Irradiation with 365 nm and 405 nm wavelength shows differences in DNA damage of swine pancreatic islets.

INTRODUCTION: 3D printing is being used more extensively in modern biomedicine. One of the problems is selecting a proper crosslinking method of bioprinted material. Amongst currently used techniques we can distinguish: physical crosslinking (e.g. Ca2+ and Sr2+) and chemical crosslinking-the UV light crosslinking causing the biggest discussion. UV radiation is selectively absorbed by DNA, mainly in the UV-B region but also (to some extent) in UV-A and UV-C regions. DNA excitement results in typical photoproducts. The amount of strand breaks may vary depending on the period of exposition, it can also differ when cells undergo incubation after radiation. AIM: The aim of this study was to show whether and how the time of irradiation with 405 nm and 365 nm wavelengths affect DNA damage in cell lines and micro-organs (pancreatic islets). MATERIALS AND METHODS: The degree of DNA damage caused by different wavelengths of radiation (405 nm and 365 nm) was evaluated by a comet assay. The test was performed on fibroblasts, alpha cells, beta cells and porcine pancreatic islets after 24 hours incubation period. Samples without radiation treatment were selected as a control group. Results analysis consisted of determining the percent of cells with damaged DNA and the tail intensity evaluation. RESULTS: The degree of DNA damage in pancreatic islets after exposure to 405 nm wavelength oscillated between 2% and 6% depending on the tested time period (10 - 300 seconds). However, treating islets using 365 nm wavelength resulted in damage up to 50%. This clearly shows significantly less damage when using 405 nm wavelength. Similar results were obtained for the tested cell lines. CONCLUSIONS: Crosslinking with 405 nm is better for pancreatic islets than crosslinking with 365 nm UV light.

The combined antibacterial and anticancer properties of nano Ce-containing Mg-phosphate ceramic.

AIM: This paper was mainly aimed at synthesis of Ce-containing nano-Mg-phosphate ceramic as a multifunctional material. MATERIALS AND METHODS: Two ceramics based on Mg3(PO4)2 and Ce0.2Mg2.8(PO4)2 formulas (MP and MP-C, respectively) were synthesized. The synthesized powders were characterized by XRD, TEM, Zeta potential, and FTIR. Also, their dissolution behavior was tested in Tris-HCl buffer solution. Moreover, the antimicrobial efficacy was evaluated against gram-positive bacteria (Bacillus sphaericus MTCC 511 &Staphylococcus aureus MTCC 87) and gram-negative bacteria (Enterobacter aerogenes MTCC 111 &Pseudomonas aeruginosa MTCC 1034) using dick diffusion assay and microdilution method. Furthermore, the cell viability test was performed for the ceramics on Vero cells (African green monkey kidney cells), and their antitumor activity was determined by PC3 cell line (prostatic cancer). Also, the cellular uptake was determined by the flow cytometry. KEY FINDINGS: The results showed that the substitution of Mg by Ce decreased the particle size from 40 to 90 nm for MP sample to 2-10 nm for MP-C sample and increased the degradation rate. Both samples showed excellent antimicrobial activities. Moreover, MP demonstrated more cell viability than MP-C on Vero cells at high concentrations, whereas, MP-C showed more antitumor activity on PC3 cells than MP sample. Moreover, MP-C showed a higher cell uptake than MP due to its smaller size and more negative charge. SIGNIFICANCE: Mg-phosphate ceramic can be used in this study successfully as a delivery system for cerium ions and showed a high antitumor activity, which makes it highly recommended as safe and effective cancer treatment materials.

Engineering biological gradients.

Biological gradients profoundly influence many cellular activities, such as adhesion, migration, and differentiation, which are the key to biological processes, such as inflammation, remodeling, and tissue regeneration. Thus, engineered structures containing bioinspired gradients can not only support a better understanding of these phenomena, but also guide and improve the current limits of regenerative medicine. In this review, we outline the challenges behind the engineering of devices containing chemical-physical and biomolecular gradients, classifying them according to gradient-making methods and the finalities of the systems. Different manufacturing processes can generate gradients in either in-vitro systems or scaffolds, which are suitable tools for the study of cellular behavior and for regenerative medicine; within these, rapid prototyping techniques may have a huge impact on the controlled production of gradients. The parallel need to develop characterization techniques is addressed, underlining advantages and weaknesses in the analysis of both chemical and physical gradients.

Computer aided design of architecture of degradable tissue engineering scaffolds.

One important factor affecting the process of tissue regeneration is scaffold stiffness loss, which should be properly balanced with the rate of tissue regeneration. The aim of the research reported here was to develop a computer tool for designing the architecture of biodegradable scaffolds fabricated by melt-dissolution deposition systems (e.g. Fused Deposition Modeling) to provide the required scaffold stiffness at each stage of degradation/regeneration. The original idea presented in the paper is that the stiffness of a tissue engineering scaffold can be controlled during degradation by means of a proper selection of the diameter of the constituent fibers and the distances between them. This idea is based on the size-effect on degradation of aliphatic polyesters. The presented computer tool combines a genetic algorithm and a diffusion-reaction model of polymer hydrolytic degradation. In particular, we show how to design the architecture of scaffolds made of poly(DL-lactide-co-glycolide) with the required Young's modulus change during hydrolytic degradation.

Observations of mineralised tissues of teeth in X-ray micro-computed tomography.

BACKGROUND: The one of the most recent imaging technology is X-ray microtomography which allows non-invasive three-dimensional visualisation of structures. It also offers the opportunity to conduct a comprehensive quantitative analysis of the tested objects such as measuring the shares of the various phases, determining the material density and distribution of the size of pores and particles. The aim of the paper was to present an overview on the applicability and relevance of X-ray microtomography in the study of mineralised tissues of the teeth. MATERIALS AND METHODS: The article is based on the most recent and significant literature and own observations. RESULTS: The use of X-ray microtomography in dentistry has recently increased and includes, inter alia, the assessment of the density of minerals in enamel and dentin, the detection of demineralisation in an artificially and a naturally induced caries, the automatic measurement of the depth of cavities in dentin, the measurement of the amount of removed dentin in preparation of carious lesions by various methods, the assessment of microleakage around fillings and fissure sealants, cortical bone density measurement, evaluation of root canal morphology, comparison of the accuracy of root canal working and filling by various methods. CONCLUSIONS: X-ray microtomography offers within the analysis of mineralised tissues - complex structures of bone, teeth and biomedical materials, turn out to be indispensable since it opens new opportunities for cognitive and implementation research.

Ternary composite scaffolds with tailorable degradation rate and highly improved colonization by human bone marrow stromal cells.

Poly(ε-caprolactone), PCL, is of great interest for fabrication of biodegradable scaffolds due to its high compatibility with various manufacturing techniques, especially Fused Deposition Modeling (FDM). However, slow degradation and low strength make application of PCL limited only to longer-term bioresorbable and non-load bearing implants. To overcome latter drawbacks, ternary PCL-based composite fibrous scaffolds consisting of 70-95 wt % PCL, 5 wt % Tricalcium Phosphate (TCP) and 0-25 wt % poly(lactide-co-glycolide) (PLGA) were fabricated using FDM. In the present study, the effect of composition of the scaffolds on their mechanical properties, degradation kinetics, and surface properties (wettability, surface energy, and roughness) was investigated and correlated with response of human bone marrow mesenchymal stromal cells (HBMC). The presence of PLGA increased degradation kinetics, surface roughness and significantly improved scaffold colonization. Of the evaluated surface properties only the wettability was correlated with the surface area colonized by HBMC. This study demonstrates that introduction of PLGA into PCL-TCP binary composite could largely abolish the disadvantages of the PCL matrix and improve biocompatibility by increasing wettability and polar interactions rather than surface roughness. Additionally, we showed great potential of multicellular spheroids as a sensitive in vitro tool for detection of differences in chemistry of 3D scaffolds.

Tailored degradation of biocompatible poly(3-hydroxybutyrate-co-3-hydroxyvalerate)/calcium silicate/poly(lactide-co-glycolide) ternary composites: an in vitro study.

Biodegradable materials, which are currently available for bone tissue regeneration, still have limitations regarding their degradation rate, mechanical stability and/or biological response. Thus, a novel generation of materials for bioactive bone scaffolds is needed that triggers hydroxyapatite formation and can be tailored to suit application-specific requirements. In this study we developed ternary bioactive composite materials composed of poly(3-hydroxybutyrate-co-3-hydroxyvalerate), calcium silicate and poly(lactide-co-glycolide) (PHBV/CS/PLGA), which merged the good bioactivity of CS/PHBV composite and the improved degradation velocity of PHBV/PLGA blend. Bioactive character of all composites was proven by formation of hydroxyapatite-like crystals after already one week of incubation in simulated body fluid. Addition of PLGA significantly increased initial ultimate tensile strength (UTS0) and Young's modulus of the ternary composites from 14.3±1.1 MPa (binary composite) to 22.3±2.6 MPa and 1.23±0.05 GPa up to 1.64±0.14 GPa, respectively. Furthermore the degradation rate (measured as a decrease of UTS during degradation) could be successfully tailored and was in range of -0.033 UTS0 to -0.118 UTS0 MPa/week. The bioacceptance of the materials was proven in vitro using 2-D (conventional setup) and 3-D (multicellular spheroids) human bone marrow stromal cell cultures.

Biological characterization of woven fabric using two- and three-dimensional cell cultures.

The integration and long-term functional retention of tissue implants are both strongly linked to the implant material characteristics. As a first approach, the cytocompatibility and bioactivity of such materials are evaluated using in vitro-based cell culture models. Typically, in vitro bioactivity is assessed by seeding single cells onto the test material to evaluate certain parameters such as cell adhesion, survival, proliferation, and functional differentiation. Probably, due to the reduction from three dimensional (3D) toward the two dimensional (2D) situation the data obtained from 2D culture models falls short of predicting the in vivo behavior of the biomaterial in question. In this study, a three dimensional (3D) in vitro cell culture model was applied to evaluate the bioactivity of well characterized fiber-based scaffolds using scaffold colonization as a bioactivity indicator. Cell behavior in this culture model was evaluated against a classical comparable, 2D cell culture system using polyethylene terephthalat and polyamide 6.6 fabrics. By using the 3D culture model, however, differences in cell population performance as a function of fiber diameter and mesh angle were evident. The use of 3D cell culture model clearly outperformed typical cell culture setup as means to evaluate cell population-scaffold interaction.

Evolutionary design of bone scaffolds with reference to material selection.

The favourable scaffold for bone tissue engineering should have desired characteristic features, such as adequate mechanical strength and three-dimensional open porosity, which guarantee a suitable environment for tissue regeneration. In fact, the design of such complex structures like bone scaffolds is a challenge for investigators. One of the aims is to achieve the best possible mechanical strength-degradation rate ratio. In this paper we attempt to use numerical modelling to evaluate material properties for designing bone tissue engineering scaffold fabricated via the fused deposition modelling technique. For our studies the standard genetic algorithm was used, which is an efficient method of discrete optimization. For the fused deposition modelling scaffold, each individual strut is scrutinized for its role in the architecture and structural support it provides for the scaffold, and its contribution to the overall scaffold was studied. The goal of the study was to create a numerical tool that could help to acquire the desired behaviour of tissue engineered scaffolds and our results showed that this could be achieved efficiently by using different materials for individual struts. To represent a great number of ways in which scaffold mechanical function loss could proceed, the exemplary set of different desirable scaffold stiffness loss function was chosen.

Comparison of bacterial adhesion and cellular proliferation on newly developed three-dimensional scaffolds manufactured by rapid prototyping technology.

Scaffolds used in the field of tissue engineering should facilitate the adherence, spreading, and ingrowth of cells as well as prevent microbial adherence. For the first time, this study simultaneously deals with microbial and tissue cell adhesion to rapid prototyping-produced 3D-scaffolds. The cell growth of human osteosarcoma cells (CAL-72) over a time period of 3-11 days were examined on three scaffolds (PLGA, PLLA, PLLA-TCP) and compared to the adhesion of salivary microorganisms and representative germs of the oral flora (Porphyromonas gingivalis, Prevotella nigrescens, Candida albicans, Enterococcus faecalis, Streptococcus mutans, and Streptococcus sanguinis). Scanning electron microscopy (SEM), cell proliferation measurements, and determination of the colony forming units (CFU) were performed. The cell proliferation rates on PLLA and PLLA-TCP after 3, 7, and 11 days of cultivation were higher than on PLGA. On day 3 the proliferation rates on PLLA and PLLA-TCP, and on day 5 on PLLA-TCP, proved to be significantly higher compared to that of the control (culture plate). The strain which showed the most CFUs on all of the investigated scaffolds was P. gingivalis, followed by E. faecalis. No significant CFU differences were determined examining P. gingivalis among the biomaterials. In contrast, E. faecalis was significantly more adherent to PLGA and PLLA compared to PLLA-TCP. The lowest CFU values were seen with C. albicans and P. nigrescens. Salivary born aerobic and anaerobic microorganisms adhered significantly more to PLGA compared to PLLA-TCP. These results supported by SEM point out the high potential of PLLA-TCP in the field of tissue engineering.

Bone ingrowth simulation for a concept glenoid component design.

Glenoid component loosening is the major problem of total shoulder arthroplasty. It is possible that uncemented component may be able to achieve superior fixation relative to cemented component. One option for uncemented glenoid is to use porous tantalum backing. Bone ingrowth into the porous backing requires a degree of stability to be achieved directly post-operatively. This paper investigates the feasibility of bone ingrowth with respect to the influence of primary fixation, elastic properties of the backing and friction at the bone prosthesis interface. Finite element models of three glenoid components with different primary fixation configurations are created. Bone ingrowth into the porous backing is modelled based on the magnitude of the relative interface micromotions and mechanoregulation of the mesenchymal stem cells that migrated via the bonded part of the interface. Primary fixation had the most influence on bone ingrowth. The simulation showed that its major role was not to firmly interlock the prosthesis, but rather provide such a distribution of load, that would result in reduction of the peak interface micromotions. Should primary fixation be provided, friction has a secondary importance with respect to bone ingrowth while the influence of stiffness was counter intuitive: a less stiff backing material inhibits bone ingrowth by higher interface micromotions and stimulation of fibrous tissue formation within the backing.

Translational stiffness of the replaced shoulder joint.

Results after a total shoulder arthroplasty in rheumatoid patients are poor, indicated by loosening of especially the glenoid component, bad joint functionality and the possibility of a joint dislocation. The failure mechanisms behind this are multiple, including patient, surgical and design factors. These results must be improved. At present, the optimal geometrical prosthesis component design, focused on joint conformity and constraint, still has to be investigated. Proper understanding of the effect of geometrical design parameters on the theoretical relationship between joint translations and joint forces may contribute to improved designs. The main objective of this study is to theoretically describe this relationship and to investigate the joint translational stiffness, which can be used to investigate the effect of design parameters on joint motion. Joint translational stiffness is the gradient of the subluxation force with respect to the humeral head displacement. For this static analysis a potential field is introduced, as the result of a joint compressive force (muscle forces) and a subluxation force (external forces). The positive and negative stiffness during articulation inside and subluxation outside the glenoid cavity, lead to stable and unstable equilibrium joint positions, respectively. A most lateral position of the humeral head centre coincides with a zero subluxation force; at this position the humerus is dislocated and a restoring force is needed to relocate the humeral head. Joint conformity and compression force influence the joint translational stiffness during articulation inside the glenoid cavity, whereas during articulating outside the glenoid cavity this is influenced by the joint compression force and humeral radius of curvature. The glenoid radius of curvature influences the contact point and, in combination with the glenoid superior-inferior chord length, it also influences the constraintness angle, which influences the maximum allowable subluxation load to prevent a joint dislocation. This constraintness angle together with the joint conformity also influences maximum joint translations before articulation outside the glenoid cavity. Furthermore, the sign of the joint translational stiffness determines the stability of shoulder motion, which is stable and unstable if this stiffness is positive and negative, respectively.

Contact stresses in the glenoid component in total shoulder arthroplasty.

Several studies of retrieved glenoid components from total shoulder arthroplasty show an erosion of the rim, surface irregularities, component fracture and wear resulting from polyethylene deformation in vivo. Particles resulting from polyethylene wear might be one of the reasons for the very high rate of glenoid component loosening found clinically. Because wear can be the result of high contact stresses, the aim of this study is to find out whether or not contact stresses are high enough to cause wear of the glenoid component and what influence the component type and geometry have on polyethylene contact stresses for different humerus abduction angles. Elasticity theory is used in a parametric study of contact stresses in several glenoid component designs. A finite element method is used to confirm the accuracy of the analytical solution. The analysis shows that the peak stress generated in glenoid components under conditions of normal living can be as high as 25 MPa; since this exceeds the polyethylene yield strength, wear and also cold flow of the components can be expected. It is predicted that more conforming components have lower contact stresses, which might result in lower wear rate and less cold flow. It is also found that a metal-backed component promotes higher contact stresses than an all-polyethylene component with the same total thickness, therefore it can be expected that metal-backed components have inferior wear properties.

The anatomic features of the radial head and their implication for prosthesis design.

OBJECTIVE: To establish the anatomical features of the radial head of an average normal human being and to verify the hypothesis that no significant difference exists between the geometry of the left and right normal radial heads. DESIGN: 17 proximal ends of the radius from the left and right forearms of fresh male (average age 50 and range 40-70) cadavers were measured. BACKGROUND: A reconstruction of anatomical features of the normal bone is important for prosthesis design. METHODS: A morphologic study of the radial head was performed using a co-ordinate measuring machine integrated with a computer aided design system. For comparative purposes, a statistical analysis including linear regression and correlation was performed. RESULTS: The maximum diameter (mean 23.36 mm (SD, 1.14 mm)) and height (mean 10.14 mm (SD, 1.38 mm)) of the radial head as well as the depth (mean 1.92 mm (SD, 0.32 mm)) and maximum radius (mean 20.27 mm (SD, 4.61 mm)) of the concave articulate surface are the most important anatomical features, which should be implicated in prosthesis design. The inclinations mean 2.50 degrees (SD, 0.41 degrees ) and mean 9.50 degrees (SD, 0.52 degrees ) and shift (mean 1.71 mm (SD, 0.35 mm)) of the radial head relative to its neck should also be taken into account in prosthetic design. CONCLUSIONS: The results of the study showed that "left is equal to right" (no significant differences between sides were obtained, for probability value P>0.05). RELEVANCE: This paper describes a morphological study of the proximal radius. The results can be used to reconstruct the geometry of the injured radial head based on the obtained geometric features of the contra-lateral side. These results can be also used to design radial head prosthesis.