Future Proteins: Sustainable Diets for Tenebrio molitor Rearing Composed of Food By-Products.

Since the human population is continuously growing, sufficient food with low environmental impact is required. Especially, the challenge of providing proteins will deepen and insects can contribute to a more sustainable and efficient source of protein for human consumption. Tenebrio molitor larvae are highly nutritious and rearing mealworms is more environmentally friendly compared to the production of traditional livestock meat. To use T. molitor as a more sustainable alternative to conventional proteins, it is essential to apply diets from a local and sustainable source. Therefore, the objective of this study was to find local by-products or leftovers which can be used in mass production of larvae as a main substrate. Feeding trials investigating twenty-nine different substrates were conducted to evaluate larval growth performance and adult reproduction by determining development times, survival rates, biomass, and fecundity. Several suitable by-products were identified that can be used in high quantities as single component diet for T. molitor rearing, revealing a high survival rate, short development time, high mean total biomass, and successful breeding. The most successful substrate-malt residual pellets-was found to be an alternative to the most used substrate, wheat bran. Furthermore, corn germ meal, sweet chestnuts, bread remains, soybeans, sweet potatoes, and wheat germs have been discovered to be suitable diets for T. molitor. Moreover, the findings of this study contribute towards using several substrates as supplements.

Liking and Description of Pasta Sauces with Varying Mealworm Content.

Entomophagy is directly connected with culture, explaining why it is commonly rejected in Western countries. Due to increased meat consumption in recent years with its associated negative impacts on health and sustainability, the development of products based on alternative protein sources has become urgent. The larval form of Tenebrio molitor (mealworm) has the potential to substitute meat as it requires less resources and produces less emissions compared to other forms of meat production. Therefore, in this project we have aimed to develop pasta sauces with differing mealworm contents based on a common meat sauce and to test the acceptance with 91 consumers in Austria. Three sauces (100% mealworm, 50% mealworm and 50% meat, 100% meat) were developed and tested using a 9-point hedonic scale for acceptance, and the CATA (Check-All-That-Apply) method was integrated to also receive descriptive information. The analysis of the liking data revealed that the liking for the hybrid sauce with meat and mealworm content was comparable to the meat sauce (6.9 ± 1.8. vs. 6.5 ± 1.8, p > 0.05). Less liked was the sauce with the highest mealworm content (5.7 ± 1.8, p < 0.05). The CATA analysis demonstrated the strongest positive effects on the mean in terms of how much the products were liked for the attribute "fleshy" (0.8). On the other hand, the attributes "brownish" (-0.9) or "mushy" (-1.0) had the strongest negative effects on the mean of the liking of products. We have seen that meat cannot be substituted by mealworm immediately and completely. The results suggest a stepwise substitution and the further adaptation of products regarding the (negative and positive effecting) attributes to increase consumer acceptance.

Tenebrio molitor (Linnaeus, 1758): Microbiological Screening of Feed for a Safe Food Choice.

As a result of the increasing focus on alternative protein sources which are ideally still sustainable, the yellow mealworm, Tenebrio molitor, has come into focus. To verify its suitability as a food source in relation to human health, an analysis of the microbiome of larvae of T. molitor is pertinent. Subsequently, the focus of this study was, on the one hand, to analyze the influence of the substrate on the microbial load of the larvae microbiome, and, on the other hand, to determine which processing methods ensure the risk-free consumption of mealworms. For this purpose, mealworms were grown on 10 different substrates derived from by-products of food production (malt residual pellets, corn germ meal, chestnut breakage and meal, wheat bran, bread remains, draff, nettle, hemp seed oil cake, oyster mushrooms with coffee grounds, pumpkin seed oil cake) and microbial loads were analyzed using different selective media. Further starvation/defecation and heating (850 W for 10 min) methods were used to investigate how the reduction of microorganisms is enabled by these methods. The results showed that there was no significant relationship between the microbial load of the substrate and the mealworm. Starvation and defecation led to a lower stock of microorganisms. Heating led to a significant microbial reduction in non-defecated mealworms. The group of defecated and heated mealworms showed no detectable microbial load. In conclusion, firstly, the choice of substrate showed no effect on the microbial load of larvae of Tenebrio molitor and secondly, heating and starvation allow risk-free consumption. This study makes an important contribution for evaluating the safety of mealworms as a sustainable protein source in human nutrition.

Use Them for What They Are Good at: Mealworms in Circular Food Systems.

Future food systems must provide more food produced on less land with fewer greenhouse gas emissions if the goal is to keep planetary boundaries within safe zones. The valorisation of agricultural and industrial by-products by insects is an increasingly investigated strategy, because it can help to address resource scarcities and related environmental issues. Thus, insects for food and feed have gained increasing attention as a sustainable protein production strategy in circular food systems lately. In this article, we provide an overview on by-products, which have already been fed to T. molitor (mealworms), a common edible insect species. In addition, we investigate other by-products in Austria, which can be suitable substrates for T. molitor farming. We also provide an overview and discuss different perspectives on T. molitor and link it with the circular economy concept. We identify several future research fields, such as more comprehensive feeding trials with other by-products, feeding trials with mealworms over several generations, and the development of a standardized framework for insect rearing trials. In addition, we argue that due to their ability to convert organic by-products from agricultural and industrial processes into biomass in an efficient way, T. molitor can contribute towards resource-efficient and circular food and feed production. However, several hurdles, such as legal frameworks, need to be adapted, and further research is needed to fully reap the benefits of mealworm farming.

Assessing the osteogenic potential of zirconia and titanium surfaces with an advanced in vitro model.

OBJECTIVES: In recent years, zirconia dental implants have gained increased attention especially for patients with thin gingival biotypes or patients seeking metal-free restoration. While physical and chemical material surface properties govern the blood-material interaction and subsequent osseointegration processes, the organizational principles underlying the interplay of biochemical and biophysical cues are still not well understood. Therefore, this study investigated how the interaction of a microstructured zirconia surface with blood influences its osseointegration potential compared to microstructured titanium with or without additional nanostructures. METHODS: Microstructured zirconia and micro- (and nano)structured titanium surfaces were fabricated via sandblasting followed by acid etching and their topographical as well as physico-chemical features were thoroughly characterized. Following, an advanced in vitro approach mimicking the initial blood interaction of material surfaces upon implantation was applied. Fibrinogen adsorption, human blood coagulation as well as their influence on cell fate decisions of primary human bone and progenitor cells (HBC) were studied. RESULTS: Obtained surface micro- and nanostructures on titanium surfaces were sharp with rugged peaks whereas zirconia surfaces were less rough with structures being shallower, more round and granular. Compared to titanium surfaces, the zirconia surface showed increased fibrinogen adsorption, higher levels of total accessible fibrinogen γ-chain moieties yielding in increased platelet adhesion and activation and consequently thrombogenicity. Mineralization of HBC on microstructured surfaces was significantly higher on zirconia than on titanium, but was significantly lower compared to titanium surfaces with nanostructures. SIGNIFICANCE: This study provides insights into blood-material interaction and subsequent cellular events that are important for implant surface development.

Surface modification of ultrafine-grained titanium: Influence on mechanical properties, cytocompatibility, and osseointegration potential.

OBJECTIVE: The main objective of this study was to demonstrate that dental implants made from ultrafine-grain titanium (UFG-Ti) can be created that replicate state of the art surfaces of standard coarse-grain titanium (Ti), showing excellent cytocompatibility and osseointegration potential while also providing improved mechanical properties. MATERIAL AND METHODS: UFG-Ti was prepared by continuous equal channel angular processing (ECAP), and surfaces were treated by sandblasting and acid etching. Mechanical properties (tensile and fatigue strength), wettability, and roughness parameters were evaluated. Human trabecular bone-derived osteoblast precursor cells (HBCs) were cultured on all samples to examine cytocompatibility and mineralization after 4 and 28 days, respectively. Biomechanical pull-out measurements were performed in a rabbit in vivo model 4 weeks after implantation. RESULTS: Both yield and tensile strength as well as fatigue endurance were higher for UFG-Ti compared to Ti by 40%, 45%, and 34%, respectively. Fatigue endurance was slightly reduced following surface treatment. Existing surface treatment protocols could be applied to UFG-Ti and resulted in similar roughness and wettability as for standard Ti. Cell attachment and spreading were comparable on all samples, but mineralization was higher for the surfaces with hydrophilic treatment with no significant difference between UFG-Ti and Ti. Pull-out tests revealed that osseointegration of surface-treated UFG-Ti was found to be similar to that of surface-treated Ti. CONCLUSION: It could be demonstrated that existing surface treatments for Ti can be translated to UFG-Ti and, furthermore, that dental implants made from surface-treated UFG-Ti exhibit superior mechanical properties while maintaining cytocompatibility and osseointegration potential.

Comparable responses of osteoblast lineage cells to microstructured hydrophilic titanium-zirconium and microstructured hydrophilic titanium.

OBJECTIVES: Although titanium (Ti) is commonly used for dental implants, Ti alloy materials are being developed to improve their physical material properties. Studies indicate that osteoblast differentiation and maturation of human mesenchymal stem cells (MSCs) and normal human osteoblasts (NHOsts) respond to microstructured Ti and titanium-aluminum-vanadium (Ti6Al4V) surfaces in a similar manner. The goal of this study was to determine whether this is the case for osteoblast lineage cells grown on microstructured TiZr surfaces and whether their response is affected by surface nanotexture and hydrophilicity. MATERIALS AND METHODS: Grade 4 Ti and TiZr (13-17% Zr) disks were modified by large grit sand-blasting and acid-etching with storage in saline solution, resulting in a complex microstructured and hydrophilic surface corresponding to the commercially available implants SLActive® and Roxolid® SLActive® (Institut Straumann AG, Basel, Switzerland). The subsequent Ti modSLA and TiZr modSLA surfaces were characterized and osteogenic markers were measured. RESULTS: Evaluation of physical parameters revealed that the fabrication method was capable of inducing a microstructured and hydrophilic surface on both the Ti and TiZr disks. Overall, the surfaces were similar, but differences in nanostructure morphology/density and surface chemistry were detected. On Ti modSLA and TiZr modSLA, osteoblastic differentiation and maturation markers were enhanced in both MSCs and NHOsts, while inflammatory markers decreased compared with TCPS. CONCLUSIONS: These results indicate a similar positive cell response of MSCs and NHOsts when cultured on Ti modSLA and TiZr modSLA. Both surfaces were hydrophilic, indicating the importance of this property to osteoblast lineage cells.

Chemically modified titanium-zirconium implants in comparison with commercially pure titanium controls stimulate the early molecular pathways of bone healing.

OBJECTIVES: Titanium-zirconium (TiZr) has been proposed as a mechanically stronger alternative to commercially pure titanium for oral and orthopaedic implants. However, not much is known on the osseointegration kinetics of TiZr surfaces. In this study, we aimed to identify the genetic response of bone around TiZr implants compared to pure Ti. MATERIAL AND METHODS: Microtextured and hydrophilic TiZr implants (tests) and cpTi implants grade IV (controls) were placed in the tibia of 30 New Zealand white rabbits. At 2, 4 and 12 weeks, the implants were subjected to removal torque test (RTQ). The expression of a panel of genes involved in the process of osseointegration was measured in the bone around the test and control implants by means of quantitative real-time polymerase chain reaction (PCR) and compared to the control samples. RESULTS: The controls yielded statistically significant higher RTQ at 4 weeks, but the RTQ of the tests had a larger increase between 4 and 12 weeks, when both groups reached similar values. The gene expression analysis showed that all selected markers for bone formation, bone remodelling and cytokines were significantly upregulated around TiZr implants after 2 weeks. After 4 weeks of healing, two bone formation markers were significantly more expressed in the test samples, while at 12 weeks, the expression of all genes was similar in the two groups. CONCLUSIONS: TiZr implants showed comparable biomechanical outcomes to cpTi up to 12 weeks of healing. However, at early healing stages, they showed a significant upregulation of osteogenesis and osteoclastogenesis markers.

Proliferation, behavior, and differentiation of osteoblasts on surfaces of different microroughness.

OBJECTIVES: Titanium surface roughness is recognized as an important parameter influencing osseointegration. However, studies concerning the effect of well-defined surface topographies of titanium surfaces on osteoblasts have been limited in scope. In the present study we have investigated how Ti surfaces of different micrometer-scale roughness influence proliferation, migration, and differentiation of osteoblasts in-vitro. METHODS: Titanium replicas with surface roughnesses (Ra) of approximately 0, 1, 2, and 4µm were produced and MG-63 osteoblasts were cultured on these surfaces for up to 5 days. The effect of surface micrometer-scale roughness on proliferation, migration in time-lapse microscopy experiments, as well as the expression of alkaline phosphatase, osteocalcin, vascular-endothelial growth factor (VEGF), osteoprotegerin (OPG), and receptor activator of nuclear factor kappa-B ligand (RANKL) were investigated. RESULTS: Proliferation of MG-63 cells was found to decrease gradually with increasing surface roughness. However, the highest expression of alkaline phosphatase, osteocalcin and VEGF was observed on surfaces with Ra values of approximately 1 and 2µm. Further increase in surface roughness resulted in decreased expression of all investigated parameters. The cell migration speed measured in time-lapse microscopy experiments was significantly lower on surfaces with a Ra value of about 4µm, compared to those with lower roughness. No significant effect of surface roughness on the expression of OPG and RANKL was observed. SIGNIFICANCE: Thus, surfaces with intermediate Ra roughness values of 1-2µm seem to be optimal for osteoblast differentiation. Neither proliferation nor differentiation of osteoblasts appears to be supported by surfaces with higher or lower Ra values.

Osteogenic response of human MSCs and osteoblasts to hydrophilic and hydrophobic nanostructured titanium implant surfaces.

Microstructured implant surfaces created by grit blasting and acid etching titanium (Ti) support osseointegration. This effect is further enhanced by storing in aqueous solution to retain hydrophilicity, but this also leads to surface nanostructure formation. The purpose of this study was to assess the contributions of nanostructures on the improved osteogenic response of osteoblast lineage cells to hydrophilic microstructured Ti. Human mesenchymal stem cells (MSCs) and normal human osteoblasts (NHOsts) were cultured separately on non-nanostructured/hydrophobic (SLA), nanostructured/hydrophilic (modSLA), or nanostructured/hydrophobic (SLAnano) Ti surfaces. XPS showed elevated carbon levels on SLA and SLAnano compared to modSLA. Contact angle measurements indicated only modSLA was hydrophilic. Confocal laser microscopy revealed minor differences in mean surface roughness. SEM showed the presence of nanostructures on modSLA and SLAnano. MSCs and NHOst cells exhibited similar morphology on the substrates and osteoblastic differentiation and maturation were greatest on modSLA. These results suggest that when the appropriate microstructure is present, hydrophilicity may play a greater role in stimulating MSC and NHOst osteoblastic differentiation and maturation than the presence of nanostructures generated during storage in an aqueous environment. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 3137-3148, 2016.

Microstructure and mechanical properties of Ti-15Zr alloy used as dental implant material.

Ti-Zr alloys have recently started to receive a considerable amount of attention as promising materials for dental applications. This work compares mechanical properties of a new Ti-15Zr alloy to those of commercially pure titanium Grade4 in two surface conditions - machined and modified by sand-blasting and etching (SLA). As a result of significantly smaller grain size in the initial condition (1-2µm), the strength of Ti-15Zr alloy was found to be 10-15% higher than that of Grade4 titanium without reduction in the tensile elongation or compromising the fracture toughness. The fatigue endurance limit of the alloy was increased by around 30% (560MPa vs. 435MPa and 500MPa vs. 380MPa for machined and SLA-treated surfaces, respectively). Additional implant fatigue tests showed enhanced fatigue performance of Ti-15Zr over Ti-Grade4.

Biomechanical and Histomorphometrical Evaluation of TiZr Alloy Implants: An in vivo Study in the Rabbit.

BACKGROUND: Clinically, there is a demand for mechanically stronger alloyed implants; however, not much evidence exists with regard to these materials. PURPOSE: To test the osseointegration property of TiZr1317 implants in a rabbit model. MATERIALS AND METHODS: Hydrophilic titanium-zirconium alloy (TiZr1317) implants with sand-blasted and acid-etched surface (test) and hydrophilic cpTi implants with the same treatment (control) were placed pairwise in the hind limbs (two in each tibia and one in each femur) of 36 Swedish lop-eared rabbits. After 2, 4, and 12 weeks (n = 12/time point), the bone samples were subjected to removal torque (RTQ, proximal tibia and femur) and histologic/histomorphometric (distal tibia) testings. RESULTS: The control presented significantly higher RTQ than the test at 2 weeks (55 vs 36 Ncm). No differences were observed for other time points. The test presented higher mean BIC than the control (19.25 vs 13.89 %) at 4 weeks; however, there were no statistical differences for the following time point tested in vivo.The new bone area was significantly higher for the test at 4 weeks in the marrow areas. CONCLUSION: The TiZr1317 implants presented comparable biologic outcomes to that of the cpTi implants through a 12-week evaluation period.

Enhanced differentiation of human osteoblasts on Ti surfaces pre-treated with human whole blood.

Early and effective integration of a metal implant into bone tissue is of crucial importance for its long-term stability. While different material properties including surface roughness and wettability but also initial blood-implant surface interaction are known to influence this osseointegration, implications of the latter process are still poorly understood. In this study, early interaction between blood and the implant surface and how this affects the mechanism of osseointegration were investigated. For this, blood coagulation on a micro-roughened hydrophobic titanium (Ti) surface (SLA-H(phob)) and on a hydrophilic micro-roughened Ti surface with nanostructures (SLActive-H(phil)NS), as well as the effects of whole human blood pre-incubation of these two surfaces on the differentiation potential of primary human bone cells (HBC) was assessed. Interestingly, pre-incubation with blood resulted in a dense fibrin network over the entire surface on SLActive-H(phil)NS but only in single patches of fibrin and small isolated fibre complexes on SLA-H(phob). On SLActive-H(phil)NS, the number of HBCs attaching to the fibrin network was greatly increased and the cells displayed enhanced cell contact to the fibrin network. Notably, HBCs displayed increased expression of the osteogenic marker proteins alkaline phosphatase and collagen-I when cultivated on both surfaces upon blood pre-incubation. Additionally, blood pre-treatment promoted an earlier and enhanced mineralization of HBCs cultivated on SLActive-H(phil)NS compared to SLA-H(phob). The results presented in this study therefore suggest that blood pre-incubation of implant surfaces mimics a more physiological situation, eventually providing a more predictive in vitro model for the evaluation of novel bone implant surfaces.

The role of nanostructures and hydrophilicity in osseointegration: In-vitro protein-adsorption and blood-interaction studies.

Protein adsorption and blood coagulation play important roles in the early stages of osseointegration and are strongly influenced by surface properties. We present a systematic investigation of the influence of different surface properties on the adsorption of the blood proteins fibrinogen and fibronectin and the degree of early blood coagulation. Experiments on custom-made and commercially available, microroughened hydrophobic titanium (Ti) surfaces (Ti SLA-Hphob ), hydrophilic (Hphil ) microroughened Ti surfaces with nanostructures (Ti SLActive-Hphil NS), and on bimetallic Ti zirconium alloy (TiZr, Roxolid®) samples were performed, to study the biological response in relation to the surface wettability and the presence of nanostructures (NS). Protein adsorption on the different substrates showed a highly significant effect of surface NS. Hydrophilicity alone did not significantly enhance protein adsorption. Overall, the combination of NS and hydrophilicity led to the highest adsorption levels; independent of whether Ti or TiZr were used. Hydrophilicity induced a strong effect on blood coagulation, whereas the effect of NS alone was weak. The combination of both surface characteristics led to early and most pronounced blood-coagulation. Therefore, nanostructured, hydrophilic Ti and TiZr surfaces may perform better in terms of osseointegration due to continuous protein adsorption and the formation of a layer of blood components on the implant surface.

The angiogenic behaviors of human umbilical vein endothelial cells (HUVEC) in co-culture with osteoblast-like cells (MG-63) on different titanium surfaces.

OBJECTIVES: Interaction between osteogenesis and angiogenesis plays an important role in implant osseointegration. In the present study we investigated the influence of titanium surface properties on the angiogenic behaviors of endothelial cells grown in direct contact co-culture with osteoblasts. METHODS: Human umbilical vein endothelial cells (HUVECs) and osteoblast-like cells (MG-63 cells) were grown in direct co-culture on the following titanium surfaces: acid-etched (A), hydrophilic A (modA), coarse-gritblasted and acid-etched (SLA) and hydrophilic SLA (SLActive). Cell proliferation was evaluated by cell counting combined with flow cytometry. The expression of von Willebrand Factor (vWF), thrombomodulin (TM), endothelial cell protein C receptor (EPCR), E-Selectin, as well as vascular endothelial growth factor (VEGF) receptors Flt-1 and KDR in HUVECs and VEGF in MG-63 were measured by qPCR. The dynamic behavior of endothelial cells was recorded by time-lapse microscopy. RESULTS: Proliferation of HUVECs was highest on A, followed by SLA, modA and SLActive surfaces. The expression of vWF, TM, EPCR, E-Selectin and Flt-1 in HUVECs was significantly higher on A than on all other surfaces. The expression of KDR in HUVECs grown on A surface was below detection limit. VEGF expression in MG-63 cells was significantly higher on SLActive vs SLA and modA vs A surfaces. Time-lapse microscopy revealed that HUVECs moved quickest and formed cell clusters earlier on A surface, followed by SLA, modA and SLActive surface. CONCLUSIONS: In co-culture conditions, proliferation and expression of angiogenesis associated genes in HUVECs are promoted by smooth hydrophobic Ti surface, which is in contrast to previous mono-culture studies.

Nanostructures and hydrophilicity influence osseointegration: a biomechanical study in the rabbit tibia.

OBJECTIVE: Implant surface properties have long been identified as an important factor to promote osseointegration. The importance of nanostructures and hydrophilicity has recently been discussed. The aim of this study was to investigate how nanostructures and wettability influence osseointegration and to identify whether the wettability, the nanostructure or both in combination play the key role in improved osseointegration. MATERIALS AND METHODS: Twenty-six adult rabbits each received two Ti grade 4 discs in each tibia. Four different types of surface modifications with different wettability and nanostructures were prepared: hydrophobic without nanostructures (SLA), with nanostructures (SLAnano); hydrophilic with two different nanostructure densities (low density: pmodSLA, high density: SLActive). All four groups were intended to have similar chemistry and microroughness. The surfaces were evaluated with contact angle measurements, X-ray photoelectron spectroscopy, scanning electron microscopy, atomic force microscopy and interferometry. After 4 and 8 weeks healing time, pull-out tests were performed. RESULTS: SLA and SLAnano were hydrophobic, whereas SLActive and pmodSLA were super-hydrophilic. No nanostructures were present on the SLA surface, but the three other surface modifications clearly showed the presence of nanostructures, although more sparsely distributed on pmodSLA. The hydrophobic samples showed higher carbon contamination levels compared with the hydrophilic samples. After 4 weeks healing time, SLActive implants showed the highest pull-out values, with significantly higher pull-out force than SLA and SLAnano. After 8 weeks, the SLActive implants had the highest pull-out force, significantly higher than SLAnano and SLA. CONCLUSIONS: The strongest bone response was achieved with a combination of wettability and the presence of nanostructures (SLActive).

Spontaneously formed nanostructures on titanium surfaces.

OBJECTIVES: The aim was to investigate the evolution of nanostructures on the SLActive surface, as a function of time, storage conditions, material dependence and to identify the step in which the reorganization of the outermost titanium oxide layer into well defined nanostructures takes place. MATERIAL AND METHODS: Titanium grade 2 discs were surface modified in seven different modes; (1) SLA (sand blasted, large grit, acid etched) protocol. (2) SLActive protocol (SLA stored in 0.9% NaCl solution), (3) SLActive, but stored in water instead of 0.9% NaCl solution, (4) pmod SLA: SLA discs subjected to oxygen plasma cleaning and stored in 0.9% NaCl solution, (5) SLAnano: SLActive discs aged for several months and then dried, (6) Mod A: same etching procedure and storage as for SLActive, but no sand blasting prior to etching, (7) pmod P: the discs were polished, oxygen plasma cleaned and stored in 0.9% NaCl solution. In addition TiZr alloy discs were prepared like the Ti SLActive samples. The surfaces were evaluated with SEM, interferometry, contact angle measurements and XPS. RESULTS: The samples stored dry were hydrophobic whereas the discs stored in liquid were hydrophilic. The evolution of nanostructures took 2 weeks, thereafter they were stable over time. The nanostructures occured after storage both in water and NaCl solution. Nanostructures were formed on Ti and TiZr although the morphology and distribution was quite different between the two materials. CONCLUSIONS: Acid etching in conjunction with storage in aqueous solution is responsible for the reorganization of the outermost titanium oxide layer into well defined nanostructures.

Chiral distortion of confined ice oligomers (N = 5,6).

Ice nuclei have been studied on the hexagonal boron nitride nanomesh (h-BN/Rh(111)), a template with 2 nm wide molecule traps. Scanning tunneling microscopy shows confined clusters, where oligomers with three protrusions are particularly abundant. Together with local barrier height dI/dz maps, it is found that the dipoles of the water molecules arrange in a homodrome, which is consistent with density functional theory calculations. Hydrogen bonds toward the substrate identify h-BN/Rh(111) to be hydrophilic. The substrate distorts the hexamers (n = 6) and possibly pentamers (n = 5), where the experimentally observed footprints of the three protrusions appear more chiral than expected.

The effects of combined micron-/submicron-scale surface roughness and nanoscale features on cell proliferation and differentiation.

Titanium (Ti) osseointegration is critical for the success of dental and orthopedic implants. Previous studies have shown that surface roughness at the micro- and submicro-scales promotes osseointegration by enhancing osteoblast differentiation and local factor production. Only relatively recently have the effects of nanoscale roughness on cell response been considered. The aim of the present study was to develop a simple and scalable surface modification treatment that introduces nanoscale features to the surfaces of Ti substrates without greatly affecting other surface features, and to determine the effects of such superimposed nano-features on the differentiation and local factor production of osteoblasts. A simple oxidation treatment was developed for generating controlled nanoscale topographies on Ti surfaces, while retaining the starting micro-/submicro-scale roughness. Such nano-modified surfaces also possessed similar elemental compositions, and exhibited similar contact angles, as the original surfaces, but possessed a different surface crystal structure. MG63 cells were seeded on machined (PT), nano-modified PT (NMPT), sandblasted/acid-etched (SLA), and nano-modified SLA (NMSLA) Ti disks. The results suggested that the introduction of such nanoscale structures in combination with micro-/submicro-scale roughness improves osteoblast differentiation and local factor production, which, in turn, indicates the potential for improved implant osseointegration in vivo.