Engineering T cells to express chimeric antigen receptors (CARs) specific for antigens on hematological cancers has yielded remarkable clinical responses, but with solid tumors, benefit has been more ...limited. This may reflect lack of suitable target antigens, immune evasion mechanisms in malignant cells, and/or lack of T cell infiltration into tumors. An alternative approach, to circumvent these problems, is targeting the tumor vasculature rather than the malignant cells directly. CLEC14A is a glycoprotein selectively overexpressed on the vasculature of many solid human cancers and is, therefore, of considerable interest as a target antigen. Here, we generated CARs from 2 CLEC14A-specific antibodies and expressed them in T cells. In vitro studies demonstrated that, when exposed to their target antigen, these engineered T cells proliferate, release IFN-γ, and mediate cytotoxicity. Infusing CAR engineered T cells into healthy mice showed no signs of toxicity, yet these T cells targeted tumor tissue and significantly inhibited tumor growth in 3 mouse models of cancer (Rip-Tag2, mPDAC, and Lewis lung carcinoma). Reduced tumor burden also correlated with significant loss of CLEC14A expression and reduced vascular density within malignant tissues. These data suggest the tumor vasculature can be safely and effectively targeted with CLEC14A-specific CAR T cells, offering a potent and widely applicable therapy for cancer.
This paper presents a study on graphene platelet (GPL)-reinforced alumina (Al2O3) ceramic composites and the relationships between the loading of GPL and both mechanical properties and in vitro ...biocompatibility. Al2O3 powders with different GPL contents were prepared and sintered using a gas protected pressure-less furnace. The examination of the results shows the density of the composites varying from 99.2% to 95.6% with the loading of GPL from 0.75 to 1.48 vol %. Raman studies show that moderate agglomerations of GPLs occur during the ball milling process and graphitic defects were produced during the high temperature processing. Mechanical properties of the Al2O3 matrix are significantly improved by adding GPLs. A maximum increase of approximately 60% in flexural strength and 70% in fracture toughness are achieved by introducing 0.75 vol % GPLs. In the biocompatibility tests, it was found that cells directly seeding on top of GPL/Al2O3 samples showed better initial attachment (3 h after seeding) and viability (3 days after incubation) than the monolithic Al2O3, indicating that the GPL/Al2O3 composites have comparable or more favorable biocompatibility. The excellent mechanical and biomedical properties of the GPL/Al2O3 composites may enable them to be applied to a wide range of engineering and biomedical applications.
The generation of retinal organoids from human pluripotent stem cells (hPSC) is now a well-established process that in part recapitulates retinal development. However, hPSC-derived photoreceptors ...that exhibit well-organized outer segment structures have yet to be observed. To facilitate improved inherited retinal disease modeling, we determined conditions that would support outer segment development in maturing hPSC-derived photoreceptors. We established that the use of antioxidants and BSA-bound fatty acids promotes the formation of membranous outer segment-like structures. Using new protocols for hPSC-derived retinal organoid culture, we demonstrated improved outer segment formation for both rod and cone photoreceptors, including organized stacked discs. Using these enhanced conditions to generate iPSC-derived retinal organoids from patients with X-linked retinitis pigmentosa, we established robust cellular phenotypes that could be ameliorated following adeno-associated viral vector-mediated gene augmentation. These findings should aid both disease modeling and the development of therapeutic approaches for the treatment of photoreceptor disorders.
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•Antioxidants and lipids are required for the formation of organized outer segments•Both rod and cone hPSC-derived photoreceptors generate well-formed outer segments•Improved conditions provide a robust model of X-linked retinitis pigmentosa type 3•Enhanced segment formation permits the evaluation of therapeutic interventions
In this article, Ali and colleagues demonstrate the generation of hPSC-derived photoreceptors, bearing organized outer segments. Using these enhanced conditions, they report robust cellular phenotypes in an iPSC-based model of X-linked retinitis pigmentosa, which were ameliorated with gene supplementation. The findings presented will aid disease modeling of human photoreceptor-specific disorders and the development of novel therapeutic approaches.
The adaptive foam reticulation technique combines the foam reticulation and freeze casting methodologies of fabricating bone reparative scaffolds to offer a potential alternative to autografts. For ...the first time this paper studies the effect of processing on the mechanical properties and in-vitro cell growth of controllably generating a hierarchical structure of macro- (94 ± 6 to 514 ± 36 μm) and microporosity (2–30 μm) by the inclusion of camphene as a porogen during processing. Scaffolds were produced with porogen additions of 0–25 wt%. Porosity values of the structures of 85–96% were determined using the Archimedes technique and verified using X-ray Computed Tomography. The strength of the hydroxyapatite scaffolds, 5.70 ± 1.0 to 159 ± 61 kPa, correlated to theoretically determined values, 3.71 ± 0.8 to 134 ± 12 kPa, calculated by the novel incorporation of a shape factor into a standard equation. Fibroblast (3T3) and pre-osteoblast (MC3T3) cell growth was found to be significantly (P < 0.005) improved using 25 wt% porogen. This was supported by increased levels of alkaline phosphatase and was thought to result from greater dissolution as quantified by increased calcium levels in incubating media. The combination of these properties renders adaptive foam reticulation-fabricated scaffolds suitable for non-structural bone regenerative applications in non-load bearing bone defects.
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•Hydroxyapatite scaffolds were fabricated using adaptive foam reticulation technique.•Total porosity was determined to be 86–95%, with strut microporosity of 23–51%.•Porogen inclusion affected mechanical strength for higher porosity structures.•The inclusion of a porogen significantly increased 3T3 and MC3T3 cell growth.•This indicates that adaptive foam reticulation offers a viable method of producing bone implants.
Cider and/or beer lees has normally either been used as low cost animal feed or been disposed of at great cost. A higher value use for the yeast was therefore sought. This has been developed with the ...use of environmentally friendly subcritical water extraction. Results have shown that the extract contains anti-oxidant activity using two separate anti-oxidant assays, with a large improvement in activity above a process temperature of 200 °C. This is due in large part to an increase in the concentration of phenolic compounds in the extract. As a result of this, a refined extract was produced using supercritical CO2 that improved anti-oxidant activity compared to the crude extract. The anti-oxidant activity of the 200°C and refined extract has also been demonstrated using the comet assay in cells with the performance of the extracts being comparable to that of Trolox. The rheological stability of a number of cosmetic formulations incorporating the extract has also been tested with 2 of the 4 formulations being stable. However, colour change issues have been observed with all four formulations tested. Overall, a novel and biologically effective extract has been produced using data from anti-oxidant assays to improve activity of the extract.
Tissue engineering offers a promising alternative to the use of autografts in the treatment of ligament injuries. However, current approaches using only biodegradable materials have insufficient ...mechanical strength for load bearing applications. In this research, hybrid bio-artificial ligaments were fabricated using a combination of a titanium alloy spring and a fibrin gel/fibroblast construct. The ends of the ligament prosthesis were incorporated into brushite cement anchors to allow fusion with the host bone. Cell attachment to the titanium spring was examined using scanning electron microscopy and fluorescent staining of cells. The unreinforced constructs were observed to fail at the anchor-ligament junction, while the titanium spring reinforcement was found to assist in even transmission of the load to the ligament, and hence to provide a means of load sharing between the biological construct and the spring. As a result, the reinforced construct failed primarily in the soft tissue region. The good load distribution features from the mechanical data was attributed to the good cellular level adhesion to, and alignment along the coiling of, the length of the spring reinforcement. Incorporation of a biocompatible reinforcement in conjunction with a tissue engineered construct gave improved load distribution, reducing stress concentrations, and significantly increased the ultimate strength at failure. The results suggest that the hybrid approach used here shows promise in developing improved therapies for connective tissue injuries.
Cider and/or beer lees has normally either been used as low cost animal feed or been disposed of at great cost. A higher value use for the yeast was therefore sought. This has been developed with the ...use of environmentally friendly subcritical water extraction. Results have shown that the extract contains anti-oxidant activity using two separate anti-oxidant assays, with a large improvement in activity above a process temperature of 200 °C. This is due in large part to an increase in the concentration of phenolic compounds in the extract. As a result of this, a refined extract was produced using supercritical CO2 that improved anti-oxidant activity compared to the crude extract. The anti-oxidant activity of the 200°C and refined extract has also been demonstrated using the comet assay in cells with the performance of the extracts being comparable to that of Trolox. The rheological stability of a number of cosmetic formulations incorporating the extract has also been tested with 2 of the 4 formulations being stable. However, colour change issues have been observed with all four formulations tested. Overall, a novel and biologically effective extract has been produced using data from anti-oxidant assays to improve activity of the extract.
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