Marine spongin incorporation into Biosilicate® for tissue engineering applications: An in vivo study.

Biomaterials and bone grafts, with the ability of stimulating tissue growth and bone consolidation, have been emerging as very promising strategies to treat bone fractures. Despite its well-known positive effects of biosilicate (BS) on osteogenesis, its use as bone grafts in critical situations such as bone defects of high dimensions or in non-consolidated fractures may not be sufficient to stimulate tissue repair. Consequently, several approaches have been explored to improve the bioactivity of BS. A promising strategy to reach this aim is the inclusion of an organic part, such as collagen, in order to mimic bone structure. Thus, the present study investigated the biological effects of marine spongin (SPG)-enriched BS composites on the process of healing, using a critical experimental model of cranial bone defect in rats. Histopathological and immunohistochemistry analyzes were performed after two and six weeks of implantation to investigate the effects of the material on bone repair (supplemental material-graphical abstract). Histological analysis demonstrated that for both BS and BS/SPG, similar findings were observed, with signs of material degradation, the presence of granulation tissue along the defect area and newly formed bone into the area of the defect. Additionally, histomorphometry showed that the control group presented higher values for Ob.S/BS (%) and for N.Ob/T.Ar (mm2) (six weeks post-surgery) compared to BS/SPG and higher values of N.Ob/T.Ar (mm2) compared to BS (two weeks post-surgery). Moreover, BS showed higher values for OV/TV (%) compared to BS/SPG (six weeks post-surgery). Also, VEGF immunohistochemistry was increased for BS (two weeks post-surgery) and for BS/SPG (six weeks) compared to CG. TGFb immunostaining was higher for BS compared to CG. The results of this study demonstrated that the BS and BS/SPG scaffolds were biocompatible and able to support bone formation in a critical bone defect in rats. Moreover, an increased VEGF immunostaining was observed in BS/SPG.

Effect of 830-nm laser phototherapy on olfactory neuronal ensheathing cells grown in vitro on novel bioscaffolds.

BACKGROUND: The purpose of this study was to analyze olfactory ensheathing cell (OEC) proliferation and growth on Biosilicate and collagen bioscaffolds, and to determine whether the application of laser phototherapy would result in increased OEC proliferation on the scaffolds. The use of bioscaffolds is considered a promising strategy in a number of clinical applications where tissue healing is suboptimal. As in vitro OEC growth is a slow process, laser phototherapy could be useful to stimulate proliferation on bioscaffolds. METHODS: OEC cells were seeded on the Biosilicate and collagen scaffolds. Seeded scaffolds were irradiated with a single exposure of 830-nm laser. Nonirradiated seeded scaffolds acted as negative controls. Cell proliferation was assessed 7 days after irradiation. RESULTS: OECs were successfully grown on discs composed of a glass-ceramic and collagen composite. Laser irradiation produced a 32.7% decrease and a 13.2% increase in OEC proliferation on glass-ceramic discs and on collagen scaffolds, respectively, compared with controls. Laser phototherapy resulted in a reduction in cell growth on the Biosilicate scaffolds and an increase in cell proliferation on collagen scaffolds. CONCLUSIONS: These results were probably due to the nature of the materials. Future research combining laser phototherapy and glass-ceramic scaffolds should take into account possible interactions of the laser with matrix compounds.

Effects of biosilicate(®) scaffolds and low-level laser therapy on the process of bone healing.

OBJECTIVE: This study aimed to investigate the in vivo tissue performance of the association of Biosilicate(®) scaffolds and low-level laser therapy (LLLT) in a tibial bone defects model in rats. BACKGROUND DATA: Many studies have been demonstrating the osteogenic potential of Biosilicate and LLLT. However, there is a need to investigate the effects of both treatments for bone consolidation. METHODS: The animals were divided into control group (CG), Biosilicate scaffold group (BG), and Biosilicate scaffolds plus LLLT group (BLG). Animals were euthanized after 15, 30, and 45 days post-injury. RESULTS: The histological analysis revealed that all the experimental groups showed inflammatory infiltrate and granulation tissue, at the area of the defect at day 15. After 30 days, CG still showed granulation tissue and bone ingrowth. Both Biosilicate groups presented newly formed bone and interconected trabeculae. At 45 days, CG showed immature newly formed bone. A more mature newly formed bone was observed in BG and BLG. On day 15, BG demonstrated a statistically higher expression of cyclooxygenase (COX)-2 compared with CG and BLG. No statistically significant difference was observed in COX-2 immunoexpression among the groups at 30 and 45 days. Similar expression of bone morphogenetic protein (BMP)-9 was demonstrated for all experimental groups at 15 and 30 days. At 45 days, the BMP-9 immunoexpression was statistically upregulated in the BLG compared with the CG and BG. No statistically significant difference was observed in the receptor activator of nuclear factor kappa-B ligand (RANKL) immunoexpression among the groups in all periods evaluated. Biosilicate groups presented a decrease in biomechanical properties compared with CG at 30 and 45 days post-surgery. CONCLUSIONS: Our findings suggest that Biosilicate presented osteogenic activity, accelerating bone repair. However, laser therapy was not able to enhance the bioactive properties of the Biosilicate.

The influence of phosphorus precursors on the synthesis and bioactivity of SiO2-CaO-P 2O 5 sol-gel glasses and glass-ceramics.

Bioactive glasses and glass-ceramics of the SiO(2)-CaO-P(2)O(5) system were synthesised by means of a sol-gel method using different phosphorus precursors according to their respective rates of hydrolysis-triethylphosphate (OP(OC(2)H(5))(3)), phosphoric acid (H(3)PO(4)) and a solution prepared by dissolving phosphorus oxide (P(2)O(5)) in ethanol. The resulting materials were characterised by differential scanning calorimetry and thermogravimetry, X-ray diffraction, Fourier transform infrared spectroscopy, scanning electron microscopy coupled with energy dispersive X-ray spectroscopy and by in vitro bioactivity tests in acellular simulated body fluid. The different precursors significantly affected the main steps of the synthesis, beginning with the time required for gel formation. The most striking influence of these precursors was observed during the thermal treatments at 700-1,200 °C that were used to convert the gels into glasses and glass-ceramics. The samples exhibited very different mineralisation behaviours; especially those prepared using the phosphoric acid, which had a reduced onset temperature of crystallisation and an increased resistance to devitrification. However, all resulting materials were bioactive. The in vitro bioactivity of these materials was strongly affected by the heat treatment temperature. In general, their bioactivity decreased with increasing treatment temperature. For crystallised samples obtained above 900 °C, the bioactivity was favoured by the presence of two crystalline phases: wollastonite (CaSiO(3)) and tricalcium phosphate (α-Ca(3)(PO(4))(2)).

Biosilicate® and low-level laser therapy improve bone repair in osteoporotic rats.

The aim of this study was to investigate the effects of a novel bioactive material (Biosilicate®) and low-level laser therapy (LLLT) on bone fracture consolidation in osteoporotic rats. Forty female Wistar rats were submitted to ovariectomy (OVX) to induce osteopenia. Eight weeks after surgery, the animals were randomly divided into four groups of 10 animals each: a bone defect control group (CG); a bone defect filled with Biosilicate group (BG); a bone defect filled with Biosilicate and irradiated with LLLT at 60 J/cm(2) group (BG60); and a bone defect filled with Biosilicate and irradiated with LLLT at 120 J/cm(2) group (BG120). Bone defects were surgically performed on both tibias. The size of particle used for Biosilicate was 180-212 µm. Histopathological analysis showed that bone defects were predominantly filled with the biomaterial in specimens treated with Biosilicate. LLLT with either 60 or 120 J/cm(2) was able to increase collagen, Cbfa-1, VGEF and COX-2 expression in the circumjacent cells of the biomaterial. A morphometric analysis revealed that the Biosilicate + laser groups showed a higher amount of newly formed bone. Our results indicate that laser therapy improves bone repair process in contact with Biosilicate as a result of increasing bone formation, as well as COX-2 and Cbfa-1 immunoexpression, angiogenesis and collagen deposition in osteoporotic rats.

Effect of 830 nm laser phototherapy on osteoblasts grown in vitro on Biosilicate scaffolds.

OBJECTIVE: The purpose of this study was (i) to develop a method for successfully seeding osteoblasts onto a glass-ceramic scaffold designed for use in clinical settings, and (ii) to determine whether the application of laser phototherapy at 830 nm would result in osteoblast proliferation on the glass-ceramic scaffold. BACKGROUND: The use of bioscaffolds is considered a promising strategy for a number of clinical applications where tissue healing is sub-optimal. As in vitro osteoblast growth is a slow process, laser phototherapy could be used to stimulate osteoblast proliferation on bioscaffolds. METHODS: A methodology was developed to seed an osteoblastic (MC3T3) cell line onto a novel glass-ceramic scaffold. Seeded scaffolds were irradiated with a single exposure of 830 nm laser at 10 J/cm(2) (at diode). Non-irradiated seeded scaffolds acted as negative controls. Cell proliferation was assessed seven days after irradiation. RESULTS: Osteoblastic MC3T3 cells were successfully grown on discs composed of a glass-ceramic composite. Laser irradiation produced a 13% decrease in MC3T3 cell proliferation on glass-ceramic discs (mean +/- SD = 0.192 +/- 0.002) compared with control (non-irradiated) discs (mean +/-SD = 0.22 +/- 0.002). CONCLUSIONS: Despite successful seeding of bioscaffolds with osteoblasts, laser phototherapy resulted in a reduction in cell growth compared to non-irradiated controls. Future research combining laser phototherapy and glass-ceramic scaffolds should take into account possible interactions of the laser with matrix compounds.