation has been used. Then the old data have been systematized, reanalyzed and reconsidered from the modern points of view. The reported historical findings clearly demonstrate that many famous scientists of the past contributed to the subject. Furthermore, the significant quantity of the modern scientific facts, ideas and experimental approaches appear to have been known for very many decades and, in fact, a good deal of the recent investigations on calcium orthophosphates is just either a further development of the earlier studies or a rediscovery of the already forgotten knowledge.

Calcium orthophosphates in dentistry.

Dental caries, also known as tooth decay or a cavity, remains a major public health problem in the most communities even though the prevalence of disease has decreased since the introduction of fluorides for dental care. Therefore, biomaterials to fill dental defects appear to be necessary to fulfill customers' needs regarding the properties and the processing of the products. Bioceramics and glass-ceramics are widely used for these purposes, as dental inlays, onlays, veneers, crowns or bridges. Calcium orthophosphates belong to bioceramics but they have some specific advantages over other types of bioceramics due to a chemical similarity to the inorganic part of both human and mammalian bones and teeth. Therefore, calcium orthophosphates (both alone and as components of various formulations) are used in dentistry as both dental fillers and implantable scaffolds. This review provides brief information on calcium orthophosphates and describes in details current state-of-the-art on their applications in dentistry and dentistry-related fields. Among the recognized dental specialties, calcium orthophosphates are most frequently used in periodontics; however, the majority of the publications on calcium orthophosphates in dentistry are devoted to unspecified "dental" fields.

Calcium Orthophosphate-Based Bioceramics.

Various types of grafts have been traditionally used to restore damaged bones. In the late 1960s, a strong interest was raised in studying ceramics as potential bone grafts due to their biomechanical properties. A bit later, such synthetic biomaterials were called bioceramics. In principle, bioceramics can be prepared from diverse materials but this review is limited to calcium orthophosphate-based formulations only, which possess the specific advantages due to the chemical similarity to mammalian bones and teeth. During the past 40 years, there have been a number of important achievements in this field. Namely, after the initial development of bioceramics that was just tolerated in the physiological environment, an emphasis was shifted towards the formulations able to form direct chemical bonds with the adjacent bones. Afterwards, by the structural and compositional controls, it became possible to choose whether the calcium orthophosphate-based implants remain biologically stable once incorporated into the skeletal structure or whether they were resorbed over time. At the turn of the millennium, a new concept of regenerative bioceramics was developed and such formulations became an integrated part of the tissue engineering approach. Now calcium orthophosphate scaffolds are designed to induce bone formation and vascularization. These scaffolds are often porous and harbor different biomolecules and/or cells. Therefore, current biomedical applications of calcium orthophosphate bioceramics include bone augmentations, artificial bone grafts, maxillofacial reconstruction, spinal fusion, periodontal disease repairs and bone fillers after tumor surgery. Perspective future applications comprise drug delivery and tissue engineering purposes because calcium orthophosphates appear to be promising carriers of growth factors, bioactive peptides and various types of cells.

Self-setting calcium orthophosphate formulations.

In early 1980s, researchers discovered self-setting calcium orthophosphate cements, which are bioactive and biodegradable grafting bioceramics in the form of a powder and a liquid. After mixing, both phases form pastes, which set and harden forming either a non-stoichiometric calcium deficient hydroxyapatite or brushite. Since both of them are remarkably biocompartible, bioresorbable and osteoconductive, self-setting calcium orthophosphate formulations appear to be promising bioceramics for bone grafting. Furthermore, such formulations possess excellent molding capabilities, easy manipulation and nearly perfect adaptation to the complex shapes of bone defects, followed by gradual bioresorption and new bone formation. In addition, reinforced formulations have been introduced, which might be described as calcium orthophosphate concretes. The discovery of self-setting properties opened up a new era in the medical application of calcium orthophosphates and many commercial trademarks have been introduced as a result. Currently such formulations are widely used as synthetic bone grafts, with several advantages, such as pourability and injectability. Moreover, their low-temperature setting reactions and intrinsic porosity allow loading by drugs, biomolecules and even cells for tissue engineering purposes. In this review, an insight into the self-setting calcium orthophosphate formulations, as excellent bioceramics suitable for both dental and bone grafting applications, has been provided.

Biphasic, triphasic and multiphasic calcium orthophosphates.

Biphasic, triphasic and multiphasic (polyphasic) calcium orthophosphates have been sought as biomaterials for reconstruction of bone defects in maxillofacial, dental and orthopedic applications. In general, this concept is determined by advantageous balances of more stable (frequently hydroxyapatite) and more resorbable (typically tricalcium orthophosphates) phases of calcium orthophosphates, while the optimum ratios depend on the particular applications. Therefore, all currently known biphasic, triphasic and multiphasic formulations of calcium orthophosphate bioceramics are sparingly soluble in water and, thus, after being implanted they are gradually resorbed inside the body, releasing calcium and orthophosphate ions into the biological medium and, hence, seeding new bone formation. The available formulations have already demonstrated proven biocompatibility, osteoconductivity, safety and predictability in vitro, in vivo, as well as in clinical models. More recently, in vitro and in vivo studies have shown that some of them might possess osteoinductive properties. Hence, in the field of tissue engineering biphasic, triphasic and multiphasic calcium orthophosphates represent promising biomaterials to construct various scaffolds capable of carrying and/or modulating the behavior of cells. Furthermore, such scaffolds are also suitable for drug delivery applications. This review summarizes the available information on biphasic, triphasic and multiphasic calcium orthophosphates, including their biomedical applications. New formulations are also proposed.

Calcium orthophosphates and human beings: a historical perspective from the 1770s until 1940.

The historical development of a scientific knowledge on calcium orthophosphates from the 1770s until 1940 is described. Many forgotten and poorly known historical facts and approaches have been extracted from old publications and then they have been analyzed, systematized and reconsidered from the modern point of view. The chosen time scale starts with the earliest available studies of 1770s (to the best of my findings, calcium orthophosphates had been unknown before), passes through the entire 19th century and finishes in 1940, because since then the amount of publications on calcium orthophosphates rapidly increases and the subject becomes too broad. Furthermore, since publications of the second half of the 20th century are easily accessible, a substantial amount of them have already been reviewed by other researchers. The reported historical findings clearly demonstrate that the substantial amount of the scientific facts and experimental approaches have been known for very many decades and, in fact, the considerable quantity of relatively recent investigations on calcium orthophosphates is just either a further development of the earlier studies or a rediscovery of the already forgotten knowledge.

Calcium orthophosphate coatings, films and layers.

In surgical disciplines, where bones have to be repaired, augmented or improved, bone substitutes are essential. Therefore, an interest has dramatically increased in application of synthetic bone grafts. As various interactions among cells, surrounding tissues and implanted biomaterials always occur at the interfaces, the surface properties of the implants are of the paramount importance in determining both the biological response to implants and the material response to the physiological conditions. Hence, a surface engineering is aimed to modify both the biomaterials, themselves, and biological responses through introducing desirable changes to the surface properties of the implants but still maintaining their bulk mechanical properties. To fulfill these requirements, a special class of artificial bone grafts has been introduced in 1976. It is composed of various mechanically stable (therefore, suitable for load bearing applications) biomaterials and/or bio-devices with calcium orthophosphate coatings, films and layers on their surfaces to both improve interactions with the surrounding tissues and provide an adequate bonding to bones. Many production techniques of calcium orthophosphate coatings, films and layers have been already invented and new promising techniques are continuously investigated. These specialized coatings, films and layers used to improve the surface properties of various types of artificial implants are the topic of this review.

Dissolution mechanism of calcium apatites in acids: A review of literature.

Eight dissolution models of calcium apatites (both fluorapatite and hydroxyapatite) in acids were drawn from the published literature, analyzed and discussed. Major limitations and drawbacks of the models were conversed in details. The models were shown to deal with different aspects of apatite dissolution phenomenon and none of them was able to describe the dissolution process in general. Therefore, an attempt to combine the findings obtained by different researchers was performed which resulted in creation of the general description of apatite dissolution in acids. For this purpose, eight dissolution models were assumed to complement each other and provide the correct description of the specific aspects of apatite dissolution. The general description considers all possible dissolution stages involved and points out to some missing and unclear phenomena to be experimentally studied and verified in future. This creates a new methodological approach to investigate reaction mechanisms based on sets of affine data, obtained by various research groups under dissimilar experimental conditions.

Biocomposites and hybrid biomaterials based on calcium orthophosphates.

The state-of-the-art of biocomposites and hybrid biomaterials based on calcium orthophosphates that are suitable for biomedical applications is presented in this review. Since these types of biomaterials offer many significant and exciting possibilities for hard tissue regeneration, this subject belongs to a rapidly expanding area of biomedical research. Through successful combinations of the desired properties of matrix materials with those of fillers (in such systems, calcium orthophosphates might play either role), innovative bone graft biomaterials can be designed. Various types of biocomposites and hybrid biomaterials based on calcium orthophosphates, either those already in use or being investigated for biomedical applications, are extensively discussed. Many different formulations, in terms of the material constituents, fabrication technologies, structural and bioactive properties as well as both in vitro and in vivo characteristics, have already been proposed. Among the others, the nanostructurally controlled biocomposites, those containing nanodimensional compounds, biomimetically fabricated formulations with collagen, chitin and/or gelatin as well as various functionally graded structures seem to be the most promising candidates for clinical applications. The specific advantages of using biocomposites and hybrid biomaterials based on calcium orthophosphates in the selected applications are highlighted. As the way from the laboratory to the hospital is a long one, and the prospective biomedical candidates have to meet many different necessities, this review also examines the critical issues and scientific challenges that require further research and development.