β-TCP Powder Research Articles

Evaluation of europium-doped HA/β-TCP ratio fluorescence in biphasic calcium phosphate nanocomposites controlled by the pH value during the synthesis

Europium-doped hydroxyapatite (HA), beta-tricalcium phosphate (β-TCP) and biphasic phosphate nanopowders were synthesized by co-precipitation method and their crystal structures and fluorescence properties were investigated depending on the pH of the starting solution. In the range of pH 6–10, HA and β-TCP phases coexist. The β-TCP proportion increased as the pH of the solution decreased, while HA yields decreased. At pH below 6, monophasic β-TCP powder was obtained after thermal treatment. In particular, HA and β-TCP can be used as luminescent materials when activated by Eu3+ ions in substitution of Ca2+ ions. Herein, the Eu3+ ions doped HA and β-TCP phase composition were analyzed in order to investigate the fluorescence emission of the HA, β-TCP and biphasic compounds. Eu-doped HA exhibited a red-orange emission at 575nm with several minor peaks at 610–640nm, while Eu-doped β-TCP had an unexpected strong red emission at 610–620nm and a secondary band at 590–600nm. In fact, the Eu:β-TCP integrated emission area is almost 20-fold higher than Eu:HA for the same europium ion concentration. These results demonstrate the potential of Eu:β-TCP as biomarker for medical applications, as drug release and targeting based on their luminescent properties.

Zinc-releasing calcium phosphate cements for bone substitute materials

The goal of the present study was to develop zinc-doped calcium phosphate bone cements (CPCs-Zn) for functionalized bone grafts applications. An aqueous co-precipitation process for synthesis of β-tricalcium phosphate (β-TCP) powder samples with Zn addition ranging from 0wt% up to 1.2wt% was proposed as an alternative approach to produce CPCs-Zn, instead of mechanical mixing of powders. Real-time monitoring of structural transformations and kinetics of CPCs-Zn (Zn=0wt%, 0.6wt% and 1.2wt%) was carried out by the Energy Dispersive X-Ray Diffraction technique. The microstructure development in CPCs-Zn was investigated by Scanning Electron Microscopy. Zn-ion release from CPC-Zn 0.6wt% and CPC-Zn 1.2wt% cements, measured by the Atomic Emission Spectroscopy, corresponded to the average values of 0.30±0.01 and 0.10±0.01µg/L, respectively. The results of antibacterial tests proved the inhibitory effect towards pathogenic Escherichia coli only for the CPC-Zn 0.6wt% cement formulation.

Indirect fabrication of hydroxyapatite/β-tricalcium phosphate scaffold for osseous tissue formation using additive manufacturing technology

Patient specific HA/β-TCP scaffold were fabricated using an indirect fabrication approach which involves the application of rapid prototyping technology. β-TCP powder with 10, 20 and 30 % HA concentrations were analyzed for different sintering temperatures. Scaffolds were evaluated by Scanning Electron Microscopy, X-ray diffraction, compression tests, Fourier transform infrared spectroscopy and cytotoxicity tests using human osteosarcoma cell line. Results include HA/β-TCP scaffold porosity, volumetric shrinkage, mechanical properties, bonding, structural phase change and cytotoxicity.

Synthesis and characterization of sintered beta-tricalcium phosphate: A comparative study on the effect of preparation route

Beta-tricalcium phosphate (β-TCP) was prepared by three different routes namely, wet chemical coprecipitation, sol-gel and solution combustion synthesis. The synthesized powders were calcined at different temperatures and characterized for phase evolution study, thermal analysis, Fourier transform infrared (FTIR) spectroscopy, microstructural study for comparative analysis. The optimal thermal treatment required to prepare pure β-TCP powders was determined and after calcination of the synthesize powders prepared by different routes, pure β-TCP was obtained. The sintering temperature required to prepare fully dense β-TCP completely free from α-form was identified. The powders were then used to make dense and compact bodies sintered at 1200 and 1250°C. The sintering behaviour of the dense bodies was analysed using dilatometry, densification and microstructural study. It was found that the pellet prepared from powder synthesized via co-precipitation route attained maximum density compared to the pellets prepared from powders synthesized via sol-gel and solution combustion route.

Properties of calcium phosphates ceramic composites derived from natural materials

In this study, ceramics containing mixed phases of hydroxyapatite/beta-tricalcium phosphate (HA/β-TCP) were fabricated by a solid-state reaction technique. The HA powder was synthesized from cockle shells while the β-TCP powder was synthesized from egg shells. Pure HA and β-TCP fine powders were successfully obtained. The HA and β-TCP were mixed and subjected to a thermal treatment up to 1100°C. To form the mixed phase ceramics, the resulting powders were sintered at 1350°C. Effects of HA concentration on the properties of the studied ceramic were investigated. X-ray diffraction analysis revealed that all samples presented multiphase of calcium phosphate compounds. Average grain size of the ceramics decreased with the HA additive content. The 75wt% HA ceramic showed the maximum hardness value (5.5GPa) which is high when compared with many calcium phosphate ceramics. In vitro bioactivity test indicated that apatite forming increased with the HA additive content. To increase antibacterial activity, selected ceramics were coated with AgNO3. Antibacterial test suggested that an Ag compound coating on the ceramics could improve the antibacterial ability of the studied ceramics. In addition, the antibacterial ability for the Ag coated ceramics depended on the porosity of the ceramics.

Effect of Stirring and Aging Time on the Formation of β-Tricalcium Phosphate (β-TCP) Powder

In this research, the effect of stirring and aging time on the formation of β-tricalcium phosphate (β-TCP) powder was studied. β-TCP powder was synthesized using calcium nitrate tetrahydrate (Ca(NO3)2.4H2O) (0.6M) and diammonium hydrogen phosphate (NH4)2HPO4) (0.4M) via wet precipitation method. The mixture was stirred with different duration (1, 3, 5 and 7 hours) then centrifuged before washed with distilled water (twice) and ethanol followed by drying in oven (80°C, 24 hours). The cake was ground to form powder. The as prepared powder was analyzed using thermo-gravimetric (TGA) to determine the suitable calcinations temperature. TGA results show that the proper calcinations temperature was 800°C. The formation of β-TCP was characterized using X-ray Diffraction (XRD) analysis. Sample with optimum formation of β-TCP phase will choose for further study on the effect of aging time (0.5, 1, 20 and 24 hours). XRD analysis confirmed that sample stirred for 7 hours and aging for 24 hours produced β-TCP as major phase. Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM) proved that β-TCP powder form as agglomerated particles

Biotin-avidin mediates the binding of adipose-derived stem cells to a porous β-tricalcium phosphate scaffold: Mandibular regeneration.

The present study aimed to investigate the properties of a promising bone scaffold for bone repair, which consisted of a novel composite of adipose-derived stem cells (ADSCs) attached to a porous β-tricalcium phosphate (β-TCP) scaffold with platelet-rich plasma (PRP). The β-TCP powder was synthesized and its composition was determined using X-ray diffraction and Fourier transform infrared spectroscopy. The surface morphology and microstructure of the fabricated porous β-TCP scaffold samples were analyzed using light and scanning electron microscopy, and their porosity and compressive strength were also evaluated. In addition, the viability of rabbit ADSCs incubated with various concentrations of the β-TCP extraction fluid was analyzed. The rate of attachment and the morphology of biotinylated ADSCs (Bio-ADSCs) on avidin-coated β-TCP (Avi-β-TCP), and untreated ADSCs on β-TCP, were compared. Furthermore, in vivo bone-forming abilities were determined following the implantation of group 1 (Bio-ADSCs/Avi-β-TCP) and group 2 (Bio-ADSCs/Avi-β-TCP/PRP) constructs using computed tomography, and histological osteocalcin (OCN) and alkaline phosphatase (ALP) expression analyses in a rabbit model of mandibulofacial defects. The β-TCP scaffold exhibited a high porosity (71.26±0.28%), suitable pore size, and good mechanical strength (7.93±0.06 MPa). Following incubation with β-TCP for 72 h, 100% of viable ADSCs remained. The avidin-biotin binding system significantly increased the initial attachment rate of Bio-ADSCs to Avi-β-TCP in the first hour (P<0.01). Following the addition of PRP, group 2 exhibited a bony-union and mandibular body shape, newly formed bone and increased expression levels of OCN and ALP in the mandibulofacial defect area, as compared with group 1 (P<0.05). The results of the present study suggested that the novel Bio-ADSCs/Avi-β-TCP/PRP composite may have potential application in bone repair and bone tissue engineering.

3D Porous Architecture of Stacks of β-TCP Granules Compared with That of Trabecular Bone: A microCT, Vector Analysis, and Compression Study.

The 3D arrangement of porous granular biomaterials usable to fill bone defects has received little study. Granular biomaterials occupy 3D space when packed together in a manner that creates a porosity suitable for the invasion of vascular and bone cells. Granules of beta-tricalcium phosphate (β-TCP) were prepared with either 12.5 or 25 g of β-TCP powder in the same volume of slurry. When the granules were placed in a test tube, this produced 3D stacks with a high (HP) or low porosity (LP), respectively. Stacks of granules mimic the filling of a bone defect by a surgeon. The aim of this study was to compare the porosity of stacks of β-TCP granules with that of cores of trabecular bone. Biomechanical compression tests were done on the granules stacks. Bone cylinders were prepared from calf tibia plateau, constituted high-density (HD) blocks. Low-density (LD) blocks were harvested from aged cadaver tibias. Microcomputed tomography was used on the β-TCP granule stacks and the trabecular bone cores to determine porosity and specific surface. A vector-projection algorithm was used to image porosity employing a frontal plane image, which was constructed line by line from all images of a microCT stack. Stacks of HP granules had porosity (75.3 ± 0.4%) and fractal lacunarity (0.043 ± 0.007) intermediate between that of HD (respectively 69.1 ± 6.4%, p < 0.05 and 0.087 ± 0.045, p < 0.05) and LD bones (respectively 88.8 ± 1.57% and 0.037 ± 0.014), but exhibited a higher surface density (5.56 ± 0.11 mm2/mm3 vs. 2.06 ± 0.26 for LD, p < 0.05). LP granular arrangements created large pores coexisting with dense areas of material. Frontal plane analysis evidenced a more regular arrangement of β-TCP granules than bone trabecule. Stacks of HP granules represent a scaffold that resembles trabecular bone in its porous microarchitecture.

Microstructure, physical properties, and bone regeneration effect of the nano-sized β-tricalcium phosphate granules

The nano-sized β-tricalcium phosphate granules for the practical application of bone graft substitutes could be prepared from the wet chemically precipitated β-TCP powders by the liquid–solid mixture route and by controlling the pH of mixture solution to 7.5 within a shorter processing time. The phase purity of prepared β-TCP granules was higher above 99% and their particle sizes ranged from 200 to 650nm. Also, their average compressive strength was higher at 2.22MPa. It is considered that the phase purity, particle refinement, and mechanical compressive strength of β-TCP granules could be significantly improved through the β-TCP powders synthesized through the liquid–solid mixture precipitation at pH of 7.5. Meanwhile both the porosity and the specific surface area positively associated with the osteoconductivity for bone regeneration were higher at 75% and 2.50m2/g respectively due to the nano-sized particles of porous β-TCP granules. Furthermore, the histological analysis in beagle mandibular defect showed that β-TCP granules demonstrated remarkable bone regeneration effectcompared with that of the non-treatment group, indicating the increased new formation of bone (except for callus) (48.42±6.57%) and rapid resorption (69.49±2.40%) without toxicologically significant changes at 12weeks after implantation.

Pulse electric current sintering of hydroxyapatite/β-tricalcium phosphate composites

Hydroxyapatite (HA)/β-tricalcium phosphate (β-TCP) composites attract attentions as bone implant materials. As one of the fabrication method of HA/β-TCP is mixing of HA and β-TCP powder in advance of sintering. This method enables to control the ratio of content of β-TCP easier. However, it is difficult to obtain dense composites. In this study, we focused on pulse electric current sintering (PECS) to obtain dense HA/β-TCP composites. The sinterability is evaluated with relative density and grain size measurements. Composition of sintered body was also characterized by X-ray diffraction. In comparison with pressureless sintering, PECS increased relative density of the composites without grain growth. In HA/β-TCP sintered by PECS, the phase transformation from β-TCP to α-TCP was promoted. This is due to higher thermal energy by spark discharge during PECS. On the other hand, sintering additives (MgO) inhibited phase transformation. It was suggested that sinterability of HA/β-TCP composites was improved by PECS.

Analysis of β-tricalcium phosphate granules prepared with different formulations by nano-computed tomography and scanning electron microscopy.

Among biomaterials used for filling bone defects, beta-tricalcium phosphate (β-TCP) is suitable in non-bearing bones, particularly in dental implantology, oral and maxillofacial surgery. When β-TCP granules are placed in a bone defect, they occupy the void 3D volume. Little is known about the 3D arrangement of the granules, which depends on the nature and size of the granules. The aim of this study was to examine the 3D architecture of porous β-TCP granules. Granules were prepared with different concentrations of β-TCP powder in slurry (10, 11, 15, 18, 21, and 25g of β-TCP powder in distilled water). Granules were prepared by the polyurethane foam method. They were analyzed by nano-computed tomography (nanoCT) and compared with scanning electron microscopy (SEM). Commercial granules of hydroxyapatite-β-TCP prepared by the same methodology were also used. The outer and inner architectures of the granules were shown by nanoCT which evidenced macroporosity, internal porosity and microporosity between the sintered grains. Macroporosity was reduced at high concentration and conversely, numerous concave surfaces were observed. Internal porosity, related to the sublimation of the polyurethane foam, was present in all the granules. Microporosity at the grain joints was evidenced by SEM and on 2D nanoCT sections. Granules presented a heterogeneous aspect due to the different mineralization degree of the sintered powder grains in the β-TCP granules; the difference between hydroxyapatite and β-TCP was also evidenced. NanoCT is an interesting method to analyze the fine morphology of biomaterials with a resolution close to synchrotron and better than microcomputed tomography.

Effects of high-energy ball-milling on injectability and strength of β-tricalcium-phosphate cement

Calcium phosphate cement (CPC) offers many advantages as a bone-substitution material. The objective of this study is to develop a new CPC that simultaneously exhibits fine injectability, a short setting time, and high strength. β-tricalcium phosphate (β-TCP, control) powder was ball-milled for 24h to produce a new cement powder. The modified β-TCP after 24h milling (mβ-TCP-24h) exhibited excellent injectability even 1h after mixing. The mechanical properties of the set cement (compact) were evaluated using compressive strength (CS) and diametral tensile strength (DTS) testing. The CS and DTS values of the mβ-TCP-24h compacts were 8.02MPa and 2.62MPa, respectively, at 5h after mixing, and were 49.6MPa and 7.9MPa, respectively, at 2 weeks after mixing. All the CS and DTS values of the mβ-TCP-24h compacts were significantly higher than those of the control for the same duration after mixing. These results suggest that the mechano-chemically modified β-TCP powder dissolves rapidly and accelerates hydroxyapatite precipitation, which successfully shortens the cement setting time and enhances the strength. This study supports that mβ-TCP-24h is a promising candidate for use in injectable CPCs with improved strength.

Microwave-assisted synthesis of high purity β-tricalcium phosphate crystalline powder from the waste of Green mussel shells (Perna canaliculus)

Beta-tricalcium phosphate (β-TCP) was successfully synthesized using the waste of Green mussel shells, Perna canaliculus. Calcined mussel shells and phosphoric acid were mixed in 1.5 Ca/P molar ratio and subjected to microwave irradiation (1100W) for 30min and subsequently calcined at 750°C. The synthesized powder was chemically, compositionally and structurally characterized and was found to be very similar to a commercial β-TCP. Furthermore, the obtained powder was stable up to 1000°C and lost only 2% of its weight. Its toxic metallic contents (e.g. Cd, Pb and As) were lower than standard limits for biogenic calcium phosphate for medical application. The synthesized β-TCP powder shows spherical morphology having diameter in the range of 100–150nm and Ca/P molar ratio of 1.49, which is close to the stoichiometric ratio. The results obtained in this study showed that pure β-TCP can be produced from waste mussel shells in a simple and fast way using microwave irradiation.

Preparation of α-Tricalcium Phosphate Powders Surface-modified with Inositol Phosphate for Cement Fabrication

We have previously developed biodegradable β-tricalcium phosphate (β-TCP) cement based on the chelate-setting mechanism of inositol phosphate (IP6). The β-TCP cement powder for the cement fabrication was prepared via a novel powder preparation process, in which the starting β-TCP powders were prepared by simultaneous ball-milling and surface-modification in the IP6 solution. In the present study, the novel powder preparation process was applied to an α-TCP powder, and effect of milling time and beads size for ball-milling on the material properties of the α-TCP powders was investigated. The α-TCP powder ball-milled in 1000 ppm IP6 solution for 4 h with 2 mm-diameter beads was composed of single phase α-TCP with the smallest particle size of 2.2 µm. Dissolution of 4 h-milled α-TCP powder was approximately twice higher than that of starting α-TCP powder before ball-milling. The α-TCP powder with high dissolution property prepared via the novel powder preparation process is potential candidate for fabrication of the chelate-setting cement.

Preparation of α-Tricalcium Phosphate Powders Surface-Modified with Inositol Phosphate for Cement Fabrication

We have previously developed biodegradable β-tricalcium phosphate (β-TCP) cement based on the chelate-setting mechanism of inositol phosphate (IP6). The β-TCP cement powder for the cement fabrication was prepared via a novel powder preparation process, in which the starting β-TCP powders were prepared by simultaneous ball-milling and surface-modification in the IP6 solution. In the present study, the novel powder preparation process was applied to an α-TCP powder, and effect of milling time and beads size for ball-milling on the material properties of the α-TCP powders was investigated. The α-TCP powder ball-milled in 1000 ppm IP6 solution for 4 h with 2 mm-diameter beads was composed of single phase α-TCP with the smallest particle size of 2.2 µm. Dissolution of 4 h-milled α-TCP powder was approximately twice higher than that of starting α-TCP powder before ball-milling. The α-TCP powder with high dissolution property prepared via the novel powder preparation process is potential candidate for fabrication of the chelate-setting cement.

Biocompatibility of Silver-Containing Calcium-Phosphate Cements with Anti-Bacterial Properties

We have previously synthesized silver-containing hydroxyapatite (Ag-HAp) powders by an ultrasonic spray-pyrolysis (USSP) technique. On the other hand, we have successfully fabricated novel calcium-phosphate cements (CPCs) composed of mainly β-tricalcium phosphate (β-TCP) phase with anti-washout property (hereafter, β-TCP cement), which was set on the basis of chelate-bonding ability of inositol phosphate (IP6). In this study, we developed novel CPCs with both anti-bacterial and anti-washout properties by adding the Ag-HAp powder into the above β-TCP cements, and examined their anti-bacterial property and cytotoxicity. The Ag-HAp powders with Ag contents of 0, 2, and 5 mol% as a nominal composition were synthesized by an USSP technique. The raw powder for β-TCP cement was prepared by ball-milling the commercially-available β-TCP powder in the IP6 solution. The Ag-HAp/β-TCP powders were prepared by mixing Ag-HAp powder and β-TCP cement powder at a ratio of 25:75 in mass. The Ag-HAp/β-TCP cement was fabricated by mixing the above-mentioned Ag-HAp/β-TCP powder and 2.5 mass% Na2HPO4 solution at a powder/liquid ratio of 1/0.3 [g/cm3]. The anti-bacterial property of resulting cements was evaluated using Staphylococcus aureus by biofilm formation test. The Ag-HAp/β-TCP cements containing 2 and 5 mol% Ag showed strong anti-bacterial property among examined specimens. Furthermore, the cytotoxicity of Ag+ ion eluted from these cements was also examined using osteoblastic MC3T3-E1 cells and Transwell® kit. The relative cell viability cultured on each Ag-containing cement specimen was over 80 %, compared with the control (polystyrene plate). These results demonstrate that the present Ag-HAp/β-TCP cements containing 2 mol% Ag are promising one of the candidates as CPCs with both anti-bacterial property and biocompatibility.

Effect of MgO addition on solid state synthesis and thermal behavior of beta-tricalcium phosphate

In the present work the effect of Mg2+ ion doping (1–14mol%) on the solid state synthesis of beta-tricalcium phosphate (β-TCP) powder and its thermal behaviour is investigated. The powders are synthesized at temperatures between 1000 and 1300°C for 2h, starting from brushite (hydrated calcium hydrogen phosphate—CaHPO4·2H2O) and calcium carbonate (Ca CO3). The maximum substitution of Mg2+ ion in β-TCP lattice decreases from >14mol% to less than 8mol% as the synthesis temperature increases from 1000 to 1300°C. It is also found that already in 1mol% MgO doped powders the β-TCP phase is stabilized at temperatures higher than 1500°C.

Inhibition of phase transformation from β- to α-tricalcium phosphate with addition of poly (L-lactic acid) in selective laser sintering

Purpose – The paper aims to fabricate an α-tricalcium phosphate (TCP) scaffold with an interconnected porous structure via selective laser sintering (SLS). To inhibit the phase transformation from β- to α-TCP in fabrication process of porous scaffolds, a small amount (1 weight per cent) of poly (L-lactic acid) (PLLA) is added into β-TCP powder to introduce the transient liquid phase. Design/methodology/approach – The paper opted for the transient liquid phase of melting PLLA to decrease the sintering temperature in SLS. Meanwhile, the densification of β-TCP is enhanced with a combined effect of the capillary force caused by melting PLLA and the surface energy of β-TCP particles. Moreover, the PLLA will gradually decompose and completely disappear with laser irradiation. Findings – The testing results show the addition of PLLA enables the scaffolds to achieve a higher β-TCP content of 77 ± 1.49 weight per cent compared with the scaffold sintered from β-TCP powder (60 ± 1.65 weight per cent), when the laser energy density is 0.4 J/mm2. The paper provides the mechanism of PLLA inhibition on the phase transformation from β- to α-TCP. And the optimum sintering parameters are obtained based on experimental results, which are used to prepare a TCP scaffold with an interconnected porous structure via SLS. Research limitations/implications – This paper shows that the laser energy density is an important sintering parameter that can provide the means to control the micro-porous structure of the scaffold. If the laser energy density is too low, the densification is not enough. On the other hand, if the laser energy density is too high, the microcracks are observed which are attributed to the volume expansion during the phase transformation from β- to α-TCP. Therefore, the laser energy density must be optimized. Originality/value – The paper provides a feasible method for fabricating TCP artificial bone scaffold with good biological and mechanical properties.

Synthesis, structure, thermal stability, mechanical and antibacterial behaviour of lanthanum (La3 +) substitutions in β-tricalciumphosphate

Five different concentrations of lanthanum (La3+) substituted β-tricalcium phosphate [β-TCP, β-Ca3(PO4)2] were formed through aqueous precipitation technique and the results were compared with stoichiometric β-TCP. All the La3+ substituted β-TCP powders were characterized using XRD, FT-IR, XRF, Raman spectroscopy and Rietveld refinement of the XRD data. The results from the investigation confirmed the presence of La3+ in rhombohedral β-TCP structure. The substitution of higher sized of La3+ led to the considerable enhancement in lattice parameters of β-TCP crystal structure and La3+ was found to have occupied the eight fold coordinated Ca (3) site of β-TCP structure. La3+ occupancy at the Ca (3) site resulted in the significant distortions of the associated PO4 tetrahedra, which were supported by the Raman and FT-IR spectroscopic techniques. La3+ presence in the crystal lattice of β-TCP also led to the delay in allotropic phase transformation of β-TCP to α-TCP till 1300°C, thus signifying the good thermal stability of La3+ substituted β-TCP powders. The antibacterial efficiency of La3+ substituted β-TCP powders was confirmed from the in vitro tests done on microbes such as Staphylococcus aureus and Escheria coli. Further, the presence of La3+ in the crystal lattice of β-TCP did not affect the hardness and Young's modulus values of β-TCP.

Effects of Proliferation and Differentiation of Mesenchymal Stem Cells on Compressive Mechanical Behavior of Collagen/β-TCP Composite Scaffold

The primary aim of this study is to characterize the effects of cell culture on the compressive mechanical behavior of the collagen/β-tricalcium phosphate (TCP) composite scaffold. The composite and pure collagen scaffolds were fabricated by the solid–liquid phase separation technique and the subsequent freeze-drying method. Rat bone marrow mesenchymal stem cells (rMSCs) were then cultured in these scaffolds up to 28 days. Compression test of the scaffolds with rMSCs were conducted periodically. Biological properties such as cell number, alkaline phosphatase (ALP) activity, and gene expressions of osteogenetic bone markers were evaluated during cell culture. The microstructural changes in the scaffolds during cell culture were also examined using a scanning electron microscope. The compressive elastic modulus was then correlated with those of the biological properties and microstructures to understand the mechanism of variational behavior of the macroscopic elastic property. The composite scaffold exhibited higher ALP activity and more active generation of osteoblastic markers than the collagen scaffold, indicating that β-TCP can activate the differentiation of rMSCs into osteoblasts and extracellular matrix (ECM) formation such as type I collagen and the following mineralization. The variational behavior of the compressive modulus of the composite scaffold was affected by both the material degradation and the proliferation of cells and the ECM formation. In the first stage, the modulus of the composite scaffold tended to increase due to cell proliferation and the following formation of network structure. In the second stage, the modulus tended to decrease because the material degradation such as ductile deformation of collagen and decomposition of β-TCP were more effective on the property than the ECM formation. In the third stage, active calcification by formation and growth of mineralized nodules resulted in the recovery of modulus. It is concluded that the introduction of β-TCP powder into the porous collagen matrix is very effective to improve the mechanical and biological properties of collagen scaffold prepared for bone tissue engineering. Furthermore, the compressive modulus of the composite scaffold is strongly affected by the material degradation and the ECM formation by stem cells under in vitro culture condition.