ALBERT

Description: Two dense biphasic ceramics, with a hypereutectic composition of 30 wt % CaSiO3–70 wt % Ca3(PO4)2, were synthesized by a solid-state reaction of homogeneous pressed combinations of previously synthesized synthetic CaSiO3 and Ca3(PO4)2 powders. The objective was to produce a dense structure to generate large enough in situ pores for the ceramic to be used in tissue engineering. To develop such a structure, two grain sizes of CaSiO3 were used (63–100 µm and 100–150 μm) and some of their properties were studied in vitro, as they are relevant for tissue engineering. X-ray diffraction analysis, μ-Raman spectroscopy, diametrical compression test, and scanning electron microscopy with elemental mapping showed a coarse-grained homogeneous microstructure for the materials, which consisted of wollastonite (α-CaSiO3) and tricalcium phosphate (α-Ca3(PO4)2), with adequate mechanical properties for implantation. In vitro bioactivity was evaluated in simulated body fluid (SBF) by exploring a hydroxyapatite (HA)-like formation. The results showed that tricalcium phosphate grains dissolved more preferentially than those of wollastonite, but not fast enough to leave a pore before the surface was coated with an HA-like layer after soaking only for three days. Biocompatibility was evaluated by in vitro cell experiments, which showed cell proliferation, adhesion, and spreading on the ceramic surface. This ceramic is expected to be used as a bone graft substitute.

Description: The characterization process of a new porous Nurse’s A ceramic and the physico chemical nature of the remodeled interface between the implant and the surrounding bone were studied after in vivo implantation. Scaffolds were prepared by a solid-state reaction and implanted in New Zealand rabbits. Animals were sacrificed on days 15, 30, and 60. The porous biomaterial displayed biocompatible, bioresorbable, and osteoconductive capacity. The degradation processes of implants also encouraged osseous tissue ingrowths into the material’s pores, and drastically changed the macro- and microstructure of the implants. After 60 healing days, the resorption rates were 52.62% ± 1.12% for the ceramic and 47.38% ± 1.24% for the residual biomaterial. The elemental analysis showed a gradual diffusion of the Ca and Si ions from the materials into the newly forming bone during the biomaterial’s resorption process. The energy dispersive spectroscopy (EDS) analysis of the residual ceramic revealed some particle categories with different mean Ca/P ratios according to size, and indicated various resorption process stages. Since osteoconductive capacity was indicated for this material and bone ingrowth was possible, it could be applied to progressively substitute an implant.

Description: This aim of this research was to develop a novel ceramic scaffold to evaluate the response of bone after ceramic implantation in New Zealand (NZ) rabbits. Ceramics were prepared by the polymer replication method and inserted into NZ rabbits. Macroporous scaffolds with interconnected round-shaped pores (0.5–1.5 mm = were prepared). The scaffold acted as a physical support where cells with osteoblastic capability were found to migrate, develop processes, and newly immature and mature bone tissue colonized on the surface (initially) and in the material’s interior. The new ceramic induced about 62.18% ± 2.28% of new bone and almost complete degradation after six healing months. An elemental analysis showed that the gradual diffusion of Ca and Si ions from scaffolds into newly formed bone formed part of the biomaterial’s resorption process. Histological and radiological studies demonstrated that this porous ceramic scaffold showed biocompatibility and excellent osteointegration and osteoinductive capacity, with no interposition of fibrous tissue between the implanted material and the hematopoietic bone marrow interphase, nor any immune response after six months of implantation. No histological changes were observed in the various organs studied (para-aortic lymph nodes, liver, kidney and lung) as a result of degradation products being released.

Description: In this study, the Bioeutectic® blocks were inserted into the critical size defects of eight rabbits, using both tibiae, and the physical and chemical nature of the remodeled interface between the Bioeutectic® implants and the surrounding bone were performed at four and 15 months. The results showed a new fully mineralized bone growing in direct contact with the implants. The ionic exchange, taking place at the implant interface with the body fluids was essential in the process of the implant integration through a dissolution-precipitation-transformation mechanism. The study found the interface biologically and chemically active over the 15 months implantation period. The osteoblastic cells migrated towards the interface and colonized the surface at the contact areas with the bone. The new developed apatite structure of porous morphology mimics natural bone.

Description: The subsystem Nurse’s A-phase-silicocarnotite within the system Ca3(PO4)2–Ca2SiO4 was conducted as a preliminary step toward obtaining new biomaterials with controlled microstructures. Phase composition of the resulting ceramics was studied by X-ray diffraction, differential thermal analysis, and scanning electron microscopy with attached wavelength dispersive spectroscopy. The results showed that the sub-system presents an invariant eutectoid point at 1366 ± 4 °C with a composition of 59.5 wt % Ca3(PO4)2 and 40.5 wt % Ca2SiO4, and typical eutectoid microstructure of lamellae morphology. These results are in disagreement with the previous reported data, which locate the invariant eutectoid point at 1250 ± 20 °C with a composition of 55 wt % Ca3(PO4)2 and 45 wt % Ca2SiO4. In addition, cell attachment testing showed that the new eutectoid material supported the mesenchymal stem cell adhesion and spreading, and the cells established close contact with the ceramic after 28 days of culture. These findings indicate that the new ceramic material with eutectoid microstructure of lamellae morphology possesses good bioactivity and biocompatibility and might be a promising bone implant material.

Description: Materials, Vol. 11, Pages 1580: Demineralized Bone Matrix Coating Si-Ca-P Ceramic Does Not Improve the Osseointegration of the Scaffold Materials doi: 10.3390/ma11091580 Authors: Andrés Parrilla-Almansa Nuria García-Carrillo Patricia Ros-Tárraga Carlos M. Martínez Francisco Martínez-Martínez Luis Meseguer-Olmo Piedad N. De Aza The aim of this study was to manufacture and evaluate the effect of a biphasic calcium silicophosphate (CSP) scaffold ceramic, coated with a natural demineralized bone matrix (DBM), to evaluate the efficiency of this novel ceramic material in bone regeneration. The DBM-coated CSP ceramic was made by coating a CSP scaffold with gel DBM, produced by the partial sintering of different-sized porous granules. These scaffolds were used to reconstruct defects in rabbit tibiae, where CSP scaffolds acted as the control material. Micro-CT and histological analyses were performed to evaluate new bone formation at 1, 3, and 5 months post-surgery. The present research results showed a correlation among the data obtained by micro-CT and the histomorphological results, the gradual disintegration of the biomaterial, and the presence of free scaffold fragments dispersed inside the medullary cavity occupied by hematopoietic bone marrow over the 5-month study period. No difference was found between the DBM-coated and uncoated implants. The new bone tissue inside the implants increased with implantation time. Slightly less new bone formation was observed in the DBM-coated samples, but it was not statistically significant. Both the DBM-coated and the CSP scaffolds gave excellent bone tissue responses and good osteoconductivity.

Description: Materials, Vol. 11, Pages 1349: Preclinical Studies of the Biosafety and Efficacy of Human Bone Marrow Mesenchymal Stem Cells Pre-Seeded into β-TCP Scaffolds after Transplantation Materials doi: 10.3390/ma11081349 Authors: Mar Gonzálvez-García Carlos M. Martinez Victor Villanueva Ana García-Hernández Miguel Blanquer Luis Meseguer-Olmo Ricardo E. Oñate Sánchez José M. Moraleda Francisco Javier Rodríguez-Lozano Background: Cell-Based Therapies (CBT) constitute a valid procedure for increasing the quantity and quality of bone in areas with an inadequate bone volume. However, safety and efficacy should be investigated prior to clinical application. The objective of this study was to evaluate the biodistribution, safety and osteogenic capacity of bone marrow-derived human mesenchymal stem cells (hBMMSCs) pre-seeded into β-tricalcium phosphate (TCP) and implanted into NOD/SCID mice at subcutaneous and intramuscular sites. Methods: hBMMSCs were isolated, characterized and then cultured in vitro on a porous β-TCP scaffold. Cell viability and attachment were analyzed and then hBMMSCs seeded constructs were surgically placed at subcutaneous and intramuscular dorsal sites into NOD/SCID mice. Acute and subchronic toxicity, cell biodistribution and efficacy were investigated. Results: There were no deaths or adverse events in treated mice during the 48-hour observation period, and no toxic response was observed in mice. In the 12-week subchronic toxicity study, no mortalities, abnormal behavioral symptoms or clinical signs were observed in the saline control mice or the hBMMSCs/β-TCP groups. Finally, our results showed the bone-forming capacity of hBMMSCs/β-TCP since immunohistochemical expression of human osteocalcin was detected from week 7. Conclusions: These results show that transplantation of hBMMSCs/β-TCP in NOD/SCID mice are safe and effective, and might be applied to human bone diseases in future clinical trials.

Description: The aim of the study is to determine the existing correlation between high-resolution 3D imaging technique obtained through Micro Computed Tomography (mCT) and histological-histomorphometric images to determine in vivo bone osteogenic behavior of bioceramic scaffolds. A Ca-Si-P scaffold ceramic doped and non-doped (control) with a natural demineralized bone matrix (DBM) were implanted in rabbit tibias for 1, 3, and 5 months. A progressive disorganization and disintegration of scaffolds and bone neoformation occurs, from the periphery to the center of the implants, without any differences between histomorphometric and radiological analysis. However, significant differences (p 〈 0.05) between DMB-doped and non-doped materials where only detected through mathematical analysis of mCT. In this way, average attenuation coefficient for DMB-doped decreased from 0.99 ± 0.23 Hounsfield Unit (HU) (3 months) to 0.86 ± 0.32 HU (5 months). Average values for non-doped decreased from 0.86 ± 0.25 HU (3 months) to 0.66 ± 0.33 HU. Combination of radiological analysis and mathematical mCT seems to provide an adequate in vivo analysis of bone-implanted biomaterials after surgery, obtaining similar results to the one provided by histomorphometric analysis. Mathematical analysis of Computed Tomography (CT) would allow the conducting of long-term duration in vivo studies, without the need for animal sacrifice, and the subsequent reduction in variability.

Description: The purpose of this study was to evaluate the bioactivity and cell response of a well-characterized Nurse’s A-phase (7CaO·P2O5·2SiO2) ceramic and its effect compared to a control (tissue culture polystyrene-TCPS) on the adhesion, viability, proliferation, and osteogenic differentiation of ahMSCs in vitro. Cell proliferation (Alamar Blue Assay), Alizarin Red-S (AR-s) staining, alkaline phosphatase (ALP) activity, osteocalcin (OCN), and collagen I (Col I) were evaluated. Also, field emission scanning electron microscopy (FESEM) images were acquired in order to visualise the cells and the topography of the material. The proliferation of cells growing in a direct contact with the material was slower at early stages of the study because of the new environmental conditions. However, the entire surface was colonized after 28 days of culture in growth medium (GM). Osteoblastic differentiation markers were significantly enhanced in cells growing on Nurse’s A phase ceramic and cultured with osteogenic medium (OM), probably due to the role of silica to stimulate the differentiation of ahMSCs. Moreover, calcium nodules were formed under the influence of ceramic material. Therefore, it is predicted that Nurse’s A-phase ceramic would present high biocompatibility and osteoinductive properties and would be a good candidate to be used as a biomaterial for bone tissue engineering.

Description: A new type of bioceramic with osteogenic properties, suitable for hard tissue regeneration, was synthesised. The ceramic was designed and obtained in the Nurse’s A-phase-silicocarnotite subsystem. The selected composition was that corresponding to the eutectoid 28.39 wt % Nurse’s A-phase-71.61 wt % silicocarnotite invariant point. We report the effect of Nurse’s A-phase-silicocarnotite ceramic on the capacity of multipotent adult human mesenchymal stem cells (ahMSCs) cultured under experimental conditions, known to adhere, proliferate and differentiate into osteoblast lineage cells. The results at long-term culture (28 days) on the material confirmed that the undifferentiated ahMSCs cultured and in contact with the material surface adhered, spread, proliferated, and produced a mineralised extracellular matrix on the studied ceramic, and finally acquired an osteoblastic phenotype. These findings indicate that it underwent an osteoblast differentiation process. All these findings were more significant than when cells were grown on plastic, in the presence and absence of this osteogenic supplement, and were more evident when this supplement was present in the growth medium (GM). The ceramic evaluated herein was bioactive, cytocompatible and capable of promoting the proliferation and differentiation of undifferentiated ahMSCs into osteoblasts, which may be important for bone integration into the clinical setting.