Structural Basis for Molecular Recognition of Cannabinoids by Inhibitory Cys-Loop Channels.

Cannabis sativa has a long history of medicinal use, dating back to ancient times. This plant produces cannabinoids, which are now known to interact with several human proteins, including Cys-loop receptors for glycine (GlyR) and gamma-aminobutyric acid (GABAAR). As these channels are the primary mediators of inhibitory signals, they contribute to the diverse effects of cannabinoids on the nervous system. Evidence suggests that cannabinoid binding sites are located within the transmembrane domain, although their precise location has remained undetermined for over a decade. The process of identification of the binding site and the computational approaches employed are the main subjects of this Perspective, which includes an analysis of the most recently resolved cryo-EM structures of zebrafish GlyR bound to Δ9-tetrahydrocannabinol and the THC-GlyR complex obtained through molecular dynamics simulations. With this work, we aim to contribute to guiding future studies investigating the molecular basis of cannabinoid action on inhibitory channels.

Molecular determinants of tetrahydrocannabinol binding to the glycine receptor.

The recognition of Cannabis as a source of new compounds suitable for medical use has attracted strong interest from the scientific community in its research, and substantial progress has accumulated regarding cannabinoids' activity; however, a thorough description of their molecular mechanisms of action remains a task to complete. Highlighting their complex pharmacology, the list of cannabinoids' interactors has vastly expanded beyond the canonical cannabinoid receptors. Among those, we have focused our study on the glycine receptor (GlyR), an ion channel involved in the modulation of nervous system responses, including, to our interest, sensitivity to peripheral pain. Here, we report the use of computational methods to investigate possible binding modes between the GlyR and Δ9 -tetrahydrocannabinol (THC). After obtaining a first pose for the THC binding from a biased molecular docking simulation and subsequently evaluating it by molecular dynamic simulations, we found a dynamic system with an identifiable representative binding mode characterized by the specific interaction with two transmembrane residues (Phe293 and Ser296). Complementarily, we assessed the role of membrane cholesterol in this interaction and positively established its relevance for THC binding to GlyR. Lastly, the use of restrained molecular dynamics simulations allowed us to refine the description of the binding mode and of the cholesterol effect. Altogether, our findings contribute to the current knowledge about the GlyR-THC mode of binding and propose a new starting point for future research on how cannabinoids in general, and THC in particular, modulate pain perception in view of its possible clinical applications.

The dangers of reused personal protective equipment: healthcare workers and workstation contamination.

BACKGROUND: Personal protective equipment (PPE) is essential to protect healthcare workers (HCWs). The practice of reusing PPE poses high levels of risk for accidental contamination by HCWs. Scarce medical literature compares practical means or methods for safe reuse of PPE while actively caring for patients. METHODS: In this study, observations were made of 28 experienced clinical participants performing five donning and doffing encounters while performing simulated full evaluations of patients with coronavirus disease 2019. Participants' N95 respirators were coated with a fluorescent dye to evaluate any accidental fomite transfer that occurred during PPE donning and doffing. Participants were evaluated using blacklight after each doffing encounter to evaluate new contamination sites, and were assessed for the cumulative surface area that occurred due to PPE doffing. Additionally, participants' workstations were evaluated for contamination. RESULTS: All participants experienced some contamination on their upper extremities, neck and face. The highest cumulative area of fomite transfer risk was associated with the hook and paper bag storage methods, and the least contamination occurred with the tabletop storage method. Storing a reused N95 respirator on a tabletop was found to be a safer alternative than the current recommendation of the US Centers for Disease Control and Prevention to use a paper bag for storage. All participants donning and doffing PPE were contaminated. CONCLUSION: PPE reusage practices pose an unacceptably high level of risk of accidental cross-infection contamination to healthcare workers. The current design of PPE requires complete redesign with improved engineering and usability to protect healthcare workers.

Multi-material cellular structured orthopedic implants design: In vitro and bio-tribological performance.

In this study, Selective Laser Melting (SLM) was used to produce mono-material Ti64Al4V- and NiTi-cubic cellular structures with an open-cell size and wall thickness of 500 µm and 100 µm, respectively. Bioactive beta-tricalcium phosphate (ßTCP) and polymer poly-ether-ether ketone (PEEK) were used to fill the produced structures open-cells, thus creating multi-material components. These structures were characterized in vitro in terms of cell viability, adhesion, differentiation and mineralization. Also, bio-tribological experiments were performed against bovine plate to mimic the moment of implant insertion. Results revealed that metabolic activity and mineralization were improved on SLM mono-material groups, when compared to the control group. All cell metrics were improved with the addition of PEEK, conversely to ßTCP where no significant differences were found. These results suggest that the proposed solutions can be used to improve implants performance.

Neuroendocrine pathways at risk? Simvastatin induces inter and transgenerational disruption in the keystone amphipod Gammarus locusta.

The primary focus of environmental toxicological studies is to address the direct effects of chemicals on exposed organisms (parental generation - F0), mostly overlooking effects on subsequent non-exposed generations (F1 and F2 - intergenerational and F3 transgenerational, respectively). Here, we addressed the effects of simvastatin (SIM), one of the most widely prescribed human pharmaceuticals for the primary treatment of hypercholesterolemia, using the keystone crustacean Gammarus locusta. We demonstrate that SIM, at environmentally relevant concentrations, has significant inter and transgenerational (F1 and F3) effects in key signaling pathways involved in crustaceans' neuroendocrine regulation (Ecdysteroids, Catecholamines, NO/cGMP/PKG, GABAergic and Cholinergic signaling pathways), concomitantly with changes in apical endpoints, such as depressed reproduction and growth. These findings are an essential step to improve hazard and risk assessment of biological active compounds, such as SIM, and highlight the importance of studying the transgenerational effects of environmental chemicals in animals' neuroendocrine regulation.

Bos taurus haplotypes segregating in Nellore (Bos indicus) cattle.

Brazil is the largest exporter of beef in the world, and most of that beef derives from Nellore cattle. Although considered a zebu breed (Bos indicus), the history of Nellore cattle in Brazil is marked by the importation of bulls from India, the use of a Creole taurine (Bos taurus) maternal lineage to quickly expand the herds and backcrossing to Nellore bulls to recover zebu ancestry. As a consequence, the current Brazilian Nellore population carries an average taurine ancestry of approximately 1%. Although that percentage seems small, some taurine variants deviate substantially from that average, with the better-known cases being the PLAG1-Q haplotype involved with body size variation and the Guarani (PG ) polled variant producing hornless animals. Here, we report taurine haplotypes in 9074 Nellore animals genotyped for 539 657 imputed SNP markers. Apart from PLAG1-Q and PG , our analysis further revealed common taurine haplotypes (>3%) spanning genes related to immunity, growth, reproduction and hair and skin phenotypes. Using data from 22 economically important traits, we showed that many of the major QTL previously reported in the breed are at least partially driven by taurine haplotypes. As B. taurus and B. indicus haplotypes are highly divergent, presenting widely different sets of functional variants, our results provide promising targets for future scrutiny in Nellore cattle.

Additive Manufacturing Tools to Improve the Performance of Chromatographic Approaches.

Chromatography is widely applied industrially. However, some limitations are associated with its common supports, and the impossibility to fully control their structural features is particularly restrictive. Additive manufacturing (AM) is emerging as a fast, highly precise, and reproducible technology for producing chromatographic supports that can improve its performance.

Nuclear maturation kinetics and in vitro fertilization of immature bovine oocytes injected into pre-ovulatory follicles.

The maturation kinetics and in vitro fertilization of immature bovine oocytes injected by the intra-follicular oocyte injection (IFOT) technique into pre-ovulatory follicles of previously synchronized cows were evaluated. In Experiment 1, grade I, II and III cumulus-oocyte complexes (COCs) were randomly distributed to one of three Groups: Matvitro22 (COCs matured in vitro for 22 h), MatFol20 and MatFol28 (COCs matured in vivo after being injected into a pre-ovulatory follicle of previously synchronized cows for 19.8 ± 0.1 h and 28.3 ± 0.1 h, respectively). Cows received 12.5 mg of LH (Lutropin, Bioniche, Canada) at the time of IFOT in the MatFol20 Group or 10 h after IFOT in the MatFol28 Group. MatFol20 and MatFol28 COCs were aspirated approximately 20 h after the LH injection for nuclear maturation kinetics and recovery rate assessment. In Experiment 2, grade I, II, and III COCs were randomly distributed into two Groups: Matvitro22 Group, COCs were matured and fertilized in vitro, and MatFol20 Group, COCs were matured as in the MatFol20 Group in Experiment 1, but COCs were fertilized in vitro. Putative zygotes were classified as fertilized, unfertilized or polyspermic. In Experiment 1, the recovery rate was lower (P < 0.001) in the MatFol20 Group (52.9%, 91/172) compared with MatFol28 (72.9%, 113/155). Rate of oocytes in germinal vesicle stage, metaphase I, anaphase I and telophase I were similar among Groups. However, oocytes matured in vivo for 28.3 h had lower rate of metaphase II (P = 0.001) and greater rates of degenerated (P = 0.001) and parthenogenetically activated (P = 0.001) oocytes. In experiment 2, the rates of polyspermy and degenerated were similar between Groups. However, the rate of fertilized oocytes was greater (P = 0.05) in oocytes in the MatFol20 Group. It is concluded that oocyte in vivo maturation for 19.8 h after IFOT does not compromise the nuclear maturation kinetics and increases in vitro fertilization rates. However, the extra 10 h of intra-follicular incubation time decreased oocyte viability.

Total Energy Expenditure and Functional Status in Older Adults: A Doubly Labelled Water Study.

OBJECTIVES: Higher total energy expenditure in free living conditions, regardless of any activity, has been strongly associated with a lower risk of mortality in healthy older adults. Also, a good performance in physical and functional tests is a marker of good functional prognosis. However, it is not yet clear what is the association between total energy expenditure and the performance in physical and functional tests. The objective of this study was to verify the association between the total energy expenditure of older adults measured by doubly labelled water and the performance in functional tests. DESIGN: Cross-sectional study. SETTINGS AND PARTICIPANTS: Fifty-six older people were recruited from health services linked to the participating institutions. MEASUREMENTS: Socio-demographic, anthropometric and clinical characteristics were assessed through the application of a structured questionnaire. Body composition was evaluated by isotopic dilution of deuterium oxide and functional status was assessed by the gait speed test, 6-minute walk test and handgrip strength. Total energy expenditure (GET) was assessed using the doubly labelled water method and the physical activity profile was verified using an activity monitor based on accelerometery. RESULTS: The results showed that the highest total energy expenditure correlated with the best performance in the gait speed tests (r = 0.266; p = 0.047), 6-minute walk test (r = 0.424; p = 0.001) and maximum handgrip strength (r = 0.478; p = 0.000). Multivariate regression analysis in a model adjusted for sex and fat-free mass revealed an association between total energy expenditure and the 6-minute walk test (ß = 1.790; t = 2.080; p = 0.044) and the number of sedentary events ( ß = 6.389; t = 2.147; p = 0.038). CONCLUSION: The results of this study suggest that, in clinical practice, older individuals with lower gait speed, worse performance in the 6-minute walk test and lower handgrip strength, may have lower total energy expenditure, being the stimulus for its increase important for the prevention of possible problems related to low energy expenditure.

Pyroligneous acid from Mimosa tenuiflora and Eucalyptus urograndis as an antimicrobial in dairy goats.

AIMS: To evaluate the applicability of the Mimosa tenuiflora and Eucalyptus urograndis pyroligneous acids (PAs) as alternative antiseptics in dairy goats. METHODS AND RESULTS: Cytotoxicity was evaluated in vitro using bacteria, as well as in vivo using goats, and the influence of PAs on the physicochemical parameters of fresh milk were examined. The cytotoxicity of PAs was evaluated in terms of morphology, cell viability and metabolic activity of goat tegumentary cells. The PA of M. tenuiflora had results similar to those of 2% iodine. For the in vitro tests, strains of Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa were used with the well technique, demonstrating inhibition halos greater than 9 mm. In the in vivo test, 15 animals were used per phase of the experiment, and the plate counting technique showed that there was antiseptic action of both extracts, with emphasis on the M. tenuiflora PA. Physicochemical analysis of the milk showed that neither PAs interfered with its physical-chemical parameters. CONCLUSIONS: The PA of M. tenuiflora presented potential as an alternative antiseptic in dairy goats. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates the use of PA as an antimicrobial agent in animals.

Selective Laser Melting of Ti6Al4V sub-millimetric cellular structures: Prediction of dimensional deviations and mechanical performance.

Ti6Al4V sub-millimetric cellular structures arise as promising solutions concerning the progress of conventional orthopedic implants due to its ability to address a combination of mechanical, physical and topological properties. Such ability can improve the interaction between implant materials and surrounding bone leading to long-term successful orthopedic implants. Selective Laser Melting (SLM) capability to produce high quality Ti6Al4V porous implants is in great demand towards orthopedic biomaterials. In this study, Ti6Al4V cellular structures were designed, modeled, SLM produced and characterized targeting orthopedic implants. For that purpose, a set of tools is proposed to overcome SLM limited accuracy to produce porous biomaterials with desired dimensions and mechanical properties. Morphological analyses were performed to evaluate the dimensional deviations noticed between the model CAD and the SLM produced structures. Tensile tests were carried out to estimate the elastic modulus of the Ti6Al4V cellular structures. The present work proposes a design methodology showing the linear correlations found for the dimensions, the porosity and the elastic modulus when comparing the model CAD designs with Ti6Al4V structures by SLM.

Engineering the elastic modulus of NiTi cellular structures fabricated by selective laser melting.

Nickel-titanium (NiTi) cellular structures are a very promising solution to some issues related to orthopaedic implant failure. These structures can be designed and fabricated to simultaneously address a combination of mechanical and physical properties, such as elastic modulus, porosity, wear and corrosion resistance, biocompatibility and appropriate biological environment. This ability can enhance the modest interaction currently existing between metallic dense implants and surrounding bone tissue, allowing long-term successful orthopaedic implants. For that purpose, NiTi cellular structures with different levels of porosity intended to reduce the elastic modulus were designed, modelled, selective laser melting (SLM) fabricated and characterized. Significant differences were found between the CAD design and the SLM-produced NiTi structures by performing systematic image analysis. This work proposes designing guidelines to anticipate and correct the systematic differences between CAD and produced structures. Compressive tests were carried out to estimate the elastic modulus of the produced structures and finite element analyses were performed, for comparison purposes. Linear correlations were found for the dimensions, porosity, and elastic modulus when comparing the CAD design with the SLM structures. The produced NiTi structures exhibit elastic moduli that match that of bone tissue, which is a good indication of the potential of these structures in orthopaedic implants.

Transgenerational inheritance of chemical-induced signature: A case study with simvastatin.

The hypothesis that exposure to certain environmental chemicals during early life stages may disrupt reproduction across multiple non-exposed generations has significant implications for understanding disease etiology and adverse outcomes. We demonstrate here reproductive multi and transgenerational effects, at environmentally relevant levels, of one of the most prescribed human pharmaceuticals, simvastatin, in a keystone species, the amphipod Gammarus locusta. The transgenerational findings has major implications for hazard and risk assessment of pharmaceuticals and other contaminants of emerging concern given that transgenerational effects of environmental chemicals are not addressed in current hazard and risk assessment schemes. Considering that the mevalonate synthesis, one of the key metabolic pathways targeted by simvastatin, is highly conserved among metazoans, these results may also shed light on the potential transgenerational effects of simvastatin on other animals, including humans.

Composition of semen and foreskin mucosa aerobic microbiota and its impact on sperm parameters of captive collared peccaries (Pecari tajacu).

AIM: To evaluate the bacterial composition of collared peccary semen and foreskin mucosa, and to verify the sensitivity of isolates to antimicrobials used in semen conservation and to Aloe vera gel, which is an alternative external cryoprotectant. METHODS AND RESULTS: Nine foreskin mucosa and ejaculate samples from adult animals were used. Sperm characteristics and bacterial load were evaluated in fresh semen. The preputial mucosa and semen bacterial isolates were identified and tested against five concentrations of each antimicrobial (streptomycin-penicillin and gentamicin) and A. vera gel. Corynebacterium sp. and Staphylococcus sp. were isolated in greater numbers than others in both semen (64·10 and 20·51%, respectively) and the foreskin mucosa (60·60 and 24·25%, respectively), and ranged from 0·4 to 21 × 105 colony-forming units (CFU) per ml. The average load of Corynebacterium sp. was negatively correlated (P < 0·05) with the sperm membrane integrity (r = -0·73055) and curvilinear velocity (r = -0·69048). Streptomycin-penicillin and gentamicin inhibited most micro-organisms, and A. vera showed lower antimicrobial activity. CONCLUSION: Several Gram-positive bacteria are present in semen and foreskin mucosa of collared peccary, and the benefits of using primarily penicillin-streptomycin and gentamicin antimicrobials in the bacterial control of diluted semen of these animals are strongly indicated. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides insight into the reproductive microbiota of captive male-collared peccary. This work provides a theoretical basis to assist reproductive biotechnologies for ex situ conservation of the species.

Does supplementation of vitamin C, reduced glutathione or their association in semen extender reduce oxidative stress in bovine frozen semen? / A suplementação de vitamina C, glutationa reduzida ou sua associação no diluidor de sêmen reduz o estresse oxidativo no sêmen congelado bovino?

The aim of this study was to evaluate the addition of vitamin C, reduced glutathione and the association thereof to the bovine semen cryopreservation extender. The ejaculate from nine bulls were divided into four fractions, each corresponding to a treatment, namely: control group-semen diluted with Tris-yolk extender; vitamin C group-semen diluted in Tris-yolk extender supplemented with vitamin C (2.5mmol/mL); glutathione group-semen diluted in Tris-yolk extender supplemented with reduced glutathione (2.5mmol/mL) and associated group-semen diluted in Tris-yolk extender supplemented with vitamin C (1.25mmol/mL) and reduced glutathione (1.25mmol/mL). Afterwards, the semen was packed into French straws and submitted to cryopreservation using automated equipment. After cryopreservation, the semen was thawed and evaluated considering sperm motility, morphology, plasma membrane, acrosome, mitochondrial potential and oxidative stress, as well as the thermo resistance test. Extender's supplementation with the association of vitamin C and reduced glutathione showed benefic effects on sperm motility and preservation of plasma and acrosomal membranes during semen cryopreservation, being also the group that showed higher values of reactive oxygen species. Thus, the association of both antioxidants contributed to the preservation of sperm cells in every analyzed characteristic, suggesting its use on bovine semen cryopreservation.(AU)

O objetivo deste estudo foi avaliar a adição de vitamina C, glutationa reduzida e sua associação ao diluidor de criopreservação de sêmen bovino. O ejaculado de nove touros foi dividido em quatro frações, cada uma correspondendo a um tratamento, a saber: grupo controle - sêmen diluído em Tris-gema; grupo vitamina C - sêmen diluído em Tris-gema, suplementado com vitamina C (2,5mmol/mL); grupo glutationa - sêmen diluído em Tris-gema, suplementado com glutationa reduzida (2,5mmol/mL) e grupo sêmen associado - diluído em Tris-gema, suplementado com vitamina C (1,25mmol/mL) e glutationa reduzida (1,25mmol/mL ). Posteriormente, o sêmen foi envasado em palhetas francesas e submetido à criopreservação por meio de equipamento automatizado. Após a criopreservação, o sêmen foi descongelado e avaliado quanto à motilidade espermática, à morfologia, à membrana plasmática, ao acrossoma, ao potencial mitocondrial e ao estresse oxidativo, bem como pelo teste de resistência térmica. A suplementação de extensor com a associação de vitamina C e glutationa reduzida mostrou efeitos benéficos sobre a motilidade espermática e a preservação das membranas plasmática e acrossomal durante a criopreservação de sêmen, sendo também o grupo que apresentou maiores valores de espécies reativas de oxigênio. Assim, a associação de ambos os antioxidantes contribuiu para a preservação dos espermatozóides em todas as características analisadas, sugerindo sua utilização na criopreservação de sêmen bovino.(AU)

Comparative study of osteoarthritis induced by monoiodoacetate and papain in rabbit temporomandibular joints: macroscopic and microscopic analysis.

BACKGROUND: Osteoarthritis (OA) is a chronic, progressive, degenerative pathology. Inducing OA in an animal model is useful for studying the pathology and testing the effectiveness of new treatments. The object of the present study was to determine the macroscopic and microscopic changes occurring in rabbit temporomandibular joints (TMJ) at 15, 30 and 45 days after induction of OA by monoiodoacetate (MIA) and papain. MATERIALS AND METHODS: Twenty two male rabbits were used in the experiment, divided into three groups: a control group (n = 4) and two experimental groups, MIA (n = 9) and papain (n = 9). The progress of the disease was analysed at 15, 30 and 45 days after induction of OA. Morphological and histological analyses were carried out of the joint disc and the mandibular condyle. RESULTS: The most evident changes were expressed in the condyle and disc of joints with OA induced by MIA. The condyles presented deformation, fissures and loss of joint surface, the chondrocytes lost their morphology and organisation. In more advanced stages there was loss of the mid zone of the joint disc. CONCLUSIONS: The effects of papain were associated with condyle deformation, disorientation of the chondrocytes in the middle layer, and proliferation in deep zones; there was also an increase in the extracellular matrix. Both inductors generated changes in the TMJ and its joint surfaces; MIA was more effective and coincided more closely with the classic signs of the evolution of OA.

Structural Insights into the Ligand Binding Domain of the Glucocorticoid Receptor: A Molecular Dynamics Study.

The glucocorticoid receptor (GR) is a ligand-binding dependent transcription factor that ultimately regulates vital biological processes and inflammation response through specific gene expression control, thus representing a notable drug target to explore. Structurally, its ligand binding domain (LBD) harbors the region for the ligand-dependent transcriptional activation function 2 (AF-2), a majorly hydrophobic groove formed by residues from helices H3, H4, and H12, where the H12 position plays a critical role in AF-2 spatial conformation and GR function as a whole. However, the exact mechanisms underlying how regulatory ligands control the H12 structure and dynamics are yet to be elucidated. In this work, we have explored the correlation between ligand identity and GR LBD H12 behavior through different molecular dynamics (MD) simulations. After building diverse GR LBD systems in agonist and nonagonist states, we studied each system's response in the absence or the presence of an agonist ligand (dexamethasone) or an antagonist ligand (RU486) using classical MD simulations. We complemented them with steered MD for assessing the transition between those states and with the Umbrella Sampling method for free-energy evaluation. On the one hand, successfully obtaining fully folded nonagonist GR LBD states from the partially unfolded crystal GR LBD/RU486 underlines the role of the H1 in the GR LBD folding pathway. On the other hand, our results describe the H12 as a dynamic ensemble of conformations whose relative population is in the end determined by the interacting ligand: while dexamethasone privileges only a few poses (determined by a potential energy surface with a deep minimum), RU486 favors a wider H12 conformational amplitude, as indicated by a flatter potential landscape. By characterizing the H12 conformation in different conditions, we provide novel GR LBD models that represent potential targets for rational glucocorticoid drugs design, with the aim of accurately modulating GR activity.

Additive manufactured porous biomaterials targeting orthopedic implants: A suitable combination of mechanical, physical and topological properties.

Orthopedic implants are under incessant advancement to improve their interactions with surrounding bone tissue aiming to ensure successful outcomes for patients. A successful biological interaction between implant and surrounding bone depends on the combination of mechanical, physical and topological properties. Hence, Ti6Al4V cellular structures appear as very promising solutions towards the improvement of conventional orthopedic implants. This work addresses a set of fundamental tools that allow improving the design of Ti6Al4V cellular structures produced by Selective Laser Melting (SLM). Three-point bending tests were carried out to estimate the elastic modulus of the produced structures. Morphological analysis allowed to evaluate the dimensional differences that were noticed between the model CAD and the SLM structures. Finite element models (adjusted CAD) were constructed with the experimentally obtained dimensions to replicate the mechanical response of the SLM structures. Linear correlations were systematically found for the dimensions of the SLM structures as a function of the designed model CAD dimensions. This has also been observed for the measured porosities as a function of the designed CAD models. This data can be used in further FE analyses as design guidelines to help engineers fabricating near-net-shape SLM Ti6Al4V cellular structures. Besides, polished and sandblasted surface treatments performed on the Ti6Al4V cellular structures allowed to obtain suitable properties regarding roughness and wettability when compared to as-produced surfaces. The capillarity tests showed that all the analyzed Ti6Al4V structures are able to transport fluid along its structure. The cell viability tests demonstrate Ti6Al4V cellular structures SLM produced did not release toxic substances to the medium, indicating that these structures can assure a suitable environment for cells to proliferate and attach. This study proposes a design methodology for Ti6Al4V cellular structures, that owe suitable mechanical properties but also provide a proper combination of porosity, roughness, wettability, capillarity and cell viability, all of them relevant for orthopedic applications. A Ti6Al4V cellular structured hip implant prototype gathering the suitable features addressed in this study was successfully SLM-produced.

Composite Central Face Design-An Approach to Achieve Efficient Alginate Microcarriers.

Microparticulated drug delivery systems have been used as promising encapsulation systems for protecting drugs for in vitro and in vivo applications, enhancing its stability, providing an increased surface to volume ratio, reducing adverse effects, and hence an improvement in bioavailability. Among the studied microparticles, there is a rising interest in the research of alginate microparticles for pharmaceutical and biomedical fields confirming its potential to be used as an effective matrix for drug and cell delivery. Moreover, calcium alginate has been one of the most extensively forming microparticles in the presence of divalent cations providing prolonged drug release and suitable mucoadhesive properties. Regarding the above mentioned, in this research work, we intended to produce Ca-alginate micro-vehicles through electrospraying, presenting high encapsulation efficiency (EE%), reduced protein release across the time, reduced swelling effect, and high sphericity coefficient. To quickly achieve these characteristics and to perform an optimal combination among the percentage of alginate and CaCl2, design of Experiments was applied. The obtained model presented to be statistically significant (p-value < 0.05), with a coefficient of determination of 0.9207, 0.9197, 0.9499, and 0.9637 for each output (EE%, release, swelling, and sphericity, respectively). Moreover, the optimal point (4% of alginate and 6.6% of CaCl2) was successfully validated.