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The recapitulation of complex microenvironments that regulate cell behavior during development, disease, and wound healing is key to understanding fundamental biological processes. In vitro, multicellular morphogenesis, organoid maturation, and disease modeling have traditionally been studied using either non-physiological 2D substrates or 3D biological matrices, neither of which replicate the spatiotemporal biochemical and biophysical complexity of biology. Here, we provide a guided overview of the recent advances in the programming of synthetic hydrogels that offer precise control over the spatiotemporal properties within cellular microenvironments, such as advances in the control of cell-driven remodeling, bioprinting, or user-defined manipulation of properties (e.g., via light irradiation).

Polymethylmethacrylate bone cement is often used to reconstruct critical-sized defects generated by surgical resection of spinal metastases. Residual tumor cells after a resection can drive recurrence and destabilization. Doxorubicin (DOX) is a common chemotherapeutic drug with unwanted side-effects when administered systemically. Mesoporous silica nanoparticles are gaining attention for targeted drug delivery to bypass the negative side effects associated with systemic drug administration. We developed a nanoparticle-functionalized cement for the local release of DOX and tested its ability to suppress cancer cells. DOX was loaded onto nanoparticles which were then mixed into the cement. Drug release profiles were obtained over a period of 4 weeks. Cement constructs were incubated with 2D and 3D cultures of breast and prostate cancer cell lines, and cell metabolic activity and viability were evaluated. Cell migration and spheroid growth were assessed in collagen-coated spheroid cultures. Nanoparticles were homogenously dispersed and did not alter cement mechanical strength. A sustained DOX release profile was achieved with the addition of nanoparticles to the bone cement. The release profile of DOX from nanoparticle cement may be modified by varying the amount of the drug loaded onto the nanoparticles and the proportion of nanoparticles in the cement. Cells treated with the cement constructs showed a dose- and time-dependent inhibition. Cell migration and spheroid growth were impaired in 3D culture. We show that nanoparticles are essential for sustained DOX release from bone cement. DOX-loaded nanoparticle cement can inhibit cancer cells and impair their migration, with strong potential forin vivotranslation studies.

BACKGROUND: Real world patterns of cannabis use for health concerns are highly variable and rarely overseen by a physician. Pragmatic effectiveness studies with electronic daily diaries that capture person-specific patterns of cannabis use and health symptoms may help clarify risks and benefits. METHODS: As part of a larger, randomized trial (NCT03224468), adults (N = 181) seeking cannabis for insomnia, pain, or anxiety or depressive symptoms were randomized to obtain a medical cannabis card immediately (MCC) or a waitlist control (WLC) and completed 12-weeks of daily web-based surveys on cannabis use and sleep, pain, and depressive symptoms. RESULTS: Completion rates of daily surveys were moderate to high (median completed: 72 out of 90 days). Daily reports of cannabis use were consistent with monthly interview assessments and urinalysis. The MCC group increased cannabis use frequency in the 12 weeks following randomization, while WLC did not. Among the MCC group, self-reported sleep quality was significantly higher on cannabis use days, compared to nonuse days. The MCC group displayed long-term sleep improvements, consistent with increasing cannabis frequency. No improvements were found for pain or depressive symptoms. CONCLUSION: Cannabis use is associated with same day improvements in self-reported sleep quality, but not pain or depressive symptoms, although sleep improvements occurred in the context of increased frequency of cannabis use, raising the risk for cannabis use disorder. Daily web-based assessments of cannabis appear valid and feasible in adults seeking cannabis for health concerns, providing a flexible, complementary method for future real-world effectiveness studies with expanded and objective measures.

Osteochondritis dissecans (OCD) is a chronic disease of the articular cartilage characterized by focal lesions of subchondral bone and overlaying cartilage. Through the growing number of reports describing the high prevalence of OCD in some families, the subcategory termed familial OCD (FOCD) was established. With the development of genetic approaches such as genome-wide association studies and sequencing, aggrecan (ACAN) has been identified as one of the genes of interest associated with FOCD. Aggrecan is a crucial protein for the preservation and function of cartilage. However, due to FOCD being characterized relatively recently, there is a paucity of literature on the subject. The purpose of this review is to explore the relationship between ACAN mutations and familial OCD as well as to explore current treatment options and avenues for future research. In vitro and animal studies have shown the importance of ACAN in the preservation of cartilage. However, the only human ACAN mutation related to OCD ever identified is a V2303M mutation in the G3 domain. Multiple treatments have been superficially explored, and some options such as growth hormone (GH) and gonadotrophin-releasing hormone agonists (GnRHa) show potential. Thus, further research on FOCD in needed to identify other ACAN mutations and determine optimal treatment modalities for this patient population.

Despite the legalization and widespread use of cannabis products for a variety of medical concerns in the US, there is not yet a strong clinical literature to support such use. The risks and benefits of obtaining a medical marijuana card for common clinical outcomes are largely unknown.To evaluate the effect of obtaining a medical marijuana card on target clinical and cannabis use disorder (CUD) symptoms in adults with a chief concern of chronic pain, insomnia, or anxiety or depressive symptoms.This pragmatic, single-site, single-blind randomized clinical trial was conducted in the Greater Boston area from July 1, 2017, to July 31, 2020. Participants were adults aged 18 to 65 years with a chief concern of pain, insomnia, or anxiety or depressive symptoms. Participants were randomized 2:1 to either the immediate card acquisition group (n = 105) or the delayed card acquisition group (n = 81). Randomization was stratified by chief concern, age, and sex. The statistical analysis followed an evaluable population approach.The immediate card acquisition group was allowed to obtain a medical marijuana card immediately after randomization. The delayed card acquisition group was asked to wait 12 weeks before obtaining a medical marijuana card. All participants could choose cannabis products from a dispensary, the dose, and the frequency of use. Participants could continue their usual medical or psychiatric care.Primary outcomes were changes in CUD symptoms, anxiety and depressive symptoms, pain severity, and insomnia symptoms during the trial. A logistic regression model was used to estimate the odds ratio (OR) for CUD diagnosis, and linear models were used for continuous outcomes to estimate the mean difference (MD) in symptom scores.A total of 186 participants (mean [SD] age 37.2 [14.4] years; 122 women [65.6%]) were randomized and included in the analyses. Compared with the delayed card acquisition group, the immediate card acquisition group had more CUD symptoms (MD, 0.28; 95% CI, 0.15-0.40; P .001); fewer self-rated insomnia symptoms (MD, -2.90; 95% CI, -4.31 to -1.51; P .001); and reported no significant changes in pain severity or anxiety or depressive symptoms. Participants in the immediate card acquisition group also had a higher incidence of CUD during the intervention (17.1% [n = 18] in the immediate card acquisition group vs 8.6% [n = 7] in the delayed card acquisition group; adjusted odds ratio, 2.88; 95% CI, 1.17-7.07; P = .02), particularly those with a chief concern of anxiety or depressive symptoms.This randomized clinical trial found that immediate acquisition of a medical marijuana card led to a higher incidence and severity of CUD; resulted in no significant improvement in pain, anxiety, or depressive symptoms; and improved self-rating of insomnia symptoms. Further investigation of the benefits of medical marijuana card ownership for insomnia and the risk of CUD are needed, particularly for individuals with anxiety or depressive symptoms.ClinicalTrials.gov Identifier: NCT03224468.

The enthesis is the interfacial tissue between ligament or tendon, and bone, which connects tissues of distinctly different mechanical properties. Although ligament and enthesis injury is commonplace, the development and healing mechanisms of these tissues are yet unclear. Current models for investigating these mechanisms are primarily in vivo animal models as in vitro models have been limited. In this study, an in vitro enthesis model was developed using a modified gel aspiration-ejection (GAE) method. Continuous two-phase aligned dense collagen (ADC) hydrogels with an overlapping interface were fabricated within 2 hours. The mechanical properties of acellular two-phase ADC confirmed the continuous nature of this model, as the mechanical properties showed no significant difference compared to single-phase ADC and maintained comparable structural and mechanical characteristics of immature ligaments and unmineralized bone. Human anterior cruciate ligament (ACL) fibroblasts and human spine vertebral osteoblasts were isolated from donor tissues and were seeded to form the enthesis model. These were cultured for 14 days, at which the viability and proliferation was observed to be 85 ± 7.5% and 230 ± 52%, respectively. Histological and immunofluorescence analyses at day 14 revealed extensive cell-driven matrix remodelling, and the seeded fibroblasts and osteoblasts maintained their phenotype within their compartments of the layered co-culture model. These results demonstrate the rapid fabrication of a two-phase co-culture system that can be utilized as an in vitro model to further understand the degenerative and regenerative mechanisms within the enthesis.

Tooth sensitivity is a painful and very common problem. Often stimulated by consuming hot, cold, sweet, or acidic foods, it is associated with exposed dentin microtubules that are open to dental pulp. One common treatment for tooth hypersensitivity is the application of occlusive particles to block dentin microtubules. The primary methodology currently used to test the penetration and occlusion of particles into dentin pores relies upon dentin discs cut from extracted bovine/human teeth. However, this method is limited due to low accessibility to the raw material. Thus, there is a need for an in vitro dentin model to characterize the effectiveness of occlusive agents. Three-dimensional printing technologies have emerged that make the printing of dentin-like structures possible. This study sought to develop and print a biomaterial ink that mimicked the natural composition and structure of dentin tubules. A formulation of type I collagen (Col), nanocrystalline hydroxyapatite (HAp), and alginate (Alg) was found to be suitable for the 3D printing of scaffolds. The performance of the 3D printed dentin model was compared to the natural dentin disk by image analysis via scanning electron microscopy (SEM), both pre- and post-treatment with occlusive microparticles, to evaluate the degree of dentinal tubule occlusion. The cytocompatibility of printed scaffolds was also confirmed in vitro. This is a promising biomaterial system for the 3D printing of dentin mimics.

Background Despite it being known that subchondral bone affects the viscoelasticity of cartilage, there has been little research into the mechanical properties of osteochondral tissue as a whole system. This study aims to unearth new knowledge concerning the dynamic behaviour of human subchondral bone and how energy is transferred through the cartilage-bone interface. Methods Dynamic mechanical analysis was used to determine the frequency-dependent (1–90 Hz) viscoelastic properties of the osteochondral unit (cartilage-bone system) as well as isolated cartilage and bone specimens extracted from human femoral heads obtained from patients undergoing total hip replacement surgery, with a mean age of 78 years (N = 5, n = 22). Bone mineral density (BMD) was also determined for samples using micro-computed tomography as a marker of tissue health. Results Cartilage storage and loss moduli along with bone storage modulus were found to increase logarithmically (p p − 1, while the loss stiffness was determined to be frequency-independent at 78.84 ± 2.48 N.mm− 1. Notably, a statistically significant (p p = 0.09). Conclusions The viscoelastic properties of the cartilage-bone core were significantly different to the tissues in isolation (p

Study Design An observational descriptive study based on a single cohort of patients. Objective To determine whether spinal facet osteoblasts at the curve apex display a different phenotype to osteoblasts from outside the curve in adolescent idiopathic scoliosis (AIS) patients. Summary of Background Data Intrinsic differences in the phenotype of spinal facet bone tissue and in spinal osteoblasts have been implicated in the pathology of AIS. However, no study has compared the phenotype of facet osteoblasts at the curve apex compared with outside the curve in AIS patients. Methods Facet spinal tissue was collected perioperatively from three sites, the concave and convex side at the curve apex and from outside the curve (noncurve) from three AIS female patients aged 13–16 years. Spinal tissue was analyzed by micro–computed tomography to determine bone mineral density (BMD) and trabecular structure. Primary osteoblasts were cultured from concave, convex, and noncurve facet bone chips. The phenotype of osteoblasts was determined by assessment of cellular proliferation, cellular metabolism (alkaline phosphatase and Seahorse Analyzer), bone nodule mineralization (Alizarin red assay), and the mRNA expression of Wnt signaling genes (quantitative reverse transcriptase polymerase chain reaction). Results Convex facet tissue exhibited greater BMD and trabecular thickness, compared with concave facet tissue. Osteoblasts at the convex side of the curve apex exhibited a significantly higher proliferative and metabolic phenotype and a greater capacity to form mineralized bone nodules, compared with concave osteoblasts. mRNA expression of SKP2 was significantly greater in both concave and convex osteoblasts, compared with noncurve osteoblasts. The expression of SFRP1 was significantly downregulated in convex osteoblasts, compared with either concave or noncurve. Conclusions Intrinsic differences that affect osteoblast function are exhibited by spinal facet osteoblasts at the curve apex in AIS patients. Level of Evidence Level IV, Prognostic.

The current gold standard technique for treatment of anterior cruciate ligament (ACL) injury is reconstruction with autograft. These treatments have a relatively high failure and re-tear rate. To overcome this, tissue engineering and additive manufacturing are being used to explore the potential of 3D scaffolds as autograft substitutes. However, mechanically optimal polymers for this have yet to be identified. Here, we use 3D printing technology and various materials with the aim of fabricating constructs better matching the mechanical properties of the native ACL. A fused deposition modeling (FDM) 3D printer was used to microfabricate dog bone-shaped specimens from six different polymers&mdash, PLA, PETG, Lay FOMM 60, NinjaFlex, NinjaFlex-SemiFlex, and FlexiFil&mdash, at three different raster angles. The tensile mechanical properties of these polymers were determined from stress&ndash, strain curves. Our results indicate that no single material came close enough to successfully match reported mechanical properties of the native ACL. However, PLA and PETG had similar ultimate tensile strengths. Lay FOMM 60 displayed a percentage strain at failure similar to reported values for native ACL. Furthermore, raster angle had a significant impact on some mechanical properties for all of the materials except for FlexiFil. We therefore conclude that while none of these materials alone is optimal for mimicking ACL mechanical properties, there may be potential for creating a 3D-printed composite constructs to match ACL mechanical properties. Further investigations involving co-printing of stiff and elastomeric materials must be explored.

The current gold standard technique for treatment of anterior cruciate ligament (ACL) injury is reconstruction with autograft. These treatments have a relatively high failure and re-tear rate. To overcome this, tissue engineering and additive manufacturing are being used to explore the potential of 3D scaffolds as autograft substitutes. However, mechanically optimal polymers for this have yet to be identified. Here, we aimed to generate a functional substitute to better match the mechanical properties of the native ACL. A fused deposition modeling (FDM) 3D printer was used to microfabricate specimens from six different polymers, PLA, PETG, Lay FOMM 60, NinjaFlex, NinjaFlex-SemiFlex, and FlexiFil, at three different raster angles. The tensile mechanical properties of these polymers were determined from stress-strain curves. Our results indicate that no single material came close enough to successfully match reported mechanical properties of the native ACL. However, PLA and PETG had similar ultimate tensile strengths. Lay FOMM 60 displayed a percentage strain at failure similar to reported values for native ACL. Furthermore, raster angle had a significant impact on some mechanical properties for all of the materials except for FlexiFil. We therefore conclude that while none of these materials alone is optimal for mimicking ACL properties, there may be potential for creating a 3D printed composite constructs to match ACL mechanical properties. Further investigations involving co-printing of stiff and elastomeric materials must be explored.

Improving cardiovascular fitness may buffer against age-related cognitive decline and mitigate dementia risk by staving off brain atrophy. However, it is unclear if such effects reflect factors operating in childhood (neuroselection) or adulthood (neuroprotection). Using data from 807 members of the Dunedin Study, a population-representative birth cohort, we investigated associations between cardiovascular fitness and structural brain integrity at age 45, and the extent to which associations reflected possible neuroselection or neuroprotection by controlling for childhood IQ. Higher fitness, as indexed by VO2Max, was not associated with average cortical thickness, total surface area, or subcortical gray matter volume including the hippocampus. However, higher fitness was associated with thicker cortex in prefrontal and temporal regions as well as greater cerebellar gray matter volume. Higher fitness was also associated with decreased hippocampal fissure volume. These associations were unaffected by the inclusion of childhood IQ in analyses. In contrast, a higher rate of decline in cardiovascular fitness from 26 to 45 years was not robustly associated with structural brain integrity. Our findings are consistent with a neuroprotective account of adult cardiovascular fitness but suggest that effects are not uniformly observed across the brain and reflect contemporaneous fitness more so than decline over time.

Introduction As the use of electronic cigarette (EC) continues to rise in the United States, especially among adolescents and young adults, it is necessary to better understand the factors associated with EC initiation. Specifically, it is unclear how genetic and environmental contributions influence the initiation of EC. Furthermore, the degree to which genetic and environmental influences are shared between EC initiation and conventional cigarette (CC) initiation is unknown. Methods A sample of young adult twins ages 15–20 (N = 858 individuals; 421 complete twin pairs) was used to estimate the genetic and environmental influences on the liability of initiation unique to EC and CC as well as the degree to which these factors are shared between the two. Approximately 24% of participants initiated the use of EC, 19% initiated the use of CC, and 11% initiated the dual use. Results Combined contributions of additive genetic and shared environmental influences were significant for CC (ACC = 0.19 [95% confidence interval {CI} = 0–0.79], p = 0.57; CCC = 0.42 [95% CI = 0–0.70], p = 0.13) and EC (AEC = 0.25 [95% CI = 0–0.83, p = 0.44; CEC = 0.42 [95% CI = 0–0.73], p = 0.12), whereas unique environmental influences were significant (ECC = 0.39 [95% CI = 0.18–0.57], p < 0.001; EEC = 0.32 [95% CI = 0.14–0.56], p < 0.001). Results also demonstrated a significant overlap of the unique environmental (rE = 0.87, p < 0.001) and familial influences contributing to correlation between the two phenotypes in the bivariate analysis. Conclusions These preliminary results suggest that both genes and environmental influences are potential drivers of EC initiation among adolescents and young adults. Implications This article is the first to use a sample of twin to estimate the contributions of genetic and environmental influences toward EC initiation and estimate the potential for overlapping influences with CC initiation. This study has implications for future debate about the etiology of EC and CC use with respect to potential overlapping genetic and environmental influences.

Articular cartilage is a viscoelastic tissue whose structural integrity is important in maintaining joint health. To restore the functionality of osteoarthritic joints it is vital that regenerative strategies mimic the dynamic loading response of cartilage and bone. Here, a rotating simplex model was employed to optimise the composition of agarose and gellan hydrogel constructs structured with hydroxyapatite (HA) with the aim of obtaining composites mechanically comparable to human cartilage in terms of their ability to dissipate energy. Addition of ceramic particles was found to reinforce both matrices up to a critical concentration (< 3w/v%). Beyond this, larger agglomerates were formed, as evidenced by micro computed tomography data, which acted as stress risers and reduced the ability of composites to dissipate energy demonstrated by a reduction in tan δ values. A maximum compressive modulus of 450.7±24.9 kPa was achieved with a composition of 5.8w/v% agarose and 0.5w/v% HA. Interestingly, when loaded dynamically (1-20Hz) this optimised formulation did not exhibit the highest complex modulus instead a sample with a higher concentration of mineral was identified (5.8w/v% agarose and 25w/v% HA). Thus, demonstrating the importance of examining the mechanical behaviour of biomaterials under conditions representative of physiological environments. While the complex moduli of the optimised gellan (1.0 ± 0.2MPa at 1Hz) and agarose (1.7 ± 0.2MPa at 1Hz) constructs did not match the complex moduli of healthy human cartilage samples (26.3 ± 6.5MPa at 1Hz), similar tan δ values were observed between 1 and 5Hz. This is promising since these frequencies represent the typical heel strike time of the general population. In summary, this study demonstrates the importance of considering more than just the strength of biomaterials since tissues like cartilage play a more complex role.

Microbial keratitis is a severe ocular condition and one of the most prevalent causes of corneal scarring and associated blindness worldwide. Risk factors include contact lens use, ocular trauma, ocular surface disease and immunosuppression. Initial clinical management mandates intensive (hourly or more frequent) topical administration of broad spectrum antimicrobial therapy for at least 48h, which may require hospital admission, followed by tailored therapy based on microbiological investigation and the institution of strategies to reduce inflammation and promote healing. In this work we report an ocular wound dressing which can encapsulate and give sustained release of different antibiotics. The use of this dressing would allow patients to have eye drops on a 4 hourly basis, thereby facilitating treatment compliance and reducing hospital admissions.

Introduction Electronic cigarette (e-cigarette) use prevalence is increasing among U.S. adolescents and adults but recent longitudinal data for college/university students are scarce. Furthermore, the extent that e-cigarette use is associated with the onset of cigarette smoking and the factors that lead to the uptake of e-cigarettes in college students has not been explored. Methods 3757 participants from a Mid-Atlantic university (women: 66%; White: 45%; Black: 21%; Asian: 19%; Hispanic/Latino: 6%) were surveyed in 2014 and again in 2015. Results Among participants reporting never smoking at time 1, those who had ever tried e-cigarettes or were currently using e-cigarettes (at least one use in past 30 days) were more likely to have ever tried cigarettes by time 2 relative to individuals who had not used e-cigarettes. Ever use of e-cigarettes (but not current use) also increased participants' likelihood of being current cigarette smokers at time 2. Among initial never users of e-cigarettes or cigarettes, males and ever marijuana users had an increased probability of trying e-cigarettes by time 2. Furthermore, less perseverance (an index of impulsivity) and ever use of other tobacco products increased initial never users' chances of trying both cigarettes and e-cigarettes by time 2. Conclusions Given that never-smoking participants who had tried e-cigarettes were more likely to initiate cigarette use later, limiting young adults' access to these products may be beneficial. As the long-term health implications of e-cigarette use become clearer, predictors of e-cigarette use could help identify future populations likely to use and abuse these products.

The Malvaceae family is a group of flowering plants that include approximately 244 genera, and 4225 species. Grewia mollis, and Hoheria populnea (lacebark), are examples of the Malvaceae family that are used in traditional medicine. For this study polysaccharide samples were extracted from the inner bark of Grewia mollis (unmodified (GG) and destarched grewia gum (GGDS)) and from the leaves of Hoheria populnea (lacebark polysaccharide (LB)). Wound healing properties of grewia gum and lacebark polysaccharides were investigated using 3T3 fibroblast cells cultured in supplemented DMEM. Deposition of collagen using van Gieson's stain, expression of the COL1A1 gene which encodes type I collagen using quantitative PCR, and chemotaxis using a scratch plate assay were analysed following treatment of cells with the test polysaccharides. Quantitative PCR results indicated that all three polysaccharides increased the levels of COL1A1 mRNA, with GG showing the greatest fold change. Histological staining also indicated that the fibroblasts treated with GG deposited more collagen than control cells. Additionally, scratch assay data indicated that simulated cell ‘wounds’ treated with each polysaccharide showed increased wound closure rate over a 36 h period post treatment, with GG exhibiting the greatest effect on wound closure. Analysis of the Malvaceae derived polysaccharides indicates that they could have a positive effect on mechanisms that are integral to wound healing, potentially providing greater scientific understanding behind their use in traditional medicine.

Free-form printing offers a novel biofabrication approach to generate complex shapes by depositing hydrogel materials within a temporary supportive environment. However, printed hydrogels typically lack the requisite mechanical properties and functionality of the desired tissue, limiting application and, more importantly, safety and efficacy of the implant. The study authors have developed an innovative nanoclay-based bioink to print high shape fidelity functional constructs for potential skeletal application. Laponite® (LAP) nanoclay was combined with gellan gum (GG) to generate a printable hydrogel that was highly stable in vitro, displayed limited swelling ability compared with the silicate-free control and remained stable over time. An agarose fluid gel was found to provide the requisite support for the deposition of the material ink and preservation of the printed structure before crosslinking. Printed C2C12 myoblasts remained viable and displayed extensive proliferation over 21 days in culture. Cell-laden scaffolds demonstrated functionality within 1 day of culture in vitro and that was preserved over 3 weeks. Analysis of absorption and release mechanisms from LAP-GG using model proteins (lysozyme and bovine serum albumin) demonstrated the retention capability of the clay-based materials for compound localisation and absence of burst release. Vascular endothelial growth factor ​was loaded within the agarose fluid gel and absorbed by the material ink via absorption during deposition. The 3D-printed constructs were implanted on the chorioallantoic membrane of a 10-day-old developing chick. Extensive and preferential vasculature infiltration was observed in LAP-GG–loaded vascular endothelial growth factor constructs compared with controls (p, Graphical abstract LAP-GG printing in agarose fluid gel for skeletal tissue engineering. Extrusion of LAP-GG in an agarose bed to generate three-dimensional structures (a). Low polymeric content nanosilicate bioink was used to deliver cells and preserve their viability after extrusion. Agarose fluid gel was harnessed to enable loading of compounds of interests during printing (b) in LAP-GG and GG controls. LAP-GG bioink offers the potential for enhanced compound penetration within the printed strand. LAP-GG extrusion within self-healing agarose can be used to fabricate large functional structures (c) such as 3D sleeve (i and ii) and 3D lattice (iii and iv) structures to fill critical bone defects. Cells were printed in LAP-GG (d) and viability (i and ii) and functionality (iii and iv) were assessed for skeletal tissue formation potential. Integration with chick membrane model and vascular penetration (e) of printed scaffolds implanted within the vascularised membrane of a developing chick. Scale bar: (a) 5 ​mm, (b—i and ii) 100 ​μm, (c—i and ii) 30 ​mm, (c—iii) 30 ​mm, (c—iv) 10 ​mm, (d—i) 100 ​μm, (d—iii and iv) 2 ​mm.Image 1

Additive manufacturing (AM) has emerged over the past four decades as a cost-effective, on-demand modality for fabrication of geometrically complex objects. The ability to design and print virtually any object shape using a diverse array of materials, such as metals, polymers, ceramics and bioinks, has allowed for the adoption of this technology for biomedical applications in both research and clinical settings. Current advancements in tissue engineering and regeneration, therapeutic delivery, medical device fabrication and operative management planning ensure that AM will continue to play an increasingly important role in the future of healthcare. In this review, we outline current biomedical applications of common AM techniques and materials.

BackgroundIdentifying genetic relationships between complex traits in emerging adulthood can provide useful etiological insights into risk for psychopathology. College-age individuals are under-represented in genomic analyses thus far, and the majority of work has focused on the clinical disorder or cognitive abilities rather than normal-range behavioral outcomes.MethodsThis study examined a sample of emerging adults 18–22 years of age (N = 5947) to construct an atlas of polygenic risk for 33 traits predicting relevant phenotypic outcomes. Twenty-eight hypotheses were tested based on the previous literature on samples of European ancestry, and the availability of rich assessment data allowed for polygenic predictions across 55 psychological and medical phenotypes.ResultsPolygenic risk for schizophrenia (SZ) in emerging adults predicted anxiety, depression, nicotine use, trauma, and family history of psychological disorders. Polygenic risk for neuroticism predicted anxiety, depression, phobia, panic, neuroticism, and was correlated with polygenic risk for cardiovascular disease.ConclusionsThese results demonstrate the extensive impact of genetic risk for SZ, neuroticism, and major depression on a range of health outcomes in early adulthood. Minimal cross-ancestry replication of these phenomic patterns of polygenic influence underscores the need for more genome-wide association studies of non-European populations.

Osteoarthritis (OA) is a degenerative disease that affects 25% of the world's population over fifty years of age. It is a chronic disease of the synovial joints, primarily the hip and knee. The main pathologies are degradation of the articular cartilage and changes to the subchondral bone, as a result of both mechanical wear and a locally elevated inflammatory state. This study compares the viscoelastic properties of cartilage that represents the biochemical changes in OA and age-matched healthy tissue. Further, the mechanical damage induced by this compressive loading cycle was characterised and the mechanism for it was investigated. The storage modulus of OA cartilage was shown to be significantly lower than that of healthy cartilage whilst having a higher capacity to hold water. Following mechanical testing, there was a significant increase in the surface roughness of OA cartilage. This change in surface structure occurred following a reduction in sulphated glycosaminoglycan content of the superficial region in OA, as seen by alcian blue staining and quantified by micro X-ray fluorescence. These findings are important in understanding how the chemical changes to cartilage matrix in OA influence its dynamic mechanical properties and structural integrity.Cartilage has a very specialised tissue structure which acts to resist compressive loading. In osteoarthritis (OA), there is both mechanically- and chemically-induced damage to cartilage, resulting in severe degradation of the tissue. In this study we have undertaken a detailed mechanical and chemical analysis of macroscopically undamaged OA and healthy cartilage tissue. We have demonstrated, for the first time in human tissue, that the mechanical degradation of the tissue is attributed to a chemical change across the structure. In macroscopically undamaged OA tissue, there is a reduction in the elastic response of cartilage tissue and an associated destabilisation of the matrix that leaves it susceptible to damage. Understanding this allows us to better understand the progression of OA to design better therapeutic interventions.

The purpose of this study was to address two methodological issues that have called into question whether previously reported gene–environment interaction (GxE) effects for adolescent alcohol use are ‘real’. These issues are (1) the potential correlation between the environmental moderator and the outcome across twins and (2) non-linear transformations of the behavioral outcome. Three environments that have been previously studied (peer deviance, parental knowledge, and potentially stressful life events) were examined here. For each moderator (peer deviance, parental knowledge, and potentially stressful life events), a series of models was fit to both a raw and transformed measure of monthly adolescent alcohol use in a sample that included 825 dizygotic (DZ) and 803 monozygotic (MZ) twin pairs. The results showed that the moderating effect of peer deviance was robust to transformation, and that although the significance of moderating effects of parental knowledge and potentially stressful life events were dependent on the scale of the adolescent alcohol use outcome, the overall results were consistent across transformation. In addition, the findings did not vary across statistical models. The consistency of the peer deviance results and the shift of the parental knowledge and potentially stressful life events results between trending and significant, shed some light on why previous findings for certain moderators have been inconsistent and emphasize the importance of considering both methodological issues and previous findings when conducting and interpreting GxE analyses.

A new bone augmenting material is reported, which is formed from calcium-loaded hydrogel-based spheres. On immersion of these spheres in a physiological medium, they become surrounded with a sheath of precipitate, which ruptures due to a build-up in osmotic pressure. This results in the formation of mineral tubes that protrude from the sphere surface. When brought into close contact with one another, these spheres become fused through the entanglement and subsequent interstitial mineralization of the mineral tubules. The tubular calcium phosphate induces the expression of osteogenic genes (runt-related transcription factor 2 (RUNX2), transcription factor SP7 (SP7), collagen type 1 alpha 1 (COL1A1), and bone gamma-carboxyglutamic acid-containing protein (BGLAP)) and promotes the formation of mineral nodules in preosteoblast cultures comparable to an apatitic calcium phosphate phase. Furthermore, alkaline phosphatase (ALP) is significantly upregulated in the presence of tubular materials after 10 d in culture compared with control groups (p0.001) and sintered apatite (p0.05). This is the first report of a bioceramic material that is formed in its entirety in situ and is therefore likely to provide a better proxy for biological mineral than other existing synthetic alternatives to bone grafts.

The development of new materials for clinical use is limited by an onerous regulatory framework, which means that taking a completely new material into the clinic can make translation economically unfeasible. One way to get around this issue is to structure materials that are already approved by the regulator, such that they exhibit very distinct physical properties and can be used in a broader range of clinical applications. Here, the focus is on the structuring of soft materials at multiple length scales by modifying processing conditions. By applying shear to newly forming materials, it is possible to trigger molecular reorganization of polymer chains, such that they aggregate to form particles and ribbon-like structures. These structures then weakly interact at zero shear forming a solid-like material. The resulting self-healing network is of particular use for a range of different biomedical applications. How these materials are used to allow the delivery of therapeutic entities (cells and proteins) and as a support for additive layer manufacturing of larger-scale tissue constructs is discussed. This technology enables the development of a range of novel materials and structures for tissue augmentation and regeneration.

As psychiatric genetics enters an era where gene identification is finally yielding robust, replicable genetic associations and polygenic risk scores, it is important to consider next steps and delineate how that knowledge will be applied to ultimately ameliorate suffering associated with substance use and psychiatric disorders. Much of the post-genome-wide association study discussion has focused on the potential of genetic information to elucidate the underlying biology and use this information for the development of more effective pharmaceutical treatments. In this review we focus on additional areas of research that should follow gene identification. By taking genetic findings into longitudinal, developmental studies, we can map the pathways by which genetic risk manifests across development, elucidating the early behavioral manifestations of risk, and studying how various environments and interventions moderate that risk across developmental stages. The delineation of risk across development will advance our understanding of mechanism, sex differences and risk and resilience processes in different racial/ethnic groups. Here, we review how the extant twin study literature can be used to guide these efforts. Together, these new lines of research will enable us to develop more informed, tailored prevention and intervention efforts.

Background: Genetic factors impact alcohol use behaviors and these factors may become increasingly evident during emerging adulthood. Examination of the effects of individual variants as well as aggregate genetic variation can clarify mechanisms underlying risk. Methods: We conducted genome-wide association studies (GWAS) in an ethnically diverse sample of college students for three quantitative outcomes including typical monthly alcohol consumption, alcohol problems, and maximum number of drinks in 24 hours. Heritability based on common genetic variants (h2SNP) was assessed. We also evaluated whether risk variants in aggregate were associated with alcohol use outcomes in an independent sample of young adults. Results: Two genome-wide significant markers were observed: rs11201929 in GRID1 for maximum drinks in 24 hours, with supportive evidence across all ancestry groups; and rs73317305 in SAMD12 (alcohol problems), tested only in the African ancestry group. The h2SNP estimate was 0.19 (SE=0.11) for consumption, and was non-significant for other outcomes. Genome-wide polygenic scores were significantly associated with alcohol outcomes in an independent sample. Conclusions: These results robustly identify genetic risk for alcohol use outcomes at the variant level and in aggregate. We confirm prior evidence that genetic variation in GRID1 impacts alcohol use, and identify novel loci of interest for multiple alcohol outcomes in emerging adults. These findings indicate that genetic variation influencing normative and problematic alcohol use is, to some extent, convergent across ancestry groups. Studying college populations represents a promising avenue by which to obtain large, diverse samples for gene identification.

Alcohol misuse, one of today’s greatest public health challenges, is a developmentally dynamic, complex behavior at the intersection of genetic and environmental influences. This review examines such influences from a behavior genetics perspective and discusses implications for public policy. Alcohol misuse is moderately heritable with genetic influences accounting for around 50% of its variance, but to date, few specific genes have been identified. However, numerous environmental and social factors moderate genetic risk, including parents, peers, romantic partners, family dynamics, employment, laws, and cultural influences. These moderating factors change in salience across development, and accordingly, no one-size-fits-all approach is suitable for reducing alcohol misuse at a large scale. We provide examples of some effective prevention and intervention programs and discuss a framework for using the behavior genetics evidence to inform future public policy efforts.

A method for the production of complex cell-laden structures is reported, which allows high-levels of spatial control over mechanical and chemical properties. The potential of this method for producing complicated tissues is demonstrated by manufacturing a complex hard/soft tissue interface and demonstrating that cell phenotype can be maintained over four weeks of culture.

Background The first year of university attendance represents a critical time frame for the development of alcohol use and misuse given changes in autonomy and increased access to alcohol. Prior studies have demonstrated that the establishment of drinking patterns during this period is impacted by an array of demographic, environmental, and familial factors. It is critical to consider such factors jointly, and to understand potentially differential effects on stages of alcohol use/misuse, in order to identify robust predictors that may be targeted in prevention and intervention programming. Methods As part of a longitudinal study, students at a large, public U.S. university were invited to complete online surveys that included questions related to alcohol use, emotional and behavioral health, environmental factors, sociodemographic factors, and familial environment. This study uses data from surveys administered in the fall and spring of the first year of university. We used univariate (maximum N = 7,291) and multivariate (maximum N = 4,788) logistic and linear regressions to evaluate the associations between potential risk and protective factors with 4 alcohol use outcomes: initiation, consumption, problems, and addiction resistance. Results In multivariate models, we observed associations between demographic, social/environmental, and personal-level predictors with all 4 alcohol outcomes, several of which were consistent across each stage of alcohol use. A deviant high school peer group was one of the strongest predictors of risk across outcomes. The influence of drinking motives and alcohol expectancies varied by alcohol use outcome. Externalizing characteristics were associated with increased risk across outcomes, while internalizing symptoms were associated with more problems and lower addiction resistance. Conclusions These findings underscore the complex network of factors influencing stages of alcohol use during the first year of university. Importantly, these findings demonstrate that the impact of predictors changes across stages of alcohol use/misuse, which presents opportunities for targeted prevention efforts.

OBJECTIVE The purpose of this study was to assess patterns of tobacco use across the first year of college, transitions in use, and associated predictors. METHODS The frequency of tobacco use (cigarettes, cigars, smokeless tobacco, and hookah) during the fall and spring of 4073 college students' first year at college were used as indicators in latent class (LCA) and latent transition analyses (LTA). RESULTS The LCA yielded 3 classes that represent levels of use frequency and not specific tobacco product classes: non-using, experimenting, and frequent using. The LTA results demonstrate stability in class membership from fall to spring. The most common transition was for the fall experimenters to transition out of experimentation. A series of demographic, environmental, and intrapersonal predictors were found to influence both fall class membership, and transitions from fall to spring. CONCLUSIONS Students are likely to use multiple alternative tobacco products along with cigarettes. Their frequency of use of these products is fairly stable across the first year of college. Predictors reflecting experiences during the first year of college had the greatest impact on college tobacco use, demonstrating the importance of the college experience on young adult tobacco use.