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Degradation characteristics in response to electron beam sterilization of designed and biodegradable aliphatic polyester scaffolds are relevant for clinically successful synthetic graft tissue regeneration. Scaffold degradation in vitro and in vivo were documented and correlated to the macroscopic structure and chemical design of the original polymer. The materials tested were of inherently diverse hydrophobicity and crystallinity: poly(L-lactide) (poly(LLA)) and random copolymers from L-lactide and ε-caprolactone or 1,5-dioxepan-2-one, fabricated into porous and non-porous scaffolds. After sterilization, the samples underwent hydrolysis in vitro for up to a year. In vivo, scaffolds were surgically implanted into rat calvarial defects and retrieved for analysis after 28 and 91days. In vitro, poly(L-lactide-co-1,5-dioxepan-2-one) (poly(LLA-co-DXO)) samples degraded most rapidly during hydrolysis, due to the pronounced chain-shortening reaction caused by the sterilization. This was indicated by the rapid decrease in both mass and molecular weight of poly(LLA-co-DXO). Poly(L-lactide-co-ε-caprolactone) (poly(LLA-co-CL)) samples were also strongly affected by sterilization, but mass loss was more gradual; molecular weight decreased rapidly during hydrolysis. Least affected by sterilization were the poly(LLA) samples, which subsequently showed low mass loss rate and molecular weight decrease during hydrolysis. Mechanical stability varied greatly: poly(LLA-co-CL) withstood mechanical testing for up to 182 days, while poly(LLA) and poly(LLA-co-DXO) samples quickly became too brittle. Poly(LLA-co-DXO) samples unexpectedly degraded more rapidly in vitro than in vivo. After sterilization by electron beam irradiation, the three biodegradable polymers present widely diverse degradation profiles, both in vitro and in vivo. Each exhibits the potential to be tailored to meet diverse clinical tissue engineering requirements.

This study was performed to determine the effect of commercially pure titanium surfaces blasted with TiO2 particles on the biological responses of cells derived from human mandibular bone. The morphology and attachment of those cells were investigated on turned titanium surfaces (control) and surfaces blasted with 45 μm (standard), 45–63 μm, and 63–90 μm TiO2 particles. The surfaces were analyzed in a scanning electron microscope. Based on surface analyses reported elsewhere, the turned samples had the smoothest surfaces and the roughest were those blasted with the largest particles (63–90 μm). The cell profile areas were measured using a semi-automatic interactive image analyzer. The attachment was determined as a ratio of the area of cell profiles and the total micrograph area and was expressed as percentage of attachment. Morphologically, the cells were heterogeneous. In general, the cells had spread well on all titanium surfaces, indicating good attachment to both smooth and rough surfaces. After 1, 3 and 6 h, the percentage of cell attachment did not differ significantly between the surfaces blasted with 63–90 μm and the turned surfaces, but was significantly lower on the surfaces blasted with 45 μm or 45–63 μm particles. After 24 h the surfaces blasted with 63–90 μm particles had a higher rate of cell attachment than all the other surfaces including the controls. It is concluded that attachment and growth of cells originating from human mandibular bone in vitro, are influenced by the micro-texture of the implant surface.

Our recent in vitro study demonstrated that endothelial cells (ECs) might influence the differentiation of bone marrow stromal cells (BMSCs). Therefore, the aim of this study was to describe this effect in vivo, using a rat calvarial bone defect model. BMSCs were isolated from femurs of two-donor Lewis rats and expanded in α-minimum essential medium containing 10% fetal bovine serum. One fifth of BMSCs were induced and differentiated into ECs in an Endothelial Cell Growth Medium-2 and then characterized by a flow cytometry. The remaining BMSCs were cultured in freshly prepared osteogenic stimulatory medium, containing dexamethasone, ascorbic acid and β-glycerophosphate. Either BMSCs alone (BMSC-group) or co-cultured ECs/BMSCs (CO-group) were seeded into poly(L-lactide-co-1,5-dioxepan-2-one) [poly(LLA-co-DXO)] scaffolds, cultured in spinner flasks, and then implanted into symmetrical calvarial defects prepared in recipient rats. The animals were sacrificed after 2 months. The formation of new bone was evaluated by radiography and histology and by the expression of osteogenic markers using reverse transcriptase-polymerized chain reaction (RT-PCR). To investigate vessel formation, histological staining was performed with EC's markers. The radiographical and histological results showed more rapid bone formation in the CO- than in the BMSC-group. However, the expression of EC's marker was similar on both groups by histological analysis after 2 months postoperatively. Furthermore, the CO-group exhibited greater expression of osteogenic markers as demonstrated by RT-PCR. The results are consistent with the previous in vitro findings that poly(LLA-co-DXO) scaffold might be suitable candidate for bone tissue engineering. In vivo, bone regeneration was enhanced by a construct of the polymer scaffold loaded with co-cultured cells.

In tissue engineering, the resorbable aliphatic polyester poly(L-lactide) (PLLA) is used as scaffolds in bone regeneration. Copolymers of poly(L-lactide)-co-(e-caprolactone) [poly(LLA-co-CL)] and poly(L-lactide)-co-(1,5-dioxepan-2-one) [poly(LLA-co-DXO)], with superior mechanical properties to PLLA, have been developed to be used as scaffolds, but the influence on the osteogenic potential is unclear. This in vitro study of test scaffolds of poly(LLA-co-CL) and poly(LLA-co-DXO) using PLLA scaffolds as a control demonstrates the attachment and proliferation of human osteoblast-like cells (HOB) as measured by SEM and a methylthiazol tetrazolium (MTT) colorimetric assay, and the progression of HOB osteogenesis for up to 3 weeks; expressed as synthesis of the osteoblast differentiation markers: collagen type 1 (Col 1), alkaline phosphatase, bone sialoprotein, osteocalcin (OC), osteopontin and runt related gene 2 (Runx2). Surface analysis disclosed excellent surface attachment, spread and penetration of the cells into the pores of the test scaffolds compared to the PLLA. MTT results indicated that test scaffolds enhanced the proliferation of HOBs. Cells grown on the test scaffolds demonstrated higher synthesis of Col 1 and OC and also increased bone markers mRNA expression. Compared to scaffolds of PLLA, the poly(LLA-co-CL) and poly(LLA-co-DXO) scaffolds enhanced attachment, proliferation, and expression of osteogenic markers by HOBs in vitro. Therefore, these scaffolds might be appropriate carriers for bone engineering. © 2010 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2010

A fundamental component of bone tissue engineering is an appropriate scaffold as a carrier for osteogenic cells. The aim of the study was to evaluate the response of human bone marrow stromal cells (BMSC) to scaffolds made of three biodegradable polymers: poly(L-lactide-co-e-caprolactone) (poly(LLA-co-CL)), poly(L-lactide-co-1,5dioxepan-2-one) (poly(LLA-co-DXO)), and poly(L-lactide) (poly(LLA)). Cellular response was evaluated in terms of attachment, proliferation, and differentiation. SEM disclosed earlier cell attachment and better spreading on poly(LLA-co-CL) and poly(LLA-co-DXO) scaffolds than on poly(LLA) after 1 h. At 24 h and 14 days postseeding, BMSCs had spread well, forming multiple cellular layers on the scaffolds. Cell proliferation was higher on poly(LLA-co-CL) and on poly(LLA-co-DXO) than on poly(LLA) after 1 and 7 days. Cell growth cycles of BMSC were longer on the scaffolds than on coverslips. After 7 and 14 days cultivation on scaffolds, the expression of osteogenic markers such as ALP, Col I, OPN, and Runx2 were stimulated by BMSC, which indicating that poly(LLA-co-DXO), poly(LLA-co-CL), and poly(LLA) could support the osteogenic differentiation of BMSC in vitro. Poly(LLA-co-CL) and poly(LLA-co-DXO) promoted better attachment and growth of BMSC than poly(LLA). BMSC also retained their osteogenic differentiation potential, indicating biological activity of BMSC on the scaffolds. The promising results of this in vitro study indicate that these copolymers warrant further evaluation for potential application in bone tissue engineering.

Successful attachment of the oral tissues to an endosseous dental implant relies in part on its chemical stability, nontoxic properties and maintenance of normal cell functions in the surrounding tissues. The aim of this study was therefore to investigate these 3 factors experimentally. The corrosion resistance of single crystal sapphire (alpha-Al2O3) implants was analysed with respect to the release of aluminium ions; no ions could be detected in the test solutions. The influence of single crystal sapphire on the behaviour of human epithelial cells and fibroblasts derived from biopsies of the oral mucosa, was studied. The cells were cultured in chemically defined or in low serum-containing media. Compared to the corresponding control cultures, no effects on cell morphology and growth characteristics were observed. Sapphire rods were inserted subcutaneously into rats and tissue responses were examined after 4, 8 and 12 weeks post-implantation. None or slight reactions were found in the tissues. The combined evaluation of these experimental approaches indicates that single crystal sapphire is a material well suited for biological implantation.

Objectives: This in vitro study was aimed to investigate the attachment, spreading and proliferation of human gingival fibroblasts to milled and polished non-veneered ceramic surfaces in alumina and zirconia and to ceramic surfaces veneered by two different types of porcelain baseliners. Materials and methods: Fibroblasts were cultured on discs of pressed alumina or zirconia, on discs which had been milled, on discs comprising alumina or zirconia which had been polished, on discs of alumina veneered with NobelRondo baseliner Al, on discs of zirconia veneered with Cercon-S baseliner, and on alumina or zirconia discs veneered with the above baseliners and then polished. The surfaces were analyzed using an optical interferometer and scanning electron microscopy (SEM). Cell profile areas were measured using SEM and an image analyzer. Cell attachment was determined after 3 and 24 h as a ratio of the cell profiles and the total micrograph area and was expressed as percent of attachment. MTT analyses were undertaken to determine cellular attachment after 3 h of incubation and cellular proliferation after 7 days. Results: The polished zirconia specimens had the smoothest surface in terms of average height deviation (Sa=0.03 μm): the roughest were the zirconia specimens with milled surfaces (Sa=0.36 μm). The application of the baseliners resulted in surfaces smoother than those of the non-veneered discs. The milled surfaces of both alumina and zirconia had significantly higher percentages of cell attachment and proliferation than the other surfaces whereas the milled surfaces in zirconia demonstrated better cellular attachment after 3 and 24 h of culture than the one in alumina. Fibroblasts attached and grew effectively on the surfaces veneered with NobelRondo throughout the experiments, whereas the zirconia surfaces veneered with Cercon-S had the lowest percentage of cell attachment and proliferation. Conclusions: Although the roughness of all surfaces investigated was

9 single crystal sapphire dental implants were installed bilaterally into pre-extracted areas in the lower jaw of two beagle dogs and histologically analysed after 180 days in situ. 8 implants were stable and radiographs disclosed complete bone healing. Light-, scanning- and transmission electron microscopy demonstrated that the stable implants were surrounded by a mineralizing bone boundary and a mutosa nearly free from inflammatory cell infiltrations. The 9th implant was mobile and surrounded by a non-mineralized connective tissue capsule containing bundles of collagen. The ultrastructure of the mucosa surrounding the implants closely resembled the mucosa surrounding the tooth. Histometric analysis of the alveolar bone surrounding the stable implants revealed that the value of the bone contact surface ranged from 37.1%% to 86.9% (mean value 61.8%) at the light microscopic level.

Optical tweezers were used to study the interaction and attachment of human bone cells to various types of medical implant materials. Ideally, the implant should facilitate cell attachment and promote migration of the progenitor cells in order to decrease the healing time. It is therefore of interest, in a controlled manner, to be able to monitor the cell adhesion process. Results from such studies would help foresee the clinical outcome of integrating medical implants. The interactions between two primary cell culture models, human gingival fibroblasts and bone forming human osteoblast cells, and three different implant materials, glass, titanium, and hydroxyapatite, were studied. A novel type of optical tweezers, which has a newly designed quadrant detector and a powerful 3 W laser was constructed and force calibrated using two different methods: one method in which the stiffness of the optical trap was obtained by monitoring the phase lag between the trap and the moved object when imposing a forced oscillation on the trapped object and another method in which the maximum trapping force was derived from the critical velocity at which the object escapes the trap. Polystyrene beads as well as cells were utilized for the calibrations. This is the first time that cells have been used directly for these types of force calibrations and, hence, direct measurements of forces exerted on cells can be performed, thus avoiding the difficulties often encountered when translating the results obtained from cell measurements to the calibrations obtained with reference materials. This more straightforward approach represents an advantage in comparison to established methods.

As different implant abutments are introduced to obtain a sufficient soft tissue barrier, the aim of this study was to determine the effect of three different surface modifications of densely sintered high-purity aluminium oxide on morphology, attachment and proliferation of human gingival fibroblasts. Fibroblasts were cultured on pressed aluminium oxide, milled, and then sintered to full density (1), on pressed, densely sintered (2), and on pressed, densely sintered and then polished surfaces (3). The different surfaces were analyzed using a confocal laser scanner, an atomic force microscope and a scanning electron microscope. The cell profile areas were measured using a semiautomatic interactive image analyzer and the figures were expressed as percent of attachment. The polished specimens had the smoothest surfaces and the roughest were the milled surfaces in terms of height deviation. No difference was found in the spacing between the peaks on the polished surfaces compared to the milled surfaces. Fibroblasts on the milled ceramic appeared to follow the direction of the fine irregularities on the surface. The analyses showed the polished surfaces had significantly higher percentages of initial cell attachment than the other surfaces (P

Variations in the oxide films on titanium surfaces blasted with TiO(2) particles of various sizes were analyzed after cultures with cells derived from human mandibular bone. Turned titanium surfaces and surfaces blasted with 63-90-, 106-180-, and 180-300-microm TiO(2) particles were cultured with osteoblast-like cells. The surfaces were characterized before and after the cell culture with electrochemical impedance spectroscopy (EIS). The surface chemical composition of selected samples was analyzed with X-ray photoelectron spectroscopy (XPS). EIS revealed that with respect to the turned surfaces, the effective surface area was about 5, 6, and 4 times larger on the surfaces blasted with 63-90-, 106-180-, and 180-300-microm particles, respectively. After 28 days of the cell culture, the corrosion resistance on all sample types was unaffected. The impedance characteristics suggest a considerable effect of ion incorporation and precipitation during culturing. XPS revealed that before the cell culture, a typical surface layer consisted of TiO(2). After the culture, the surface oxide film contained both phosphorus and calcium, along with large amounts of oxidized carbon (carbonate) and nitrogen. There were lower concentrations of carbon and nitrogen on the blasted surfaces. We concluded that the effective surface area was several times higher on blasted surfaces than on turned surfaces. Cells derived from human mandibular bone affected ion incorporation into the implant surface.

— The content and distribution of metallic ions on the surface of gold-plated brass screwposts (Dentatus and MecOdont) were analyzed by X-ray microanalysis. On screwposts from one of the manufacturers the protective outer layer of gold was found to be localized to specific areas of the posts, i.e. the grooves, while unprotected Cu-Zn alloy was found on the ridges. The results are discussed with regard to results on root posts published earlier. Screwposts constructed in this way would appear to be of doubtful clinical value.

– Galvanic series for different metals in different electrolytes have been published, but never for dental alloys in artificial saline solution. The material classes studied are amalgam, gold alloy, nickel alloy, chromium cobalt casting alloy and temporary crown form. The electrogalvanic potentials were measured and determined in reference to a hydrogen electrode (H2/H+) arbitrarily defined as zero. A galvanic series of the tested materials was arranged in order of their potentials.

The galvanic current densities between gold, amalgam and cobalt-chromium, three different classes of dental alloys, were determined in vitro in artificial saliva kept at 35 degrees C. The maximum current density of 200 microA/dm2 was obtained between the conventional amalgam and a type III gold alloy. Galvanic currents of lesser magnitude could also be measured between amalgams high in copper and the other alloys. No measurable current densities were obtained between gold alloys and between gold and cobalt-chromium alloy with the exception of a casting and a solder gold alloy commonly used in combination.