scholarly journals A Low-Cost Mechanical Stretching Device for Uniaxial Strain of Cells: A Platform for Pedagogy in Mechanobiology

2018 ◽  
Vol 140 (8) ◽  
Author(s):  
Hamza Atcha ◽  
Chase T. Davis ◽  
Nicholas R. Sullivan ◽  
Tim D. Smith ◽  
Sara Anis ◽  
...  
Keyword(s):  
Undergraduate Students ◽  
Low Cost ◽  
Critical Role ◽  
Cost Effective ◽  
Mechanical Stretch ◽  
Cell Types ◽  
Educational Environments ◽  
Cell Stretcher ◽  
Gear Rack

Mechanical cues including stretch, compression, and shear stress play a critical role in regulating the behavior of many cell types, particularly those that experience substantial mechanical stress within tissues. Devices that impart mechanical stimulation to cells in vitro have been instrumental in helping to develop a better understanding of how cells respond to mechanical forces. However, these devices often have constraints, such as cost and limited functional capabilities, that restrict their use in research or educational environments. Here, we describe a low-cost method to fabricate a uniaxial cell stretcher that would enable widespread use and facilitate engineering design and mechanobiology education for undergraduate students. The device is capable of producing consistent and reliable strain profiles through the use of a servomotor, gear, and gear rack system. The servomotor can be programmed to output various waveforms at specific frequencies and stretch amplitudes by controlling the degree of rotation, speed, and acceleration of the servogear. In addition, the stretchable membranes are easy to fabricate and can be customized, allowing for greater flexibility in culture well size. We used the custom-built stretching device to uniaxially strain macrophages and cardiomyocytes, and found that both cell types displayed functional and cell shape changes that were consistent with the previous studies using commercially available systems. Overall, this uniaxial cell stretcher provides a more cost-effective alternative to study the effects of mechanical stretch on cells, and can therefore, be widely used in research and educational environments to broaden the study and pedagogy of cell mechanobiology.

Development of Natural Bioactive Alkaloids: Anticancer perspective

2021 ◽  
Vol 21 ◽  
Author(s):  
Ashish Patel ◽  
Ravi Vanecha ◽  
Jay Patel ◽  
Divy Patel ◽  
Umang Shah ◽  
...  
Keyword(s):  
Anticancer Agents ◽  
Low Cost ◽  
Chemical Compounds ◽  
Cost Effective ◽  
Drug Discovery And Development ◽  
Anticancer Properties ◽  
Antimetastatic Activity ◽  
New Chemical Entities

: Cancer is a frightful disease that still poses a 'nightmare' worldwide, causing millions of casualties annually due to one of the human race's most significant healthcare challenges that requires a pragmatic treatment strategy. However, plants and plant-derived products revolutionize the field as they are quick, cleaner, eco-friendly, low-cost, effective, and less toxic than conventional treatment methods. Plants are repositories for new chemical entities and have a promising cancer research path, supplying 60% of the anticancer agents currently used. Alkaloids are important chemical compounds that serve as a rich reservoir for drug discovery and development. However, some alkaloids derived from natural herbs display anti-proliferation and antimetastatic activity on different forms of cancer, both in vitro and in vivo. Alkaloids have also been widely formulated as anticancer medications, such as camptothecin and vinblastine. Still, more research and clinical trials are required before final recommendations can be made on specific alkaloids. This review focuses on the naturally-derived bioactive alkaloids with prospective anticancer properties based on the information in the literature.

scholarly journals Gold-coated plant virus as computed tomography imaging contrast agent

2019 ◽  
Vol 10 ◽  
pp. 1983-1993 ◽  
Author(s):  
Alaa A A Aljabali ◽  
Mazhar S Al Zoubi ◽  
Khalid M Al-Batanyeh ◽  
Ali Al-Radaideh ◽  
Mohammad A Obeid ◽  
...  
Keyword(s):  
Computed Tomography ◽  
Contrast Agent ◽  
Low Cost ◽  
Cell Types ◽  
Specific Cell ◽  
Scan Time ◽  
Ct Contrast Agent ◽  
Potential Applications ◽  
Resolution Imaging

Chemical modification of the surface of viruses, both the interior and the exterior, imparts new functionalities, that have potential applications in nanomedicine. In this study, we developed novel virus-based nanomaterials as a contrast agent for computed tomography (CT) imaging in vitro. The gold-coated cowpea mosaic virus (Au-CPMV) particles were generated by the electrostatic adsorption of positively charged electrolyte on the virus capsid with the subsequent incubation and reduction of anionic gold complexes. Au-CPMV particles as a CT contrast agent offer a fast scan time (less than 2 min), low cost, and biocompatibility and allow for high-resolution imaging with ca. 150 Hounsfield units (HU). The Au-CPMV surface was further modified allowing for the incorporation of targeting molecules of specific cell types.

scholarly journals Function of Native OmtA In Vivo and Expression and Distribution of This Protein in Colonies of Aspergillus parasiticus

2002 ◽  
Vol 68 (11) ◽  
pp. 5718-5727 ◽  
Author(s):  
Li-Wei Lee ◽  
Ching-Hsun Chiou ◽  
John E. Linz
Keyword(s):  
Fusion Protein ◽  
Aspergillus Parasiticus ◽  
Polyclonal Antibodies ◽  
Critical Role ◽  
Immunohistochemical Analysis ◽  
Cell Types ◽  
Temporal Association ◽  
Thin Sections

ABSTRACT The activities of two enzymes, a 168-kDa protein and a 40-kDa protein, OmtA, purified from the filamentous fungus Aspergillus parasiticus were reported to convert the aflatoxin pathway intermediate sterigmatocystin to O-methylsterigmatocystin in vitro. Our initial goal was to determine if OmtA is necessary and sufficient to catalyze this reaction in vivo and if this reaction is necessary for aflatoxin synthesis. We generated A. parasiticus omtA-null mutant LW1432 and a maltose binding protein-OmtA fusion protein expressed in Escherichia coli. Enzyme activity analysis of OmtA fusion protein in vitro confirmed the reported catalytic function of OmtA. Feeding studies conducted with LW1432 demonstrated a critical role for OmtA, and the reaction catalyzed by this enzyme in aflatoxin synthesis in vivo. Because of a close regulatory link between aflatoxin synthesis and asexual sporulation (conidiation), we hypothesized a spatial and temporal association between OmtA expression and conidiospore development. We developed a novel time-dependent colony fractionation protocol to analyze the accumulation and distribution of OmtA in fungal colonies grown on a solid medium that supports both toxin synthesis and conidiation. OmtA-specific polyclonal antibodies were purified by affinity chromatography using an LW1432 protein extract. OmtA was not detected in 24-h-old colonies but was detected in 48-h-old colonies using Western blot analysis; the protein accumulated in all fractions of a 72-h-old colony, including cells (0 to 24 h) in which little conidiophore development was observed. OmtA in older fractions of the colony (24 to 72 h) was partly degraded. Fluorescence-based immunohistochemical analysis conducted on thin sections of paraffin-embedded fungal cells from time-fractionated fungal colonies demonstrated that OmtA is evenly distributed among different cell types and is not concentrated in conidiophores. These data suggest that OmtA is present in newly formed fungal tissue and then is proteolytically cleaved as cells in that section of the colony age.

scholarly journals Toxoplasma gondii PPM3C, a secreted protein phosphatase, affects parasitophorous vacuole effector export

2020 ◽  
Author(s):  
Joshua A. Mayoral ◽  
Tadakimi Tomita ◽  
Vincent Tu ◽  
Jennifer T. Aguilan ◽  
Simone Sidoli ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Parasitophorous Vacuole ◽  
Acute Infection ◽  
Critical Role ◽  
Cell Types ◽  
Effector Proteins ◽  
Secreted Proteins ◽  
Growth Defect ◽  
Label Free

ABSTRACTToxoplasma gondii is a highly successful parasite that infects a significant portion of the human population. As an intracellular parasite, T. gondii thrives within many different cell types due to its residence in the parasitophorous vacuole, a specialized and heavily modified compartment in which parasites divide. Within this vacuole, numerous secreted proteins facilitate functions that optimize intracellular survival. We characterized one such protein, TgPPM3C, which is predicted to contain a domain belonging to the PP2C class of serine/threonine phosphatases and is secreted by both tachyzoites and differentiating bradyzoites into the vacuolar lumen. Genetic deletion of TgPPM3C established that parasites lacking this predicted phosphatase exhibit a minor growth defect in vitro, are avirulent during acute infection in mice, and form fewer cysts in mouse brain during chronic infection. A label-free phosphoproteomic approach was utilized to identify putative TgPPM3C substrates and demonstrated several secreted proteins with altered phosphorylation status in the absence of TgPPM3C. Altered phosphorylation status was seen in MYR1, a protein essential to the process of protein effector export from the parasitophorous vacuole into the host cell, and in GRA16 and GRA28, two exported effector proteins. Defects were seen in the export of GRA16 and GRA28, but not the effector TgIST, in the TgPPM3C knockout strain. Parasites lacking TgPPM3C also exhibited defects in host c-Myc induction, a process influenced by effector export. Phosphomimetic mutations of GRA16 serine residues recapitulated export defects, implicating de-phosphorylation as an important process in facilitating the export of GRA16. These findings provide an example of the emerging critical role that phosphatases play in regulating the complex environment of the T. gondii parasitophorous vacuole.

scholarly journals Amyloid fibril-based hydrogels for high-throughput tumor spheroid modeling

2020 ◽  
Author(s):  
Namrata Singh ◽  
Komal Patel ◽  
Ambuja Navalkar ◽  
Pradeep Kadu ◽  
Debalina Datta ◽  
...  
Keyword(s):  
Drug Resistance ◽  
High Throughput ◽  
Three Dimensional ◽  
3D Cell Culture ◽  
Cancer Therapeutics ◽  
Cost Effective ◽  
Cell Types ◽  
Tumor Spheroid ◽  
Tumor Modeling

AbstractBiomaterials mimicking extracellular matrices (ECM) for three-dimensional (3D) cultures have gained immense interest in tumor modeling and in vitro organ development. Here, we introduce versatile, thixotropic amyloid hydrogels as a bio-mimetic ECM scaffold for 3D cell culture as well as high-throughput tumor spheroid formation using a drop cast method. The unique cross-β-sheet structure, sticky surface, and thixotropicity of amyloid hydrogels allow robust cell adhesion, survival, proliferation, and migration, which are essential for 3D tumor modeling with various cancer cell types. The spheroids formed show overexpression of the signature cancer biomarkers and confer higher drug resistance compared to two-dimensional (2D) monolayer cultures. Using breast tumor tissue from mouse xenograft, we showed that these hydrogels support the formation of tumor spheroids with a well-defined necrotic core, cancer-associated gene expression, higher drug resistance, and tumor heterogeneity reminiscent of the original tumor. Altogether, we have developed a rapid and cost-effective platform for generating in vitro cancer models for the screening of anti-cancer therapeutics and developing personalized medicines.

scholarly journals Estrogen Receptor α Signaling in Osteoblasts is Required for Mechanotransduction in Bone Fracture Healing

2021 ◽  
Vol 9 ◽  
Author(s):  
Lena Steppe ◽  
Benjamin Thilo Krüger ◽  
Miriam Eva Angelica Tschaffon ◽  
Verena Fischer ◽  
Jan Tuckermann ◽  
...  
Keyword(s):  
Estrogen Receptor ◽  
Fracture Healing ◽  
Critical Role ◽  
Cell Types ◽  
Estrogen Receptor Α ◽  
Whole Body ◽  
Fracture Callus ◽  
Positive Effects ◽  
Osteoblast Lineage

Biomechanical stimulation by whole-body low-magnitude high-frequency vibration (LMHFV) has demonstrated to provoke anabolic effects on bone metabolism in both non-osteoporotic and osteoporotic animals and humans. However, preclinical studies reported that vibration improved fracture healing and bone formation in osteoporotic, ovariectomized (OVX) mice representing an estrogen-deficient hormonal status, but impaired bone regeneration in skeletally healthy non-OVX mice. These effects were abolished in general estrogen receptor α (ERα)-knockout (KO) mice. However, it remains to be elucidated which cell types in the fracture callus are targeted by LMHFV during bone healing. To answer this question, we generated osteoblast lineage-specific ERα-KO mice that were subjected to ovariectomy, femur osteotomy and subsequent vibration. We found that the ERα specifically on osteoblastic lineage cells facilitated the vibration-induced effects on fracture healing, because in osteoblast lineage-specific ERα-KO (ERαfl/fl; Runx2Cre) mice the negative effects in non-OVX mice were abolished, whereas the positive effects of vibration in OVX mice were reversed. To gain greater mechanistic insights, the influence of vibration on murine and human osteogenic cells was investigated in vitro by whole genome array analysis and qPCR. The results suggested that particularly canonical WNT and Cox2/PGE2 signaling is involved in the mechanotransduction of LMHFV under estrogen-deficient conditions. In conclusion, our study demonstrates a critical role of the osteoblast lineage-specific ERα in LMHFV-induced effects on fracture healing and provides further insights into the molecular mechanism behind these effects.

Gratings on a Dish: A Scalable Cell Alignment Substrate on Optical Media

2013 ◽  
Author(s):  
Chukwuemeka G. Anene-Nzelu ◽  
Deepak Choudhury ◽  
Huipeng Li ◽  
Yi-Chin Toh ◽  
Gary Sum Huan Ng ◽  
...  
Keyword(s):  
Low Cost ◽  
C2c12 Cell ◽  
Cell Types ◽  
Cell Alignment ◽  
Optical Media ◽  
Maturation In Vitro ◽  
Optical Grating ◽  
Topographical Cues ◽  
Different Cell Types

Cell alignment by underlying topographical cues has been shown to affect important biological processes such as differentiation and functional maturation in vitro. However, the routine use of cell culture substrates with micro/nano-topographies is currently hampered by the high cost and specialized facilities required to produce these substrates. Here we present commercially available optical media as substrates for aligning cells in culture. These optical media, including CD-R, DVD-R and optical grating, allow different cell types to attach, align and grow on them. This cytoskeletal reorganization enhanced the differentiation of cardiac (H9C2), and skeletal muscle (C2C12) cell lines. These low-cost commercially available optical media can serve as scalable substrates for research or drug safety screening applications.

scholarly journals Cytotoxicity and Sterilization Resistance of Electronic Components for Disposable Smart Biomedical Devices

2019 ◽  
Vol 5 (1) ◽  
pp. 297-301
Author(s):  
Valerie M. K. Werner ◽  
Daniel Strömsdörfer ◽  
Viet Nga Bui ◽  
Niklas von Wittenburg ◽  
Markus Eblenkamp
Keyword(s):  
Low Cost ◽  
Cost Effective ◽  
Electronic System ◽  
In Vitro Cytotoxicity ◽  
Future Research ◽  
Biomedical Devices ◽  
Electronic Components ◽  
Light Emitting ◽  
Standard Components

AbstractThe design of Smart Biomedical Devices will be a defining element of future research in the context of intelligent medical devices for the Internet of Medical Things (IoMT). A prerequisite for serving the disposable market is the use of cost-effective electronic components and the highest reliability of the developed products in terms of biocompatibility and bioprotection. In the study, resistors, capacitors, and light-emitting diodes, different in their materials and construction forms, were examined. The selected types represented electronic components as they are commonly installed on electronic system from the segment of low-cost standard components. These were subjected to steam sterilization with up to 50 cycles, gamma sterilization, and a CCK-8 assay to test in vitro cytotoxicity. Functional failure could not be determined for any component. Gamma sterilization did not result in significant changes in resistance values, but in capacitors with barium titanate as dielectric. Non-cytotoxic electronic components could be identified. The results show that certain electronic standard components are suitable for disposable Smart Biomedical Devices.

scholarly journals CurvChip - chip platform for investigating cell responses to curved surface features

2018 ◽  
Vol 4 (1) ◽  
pp. 453-456
Author(s):  
Ralf Kemkemer ◽  
Kerstin Frey ◽  
Alena Fischer ◽  
Rumen Krastev
Keyword(s):  
Low Cost ◽  
Cell Types ◽  
Cell Responses ◽  
Surface Topographies ◽  
Sharp Edges ◽  
Pdms Molding ◽  
The Impact ◽  
Thermal Reflow

AbstractSurface topographies are often discussed as an important parameter influencing basic cell behavior. Whereas most in-vitro studies deal with microstructures with sharp edges, smooth, curved microscale topographies might be more relevant concerning in-vivo situations. Addressing the lack of highly defined surfaces with varying curvature, we present a topography chip system with 3D curved features of varying spacing, curvature radii as well as varying overall dimensions of curved surfaces. The CurvChip is produced by low-cost photolithography with thermal reflow, subsequent (repetitive) PDMS molding and hot embossing. The platform facilitates the systematic invitro investigation of the impact of substrate curvature on cell types like epithelial, endothelial, smooth muscle cells, or stem cells. Such investigations will not only help to further understand the mechanism of curvature sensation but may also contribute to optimize cellmaterial interactions in the field of regenerative medicine.

scholarly journals Comprehensive Evaluation of the Toxicity and Biosafety of Plasma Polymerized Nanoparticles

Nanomaterials ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 1176
Author(s):  
Praveesuda L. Michael ◽  
Yuen Ting Lam ◽  
Juichien Hung ◽  
Richard P. Tan ◽  
Miguel Santos ◽  
...  
Keyword(s):  
Comprehensive Evaluation ◽  
Cost Effective ◽  
Cell Types ◽  
Repeated Injection ◽  
In Vivo Models ◽  
Intravenous Injections ◽  
Cost Effective Alternative ◽  
High Doses

The rapid growth of nanoparticle-based therapeutics has underpinned significant developments in nanomedicine, which aim to overcome the limitations imposed by conventional therapies. Establishing the safety of new nanoparticle formulations is the first important step on the pathway to clinical translation. We have recently shown that plasma-polymerized nanoparticles (PPNs) are highly efficient nanocarriers and a viable, cost-effective alternative to conventional chemically synthesized nanoparticles. Here, we present the first comprehensive toxicity and biosafety study of PPNs using both established in vitro cell models and in vivo models. Overall, we show that PPNs were extremely well tolerated by all the cell types tested, significantly outperforming commercially available lipid-based nanoparticles (lipofectamine) used at the manufacturer’s recommended dosage. Supporting the in vitro data, the systemic toxicity of PPNs was negligible in BALB/c mice following acute and repeated tail-vein intravenous injections. PPNs were remarkably well tolerated in mice without any evidence of behavioral changes, weight loss, significant changes to the hematological profile, or signs of histological damage in tissues. PPNs were tolerated at extremely high doses without animal mortality observed at 6000 mg/kg and 48,000 mg/kg for acute and repeated-injection regimens, respectively. Our findings demonstrate the safety of PPNs in biological systems, adding to their future potential in biomedical applications.

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