Β-tricalcium Phosphate Composites Research Articles

Fabrication of a novel biomaterial of mono-layer of halloysite nanotube-magnesium oxide/ β-tricalcium phosphate composite on implant substrate: Structural and biological properties

In the present study, a novel composite coating consisting of magnesium oxide on halloysite nanotube (HNT-MgO) and doping of zinc and strontium ions in beta-tricalcium phosphate (Mn+-β-TCP). The monolayers of biomaterials were developed by coating HNT-MgO-Mn+-β-TCP on Ti–6Al–4V alloy using the electrophoretic method. The physicochemical properties such as functional groups, phase composition, surface morphology, cross-section, and chemical composition of biomaterials were characterized by adopted analytical techniques such as Fourier transform infrared spectroscopy, X-ray diffraction analysis, Scanning electron microscopy-Energy dispersive spectroscopy, Atomic force microscopy, High-resolution transmission electron microscopy. The cytotoxic activity of the prepared composite was tested against osteosarcoma MG-63 cells using MTT assay, and AO/EB staining the results clearly showed, that by increasing the composite concentration (5–100 μg/ml) the cell viability was reduced. Further, a cell adhesion study was performed by FE-SEM analysis, which showed that the MG-63 cells could inhibit and restrict the HNT-MgO-Mn+-β-TCP monolayer composite. The composite coating confirms the significant anticancer activity against osteosarcoma cells suggesting it is a promising candidate for further studies for practical orthopedic applications.

Poly(δ-valerolactone)/Poly(ethylene-co-vinylalcohol)/β-Tri-calcium Phosphate Composite as Scaffolds: Preparation, Properties, and In Vitro Amoxicillin Release.

Two poly(δ-valerolactone)/poly(ethylene-co-vinylalcohol)/β-tricalcium phosphate (PEVAL/PDVAL/β-TCP) composites containing an equal ratio of polymer and filled with 50 and 70 wt% of β-TCP microparticles were prepared by the solvent casting method. Interconnected pores were realized using the salt leached technique, and the porosity of the resulted composites was evaluated by the scanning electron microscopy (SEM) method. The homogeneity of the hybrid materials was investigated by differential scanning calorimetry (DSC) and X-ray diffraction (XRD) analysis. The prepared materials’ SEM images showed interconnected micropores that respond to the conditions required to allow their uses as scaffolds. The porosity of each scaffold was determined from micro computed tomography (micro-CT) data, and the analysis of the mechanical properties of the prepared materials was studied through the stress-strain compressive test. The proliferation test results used human mesenchymal stem cells (MSCs) to grow and proliferate on the different types of prepared materials, reflecting that the hybrid materials were non-toxic and could be biologically acceptable scaffolds. The antibacterial activity test revealed that incorporation of amoxicillin in the specimens could inhibit the bacterial growth of S. aureus. The in vitro study of the release of amoxicillin from the PEVAL/PDVAL/amoxicillin and PEVAL/PDVAL/β-TCP/amoxicillin drug carrier systems in pH media 7.4, during eight days, gave promising results, and the antibiotic diffusion in these scaffolds obeys the Fickian model.

Influence of the material properties of a poly(D,L-lactide)/β-tricalcium phosphate composite on the processability by selective laser sintering.

Complex 3D scaffolds with interconnected pores are a promising tool for bone regeneration. Such 3D scaffolds can be manufactured by selective laser sintering (SLS) from biodegradable composite powders. However, the mechanical strength of these scaffolds is often too low for medical application. We propose that the mechanical strength of laser-sintered scaffolds can be improved through composite powders with tailored properties (e.g., suitable powder particle size and melt viscosity for SLS). To prove this, two batches of a poly(D,L-lactide) (PDLLA)/β-tricalcium phosphate (β-TCP) composite powder with 50 wt% PDLLA and 50 wt% β-TCP were synthesized. The two batches differed in polymer particle size, filler particle size, and polymer molecular weight. Both batches were processed with identical SLS process parameters to study the extent to which the material properties influence how well a PDLLA/β-TCP (50/50) composite can be processed with SLS. In the SLS process, batch 2 showed improved melting behavior due to its smaller polymer particle size (approx. 35 µm vs. 50 µm) and its lower zero-shear melt viscosity (5800 Pa∙s vs. 17,900 Pa∙s). The better melting behavior of batch 2 led to SLS test specimens with lower porosity compared to batch 1. In consequence, the batch 2 specimens exhibited a larger biaxial bending strength (62 MPa) than the batch 1 specimens did (23 MPa). We conclude that a tailored composite powder with optimized polymer particle size, filler particle size, and polymer molecular weight can increase the achievable mechanical strength of laser-sintered scaffolds.

Effects of gatifloxaine content in gatifloxacine-loaded PLGA and β-tricalcium phosphate composites on efficacy in treating osteomyelitis.

Composites of gatifloxacin (GFLX)-loaded poly (lactic-co-glycolic) acid (PLGA) and β-tricalcium phosphate (βTCP) containing 0, 1, and 10 wt % GFLX (0, 1, and 10 wt % GFLX composites), and GFLX-loaded PLGA containing 1, 5, and 10 wt % GFLX (1, 5, and 10wt % GFLX-PLGA) as controls were fabricated and characterized in vitro and in vivo. On in vitro evaluation, the 10 wt % GFLX composite released GFLX over at least 28 days in Hanks' balanced solution and exhibited clinically sufficient bactericidal activities against Streptococcus milleri and Bacteroides fragilis from 1 h to 10 days. The 0, 1, and 10 wt % GFLX composites and 10 wt % GFLX-PLGA were implanted in bone defects created by debridement of osteomyelitis lesions induced by S. milleri and B. fragilis in the mandible of rabbits (n = 5). Four weeks after implantation of the 10 wt % GFLX composite, inflammation in the debrided area disappeared in all the rabbits, while inflammation remained in all the rabbits after implantation of the 0 wt % GFLX composite and 10 wt % GFLX-PLGA, and in three rabbits after implantation of the 1 wt % GFLX composite. Bone formation appears to be less intense for the 10 wt % GFLX composite than for the 1 wt % GFLX composite probably owing to the rapid degradation of the 10 wt % GFLX composite. These findings show that the GFLX composite is effective for the local treatment of osteomyelitis.

Mechanical properties of porous β-tricalcium phosphate composites prepared by ice-templating and poly(ε-caprolactone) impregnation.

In this study ceramic scaffolds of the bioresorbable and osteoconductive bioceramic β-tricalcium phosphate (β-TCP) were impregnated with the bioresorbable and ductile polymer poly(ε-caprolactone) (PCL) to investigate the influence of the impregnation on the mechanical properties of the porous composites. The initial β-TCP scaffolds were fabricated by the ice-templating method and exhibit the typical morphology of aligned, open, and lamellar pores. This pore morphology seems to be appropriate for applications as bone replacement material. The macroporosity of the scaffolds is mostly preserved during the solution-mediated PCL impregnation as the polymer was added only in small amounts so that only the micropores of β-TCP lamellae were infiltrated and the surface of the lamellae were coated with a thin film. Composite scaffolds show a failure behavior with brittle and plastic contributions, which increase their damage tolerance, in contrast to the absolutely brittle behavior of pure β-TCP scaffolds. The energy consumption during bending and compression load was increased in the impregnated scaffolds by (a) elastic and plastic deformation of the introduced polymer, (b) drawing and formation of PCL fibrils which bridge micro- and macrocracks, and (c) friction of ceramic debris still glued together by PCL. PCL addition also increased the compressive and flexural strength of the scaffolds. An explanatory model for this strength enhancement was proposed that implicates the stiffening of cold-drawn PCL present in surface flaws and micropores.

Massive bone reconstruction with heat-treated bone graft loaded autologous bone marrow-derived stromal cells and β-tricalcium phosphate composites in canine models

Bone marrow-derived stromal cells (BMSCs) contain mesenchymal stem cells that are capable of forming various mesenchymal tissues. We hypothesized that BMSCs and β-tricalcium phosphate (β-TCP) composites would promote the remodeling of large-sized autologous devitalized bone grafts; therefore, the aim of this study was to evaluate the effects of the composites on the remodeling of autologous devitalized bone grafts. Autologous BMSCs cultured in culture medium containing dexamethasone (10(-7) M) were loaded into porous β-TCP granules under low-pressure. Theses BMSC/TCP composites were put into the bone marrow cavity of autologous heat-treated bone (femoral diaphysis, 65-mm long, 100°C, 30 min) and put back to the harvest site. In the contralateral side, β-TCP without BMSC were used in the same manner as the opposite side as the control. Treatment with the BMSC/TCP composites resulted in a significant increase in thickness, bone mineral density, and matured bone volume of the cortical bone at the center of the graft compared to the control. Histological analysis showed matured regenerated bone in the BMSC loaded group. These results indicate that BMSC/TCP composites facilitated bone regeneration and maturation at the graft site of large-sized devitalized bone. This method could potentially be applied for clinical use in the reconstruction of large bone defects such as those associated with bone tumors.

Gatifloxacine-loaded PLGA and β-tricalcium phosphate composite for treating osteomyelitis

Gatifloxacine (GFLX)-containing poly(lactide-co-glycolide) (PLGA) was introduced to the pores and surfaces of porous β-tricalcium phosphate (βTCP) granules by melt compounding whereby no toxic solvent was used. The granular composite of GFLX-loaded PLGA and βTCP released GFLX for 42 days in Hanks' balanced solution and exhibited sufficient in vitro bactericidal activity against Streptococcus milleri and Bacteroides fragilis for at least 21 days. For in vivo evaluation, the granular composite was implanted in the dead space created by the debridement of osteomyelitis lesion induced by S. milleri and B. fragilis in rabbit mandible. After a 4-week implantation, the inflammation area within the debrided area was markedly reduced accompanied with osteoconduction and vascularization in half of the rabbits, and even disappeared in one of the six rabbits without any systemic administration of antibiotics. Outside the debrided area, inflammation and sequestrum were observed but the largest of such affected areas amounted to only 0.125 times of the originally infected and debrided area. These findings showed that the granular composite was effective for the local treatment of osteomyelitis as well as an osteoconductive scaffold which supported and encouraged vascularization.

Comparison of two β-tricalcium phosphate composite grafts used for reconstruction of mandibular critical size bone defects

The aim was to compare osseointegration of blood perfused beta-tricalcium phosphate cylinders (beta-TCPB) with similar composites that were additionally loaded with cancellous bone and bone marrow (beta-TCPB/BM/CB) for mandibular reconstruction. Twelve German Black-Headed sheep with an average weight of 72.5 +/- 10 kg underwent segmental resection of the right hemi-mandible. The animals that were assigned to group A (n = 6) were reconstructed using beta-TCPB while the sheep that were assigned to group B received beta-TCPB/BM/CB grafts. Tissue quality was histologically assessed and bone-, scaffold-, cartilage- and fibrous-tissue area were estimated using semiautomated histomorphometrical software. Composite grafts that were loaded with bone marrow and cancellous bone (beta-TCPB/BM/CB) exhibited significant (p<0.01) higher amounts of bone formation than beta-TCPB. The patients that were assigned to group B achieved defect union and a high grade of bone maturation. Residual ceramic remnants were rare and disconnected. Bone maturity within group A was inferior and none of the specimens showed defect union. The defect centre was still occupied by a ceramic core. Bone and bone marrow augmented beta-tricalcium phosphate composites may qualify as a promising alternative to autograft bone for mandibular reconstruction in human and veterinary medicine.

Antibiotic-loaded poly- ε-caprolactone and porous β-tricalcium phosphate composite for treating osteomyelitis

A composite of poly- ε-caprolactone (PCL) loaded with gatifloxacine (GFLX), an antibiotic, and a β-tricalcium phosphate ( βTCP) porous ceramic body was prepared by a solvent-free process in which no toxic solvent was used. GFLX mostly retained its bactericidal property after the processing. The composite of GFLX-loaded PCL and βTCP ceramic released GFLX for 4 weeks in Hanks’ balanced solution, and had sustained bactericidal activity against Streptococcus milleri and Bacteroides fragilis for at least 1 week. The composite of the GFLX-loaded PCL and βTCP ceramic was implanted in an osteomyelitis lesion induced by S. milleri and B. fragilis in the rabbit mandible. The osteomyelitis lesion expanded in the mesial–distal direction when no composite was implanted or when the lesion was treated with debridement only. The composite of GFLX-loaded PCL and βTCP showed efficacy in controlling infection at the bone defect formed by debridement, and supported bone tissue reconstruction at the bone defect. Twelve and 50 weeks after the implantation, the inflammation even disappeared. New bone formation was observed on the surface of the composite after 4 weeks. After 50 weeks, ingrowth of bone tissues with vascular channels was observed along the PCL and βTCP interface, which indicated degradation of PCL and/or βTCP ceramic at the ceramic/polymer interface followed by replacement by bone tissues. The GFLX concentrations in the serum and soft tissues were very low. Therefore, the composite of GFLX-loaded PCL and βTCP ceramic would help arrest osteomyelitis when it is used in addition to intravenous antibiotic administration, and help new bone formation and osteoconduction.

Synthesis and Thermal Stability of Hydroxyapatite−β-Tricalcium Phosphate Composites with Cosubstituted Sodium, Magnesium, and Fluorine

The preparation of biphasic mixtures (HAP + β-TCP) of controlled ratios with the combined substitution of essential biocompatible trace elements (Na, Mg, and F) was carried out by an aqueous precipitation method. The results showed that incorporated elements have played a significant role in the thermal stability of the apatites up to 1400 °C, and that increased calcium deficiency in the apatites has led to the formation of a higher proportion of β-TCP in the biphasic mixtures. However, minor discrepancies in the structural parameters due to the incorporated trace elements and the matching of phases of the synthesized powders with respect to the stoichiometric HAP were apparent from the present investigation.

Functional Bone Engineering Usingex VivoGene Therapy and Topology-Optimized, Biodegradable Polymer Composite Scaffolds

Bone tissue engineering could provide an alternative to conventional treatments for fracture nonunion, spinal fusion, joint replacement, and pathological loss of bone. However, this approach will require a biocompatible matrix to allow progenitor cell delivery and support tissue invasion. The construct must also support physiological loads as it degrades to allow the regenerated tissue to bear an increasing load. To meet these complex requirements, we have employed topology-optimized design and solid free-form fabrication to manufacture biodegradable poly(propylene fumarate)/beta-tricalcium phosphate composites. These scaffolds were seeded with primary human fibroblasts transduced with an adenovirus expressing bone morphogenetic protein-7 and implanted subcutaneously in mice. Specimens were evaluated by microcomputed tomography, compressive testing, and histological staining. New bone was localized on the scaffold surface and closely followed its designed contours. Furthermore, the total stiffness of the constructs was retained for up to 12 weeks after implantation, as scaffold degradation and tissue invasion took place.