Search

The role of poroelasticity on the functional performance of articular cartilage has been established in the scientific literature since the 1960s. Despite the extensive knowledge on this topic there remain few attempts to design for poroelasticity and to our knowledge no demonstration of an engineered poroelastic material that approaches the physiological performance. In this paper, we report on the development of an engineered material that begins to approach physiological poroelasticity. We quantify poroelasticity using the fluid load fraction, apply mixture theory to model the material system, and determine cytocompatibility using primary human mesenchymal stem cells. The design approach is based on a fiber reinforced hydrated network and uses routine fabrication methods (electrohydrodynamic deposition) and materials (poly[ɛ-caprolactone] and gelatin) to develop the engineered poroelastic material. This composite material achieved a mean peak fluid load fraction of 68%, displayed consistency with mixture theory, and demonstrated cytocompatibility. This work creates a foundation for designing poroelastic cartilage implants and developing scaffold systems to study chondrocyte mechanobiology and tissue engineering. Poroelasticity drives the functional mechanics of articular cartilage (load bearing and lubrication). In this work we develop the design rationale and approach to produce a poroelastic material, known as a fiber reinforced hydrated network (FiHy™), that begins to approach the native performance of articular cartilage. This is the first engineered material system capable of exceeding isotropic linear poroelastic theory. The framework developed here enables fundamental studies of poroelasticity and the development of translational materials for cartilage repair. [Display omitted] [ABSTRACT FROM AUTHOR]

Magnesium is a trace element in the human body, known to have important effects on cell differentiation and the mineralisation of calcified tissues. This study aimed to synthesise highly porous Ca-Mg silicate foamed scaffolds from preceramic polymers, with analysis of their biological response. Akermanite (Ak) and wollastonite-diopside (WD) ceramic foams were obtained from the pyrolysis of a liquid silicone mixed with reactive fillers. The porous structure was obtained by controlled water release from selected fillers (magnesium hydroxide and borax) at 350 °C. The homogeneous distribution of open pores, with interconnects of modal diameters of 160–180 μm was obtained and maintained after firing at 1100 °C. Foams, with porosity exceeding 80%, exhibited compressive strength values of 1–2 MPa. In vitro studies were conducted by immersion in SBF for 21 days, showing suitable dissolution rates, pH and ionic concentrations. Cytotoxicity analysis performed in accordance with ISO10993-5 and ISO10993-12 standards confirmed excellent biocompatibility of both Ak and WD foams. In addition, MC3T3-E1 cells cultured on the Mg-containing scaffolds demonstrated enhanced osteogenic differentiation and the expression of osteogenic markers including Collagen Type I, Osteopontin and Osteocalcin, in comparison to Mg-free counterparts. The results suggest that the addition of magnesium can further enhance the bioactivity and the potential for bone regeneration applications of Ca-silicate materials. Statements of Significance Here, we show that the incorporation of Mg in Ca-silicates plays a significant role in the enhancement of the osteogenic differentiation and matrix formation of MC3T3-E1 cells, cultured on polymer-derived highly porous scaffolds. Reduced degradation rates and improved mechanical properties are also observed, compared to Mg-free counterparts, suggesting the great potential of Ca-Mg silicates as bone tissue engineering materials. Excellent biocompatibility of the new materials, in accordance to the ISO10993-5 and ISO10993-12 standard guidelines, confirms the preceramic polymer route as an efficient synthesis methodology for bone scaffolds. The use of hydrated fillers as porogens is an additional novelty feature presented in the manuscript. [ABSTRACT FROM AUTHOR]

The earthworm Eukerria saltensis can cause severe crop establishment problems in aerially sown rice grown on heavy clay soils in southern Australia. Damage occurs indirectly through destabilization of the topsoil, increased water turbidity, and mobilization of soil nutrients into the water column which leads to increased algal growth. We investigated the possibilities for chemical control of E. saltensis using laboratory bioassays and a series of field trials involving either the use of enclosures in flooded crops or soil incorporation of pesticides into rice fields during fallow periods or shortly before flooding. The four most toxic compounds in 7 day soil/water laboratory bioassays were carbofuran, acetamiprid, bendiocarb and lambda-cyhalothrin which provided corrected mortalities of 86–100% at 2 mg a.i. L −1 . Other compounds that showed some level of efficacy (corrected mortality >20% at one or more rates) were imidacloprid, esfenvalerate, thiacloprid, niclosamide and alpha-cypermethrin. Twenty-six of the 38 pesticides evaluated failed to produce mean corrected mortalities >6% at application rates of up to 2 mg a.i. L −1 . Eight trials were conducted in flooded rice crops using small stainless steel enclosures and carbofuran, thiodicarb, niclosamide and bendiocarb at rates of 1 and 2 kg a.i. ha −1 . Trials were assessed 8–14 days after chemical application. None of these treatments produced a statistically significant decrease in Eukerria biomass, although consistent downward trends in response to higher treatment rates were evident in 2 trials (one with carbofuran and one with bendiocarb). Three trials with liquid pesticides watered into fallow rice fields were conducted with carbofuran (0.5, 1.0 and 5.0 kg a.i. ha −1 ) and thiodicarb (0.94 and 1.87 kg a.i. ha −1 ) however only the 5.0 kg a.i. ha −1 carbofuran treatment provided significant ( P < 0.05) levels of control. Preflood soil applications of liquid carbofuran, thiodicarb and niclosamide (2 kg a.i. ha −1 ), granular carbofuran and granular ethoprophos (0.5–2 kg a.i. ha −1 ) also did not provide statistically significant levels of control, although the 2 kg a.i. ha −1 liquid and granular carbofuran treatments did provide moderate levels of suppression (49–84%). Although further field trials with compounds such as acetamiprid and lambda-cyhalothrin may prove valuable, our results suggest chemical control of E. saltensis may be difficult to achieve with environmentally acceptable pesticides applied at economically viable rates. Cultural approaches such as appropriate crop rotations and landforming to ensure uniformly shallow water should continue to form the basis of Eukerria management programs. [ABSTRACT FROM AUTHOR]

There is an unmet need for improved, effective tissue engineering strategies to replace or repair bone damaged through disease or injury. Recent research has focused on developing biomaterial scaffolds capable of spatially and temporally releasing combinations of bioactive growth factors, rather than individual molecules, to recapitulate repair pathways present in vivo. We have developed an ex vivo embryonic chick femur critical size defect model and applied the model in the study of novel extracellular matrix (ECM) hydrogel scaffolds containing spatio-temporal combinatorial growth factor-releasing microparticles and skeletal stem cells for bone regeneration. Alginate/bovine bone ECM (bECM) hydrogels combined with poly( d , l -lactic- co -glycolic acid) (P DL LGA)/triblock copolymer (10–30% P DL LGA–PEG–PL DL GA) microparticles releasing dual combinations of vascular endothelial growth factor (VEGF), chondrogenic transforming growth factor beta 3 (TGF-β3) and the bone morphogenetic protein BMP2, with human adult Stro-1 + bone marrow stromal cells (HBMSCs), were placed into 2 mm central segmental defects in embryonic day 11 chick femurs and organotypically cultured. Hydrogels loaded with VEGF combinations induced host cell migration and type I collagen deposition. Combinations of TGF-β3/BMP2, particularly with Stro-1 + HBMSCs, induced significant formation of structured bone matrix, evidenced by increased Sirius red-stained matrix together with collagen expression demonstrating birefringent alignment within hydrogels. This study demonstrates the successful use of the chick femur organotypic culture system as a high-throughput test model for scaffold/cell/growth factor therapies in regenerative medicine. Temporal release of dual growth factors, combined with enriched Stro-1 + HBMSCs, improved the formation of a highly structured bone matrix compared to single release modalities. These studies highlight the potential of a unique alginate/bECM hydrogel dual growth factor release platform for bone repair. [ABSTRACT FROM AUTHOR]

Current clinical treatments for skeletal conditions resulting in large-scale bone loss include autograft or allograft, both of which have limited effectiveness. In seeking to address bone regeneration, several tissue engineering strategies have come to the fore, including the development of growth factor releasing technologies and appropriate animal models to evaluate repair. Ex vivo models represent a promising alternative to simple in vitro systems or complex, ethically challenging in vivo models. We have developed an ex vivo culture system of whole embryonic chick femora, adapted in this study as a critical size defect model to investigate the effects of novel bone extracellular matrix (bECM) hydrogel scaffolds containing spatio-temporal growth factor-releasing microparticles and skeletal stem cells on bone regeneration, to develop a viable alternative treatment for skeletal degeneration. Alginate/bECM hydrogels combined with poly ( d , l -lactic- co -glycolic acid) (P DL LGA)/triblock copolymer (10–30% P DL LGA–PEG–P DL LGA) microparticles releasing VEGF, TGF-β3 or BMP-2 were placed, with human adult Stro-1+ bone marrow stromal cells, into 2 mm central segmental defects in embryonic chick femurs. Alginate/bECM hydrogels loaded with HSA/VEGF or HSA/TGF-β3 demonstrated a cartilage-like phenotype, with minimal collagen I deposition, comparable to HSA-only control hydrogels. The addition of BMP-2 releasing microparticles resulted in enhanced structured bone matrix formation, evidenced by increased Sirius red-stained matrix and collagen expression within hydrogels. This study demonstrates delivery of bioactive growth factors from a novel alginate/bECM hydrogel to augment skeletal tissue formation and the use of an organotypic chick femur defect culture system as a high-throughput test model for scaffold/cell/growth factor therapies for regenerative medicine. [ABSTRACT FROM AUTHOR]

Copper sulphate pentahydrate (CuSO 4 ·5H 2 O) is widely used for controlling Isidorella newcombi , an aquatic snail that causes substantial damage to rice crops in southeastern Australia. We conducted field trials on a Birganbigil clay loam soil that demonstrate high levels of efficacy against adult I. newcombi (95% mortality at 6.38 kg ha −1 CuSO 4 ·5H 2 O (1.14 mg Cu L −1 )). Dissolved copper fell below the detection limit (0.02 mg Cu L −1 ) between 7 and 20 d after spraying at application rates up to 2.16 mg Cu L −1 (12 kg ha −1 CuSO 4 ·5H 2 O). Total copper concentrations in the water column fell below the detection limit (0.007 mg Cu L −1 ) 7–12 d after spraying at initially applied concentrations of 0.52–1.12 mg Cu L −1 , but remained detectable (0.01–0.02 mg Cu L −1 ) until 30 days after spraying (the conclusion of monitoring) when applied at higher initial concentrations (1.18–2.16 mg Cu L −1 ). There was a strong positive correlation ( r 2 = 0.90, P < 0.001) between copper application rate and copper concentrations in surface sediments 30 d after spraying. Bioassays with immature snails using three different test soils beneath irrigation water showed that underlying soil type strongly influenced the response of snails to applied copper, with significant ( P < 0.05) differences between LC 90 values which ranged from 0.41 to 1.04 mg applied Cu L −1 . Laboratory studies showed that dissolved copper concentrations remained significantly higher ( P < 0.05) in the water column above the soil that had the most deleterious effect on copper toxicity. Dissolved organic carbon concentrations were significantly ( P < 0.05) higher in both this soil and in the overlying water in the corresponding bioassay system, and correlated more closely with LC 90 values than other water chemistry parameters such as total hardness. Our results support the ongoing use of a variable copper application rate of 6–12 kg ha −1 CuSO 4 ·5H 2 O to allow for site-specific variations in efficacy, and suggest that variations in the release of dissolved organic carbon compounds from flooded soils may be a key factor moderating copper toxicity to I. newcombi in rice fields. [ABSTRACT FROM AUTHOR]

Abstract: Articular cartilage lesions are a particular challenge for regenerative medicine strategies as cartilage function stems from a complex depth-dependent organization. Tissue engineering scaffolds that vary in morphology and function offer a template for zone-specific cartilage extracellular matrix (ECM) production and mechanical properties. We fabricated multi-zone cartilage scaffolds by the electrostatic deposition of polymer microfibres onto particulate-templated scaffolds produced with 0.03 or 1.0mm3 porogens. The scaffolds allowed ample space for chondrocyte ECM production within the bulk while also mimicking the structural organization and functional interface of cartilage’s superficial zone. Addition of aligned fibre membranes enhanced the mechanical and surface properties of particulate-templated scaffolds. Zonal analysis of scaffolds demonstrated region-specific variations in chondrocyte number, sulfated GAG-rich ECM, and chondrocytic gene expression. Specifically, smaller porogens (0.03mm3) yielded significantly higher sGAG accumulation and aggrecan gene expression. Our results demonstrate that bilayered scaffolds mimic some key structural characteristics of native cartilage, support in vitro cartilage formation, and have superior features to homogeneous particulate-templated scaffolds. We propose that these scaffolds offer promise for regenerative medicine strategies to repair articular cartilage lesions. [Copyright &y& Elsevier]

Abstract: Bioactive glasses (BG) are suitable for bone regeneration applications as they bond with bone and can be tailored to release therapeutic ions. Fluoride, which is widely recognized to prevent dental caries, is efficacious in promoting bone formation and preventing osteoporosis-related fractures when administered at appropriate doses. To take advantage of these properties, we created BG incorporating increasing levels of fluoride whilst holding their silicate structure constant, and tested their effects on human osteoblasts in vitro. Our results demonstrate that, whilst cell proliferation was highest on low-fluoride-containing BG, markers for differentiation and mineralization were highest on BG with the highest fluoride contents, a likely effect of a combination of surface effects and ion release. Furthermore, osteoblasts exposed to the dissolution products of fluoride-containing BG or early doses of sodium fluoride showed increased alkaline phosphatase activity, a marker for bone mineralization, suggesting that fluoride can direct osteoblast differentiation. Taken together, these results suggest that BG that can release therapeutic levels of fluoride may find use in a range of bone regeneration applications. [Copyright &y& Elsevier]

Abstract: Physical stimuli play a crucial role in skeletogenesis and osteochondral repair and regeneration. Although the periosteum and periosteum-derived cells offer considerable therapeutic potential, the molecular mechanisms that control their differentiation are still not fully understood. As an initial case study, this work explores the hypothesis that dynamic compression might selectively enhance chondrogenic and/or osteogenic differentiation in human periosteal cells from two donors. Donor derived human periosteal cells were expanded in monolayer culture before seeding in 3% (w/v) agarose constructs. The ability of this in vitro culture model to support cell viability, chondrogenesis, and mechanotransduction was optimised. The time course of early chondrogenic differentiation was assessed by real time RT-PCR of mRNA expression levels for bone and cartilage specific gene markers. Intermittent dynamic compression (1 Hz, 15% strain) was applied to constructs, in the presence or absence of 10 ng/ml TGF-, for up to 4 days. The combined effect of TGF- and compressive loading on the expression levels of the Sox-9, Runx-2, ALP, Collagen X, and collagen type I genes was donor dependent. A synergistic effect was noted only in donor two, with peak mRNA expression levels at 24 h, particularly Sox-9 which increased 59.0-fold. These findings suggest that the interactions between mechanical stimuli and TGF- signalling may be an important mechanotransduction pathway for human periosteal cells and that, importantly, this cellular mechanosensitivity varies between donors. [Copyright &y& Elsevier]