scholarly journals 3D Reconstruction of HvRNASET2 Molecule to Understand Its Antibacterial Role

2020 ◽  
Vol 21 (24) ◽  
pp. 9722
Author(s):  
Nicolò Baranzini ◽  
Laura Pulze ◽  
Marcella Reguzzoni ◽  
Rossella Roncoroni ◽  
Viviana Teresa Orlandi ◽  
...  
Keyword(s):  
Light Transmission ◽  
Three Dimensional ◽  
Lipoteichoic Acid ◽  
Type I ◽  
Phagocytic Cells ◽  
In Vivo Experiments ◽  
Macrophage Phagocytosis ◽  
Bacterial Aggregates

Recent studies performed on the invertebrate model Hirudo verbana (medicinal leech) suggest that the T2 ribonucleic enzyme HvRNASET2 modulates the leech’s innate immune response, promoting microbial agglutination and supporting phagocytic cells recruitment in challenged tissues. Indeed, following injection of both lipoteichoic acid (LTA) and Staphylococcus aureus in the leech body wall, HvRNASET2 is expressed by leech type I granulocytes and induces bacterial aggregation to aid macrophage phagocytosis. Here, we investigate the HvRNASET2 antimicrobial role, in particular assessing the effects on the Gram-negative bacteria Escherichia coli. For this purpose, starting from the three-dimensional molecule reconstruction and in silico analyses, the antibacterial activity was evaluated both in vitro and in vivo. The changes induced in treated bacteria, such as agglutination and alteration in wall integrity, were observed by means of light, transmission and scanning electron microscopy. Moreover, immunogold, AMPs (antimicrobial peptides) and lipopolysaccharide (LPS) binding assays were carried out to evaluate HvRNASET2 interaction with the microbial envelopes and the ensuing ability to affect microbial viability. Finally, in vivo experiments confirmed that HvRNASET2 promotes a more rapid phagocytosis of bacterial aggregates by macrophages, representing a novel molecule for counteracting pathogen infections and developing alternative solutions to improve human health.

scholarly journals The Challenging Melanoma Landscape: From Early Drug Discovery to Clinical Approval

Cells ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 3088
Author(s):  
Mariana Matias ◽  
Jacinta O. Pinho ◽  
Maria João Penetra ◽  
Gonçalo Campos ◽  
Catarina Pinto Reis ◽  
...  
Keyword(s):  
Therapeutic Potential ◽  
Drug Evaluation ◽  
Three Dimensional ◽  
Experimental Models ◽  
In Vivo Studies ◽  
Murine Models ◽  
In Vivo Experiments ◽  
Novel Therapeutic Strategies

Melanoma is recognized as the most dangerous type of skin cancer, with high mortality and resistance to currently used treatments. To overcome the limitations of the available therapeutic options, the discovery and development of new, more effective, and safer therapies is required. In this review, the different research steps involved in the process of antimelanoma drug evaluation and selection are explored, including information regarding in silico, in vitro, and in vivo experiments, as well as clinical trial phases. Details are given about the most used cell lines and assays to perform both two- and three-dimensional in vitro screening of drug candidates towards melanoma. For in vivo studies, murine models are, undoubtedly, the most widely used for assessing the therapeutic potential of new compounds and to study the underlying mechanisms of action. Here, the main melanoma murine models are described as well as other animal species. A section is dedicated to ongoing clinical studies, demonstrating the wide interest and successful efforts devoted to melanoma therapy, in particular at advanced stages of the disease, and a final section includes some considerations regarding approval for marketing by regulatory agencies. Overall, considerable commitment is being directed to the continuous development of optimized experimental models, important for the understanding of melanoma biology and for the evaluation and validation of novel therapeutic strategies.

Morphological and Functional Characteristics of Three-Dimensional Engineered Bone-Ligament-Bone Constructs Following Implantation

2009 ◽  
Vol 131 (10) ◽  
Author(s):  
Jinjin Ma ◽  
Kristen Goble ◽  
Michael Smietana ◽  
Tatiana Kostrominova ◽  
Lisa Larkin ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Type I Collagen ◽  
Three Dimensional ◽  
Functionally Graded ◽  
Neonatal Rat ◽  
Ligament Injury ◽  
Type I ◽  
Tangent Moduli

The incidence of ligament injury has recently been estimated at 400,000/year. The preferred treatment is reconstruction using an allograft, but outcomes are limited by donor availability, biomechanical incompatibility, and immune rejection. The creation of an engineered ligament in vitro solely from patient bone marrow stromal cells (has the potential to greatly enhance outcomes in knee reconstructions. Our laboratory has developed a scaffoldless method to engineer three-dimensional (3D) ligament and bone constructs from rat bone marrow stem cells in vitro. Coculture of these two engineered constructs results in a 3D bone-ligament-bone (BLB) construct with viable entheses, which was successfully used for medial collateral ligament (MCL) replacement in a rat model. 1 month and 2 month implantations were applied to the engineered BLBs. Implantation of 3D BLBs in a MCL replacement application demonstrated that our in vitro engineered tissues grew and remodeled quickly in vivo to an advanced phenotype and partially restored function of the knee. The explanted 3D BLB ligament region stained positively for type I collagen and elastin and was well vascularized after 1 and 2 months in vivo. Tangent moduli of the ligament portion of the 3D BLB 1 month explants increased by a factor of 2.4 over in vitro controls, to a value equivalent to those observed in 14-day-old neonatal rat MCLs. The 3D BLB 1 month explants also exhibited a functionally graded response that closely matched native MCL inhomogeneity, indicating the constructs functionally adapted in vivo.

scholarly journals Enhanced Regeneration of Vascularized Adipose Tissue with Dual 3D-Printed Elastic Polymer/dECM Hydrogel Complex

2021 ◽  
Vol 22 (6) ◽  
pp. 2886
Author(s):  
Soojin Lee ◽  
Hyun Su Lee ◽  
Justin J. Chung ◽  
Soo Hyun Kim ◽  
Jong Woong Park ◽  
...  
Keyword(s):  
Adipose Tissue ◽  
Adipogenic Differentiation ◽  
Three Dimensional ◽  
Human Adipose Tissue ◽  
Collagen Type I ◽  
Type I ◽  
Adipose Tissue Engineering ◽  
Decellularized Extracellular Matrix ◽  
In Vivo Experiments

A flexible and bioactive scaffold for adipose tissue engineering was fabricated and evaluated by dual nozzle three-dimensional printing. A highly elastic poly (L-lactide-co-ε-caprolactone) (PLCL) copolymer, which acted as the main scaffolding, and human adipose tissue derived decellularized extracellular matrix (dECM) hydrogels were used as the printing inks to form the scaffolds. To prepare the three-dimensional (3D) scaffolds, the PLCL co-polymer was printed with a hot melting extruder system while retaining its physical character, similar to adipose tissue, which is beneficial for regeneration. Moreover, to promote adipogenic differentiation and angiogenesis, adipose tissue-derived dECM was used. To optimize the printability of the hydrogel inks, a mixture of collagen type I and dECM hydrogels was used. Furthermore, we examined the adipose tissue formation and angiogenesis of the PLCL/dECM complex scaffold. From in vivo experiments, it was observed that the matured adipose-like tissue structures were abundant, and the number of matured capillaries was remarkably higher in the hydrogel–PLCL group than in the PLCL-only group. Moreover, a higher expression of M2 macrophages, which are known to be involved in the remodeling and regeneration of tissues, was detected in the hydrogel–PLCL group by immunofluorescence analysis. Based on these results, we suggest that our PLCL/dECM fabricated by a dual 3D printing system will be useful for the treatment of large volume fat tissue regeneration.

Three-Dimensional-Printed External Scaffolds Mitigate Loss of Volume and Topography in Engineered Elastic Cartilage Constructs

Cartilage ◽  
2021 ◽  
pp. 194760352110495
Author(s):  
Xue Dong ◽  
Ishani D. Premaratne ◽  
Jaime L. Bernstein ◽  
Arash Samadi ◽  
Alexandra J. Lin ◽  
...  
Keyword(s):  
Injection Molding ◽  
Type I Collagen ◽  
Three Dimensional ◽  
Type I ◽  
Base Area ◽  
Elastic Cartilage ◽  
3D Printed ◽  
Injection Molded

Objective: A major obstacle in the clinical translation of engineered auricular scaffolds is the significant contraction and loss of topography that occur during maturation of the soft collagen-chondrocyte matrix into elastic cartilage. We hypothesized that 3-dimensional-printed, biocompatible scaffolds would “protect” maturing hydrogel constructs from contraction and loss of topography. Design: External disc-shaped and “ridged” scaffolds were designed and 3D-printed using polylactic acid (PLA). Acellular type I collagen constructs were cultured in vitro for up to 3 months. Collagen constructs seeded with bovine auricular chondrocytes (BAuCs) were prepared in 3 groups and implanted subcutaneously in vivo for 3 months: preformed discs with (“Scaffolded/S”) or without (“Naked/N”) an external scaffold and discs that were formed within an external scaffold via injection molding (“Injection Molded/SInj”). Results: The presence of an external scaffold or use of injection molding methodology did not affect the acellular construct volume or base area loss. In vivo, the presence of an external scaffold significantly improved preservation of volume and base area at 3 months compared to the naked group ( P < 0.05). Construct contraction was mitigated even further in the injection molded group, and topography of the ridged constructs was maintained with greater fidelity ( P < 0.05). Histology verified the development of mature auricular cartilage in the constructs within external scaffolds after 3 months. Conclusion: Custom-designed, 3D-printed, biocompatible external scaffolds significantly mitigate BAuC-seeded construct contraction and maintain complex topography. Further refinement and scaling of this approach in conjunction with construct fabrication utilizing injection molding may aid in the development of full-scale auricular scaffolds.

scholarly journals Low-dose chidamide restores immune tolerance in ITP in mice and humans

Blood ◽  
2019 ◽  
Vol 133 (7) ◽  
pp. 730-742 ◽  
Author(s):  
Hong-yu Zhao ◽  
Ya-hui Ma ◽  
Da-qi Li ◽  
Tao Sun ◽  
Li-zhen Li ◽  
...  
Keyword(s):  
Immune Tolerance ◽  
Histone Deacetylase Inhibitors ◽  
Low Dose ◽  
Mononuclear Cells ◽  
Treg Cells ◽  
Peripheral Blood Mononuclear ◽  
In Vivo Experiments ◽  
Macrophage Phagocytosis

Abstract Increased macrophage phagocytosis of antibody-coated platelets, as well as decreased numbers and/or impaired function of CD4+CD25+Foxp3+ regulatory T (Treg) cells, has been shown to participate in the pathogenesis of immune thrombocytopenia (ITP). Low-dose histone deacetylase inhibitors (HDACi’s) are anti-inflammatory and immunomodulatory agents that can enhance immunosuppression in graft-versus-host disease by increasing the number and function of Foxp3+ Treg cells, but it is unclear whether they have the potential to promote immune tolerance and platelet release in ITP. In this study, we performed in vitro and in vivo experiments and found that a low-dose HDACi (chidamide) alleviated thrombocytopenia in passive and active murine models of ITP. Further, low-dose HDACi’s attenuated macrophage phagocytosis of antibody-coated platelets, stimulated the production of natural Foxp3+ Treg cells, promoted the peripheral conversion of T cells into Treg cells, and restored Treg cell suppression in vivo and in vitro. Finally, we confirmed that low-dose HDACi’s could regulate CTLA4 expression in peripheral blood mononuclear cells through modulation of histone H3K27 acetylation. Low-dose HDACi treatment in ITP could be offset by blocking the effect of CTLA4. Therefore, we propose that low-dose chidamide administration has potential as a novel treatment for ITP in the clinic.

scholarly journals Pre-aggregation of scalp progenitor dermal and epidermal stem cells activates the WNT pathway and promotes hair follicle formation in in vitro and in vivo systems

2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Yiqun Su ◽  
Jie Wen ◽  
Junrong Zhu ◽  
Zhiwei Xie ◽  
Chang Liu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Hair Follicle ◽  
Progenitor Cells ◽  
Wnt Pathway ◽  
Three Dimensional ◽  
Epidermal Cells ◽  
Type I ◽  
Epidermal Stem Cells

Abstract Background Billions of dollars are invested annually by pharmaceutical companies in search of new options for treating hair loss conditions; nevertheless, the challenge remains. One major limitation to hair follicle research is the lack of effective and efficient drug screening systems using human cells. Organoids, three-dimensional in vitro structures derived from stem cells, provide new opportunities for studying organ development, tissue regeneration, and disease pathogenesis. The present study focuses on the formation of human hair follicle organoids. Methods Scalp-derived dermal progenitor cells mixed with foreskin-derived epidermal stem cells at a 2:1 ratio aggregated in suspension to form hair follicle-like organoids, which were confirmed by immunostaining of hair follicle markers and by molecular dye labeling assays to analyze dermal and epidermal cell organization in those organoids. The hair-forming potential of organoids was examined using an in vivo transplantation assay. Results Pre-aggregation of dermal and epidermal cells enhanced hair follicle formation in vivo. In vitro pre-aggregation initiated the interactions of epidermal and dermal progenitor cells resulting in activation of the WNT pathway and the formation of pear-shape structures, named type I aggregates. Cell-tracing analysis showed that the dermal and epidermal cells self-assembled into distinct epidermal and dermal compartments. Histologically, the type I aggregates expressed early hair follicle markers, suggesting the hair peg-like phase of hair follicle morphogenesis. The addition of recombinant WNT3a protein to the medium enhanced the formation of these aggregates, and the Wnt effect could be blocked by the WNT inhibitor, IWP2. Conclusions In summary, our system supports the rapid formation of a large number of hair follicle organoids (type I aggregates). This system provides a platform for studying epithelial-mesenchymal interactions, for assessing inductive hair stem cells and for screening compounds that support hair follicle regeneration.

scholarly journals The interleaved partial active shape model (IPASM) search algorithm – towards 3D ultrasound-based bone surface reconstruction

10.29007/rbgl ◽  
2019 ◽  
Author(s):  
Benjamin Hohlmann ◽  
Klaus Radermacher
Keyword(s):  
Search Algorithm ◽  
Three Dimensional ◽  
3D Ultrasound ◽  
Three Dimensional Model ◽  
In Vivo Experiments ◽  
Distal Surface ◽  
Cheap Alternative ◽  
Orthopedic Applications

Several orthopedic applications require a three-dimensional model of the bone. Ultrasound is a radiation-free and cheap alternative to the state-of-the-art imaging modalities if its limitations in terms of image quality and viewing range can be overcome. This work presents in-vitro as well as in-vivo experiments evaluating the IPASM search, a method for combined segmentation, registration as well as extrapolation. The algorithm is capable to reconstruct the distal surface of a phantom femur with an average surface distance error of roughly 1mm in case of in-vitro as well as below 2mm for in-vivo records, even if the shape varies strongly from the initial model.

scholarly journals Experimental study of tissue-engineered urethra with collagen / chitosan composite as scaffolds

2020 ◽  
Author(s):  
Zhai Hongfeng ◽  
Qiu Changhong ◽  
Jin Jun ◽  
Shao Xin
Keyword(s):  
New Zealand ◽  
Epithelial Cells ◽  
Cell Activity ◽  
Three Dimensional ◽  
Volume Ratio ◽  
In Vivo Experiments ◽  
High Area ◽  
Chitosan Composite

AbstractIn this article we investigated the preparation of tissue-engineered urethra by using the urethral epithelial subculture cells of male New Zealand young rabbits. We inoculated the epithelial cells of urinary mucosa of male New Zealand young rabbits on collagen, chitosan and collagen chitosan composite as scaffolds to prepare tissue-engineered urethra. The results of inverted phase contrast microscope, HE staining and scanning electron microscope of three kinds of tissue-engineered urethra were compared. What’s more, we reported a new method for quantitative and rapid detection of epithelial cell activity of urinary mucosa in situ by Interactive Laser Cytometer. The collagen chitosan composite was more similar to the extracellular matrix of mammalian. Its three-dimensional porous structure had a high area volume ratio, which was conducive to cell adhesion, growth and metabolism. In vitro, the urethral epithelial cells had been cultured on collagen chitosan composite, and the tissue-engineered urethra was successfully prepared, which laid a solid foundation for further in vivo experiments.

scholarly journals Crosstalk between invadopodia and the extracellular matrix

2020 ◽  
Author(s):  
Shinji Iizuka ◽  
Ronald P. Leon ◽  
Kyle P. Gribbin ◽  
Ying Zhang ◽  
Jose Navarro ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Type I Collagen ◽  
Complex Structure ◽  
Three Dimensional ◽  
3D Culture ◽  
Model Systems ◽  
Type I ◽  
Extracellular Matrix Components

ABSTRACTThe scaffold protein Tks5α is required for invadopodia-mediated cancer invasion both in vitro and in vivo. We have previously also revealed a role for Tks5 in tumor cell growth using three-dimensional (3D) culture model systems and mouse transplantation experiments. Here we use both 3D and high-density fibrillar collagen (HDFC) culture to demonstrate that native type I collagen, but not a form lacking the telopeptides, stimulated Tks5-dependent growth, which was dependent on the DDR collagen receptors. We used microenvironmental microarray (MEMA) technology to determine that laminin, collagen I, fibronectin and tropoelastin also stimulated invadopodia formation. A Tks5α-specific monoclonal antibody revealed its expression both on microtubules and at invadopodia. High- and super-resolution microscopy of cells in and on collagen was then used to place Tks5α at the base of invadopodia, separated from much of the actin and cortactin, but coincident with both matrix metalloprotease and cathepsin proteolytic activity. Inhibition of the Src family kinases, cathepsins or metalloproteases all reduced invadopodia length but each had distinct effects on Tks5α localization. These studies highlight the crosstalk between invadopodia and extracellular matrix components, and reveal the invadopodium to be a spatially complex structure.

Vibration-Assisted Insertion of Flexible Neural Microelectrodes With Bio-Dissolvable Guides for Medical Implantation

2021 ◽  
Author(s):  
Yi Wang ◽  
Yen Yu Ian Shih ◽  
Yuan-shin Lee
Keyword(s):  
Tissue Damage ◽  
Three Dimensional ◽  
Insertion Technique ◽  
Neural Electrode ◽  
In Vivo Experiments ◽  
Neural Probe ◽  
Electrode Insertion ◽  
Neural Microelectrodes

Abstract This paper presents vibration-assisted insertion of flexible neural electrodes with bio-dissolvable guides to deliver accurate microprobe insertion with minimized tissue damage. Invasive flexible neural microprobe is an important new tool for neuromodulation and recording research for medical neurology treatment applications. Flexible neural electrode probes are susceptible to bending and buckling during surgical implantation due to the thin and flexible soft substrates. Inspired by insects in nature, a vibration-assisted insertion technique is developed for flexible neural electrode insertion to deliver accurate microprobe insertion with minimized tissue damage. A three-dimensional combined longitudinal-twisting (L&T) vibration is used to reduce the insertion friction force, and thus reducing soft tissue damage. To reduce the flexible microelectrode buckling during surgical insertion, a bio-dissolvable Polyethylene glycol (PEG) guide is developed for the enhancement of flexible neural probe stiffness. Combining these two methods, the insertion performance of the flexible neural probe is significantly improved. Both the in vitro and the in vivo experiments were conducted to validate the proposed techniques.

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