Physicochemical Properties of Inorganic and Hybrid Hydroxyapatite-Based Granules Modified with Citric Acid or Polyethylene Glycol.

This study delves into the physicochemical properties of inorganic hydroxyapatite (HAp) and hybrid hydroxyapatite-chitosan (HAp-CTS) granules, also gold-enriched, which can be used as aggregates in biomicroconcrete-type materials. The impact of granules' surface modifications with citric acid (CA) or polyethylene glycol (PEG) was assessed. Citric acid modification induced increased specific surface area and porosity in inorganic granules, contrasting with reduced parameters in hybrid granules. PEG modification resulted in a slight increase in specific surface area for inorganic granules and a substantial rise for hybrid granules with gold nanoparticles. Varied effects on open porosity were observed based on granule type. Microstructural analysis revealed increased roughness for inorganic granules post CA modification, while hybrid granules exhibited smoother surfaces. Novel biomicroconcretes, based on α-tricalcium phosphate (α-TCP) calcium phosphate cement and developed granules as aggregates within, were evaluated for compressive strength. Compressive strength assessments showcased significant enhancement with PEG modification, emphasizing its positive impact. Citric acid modification demonstrated variable effects, depending on granule composition. The incorporation of gold nanoparticles further enriched the multifaceted approach to enhancing calcium phosphate-based biomaterials for potential biomedical applications. This study demonstrates the pivotal role of surface modifications in tailoring the physicochemical properties of granules, paving the way for advanced biomicroconcretes with improved compressive strength for diverse biomedical applications.

Study on ßTCP/P(3HB) Scaffolds-Physicochemical Properties and Biological Performance in Low Oxygen Concentration.

The search for new materials for bone regenerative purposes is still ongoing. Therefore, we present a series of newly constructed composites based on ß tricalcium phosphate (ßTCP) and poly(3-hydroxybutyrate) bacteria-derived biopolymer (P(3HB)) in the form of 3D scaffolds with different pore sizes. To improve the polymer attachment to the ßTCP surface, the etching of ceramic sinters, using citric acid, was applied. As expected, pre-treatment led to the increase in surface roughness and the creation of micropores facilitating polymer adhesion. In this way, the durability and compressive strength of the ceramic-polymer scaffolds were enhanced. It was confirmed that P(3HB) degrades to 3-hydroxybutyric acid, which broadens applications of developed materials in bone tissue engineering as this compound can potentially nourish surrounding tissues and reduce osteoporosis. Moreover, to the best of our knowledge, it is one of the first studies where the impact of ßTCP/P(3HB) scaffolds on mesenchymal stem cells (MSCs), cultured in lowered (5%) oxygen concentration, was assessed. It was decided to use a 5% oxygen concentration in the culture to mimic the conditions that would be found in damaged bone in a living organism during regeneration. Scaffolds enabled cell migration and sufficient flow of the culture medium, ensuring high cell viability. Furthermore, in composites with etched ßTCP, the MSCs adhesion was facilitated by hydrophilic ceramic protrusions which reduced hydrophobicity. The developed materials are potential candidates for bone tissue regeneration. Nevertheless, to confirm this hypothesis, in vivo studies should be performed.

Effect of Gold Nanoparticles and Silicon on the Bioactivity and Antibacterial Properties of Hydroxyapatite/Chitosan/Tricalcium Phosphate-Based Biomicroconcretes.

Bioactive, chemically bonded bone substitutes with antibacterial properties are highly recommended for medical applications. In this study, biomicroconcretes, composed of silicon modified (Si-αTCP) or non-modified α-tricalcium phosphate (αTCP), as well as hybrid hydroxyapatite/chitosan granules non-modified and modified with gold nanoparticles (AuNPs), were designed. The developed biomicroconcretes were supposed to combine the dual functions of antibacterial activity and bone defect repair. The chemical and phase composition, microstructure, setting times, mechanical strength, and in vitro bioactive potential of the composites were examined. Furthermore, on the basis of the American Association of Textile Chemists and Colorists test (AATCC 100), adapted for chemically bonded materials, the antibacterial activity of the biomicroconcretes against S. epidermidis, E. coli, and S. aureus was evaluated. All biomicroconcretes were surgically handy and revealed good adhesion between the hybrid granules and calcium phosphate-based matrix. Furthermore, they possessed acceptable setting times and mechanical properties. It has been stated that materials containing AuNPs set faster and possess a slightly higher compressive strength (3.4 ± 0.7 MPa). The modification of αTCP with silicon led to a favorable decrease of the final setting time to 10 min. Furthermore, it has been shown that materials modified with AuNPs and silicon possessed an enhanced bioactivity. The antibacterial properties of all of the developed biomicroconcretes against the tested bacterial strains due to the presence of both chitosan and Au were confirmed. The material modified simultaneously with AuNPs and silicon seems to be the most promising candidate for further biological studies.

Development of highly porous calcium phosphate bone cements applying nonionic surface active agents.

A novel way of obtaining highly porous cements is foaming them with the use of nonionic surface active agents (surfactants). In this study, foamed calcium phosphate cements (fCPCs) intended for in situ use were fabricated by a surfactant-assisted foaming process. Three different surface active agents, Tween 20, Tween 80 and Tetronic 90R4, were used. The amount of surfactant, based on its critical micelle concentration and cytotoxicity as well as foaming method, was determined. It has been established that in order to avoid cytotoxic effects the concentration of all applied surfactants in the cement liquid phases should not exceed 1.25 g L-1. It was found that Tetronic 90R4 had the lowest cytotoxicity whereas Tween 20 had the highest. The influence of the type of surfactant used in the fabrication process of bioactive macroporous cement on the physicochemical and biological properties of fCPCs was studied. The obtained materials reached higher than 50 vol% open porosity and possessed compressive strength which corresponds to the values for cancellous bone. The highest porosity and compressive strength was found for the material with the addition of Tween 80. In vitro investigations proved the chemical stability and high bioactive potential of the examined materials.

Comparative study on physicochemical properties of alpha-TCP / calcium sulphate dihydrate biomicroconcretes containing chitosan, sodium alginate or methylcellulose.

PURPOSE: Recently, the attention has been drawn to complex systems - biomicroconcretes composed of a bone cement matrix and resorbable granules or microspheres. This paper presents novel bone substitutes composed of α-tricalcium phosphate (α-TCP; cement matrix), calcium sulphate dihydrate granules (GCSD; aggregates in biomicroconcrete) and various polymers (chitosan, sodium alginate, methylcellulose) used for the improvement of material properties. The aim of this work was to study α-TCP-GCSD-polymer interactions and to compare the impact of organic additives on the physicochemical properties of biomicroconcretes. METHODS: Scanning electron microscopy (SEM), mercury intrusion porosimetry (MIP), X-ray diffractometry (XRD) as well as universal testing machine (INSTRON), Gilmore apparatus and pH/ conduct-meter were used. RESULTS: The chemical bonding between α-TCP matrix and CSD granules resulted in a compressive strength appropriate for low-load bearing applications (7-12 MPa) and clinically relevant setting times (8-33 min). Biomicroconcretes consisting of sodium alginate possessed the highest mechanical strength (12 ± 2 MPa). It has also been found that the dissolution-precipitation reactions of the α-TCP were retarded with the addition of chitosan and acetic acid. This effect was not observed in the case of methylcellulose and sodium alginate. Chemical stability and bioactivity of materials were demonstrated during in vitro studies in simulated body fluid. CONCLUSIONS: Materials containing calcium sulphate-based granules were surgically handy, possessed promising physicochemical properties and are supposed to ensure desired macroporosity as well as gradual resorption in vivo. It has been demonstrated that the presence of CSD granules and polymers influenced the physicochemical properties of composites.

In vivo behavior of biomicroconcretes based on α-tricalcium phosphate and hybrid hydroxyapatite/chitosan granules and sodium alginate.

The current studies provide insights into how predictions based on results of physicochemical and in vitro tests are consistent with the results of in vivo studies. The new biomicroconcrete type materials were obtained by mixing the solid phase, composed of hybrid hydroxyapatite/chitosan granules and highly reactive α-tricalcium phosphate powder, used as the setting agent. This approach guaranteed a good adhesion of the continuous cement phase to the surface of granules. It has been demonstrated that developed biomicroconcretes are surgically handy, possessed favorable physicochemical and biological properties and can be considered as effective bone implant material. The hierarchical porosity and compressive strength (2-6 MPa) similar to cancellous bone made them suitable for low-load bearing applications. Despite the fact that final setting times of biomicroconcretes were longer than recommended in the literature (i.e., exceeded 15 min), their short cohesion time allows for a successful implantation in a rabbit femoral defect model. Histological analysis and Raman studies revealed newly formed bone tissues around the sides of implanted materials. Furthermore, the process of neovascularization and reconstruction of the bone tissue, as well as a reverse scaffolding process, was visible. No signs of inflammation or adverse tissue reactions were observed during the experiment.

Influence of Selected Surfactants on Physicochemical Properties of Calcium Phosphate Bone Cements.

The influence of the three nonionic surface active agents such as Tween 20, Tween 80, and Tetronic 90R4 on hydrolysis, setting reaction, microstructure, and mechanical properties of alpha tricalcium phosphate (α-TCP) based materials was determined. The study revealed that the addition of any of the surfactants mentioned above slightly prolonged the setting time of the tested cements (up to 5 min). On the other hand, it was found that surfactants influence the long-term hydrolysis reaction. The addition of surfactants also affected the microstructure of the final materials, especially after incubation in a simulated body fluid. Surface active agents also had an impact on mechanical behavior of the obtained cements. Sorbitan esters, Tween 20 and Tween 80, decreased compressive strength in comparison to the reference material (6.56 ± 1.59 MPa) to 3.54 ± 1.18 and 3.68 ± 1.03 MPa, respectively. Interestingly, Tetronic 90R4, never used before as an additive to calcium phosphate bone cements (CPCs) caused a 2-fold increase of this value (up to 13.28 ± 1.59 MPa). All the developed materials exhibited bioactivity in vitro. The obtained results shed new light on surfactants as CPCs additives. They should not only be considered as foaming agent or binders, but also they deserve more attention as modifiers affecting the physicochemical properties of α-TCP based materials.

Novel bioresorbable tricalcium phosphate/polyhydroxyoctanoate (TCP/PHO) composites as scaffolds for bone tissue engineering applications.

Development of new composite materials for bone tissue engineering is a constantly growing field of medicine. Therefore there is a continuous need in creating novel materials that can not only regenerate the defected tissue but also nourish it while the healing process progresses. Here we present a novel type of composite material that fulfils these requirements. The study describes creation of a composite with macroporous bioceramic core that is infiltrated with a thin biopolymer layer. The ceramic component, namely tricalcium phosphate (TCP), due to its mechanistic and bioactive properties may promote new bone creation as shown through the in vitro studies. To the best of our knowledge the coating layer was created for the first time from a representative of bacterially derived medium chain length polyhydroxyalkanoate polymers (mcl-PHA), namely polyhydroxyoctanoate (PHO). This polymer layer not only profoundly changed the stress-strain characteristics of the bioceramic foam but also released (R)-3-hydroxyacids and their dimers/trimers to the investigated environment. In the manuscript we have in depth characterised these materials employing a set of basic procedures, through 3D structure reconstruction and finishing with prolonged in vitro experiments.

Novel multicomponent organic-inorganic WPI/gelatin/CaP hydrogel composites for bone tissue engineering.

The present work focuses on the development of novel multicomponent organic-inorganic hydrogel composites for bone tissue engineering. For the first time, combination of the organic components commonly used in food industry, namely whey protein isolate (WPI) and gelatin from bovine skin, as well as inorganic material commonly used as a major component of hydraulic bone cements, namely α-TCP in various concentrations (0-70 wt%) was proposed. The results showed that α-TCP underwent incomplete transformation to calcium-deficient hydroxyapatite (CDHA) during preparation process of the hydrogels. Microcomputer tomography showed inhomogeneous distribution of the calcium phosphate (CaP) phase in the resulting composites. Nevertheless, hydrogels containing 30-70 wt% α-TCP showed significantly improved mechanical properties. The values of Young's modulus and the stresses corresponding to compression of a sample by 50% increased almost linearly with increasing concentration of ceramic phase. Incomplete transformation of α-TCP to CDHA during preparation process of composites provides them high reactivity in simulated body fluid during 14-day incubation. Preliminary in vitro studies revealed that the WPI/gelatin/CaP composite hydrogels support the adhesion, spreading, and proliferation of human osteoblast-like MG-63 cells. The WPI/gelatin/CaP composite hydrogels obtained in this work showed great potential for the use in bone tissue engineering and regenerative medicine applications.

Novel self-gelling injectable hydrogel/alpha-tricalcium phosphate composites for bone regeneration: Physiochemical and microcomputer tomographical characterization.

Mineralized hydrogels are increasingly gaining attention as biomaterials for bone regeneration. The most common mineralization strategy has been addition of preformed inorganic particles during hydrogel formation. This maintains injectability. One common form of bone cement is formed by mixing particles of the highly reactive calcium phosphate alpha-tricalcium phosphate (α-TCP) with water to form hydroxyapatite (HA). The calcium ions released during this reaction can be exploited to crosslink anionic, calcium-binding polymers such as the polysaccharide gellan gum (GG) to induce hydrogel formation. In this study, three different amounts of α-TCP particles were added to GG polymer solution to generate novel, injectable hydrogel-inorganic composites. Distribution of the inorganic phase in the hydrogel was studied by high resolution microcomputer tomography (µCT). Gelation occurred within 30 min. α-TCP converted to HA. µCT revealed inhomogeneous distribution of the inorganic phase in the composites. These results demonstrate the potential of the composites as alternatives to traditional α-TCP bone cement and pave the way for incorporation of biologically active substances and in vitro and in vivo testing. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 822-828, 2018.

How calcite and modified hydroxyapatite influence physicochemical properties and cytocompatibility of alpha-TCP based bone cements.

Nowadays successful regeneration of damaged bone tissue is a major problem of the reconstructive medicine and tissue engineering. Recently a great deal of attention has been focused on calcium phosphate cements (CPCs) as the effective bone fillers. Despite a number of studies regarding CPCs, only a few compare the physicochemical and biological properties of α-TCP based materials of various phase compositions. In our study we compared the effect of several components (calcite, hydroxyapatite doped with Mg2+, CO32- or Ag+ ions, alginate, chitosan and methylcellulose) on the physicochemical and biological properties of α-TCP-based bone cements. The influence of materials composition on their setting times, microstructure and biochemical stability in simulated body fluid was determined. A number of in vitro laboratory methods, including ICP-OES, metabolic activity test, time-lapse microscopic observation and SEM observations were performed in order to assess biocompatibility of the studied biomaterials. The positive outcome of XTT tests for ceramic extracts demonstrated that all investigated cement-type composites may be considered cytocompatible according to ISO 10993-5 standard. Results of our research indicate that multiphase cements containing MgCHA, AgHA and calcite combined with αTCP enhanced cell viability in comparison to material based only on αTCP. Furthermore materials containing chitosan and methylcellulose possessed higher cytocompatibility than those with alginate.

New approach in evaluation of ceramic-polymer composite bioactivity and biocompatibility.

Regeneration of bone defects was promoted by a novel ß-glucan/carbonate hydroxyapatite composite and characterized by Raman spectroscopy, microCT and electron microscopy. The elastic biomaterial with an apatite-forming ability was developed for bone tissue engineering and implanted into the critical-size defects of rabbits' tibiae. The bone repair process was analyzed on non-decalcified bone/implant sections during a 6-month regeneration period. Using spectroscopic methods, we were able to determine the presence of amides, lipids and assign the areas of newly formed bone tissue. Raman spectroscopy was also used to assess the chemical changes in the composite before and after the implantation process. SEM analyses showed the mineralization degree in the defect area and that the gap size decreased significantly. Microscopic images revealed that the implant debris were interconnected to the poorly mineralized inner side of a new bone tissue. Our study demonstrated that the composite may serve as a biocompatible background for collagen ingrowth and exhibits the advantages of applying Raman spectroscopy, SEM and microCT in studying these samples.

Quantitative stereological analysis of the highly porous hydroxyapatite scaffolds using X-ray CM and SEM.

BACKGROUND: Material properties of the scaffolds as well as their microstructure are vital in determining in vivo cellular response. Three-dimensional (3D), highly porous scaffolds are used in tissue engineering to provide a suitable microenvironment and to support regeneration of bone. Both pore sizes and their architecture, in particular interconnection density, impact functionality of scaffold during its biomedical applications. OBJECTIVE: In this paper a comparative study of the microstructure of highly porous hydroxyapatite scaffolds produced via gelcasting of foamed slurries and replication of polyurethane sponge were carried out. METHODS: Quantitative stereological analysis of the microstructure was conducted using transmission X-ray computed microtomography (µCT) and scanning electron microscopy (SEM). Application of the X-ray microtomography allowed obtaining the 2D cross-sectional images of examined samples, and then the 3D reflection of individual samples. RESULTS: In our studies we proved that the distribution of pores in HAp bioceramics can be controlled by selection of the manufacturing method. In the case of material produced by the gelcasting method, the porosity of the samples was about ∼78 vol.%, while for the method of replication of the porous organic matrix it was higher ∼84 vol.%. Application of gelcasting method resulted in bioceramics with the macropores ranging from 95 µm to 158 µm (the modal value of 120 µm). Furthermore, micropores of size 34 µm-60 µm - so called "windows", were observed on spherical macropores surfaces. In the case of replication of polyurethane sponge only macropores from 295 µm to 337 µm (the modal value of 300 µm) were obtained. Application of µCT and SEM give more information than classical mercury intrusion porosimetry in studies of porous bioceramics. Developed materials met the criteria for porous bone substitutes. CONCLUSIONS: The results of quantitative description of microstructure allowed determining the differences between porous hydroxyapatite bioceramics obtained via replication of porous organic matrix and gelcasting of foamed slurry. The stereological analysis demonstrated, that bioceramics prepared via gelling of foamed slurry has a lower pore size and grains (1.1-1.9 µm) than the material obtained by the method of replication of polyurethane sponge (2.1-2.3 µm). Based on morphological analysis the porosity of tested materials was determined. In the case of material produce by the gelcasting, porosity of the samples was about ∼78 vol.%, while for method of replication of the porous organic matrix the porosity was higher and constituted ∼84 vol.%. Furthermore, evaluated materials varied in porosity and the pore size distribution. It was stated that the method of gelcasting resulted in hydroxyapatite bioceramics with the macropores diameter (95-158 µm), micropores so called "windows" (34-60 µm) - observed on spherical macropores walls and micropores of size 0.6 µm-1.3 µm, which were visible in sintered areas. When the method of replication of polyurethane sponge was applied only macropores from 295 µm to 337 µm were obtained. The comparable values of shape factors such as elongation, curvature of pours boundary and convexity, confirmed that macropores in both studied series had similar shape.

Structural transformation of synthetic hydroxyapatite under simulated in vivo conditions studied with ATR-FTIR spectroscopic imaging.

Hydroxyapatite and carbonate-substituted hydroxyapatite are widely used in bone tissue engineering and regenerative medicine. Both apatite materials were embedded into recently developed ceramic/polymer composites, subjected to Simulated Body Fluid (SBF) for 30days and characterized using ATR-FTIR spectroscopic imaging to assess their behaviour and structures. The specific aim was to detect the transition phases between both types of hydroxyapatite during the test and to analyze the surface modification caused by SBF. ATR-FTIR spectroscopic imaging was successfully applied to characterise changes in the hydroxyapatite lattice due to the elastic properties of the scaffolds. It was observed that SBF treatment caused a replacement of phosphates in the lattice of non-substituted hydroxyapatite by carbonate ions. A detailed study excluded the formation of pure A type carbonate apatite. In turn, CO32- content in synthetic carbonate-substituted hydroxyapatite decreased. The usefulness of ATR-FTIR spectroscopic imaging studies in the evaluation of elastic and porous ß-glucan hydroxyapatite composites has been demonstrated.

The comparison study of bioactivity between composites containing synthetic non-substituted and carbonate-substituted hydroxyapatite.

Apatite forming ability of hydroxyapatite (HAP) and carbonate hydroxyapatite (CHAP) containing composites was compared. Two composite materials, intended for filling bone defects, were made of polysaccharide polymer and one of two types of hydroxyapatite. The bioactivity of the composites was evaluated in vitro by soaking in a simulated body fluid (SBF), and the formation of the apatite layer was determined by scanning electron microscopy with energy-dispersive spectrometer and Raman spectroscopy. The results showed that both the composites induced the formation of apatite layer on their surface after soaking in SBF. In addition, the sample weight changes and the ion concentration of the SBF were scrutinized. The results showed the weight increase for both materials after SBF treatment, higher weight gain and higher uptake of calcium ions by HAP containing scaffolds. SBF solution analysis indicated loss of calcium and phosphorus ions during experiment. All these results indicate apatite forming ability of both biomaterials and suggest comparable bioactive properties of composite containing pure hydroxyapatite and carbonate-substituted one.

Cytocompatibility of the selected calcium phosphate based bone cements: comparative study in human cell culture.

Calcium phosphate cements (CPC) are valuable bone fillers. Recently they have been also considered as the basis for drug-, growth factors- or cells-delivery systems. Broad possibilities to manipulate CPC composition provide a unique opportunity to obtain materials with a wide range of physicochemical properties. In this study we show that CPC composition significantly influences cell response. Human bone derived cells were exposed to the several well-characterized different cements based on calcium phosphates, magnesium phosphates and calcium sulfate hemihydrate (CSH). Cell viability assays, live/dead staining and real-time observation of cells in contact with the materials (time-laps) were performed. Although all the investigated materials have successfully passed a standard cytocompatibility assay, cell behavior in a direct contact with the materials varied depending on the material and the experimental system. The most recommended were the α-TCP-based materials which proved suitable as a support for cells in a direct contact. The materials which caused a decrease of calcium ions concentration in culture induced the negative cell response, however this effect might be expected efficiently compensated in vivo. All the materials consisting of CSH had negative impact on the cells. The obtained results strongly support running series of cytocompatibility studies for preclinical evaluation of bone cements.

Effect of a carbonated HAP/ß-glucan composite bone substitute on healing of drilled bone voids in the proximal tibial metaphysis of rabbits.

A novel elastic hydroxyapatite-based composite of high surgical handiness has been developed. Its potential application in orthopedics as a filler of bone defects has been studied. The biomaterial was composed of carbonated hydroxyapatite (CHAP) granules and polysaccharide polymer (ß-1,3-glucan). Cylinders of 4mm in diameter and 6mm in length were implanted into bone cavities created in the proximal metaphysis of tibiae of 24 New Zealand white rabbits. 18 sham-operated animals were used as controls. After 1, 3 or 6 months, the rabbits were euthanized, the bones were harvested and subjected to analysis. Radiological images and histological sections revealed integration of implants with bone tissue with no signs of graft rejection. Peripheral quantitative computed tomography (pQCT) indicated the stimulating effect of the biomaterial on bone formation and mineralization. Densitometry (DXA) analysis suggested that biomineralization of bones was preceded by bioresorption and gradual disappearance of porous ceramic granules. The findings suggest that the CHAP-glucan composite material enables regeneration of bone tissue and could serve as a bone defect filler.

In vivo implantation of porous titanium alloy implants coated with magnesium-doped octacalcium phosphate and hydroxyapatite thin films using pulsed laser depostion.

The use of porous titanium-based implant materials for bone contact has been gaining ground in recent years. Selective laser melting (SLM) is a rapid prototyping method by which porous implants with highly defined external dimensions and internal architecture can be produced. The coating of porous implants produced by SLM with ceramic layers based on calcium phosphate (CaP) remains relatively unexplored, as does the doping of such coatings with magnesium (Mg) to promote bone formation. In this study, Mg-doped coatings of the CaP types octacalcium phosphate and hydroxyapatite (HA) were deposited on such porous implants using the pulsed laser deposition method. The coated implants were subsequently implanted in a rabbit femoral defect model for 6 months. Uncoated implants served as a reference material. Bone-implant contact and bone volume in the region of interest were evaluated by histopathological techniques using a tri-chromatographic Masson-Goldner staining method and by microcomputed tomography (µCT) analysis of the volume of interest in the vicinity of implants. Histopathological analysis revealed that all implant types integrated directly with surrounding bone with ingrowth of newly formed bone into the pores of the implants. Biocompatibility of all implant types was demonstrated by the absence of inflammatory infiltration by mononuclear cells (lymphocytes), neutrophils, and eosinophils. No osteoclastic or foreign body reaction was observed in the vicinity of the implants. µCT analysis revealed a significant increase in bone volume for implants coated with Mg-doped HA compared to uncoated implants.

Comparative in vitro study of calcium phosphate ceramics for their potency as scaffolds for tissue engineering.

BACKGROUND: Calcium phosphate ceramics have been widely considered as scaffolds for bone tissue engineering. Selection of the best support for cultured cells, crucial for tissue engineered systems, is still required. OBJECTIVE: We examined three types of calcium phosphate compounds: α-tricalcium phosphate - the most soluble one, carbonate hydroxyapatite - chemically the most similar to the bone mineral and biphasic calcium phosphate - with the best in vivo biocompatibility in order to select the best support for osteoblastic cells for tissue engineered systems. METHODS: Human osteoblasts were tested in direct contact with both dense samples and 3D scaffolds in either static or dynamic culture. Cell viability, cell spreading, osteogenic cell capacity, and extracellular matrix production were examined. RESULTS: The obtained data indicate that biphasic calcium phosphate is the optimal cell-supporting material. In addition, dynamic culture improved cell distribution in the scaffolds, enhanced production of the extracellular matrix and promoted cells osteogenic capacity. CONCLUSIONS: Biphasic calcium phosphate should be recommended as the most suitable matrix for osteogenic cells expansion and differentiation in tissue engineered systems.

Physicochemical properties of the novel biphasic hydroxyapatite-magnesium phosphate biomaterial.

Besides high-temperature calcium phosphates (CaPs), low-temperature calcium phosphate bone cements (CPCs), due to excellent biological properties: bioactivity, biocompability and osteoconductivity, are successfully used as bone substitutes. However, some disadvantages, related mainly to their low resorption rate and poor mechanical properties result in limited range of applications of these implant materials to non-loaded places in the skeletal system. To overcome this problem, magnesium phosphate cements (MPCs) with high strength have been considered as biomaterials. The main disadvantage of MPCs is that the acid-base setting reaction is an exothermic process that must be strictly controlled to avoid tissue necrosis. In this work, a new composite bone substitute (Hydroxyapatite Magnesium Phosphate Material - HMPM) based on hydroxyapatite (HA) and magnesium phosphate cement (MPC) with sodium pyrophosphate applied as a retardant of setting reaction was obtained. Its setting time was adequate for clinical applications. Combining properties of HA and MPC has made it possible to obtain microporous (showing bimodal pore size distribution in the range of 0.005-1.700 micrometers) potential implant material showing good surgical handiness and sufficient mechanical strength. Effectiveness of sodium pyrophosphate as a retardant of exothermic setting reaction of the new cement formulation was confirmed. After setting and hardening, the material consisted of hydroxyapatite and struvite as crystalline phases. Unreacted magnesium oxide was not detected.