Comparing the Effects of Alpha-Tocopherol and Tocotrienol Isomers on Osteoblasts hFOB 1.19 Cultured on Bovine Bone Scaffold

Tocotrienol mixtures have been shown to exert anabolic actions on the skeletal system in animal studies, but it is unclear which tocotrienol isomer shows the most prominent effects. This study aims to investigate the most active tocotrienol isomers using hFOB 1.19 human osteoblasts cultured on a bovine bone scaffold. The bovine trabecular bone was sectioned, demineralised and freeze-dried to form the scaffold. hFOB 1.19 osteoblasts were cultured on the bone scaffolds in humidified condition at 37 °C and 5% carbon dioxide with vitamin E isomers (alpha-, beta-, gamma-, delta-tocotrienol and alpha-tocopherol). The cell differentiation capacity of tocotrienol isomers was investigated through morphological observation, alkaline phosphatase (ALP) activity and osteocalcin expression. Changes in the bone scaffolds were determined using histomorphometry methods. Osteoblast culture treated with gamma- and delta-tocotrienols showed a significant increase in ALP activity and osteocalcin expression. Bone structural histomorphometry analysis showed that bone scaffolds treated with gamma- and delta-tocotrienol showed significant increases in bone volume and trabecular thickness. In conclusion, gamma- and delta-tocotrienol show the most prominent bone anabolic effects by increasing osteoblast differentiation and enhancing bone microstructure.

Short-Wave Enhances Mesenchymal Stem Cells Recruitment in Fracture Healing by Increasing HIF-1 in Callus

Background: As a type of high-frequency electrotherapy, a Short-Wave can promote the fracture healing process; yet, its underlying therapeutic mechanisms remain unclear.Purpose: To observe the effect of Short-Wave therapy on mesenchymal stem cell (MSC) homing and relative mechanisms associated with fracture healing. Materials and Methods: For in vivo study, the effect of Short-Wave therapy to fracture healing was examined in stabilized femur fractures model of 40 SD rats. Radiography was used to analyze the morphology and microarchitecture of the callus. Additionally, fluorescence assays were used to analyze the GFP-labeled MSC homing after treatment in 20 nude mice with a femoral fracture. For in vitro study, osteoblast from newborn rats simulated fracture site was first irradiated by the Short-Wave; siRNA targeting HIF-1 was used to investigate the role of HIF-1. Osteoblast culture medium was then collected as chemotaxis content of MSC, and the migration of MSC from rats was evaluated using wound healing assay and trans-well chamber test. The expression of HIF-1 and its related factors were quantified by q RT-PCR, ELISA, and Western blot.Results: Our in vivo experiment indicated that Short-Wave therapy could promote MSC migration, increase local and serum HIF-1 and SDF-1 levels, induce changes in callus formation, and improve callus microarchitecture and mechanical properties, thus speeding up the healing process of the fracture site. Moreover, the in vitro results further indicated that Short-Waves therapy upregulated HIF-1 and SDF-1 expression in osteoblast and its cultured medium, as well as the expression of CXCR-4, β-catenin, F-actin and phosphorylation levels of FAK in MSC. On the other hand, the inhibition of HIF-1α was significantly restrained by the inhibition of HIF-1α in osteoblast, and it partially inhibited the migration of MSC.Conclusions: These results suggested that Short-Wave therapy could increase HIF-1 in callus, which is one of the crucial mechanisms of chemotaxis MSC homing in fracture healing.

Short-Wave Enhances MSC Recruitment in Fracture Healing from Increasing HIF-1 in Callus

Background: As a type of high-frequency electrotherapy, a Short-Wave can promote the fracture healing process; yet, its underlying therapeutic mechanisms still remain unclear.Purpose: To observe the effect of Short-Wave therapy on mesenchymal stem cell (MSC) homing and relative mechanisms associated with fracture healing. Materials and Methods: For in vivo study, the effect of Short-Wave therapy in relation to fracture healing was examined in stabilized femur fractures model of 40 SD rats. Radiography was used to analyze the morphology and micro-architecture of the callus. Additionally, fluorescence assays were used to analyze the GFP-labeled MSC homing after treatment in 20 nude mice with a femoral fracture. For in vitro study, osteoblast from newborn rats simulated fracture site was first irradiated by the Short-Wave; siRNA targeting HIF-1 was used to investigate the role of HIF-1. Osteoblast culture medium was then collected as chemotaxis content of MSC, and the migration of MSC from rats was evaluated using wound healing assay and trans-well chamber test. The expression of HIF-1 and its related factors were quantified by q RT-PCR, ELISA, and Western blot.Results: Our in vivo experiment indicated that Short-Wave therapy could promote MSC migration, increase local and serum HIF-1 and SDF-1 levels, induce changes in callus formation, and improve callus microarchitecture and mechanical properties, thus speeding up the healing process of the fracture site. Moreover, the in vitro results further indicated that Short-Waves therapy upregulated HIF-1 and SDF-1 expression in osteoblast and in the medium, as well as the expression of CXCR-4, β-catenin, F-actin and phosphorylation levels of FAK in MSC. On the other hand, the inhibition of HIF-1α was significantly restrained by the inhibition of HIF-1α in osteoblast, and it partially inhibited the migration of MSC.Conclusions: These results suggested that Short-Wave therapy could increase HIF-1 in callus, which is one of the crucial mechanisms of chemotaxis MSC homing in fracture healing.

Osteoblast Cell Response to LIPSS-Modified Ti-Implants

In the present work, the surface of Ti-6Al-7Nb samples was patterned with Laser Induced Periodic Surface Structures in order to improve biocompatibility, increase tissue ingrowth and decrease bacterial adhesion and inflammatory response for applications in dental and orthopedic implants. Polished and sandblasted disks 10 mm in diameter were treated generating LIPSS under two different sets of parameters. The surface morphology and chemistry were investigated both by secondary electrons imaging, EDS analysis and Atomic Force Microscopy. Primary rat osteoblast culture (passage 2) was used to assess cell toxicity and biocompatibility. Alamar Blue assay was used to access cell viability and proliferation on day 1, 3 and 7. The difference between cell adhesion on polished and sandblasted surface as well as between polished and LIPSS-modified surface are described and discussed.

Analysis of In Vitro Osteoblast Culture on Scaffolds for Future Bone Regeneration Purposes in Dentistry

One of the main focuses of tissue engineering is to search for tridimensional scaffold materials, complying with nature’s properties for tissue regeneration. Determining material biocompatibility is a fundamental step in considering its use. Therefore, the purpose of this study was to analyze osteoblast cell adhesion and viability on different materials to determine which was more compatible for future bone regeneration. Tridimensional structures were fabricated with hydroxyapatite, collagen, and porous silica. The bovine bone was used as material control. Biocompatibility was determined by seeding primary osteoblasts on each tridimensional structure. Cellular morphology was assessed by SEM and viability through confocal microscopy. Osteoblast colonization was observed on all evaluated materials’ surface, revealing they did not elicit osteoblast cytotoxicity. Analyses of four different materials studied with diverse compositions and characteristics showed that adhesiveness was best seen for HA and viability for collagen. In general, the results of this investigation suggest these materials can be used in combination, as scaffolds intended for bone regeneration in dental and medical fields.