Study on osteoinductive activity of biotin film by low-energy electron beam deposition.

Biotin film was prepared by low-energy electron beam deposition (LEBD). The molecular structure, chemical composition and micromorphology of the biotin film were investigated by 1HNMR, FTIR, XPS, AFM and SEM. The results showed the molecular structure of a monolayer of biotin film is fully consistent with the molecular structure of the initial biotin powders. The contact angle test showed that the biotin film exhibit good hydrophilicity. The release kinetics of biotin film was tested by UV-Vis method. It was found that the film was almost completely released in about two weeks. The cell viability of MC3T3-E1 cells on the surface of the biotin film was attaining 100.54 ± 1.7% (P < 0.05), showing excellent biocompatibility and biosafety. Titanium implant with surface of biotin film was implanted into the femoral head of rabbits as experimental group. The animals were euthanized after four weeks. Compared with the control group, mature lamellar bone formation was observed with dense trabecular bone, and the expression of Coll-I, Runx2 and BMP-2 was better. The results showed that the repair effect of bone defect in the experimental group was excellent.

Osteogenic potential evaluation of biotin combined with magnesium-doped hydroxyapatite sustained-release film.

Biotin is one of the water-soluble B-complex group of vitamins. Recent studies have found that the relative protein expression of BMP2, BSP and OPG in MC3T3-E1 cells is prominent after 14 days of co-culture with biotin film, especially for BMP2. It is also found that the rapid degradation of biotin film in vivo limits its application value. In this work, magnesium-doped hydroxyapatite (MgHA) film can form a porous network structure as a biological sustained-release film. Therefore, the multilayer (MgHA|biotin|MgHA|biotin) film was prepared by pulsed laser assisted electron beam deposition technique. The morphology, structure and properties of biotin film and multilayer film were analyzed and characterized. Also, the osteogenic effect of biotin film and multilayer film was evaluated after implantation into the femoral bone marrow cavity of SD rats. The results of micro-CT scan and 3D reconstruction showed that there were a large number of trabecular bones around the multilayer film, which was superior to biotin film in osteogenesis. Hematoxylin-eosin staining showed cancellous bone structure and intact bone marrow structure around the multilayer film, and the newly formed bone became lamellar. Masson-trichromatic staining revealed abundant osteoid and braided bone formation around the multilayer film. In conclusion, MgHA sustained release film can realize the continuous release of bioactive drugs, which provides a new route to accelerate the repair of bone defects.

Cloning Strategies for the Generation of Recombinant Capripoxvirus Through the Use of Screening and Selection Markers.

The ability to manipulate capripoxvirus through gene knockouts and gene insertions has become an increasingly valuable research tool in elucidating the function of individual genes of capripoxvirus, as well as in the development of capripoxvirus-based recombinant vaccines. The homologous recombination technique is commonly used to generate capripoxvirus knockout viruses (KO), and is based on the targeting of a particular viral gene of interest. This technique can also be used to insert a gene of interest. A protocol for the generation of a viral gene knockout is described. This technique involves the use of a plasmid which encodes the flanking sequences of the regions where the homologous recombination will occur, and will result in the insertion of an EGFP reporter gene for visualization of recombinant virus, as well as the E. coli gpt gene as a positive selection marker. If an additional gene is to be incorporated, this can be achieved by inserting a gene of interest for expression under a poxvirus promoter into the plasmid between the flanking regions for insertion. This chapter describes a protocol for generating such recombinant capripoxviruses. An alternative step for the removal of both the EGFP and gpt cassettes and an optional selection step using CRISPR technology are also described.

Generation of 5.2 fs, energy scalable blue pulses.

In this Letter, ultrashort blue pulses spanning 350-500 nm are generated by combining the broadband frequency doubling technology with the two-stage multiplate continuum (MPC) generation scheme. We prepare relatively broadband input pulses and use a two-stage configuration for MPC generation, allowing us to employ thinner and less solid plates for further spectral broadening. Therefore, the deteriorations of the spectral phase, energy conversion efficiency, and beam quality, which occur more easily for 400 nm pulses, are effectively suppressed. After fine dispersion management, we obtain clean 5.2 fs blue pulses with a root-mean-square energy stability of 0.69% over one hour and excellent beam quality. Furthermore, lower than 8% energy loss during the spectral broadening process at each stage is achieved. The overall optimized performances and energy scalability of this blue pulse, as well as the possibility of further compressing the pulse duration, are likely to motivate more strong-field research with sub-cycle time resolution in this extended wavelength range.

Formation of Porous Apatite Layer after Immersion in SBF of Fluorine-Hydroxyapatite Coatings by Pulsed Laser Deposition Improved in Vitro Cell Proliferation.

Fluorine (F), a trace element in bone tissues, promotes cell proliferation and colonization. In this study, fluorine-hydroxyapatite (F-HA) coating was manufactured onto titanium substrates using pulsed laser deposition (λ = 1024 nm). The results confirmed that the surface roughness of F-HA coatings was the highest when the deposition time was 60 min. To evaluate the biological activity of the coatings, the coatings were immersed in simulated body fluids at 37 °C for 7 days. It was clearly observed that a new apatite layer showed a micro/nano foam-like porous structure. After annealing treatment, the F-HA coating can promote the transformation of the calcium phosphate crystal phase into the HA phase. Moreover, the mass of F-HA coatings with annealing was increased by 2.3% after immersion. In vitro cytocompatibility of the coatings was evaluated; the cell adhesion and proliferation exposed to F-HA coatings were significantly stimulated and promoted when compared with the control group in the first and second weeks of co-culture (p < 0.05). Thus, F-HA coatings can potentially act as osteoinductive materials for surface functionalization of the bone tissue scaffold.