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Bioengineering, Volume 9, Issue 5 (May 2022) – 51 articles

Cover Story (view full-size image): The aim of regenerative medicine is to regrow functional tissues by manipulating exogenous or endogenous cells. Genetic engineering is one such strategy that directs cells to form the desired tissue. Among the various viral and nonviral techniques to achieve gene delivery, sonoporation has shown promising results in preclinical studies. Using ultrasound and microbubbles, sonoporation introduces genetic material to cells, leading to cell differentiation and subsequent tissue regeneration. Since it induces transient gene expression, sonoporation is considered safer compared to viral vectors, although less efficient. More research on the mode of action and the enhancement of ultrasound systems is required to optimize the therapeutic potential of sonoporation and pave its way into the clinical setting. View this paper
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20 pages, 7287 KiB  
Article
A Deep Learning Approach for the Morphological Recognition of Reactive Lymphocytes in Patients with COVID-19 Infection
by José Rodellar, Kevin Barrera, Santiago Alférez, Laura Boldú, Javier Laguna, Angel Molina and Anna Merino
Bioengineering 2022, 9(5), 229; https://doi.org/10.3390/bioengineering9050229 - 23 May 2022
Cited by 6 | Viewed by 2400
Abstract
Laboratory medicine plays a fundamental role in the detection, diagnosis and management of COVID-19 infection. Recent observations of the morphology of cells circulating in blood found the presence of particular reactive lymphocytes (COVID-19 RL) in some of the infected patients and demonstrated that [...] Read more.
Laboratory medicine plays a fundamental role in the detection, diagnosis and management of COVID-19 infection. Recent observations of the morphology of cells circulating in blood found the presence of particular reactive lymphocytes (COVID-19 RL) in some of the infected patients and demonstrated that it was an indicator of a better prognosis of the disease. Visual morphological analysis is time consuming, requires smear review by expert clinical pathologists, and is prone to subjectivity. This paper presents a convolutional neural network system designed for automatic recognition of COVID-19 RL. It is based on the Xception71 structure and is trained using images of blood cells from real infected patients. An experimental study is carried out with a group of 92 individuals. The input for the system is a set of images selected by the clinical pathologist from the blood smear of a patient. The output is the prediction whether the patient belongs to the group associated with better prognosis of the disease. A threshold is obtained for the classification system to predict that the smear belongs to this group. With this threshold, the experimental test shows excellent performance metrics: 98.3% sensitivity and precision, 97.1% specificity, and 97.8% accuracy. The system does not require costly calculations and can potentially be integrated into clinical practice to assist clinical pathologists in a more objective smear review for early prognosis. Full article
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15 pages, 3889 KiB  
Article
Poly(3-mercapto-2-methylpropionate), a Novel α-Methylated Bio-Polythioester with Rubber-like Elasticity, and Its Copolymer with 3-hydroxybutyrate: Biosynthesis and Characterization
by Lucas Vinicius Santini Ceneviva, Maierwufu Mierzati, Yuki Miyahara, Christopher T. Nomura, Seiichi Taguchi, Hideki Abe and Takeharu Tsuge
Bioengineering 2022, 9(5), 228; https://doi.org/10.3390/bioengineering9050228 - 23 May 2022
Cited by 7 | Viewed by 4471
Abstract
A new polythioester (PTE), poly(3-mercapto-2-methylpropionate) [P(3M2MP)], and its copolymer with 3-hydroxybutyrate (3HB) were successfully biosynthesized from 3-mercapto-2-methylpropionic acid as a structurally-related precursor. This is the fourth PTE of biological origin and the first to be α-methylated. P(3M2MP) was biosynthesized using an engineered Escherichia [...] Read more.
A new polythioester (PTE), poly(3-mercapto-2-methylpropionate) [P(3M2MP)], and its copolymer with 3-hydroxybutyrate (3HB) were successfully biosynthesized from 3-mercapto-2-methylpropionic acid as a structurally-related precursor. This is the fourth PTE of biological origin and the first to be α-methylated. P(3M2MP) was biosynthesized using an engineered Escherichia coli LSBJ, which has a high molecular weight, amorphous structure, and elastomeric properties, reaching 2600% elongation at break. P(3HB-co-3M2MP) copolymers were synthesized by expressing 3HB-supplying enzymes. The copolymers were produced with high content in the cells and showed a high 3M2MP unit incorporation of up to 77.2 wt% and 54.8 mol%, respectively. As the 3M2MP fraction in the copolymer increased, the molecular weight decreased and the polymers became softer, more flexible, and less crystalline, with lower glass transition temperatures and higher elongations at break. The properties of this PTE were distinct from those of previously biosynthesized PTEs, indicating that the range of material properties can be further expanded by introducing α-methylated thioester monomers. Full article
(This article belongs to the Special Issue Advances in Polyhydroxyalkanoate (PHA) Production, Volume 3)
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12 pages, 1251 KiB  
Review
Cervical and Lumbar Disc Arthroplasty: A Review of Current Implant Design and Outcomes
by Ian J. Wellington, Cameron Kia, Ergin Coskun, Barrett B. Torre, Christopher L. Antonacci, Michael R. Mancini, John P. Connors, Sean M. Esmende and Heeren S. Makanji
Bioengineering 2022, 9(5), 227; https://doi.org/10.3390/bioengineering9050227 - 23 May 2022
Cited by 4 | Viewed by 3152
Abstract
While spinal disc pathology has traditionally been treated using fusion-based procedures, recent interest in motion-preserving disc arthroplasties has grown. Traditional spinal fusion is associated with loss of motion, alteration of native spine kinematics, and increased risks of adjacent segment disease. The motion conferred [...] Read more.
While spinal disc pathology has traditionally been treated using fusion-based procedures, recent interest in motion-preserving disc arthroplasties has grown. Traditional spinal fusion is associated with loss of motion, alteration of native spine kinematics, and increased risks of adjacent segment disease. The motion conferred by disc arthroplasty is believed to combat these complications. While the first implant designs resulted in poor patient outcomes, recent advances in implant design and technology have shown promising radiographic and clinical outcomes when compared with traditional fusion. These results have led to a rapid increase in the utilization of disc arthroplasty, with rates of cervical arthroplasty nearly tripling over the course of 7 years. The purpose of this review was to discuss the evolution of implant design, the current implant designs utilized, and their associated outcomes. Although disc arthroplasty shows significant promise in addressing some of the drawbacks associated with fusion, it is not without its own risks. Osteolysis, implant migration, and the development of heterotopic ossification have all been associated with disc arthroplasty. As interest in these procedures grows, so does the interest in developing improved implant designs aimed at decreasing these adverse outcomes. Though they are still relatively new, cervical and lumbar disc arthroplasty are likely to become foundational methodologies for the treatment of disc pathology. Full article
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22 pages, 730 KiB  
Review
Applications of Photodynamic Therapy in Endometrial Diseases
by Gabriela Correia-Barros, Beatriz Serambeque, Maria João Carvalho, Carlos Miguel Marto, Marta Pineiro, Teresa M. V. D. Pinho e Melo, Maria Filomena Botelho and Mafalda Laranjo
Bioengineering 2022, 9(5), 226; https://doi.org/10.3390/bioengineering9050226 - 23 May 2022
Cited by 3 | Viewed by 2908
Abstract
Photodynamic therapy (PDT) is a medical procedure useful for several benign conditions (such as wound healing and infections) and cancer. PDT is minimally invasive, presents few side effects, good scaring, and is able to minimal tissue destruction maintaining organ anatomy and function. Endoscopic [...] Read more.
Photodynamic therapy (PDT) is a medical procedure useful for several benign conditions (such as wound healing and infections) and cancer. PDT is minimally invasive, presents few side effects, good scaring, and is able to minimal tissue destruction maintaining organ anatomy and function. Endoscopic access to the uterus puts PDT in the spotlight for endometrial disease treatment. This work systematically reviews the current evidence of PDT’s potential and usefulness in endometrial diseases. Thus, this narrative review focused on PDT applications for endometrial disease, including reports regarding in vitro, ex vivo, animal, and clinical studies. Cell lines and primary samples were used as in vitro models of cancer, adenomyosis and endometrioses, while most animal studies focused the PDT outcomes on endometrial ablation. A few clinical attempts are known using PDT for endometrial ablation and cancer lesions. This review emphasises PDT as a promising field of research. This therapeutic approach has the potential to become an effective conservative treatment method for endometrial benign and malignant lesions. Further investigations with improved photosensitisers are highly expected. Full article
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24 pages, 2187 KiB  
Review
Review of the Developments of Bacterial Medium-Chain-Length Polyhydroxyalkanoates (mcl-PHAs)
by V. Uttej Nandan Reddy, S. V. Ramanaiah, M. Venkateswar Reddy and Young-Cheol Chang
Bioengineering 2022, 9(5), 225; https://doi.org/10.3390/bioengineering9050225 - 21 May 2022
Cited by 28 | Viewed by 7143
Abstract
Synthetic plastics derived from fossil fuels—such as polyethylene, polypropylene, polyvinyl chloride, and polystyrene—are non-degradable. A large amount of plastic waste enters landfills and pollutes the environment. Hence, there is an urgent need to produce biodegradable plastics such as polyhydroxyalkanoates (PHAs). PHAs have garnered [...] Read more.
Synthetic plastics derived from fossil fuels—such as polyethylene, polypropylene, polyvinyl chloride, and polystyrene—are non-degradable. A large amount of plastic waste enters landfills and pollutes the environment. Hence, there is an urgent need to produce biodegradable plastics such as polyhydroxyalkanoates (PHAs). PHAs have garnered increasing interest as replaceable materials to conventional plastics due to their broad applicability in various purposes such as food packaging, agriculture, tissue-engineering scaffolds, and drug delivery. Based on the chain length of 3-hydroxyalkanoate repeat units, there are three types PHAs, i.e., short-chain-length (scl-PHAs, 4 to 5 carbon atoms), medium-chain-length (mcl-PHAs, 6 to 14 carbon atoms), and long-chain-length (lcl-PHAs, more than 14 carbon atoms). Previous reviews discussed the recent developments in scl-PHAs, but there are limited reviews specifically focused on the developments of mcl-PHAs. Hence, this review focused on the mcl-PHA production, using various carbon (organic/inorganic) sources and at different operation modes (continuous, batch, fed-batch, and high-cell density). This review also focused on recent developments on extraction methods of mcl-PHAs (solvent, non-solvent, enzymatic, ultrasound); physical/thermal properties (Mw, Mn, PDI, Tm, Tg, and crystallinity); applications in various fields; and their production at pilot and industrial scales in Asia, Europe, North America, and South America. Full article
(This article belongs to the Special Issue Advances in Polyhydroxyalkanoate (PHA) Production, Volume 3)
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9 pages, 2708 KiB  
Article
Influence of Sagittal Lumbopelvic Morphotypes on the Range of Motion of Human Lumbar Spine: An In Vitro Cadaveric Study
by Wei Wang, Chao Kong, Fumin Pan, Wei Wang, Xueqing Wu, Baoqing Pei and Shibao Lu
Bioengineering 2022, 9(5), 224; https://doi.org/10.3390/bioengineering9050224 - 20 May 2022
Cited by 1 | Viewed by 1988
Abstract
Background: Although spinopelvic radiographs analysis is the standard for a pathological diagnosis, it cannot explain the activities of the spine in daily life. This study investigates the correlation between sagittal parameters and spinal range of motion (ROM) to find morphological parameters with kinetic [...] Read more.
Background: Although spinopelvic radiographs analysis is the standard for a pathological diagnosis, it cannot explain the activities of the spine in daily life. This study investigates the correlation between sagittal parameters and spinal range of motion (ROM) to find morphological parameters with kinetic implications. Methods: Six L1–S1 human lumbar specimens were tested with a robotic testing device. Eight sagittal parameters were measured in the three-dimensional model. Pure moments were applied to simulate the physiological activities in daily life. Results: The correlation between sagittal parameters and the ROM was moderate in flexion and extension, but weak in lateral bending and rotation. In flexion–extension, the ROM was moderately correlated with SS and LL. SS was the only parameter correlated with the ROM under all loading conditions. The intervertebral rotation distribution showed that the maximal ROM frequently occurred at the L5–S1 segment. The minimal ROM often appeared near the apex point of the lumbar. Conclusion: Sagittal alignment mainly affected the ROM of the lumbar in flexion and extension. SS and apex may have had kinetic significance. Our findings suggest that the effect of sagittal parameters on lumbar ROM is important information for assessing spinal activity. Full article
(This article belongs to the Topic Human Movement Analysis)
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15 pages, 580 KiB  
Review
Bioengineering Approaches for Delivering Growth Factors: A Focus on Bone and Cartilage Regeneration
by Sheeba Shakoor, Eleyna Kibble and Jehan J. El-Jawhari
Bioengineering 2022, 9(5), 223; https://doi.org/10.3390/bioengineering9050223 - 20 May 2022
Cited by 6 | Viewed by 2448
Abstract
Growth factors are bio-factors that target reparatory cells during bone regeneration. These growth factors are needed in complicated conditions of bone and joint damage to enhance tissue repair. The delivery of these growth factors is key to ensuring the effectiveness of regenerative therapy. [...] Read more.
Growth factors are bio-factors that target reparatory cells during bone regeneration. These growth factors are needed in complicated conditions of bone and joint damage to enhance tissue repair. The delivery of these growth factors is key to ensuring the effectiveness of regenerative therapy. This review discusses the roles of various growth factors in bone and cartilage regeneration. The methods of delivery of natural or recombinant growth factors are reviewed. Different types of scaffolds, encapsulation, Layer-by-layer assembly, and hydrogels are tools for growth factor delivery. Considering the advantages and limitations of these methods is essential to developing regenerative therapies. Further research can accordingly be planned to have new or combined technologies serving this purpose. Full article
(This article belongs to the Special Issue Current Developments and Applications in Bone Tissue Engineering)
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15 pages, 3036 KiB  
Article
Synergistic Inorganic Carbon and Denitrification Genes Contributed to Nitrite Accumulation in a Hydrogen-Based Membrane Biofilm Reactor
by Si Pang, Bruce E. Rittmann, Chengyang Wu, Lin Yang, Jingzhou Zhou and Siqing Xia
Bioengineering 2022, 9(5), 222; https://doi.org/10.3390/bioengineering9050222 - 20 May 2022
Cited by 9 | Viewed by 2288
Abstract
Partial denitrification, the termination of NO3-N reduction at nitrite (NO2-N), has received growing interest for treating wastewaters with high ammonium concentrations, because it can be coupled to anammox for total-nitrogen removal. NO2 accumulation in the [...] Read more.
Partial denitrification, the termination of NO3-N reduction at nitrite (NO2-N), has received growing interest for treating wastewaters with high ammonium concentrations, because it can be coupled to anammox for total-nitrogen removal. NO2 accumulation in the hydrogen (H2)-based membrane biofilm reactor (MBfR) has rarely been studied, and the mechanisms behind its accumulation have not been defined. This study aimed at achieving the partial denitrification with H2-based autotrophic reducing bacteria in a MBfR. Results showed that by increasing the NO3 loading, increasing the pH, and decreasing the inorganic-carbon concentration, a nitrite transformation rate higher than 68% was achieved. Community analysis indicated that Thauera and Azoarcus became the dominant genera when partial denitrification was occurring. Functional genes abundances proved that partial denitrification to accumulate NO2 was correlated to increases of gene for the form I RuBisCo enzyme (cbbL). This study confirmed the feasibility of autotrophic partial denitrification formed in the MBfR, and revealed the inorganic carbon mechanism in MBfR denitrification. Full article
(This article belongs to the Topic Bioreactors: Control, Optimization and Applications)
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11 pages, 1867 KiB  
Article
Mechanical and Metallurgical Evaluation of 3 Different Nickel-Titanium Rotary Instruments: An In Vitro and In Laboratory Study
by Alessio Zanza, Paola Russo, Rodolfo Reda, Paola Di Matteo, Orlando Donfrancesco, Pietro Ausiello and Luca Testarelli
Bioengineering 2022, 9(5), 221; https://doi.org/10.3390/bioengineering9050221 - 20 May 2022
Cited by 6 | Viewed by 1876
Abstract
An in-depth evaluation of the mechanical and metallurgical properties of NiTi instruments is fundamental to assess their performance and to compare recently introduced instrument with widespread ones. According to this, since there are no data on this topic, the aim of the study [...] Read more.
An in-depth evaluation of the mechanical and metallurgical properties of NiTi instruments is fundamental to assess their performance and to compare recently introduced instrument with widespread ones. According to this, since there are no data on this topic, the aim of the study was to mechanically and metallurgically evaluate an instrument recently introduced into the market (ZenFlex (ZF)), by comparing it with two well-known instruments with similar characteristics: Vortex Blue (VB) and EdgeSequel Sapphire (EES). According to this, 195 instruments were selected: 65 ZF, 65 VB and 65 EES. Each group was divided in subgroups according to the mechanical tests (i.e., cyclic fatigue resistance, torsional resistance and bending ability; (n = 20)) and the metallurgical test (differential scanning calorimetry (n = 5)). A scanning electron microscopy was performed to verify the causes of fracture after mechanical tests (cyclic fatigue and torsional tests). According to results, VB showed the highest flexibility and cyclic fatigue resistance in comparison to the other instruments, with a statistically significant difference (p < 0.05). Regarding torsional resistance, EES showed the lowest value of torque at fracture, with a statistically significant difference, whilst the comparison between ZF and VB showed no statistically significant difference (p > 0.05). DSC analysis pointed out that VB had the highest austenite start and finish temperatures, followed by ESS and then ZF. ESS sample showed the highest martensite start and finish temperatures followed by VB and ZF. Considering the results, it can be concluded that VB showed the best mechanical performance during static tests in comparison to ESS and ZF. This is fundamentally due to the interaction of parameters such as instrument design and heat-treatments that are able to enhance its mechanical performance. Full article
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41 pages, 34239 KiB  
Review
MatriGrid® Based Biological Morphologies: Tools for 3D Cell Culturing
by Patrick Mai, Jörg Hampl, Martin Baca, Dana Brauer, Sukhdeep Singh, Frank Weise, Justyna Borowiec, André Schmidt, Johanna Merle Küstner, Maren Klett, Michael Gebinoga, Insa S. Schroeder, Udo R. Markert, Felix Glahn, Berit Schumann, Diana Eckstein and Andreas Schober
Bioengineering 2022, 9(5), 220; https://doi.org/10.3390/bioengineering9050220 - 20 May 2022
Cited by 4 | Viewed by 3526
Abstract
Recent trends in 3D cell culturing has placed organotypic tissue models at another level. Now, not only is the microenvironment at the cynosure of this research, but rather, microscopic geometrical parameters are also decisive for mimicking a tissue model. Over the years, technologies [...] Read more.
Recent trends in 3D cell culturing has placed organotypic tissue models at another level. Now, not only is the microenvironment at the cynosure of this research, but rather, microscopic geometrical parameters are also decisive for mimicking a tissue model. Over the years, technologies such as micromachining, 3D printing, and hydrogels are making the foundation of this field. However, mimicking the topography of a particular tissue-relevant substrate can be achieved relatively simply with so-called template or morphology transfer techniques. Over the last 15 years, in one such research venture, we have been investigating a micro thermoforming technique as a facile tool for generating bioinspired topographies. We call them MatriGrid®s. In this research account, we summarize our learning outcome from this technique in terms of the influence of 3D micro morphologies on different cell cultures that we have tested in our laboratory. An integral part of this research is the evolution of unavoidable aspects such as possible label-free sensing and fluidic automatization. The development in the research field is also documented in this account. Full article
(This article belongs to the Special Issue Analytical Approaches in 3D in vitro Systems)
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16 pages, 22378 KiB  
Article
Human-Origin iPSC-Based Recellularization of Decellularized Whole Rat Livers
by Aylin Acun, Ruben Oganesyan, Maria Jaramillo, Martin L. Yarmush and Basak E. Uygun
Bioengineering 2022, 9(5), 219; https://doi.org/10.3390/bioengineering9050219 - 19 May 2022
Cited by 10 | Viewed by 2435
Abstract
End-stage liver diseases lead to mortality of millions of patients, as the only treatment available is liver transplantation and donor scarcity means that patients have to wait long periods before receiving a new liver. In order to minimize donor organ scarcity, a promising [...] Read more.
End-stage liver diseases lead to mortality of millions of patients, as the only treatment available is liver transplantation and donor scarcity means that patients have to wait long periods before receiving a new liver. In order to minimize donor organ scarcity, a promising bioengineering approach is to decellularize livers that do not qualify for transplantation. Through decellularization, these organs can be used as scaffolds for developing new functional organs. In this process, the original cells of the organ are removed and ideally should be replaced by patient-specific cells to eliminate the risk of immune rejection. Induced pluripotent stem cells (iPSCs) are ideal candidates for developing patient-specific organs, yet the maturity and functionality of iPSC-derived cells do not match those of primary cells. In this study, we introduced iPSCs into decellularized rat liver scaffolds prior to the start of differentiation into hepatic lineages to maximize the exposure of iPSCs to native liver matrices. Through exposure to the unique composition and native 3D organization of the liver microenvironment, as well as the more efficient perfusion culture throughout the differentiation process, iPSC differentiation into hepatocyte-like cells was enhanced. The resulting cells showed significantly higher expression of mature hepatocyte markers, including important CYP450 enzymes, along with lower expression of fetal markers, such as AFP. Importantly, the gene expression profile throughout the different stages of differentiation was more similar to native development. Our study shows that the native 3D liver microenvironment has a pivotal role to play in the development of human-origin hepatocyte-like cells with more mature characteristics. Full article
(This article belongs to the Special Issue Cell-ECM Interactions for Tissue Engineering and Tissue Regeneration)
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25 pages, 9375 KiB  
Article
Oral Cells-On-Chip: Design, Modeling and Experimental Results
by Hamed Osouli Tabrizi, Abbas Panahi, Saghi Forouhi, Deniz Sadighbayan, Fatemeh Soheili, Mohammad Reza Haji Hosseini Khani, Sebastian Magierowski and Ebrahim Ghafar-Zadeh
Bioengineering 2022, 9(5), 218; https://doi.org/10.3390/bioengineering9050218 - 19 May 2022
Cited by 3 | Viewed by 2728
Abstract
Recent advances in periodontal studies have attracted the attention of researchers to the relation between oral cells and gum diseases, which is a real threat to overall human health. Among various microfabrication technologies, Complementary Metal Oxide Semiconductors (CMOSs) enable the development of low-cost [...] Read more.
Recent advances in periodontal studies have attracted the attention of researchers to the relation between oral cells and gum diseases, which is a real threat to overall human health. Among various microfabrication technologies, Complementary Metal Oxide Semiconductors (CMOSs) enable the development of low-cost integrated sensors and circuits for rapid and accurate assessment of living cells that can be employed for the early detection and control of periodontal diseases. This paper presents a CMOS capacitive sensing platform that can be considered as an alternative for the analysis of salivatory cells such as oral neutrophils. This platform consists of two sensing electrodes connected to a read-out capacitive circuitry designed and fabricated on the same chip using Austria Mikro Systeme (AMS) 0.35 µm CMOS process. A graphical user interface (GUI) was also developed to interact with the capacitive read-out system and the computer to monitor the capacitance changes due to the presence of saliva cells on top of the chip. Thanks to the wide input dynamic range (IDR) of more than 400 femto farad (fF) and high resolution of 416 atto farad (aF), the experimental and simulation results demonstrate the functionality and applicability of the proposed sensor for monitoring cells in a small volume of 1 µL saliva samples. As per these results, the hydrophilic adhesion of oral cells on the chip varies the capacitance of interdigitated electrodes (IDEs). These capacitance changes then give an assessment of the oral cells existing in the sample. In this paper, the simulation and experimental results set a new stage for emerging sensing platforms for testing oral samples. Full article
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12 pages, 840 KiB  
Review
Application of Sygen® in Diabetic Peripheral Neuropathies—A Review of Biological Interactions
by Marcelo Amaral Coelho, Madhan Jeyaraman, Naveen Jeyaraman, Ramya Lakshmi Rajendran, André Atsushi Sugano, Tomas Mosaner, Gabriel Silva Santos, João Vitor Bizinotto Lana, Anna Vitória Santos Duarte Lana, Lucas Furtado da Fonseca, Rafael Barnabé Domingues, Prakash Gangadaran, Byeong-Cheol Ahn and José Fábio Santos Duarte Lana
Bioengineering 2022, 9(5), 217; https://doi.org/10.3390/bioengineering9050217 - 18 May 2022
Cited by 2 | Viewed by 3110
Abstract
This study investigates the role of Sygen® in diabetic peripheral neuropathy, a severe disease that affects the peripheral nervous system in diabetic individuals. This disorder often impacts the lower limbs, causing significant discomfort and, if left untreated, progresses into more serious conditions [...] Read more.
This study investigates the role of Sygen® in diabetic peripheral neuropathy, a severe disease that affects the peripheral nervous system in diabetic individuals. This disorder often impacts the lower limbs, causing significant discomfort and, if left untreated, progresses into more serious conditions involving chronic ulcers and even amputation in many cases. Although there are management strategies available, peripheral neuropathies are difficult to treat as they often present multiple causes, especially due to metabolic dysfunction in diabetic individuals. Gangliosides, however, have long been studied and appreciated for their role in neurological diseases. The monosialotetrahexosylganglioside (GM1) ganglioside, popularly known as Sygen, provides beneficial effects such as enhanced neuritic sprouting, neurotrophism, neuroprotection, anti-apoptosis, and anti-excitotoxic activity, being particularly useful in the treatment of neurological complications that arise from diabetes. This product mimics the roles displayed by neurotrophins, improving neuronal function and immunomodulation by attenuating exacerbated inflammation in neurons. Furthermore, Sygen assists in axonal stabilization and keeps nodal and paranodal regions of myelin fibers organized. This maintains an adequate propagation of action potentials and restores standard peripheral nerve function. Given the multifactorial nature of this complicated disorder, medical practitioners must carefully screen the patient to avoid confusion and misdiagnosis. There are several studies analyzing the role of Sygen in neurological disorders. However, the medical literature still needs more robust investigations such as randomized clinical trials regarding the administration of this compound for diabetic peripheral neuropathies, specifically. Full article
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14 pages, 1839 KiB  
Article
3D Cephalometric Normality Range: Auto Contractive Maps (ACM) Analysis in Selected Caucasian Skeletal Class I Age Groups
by Marco Farronato, Giuseppe Baselli, Benedetta Baldini, Gianfranco Favia and Gianluca Martino Tartaglia
Bioengineering 2022, 9(5), 216; https://doi.org/10.3390/bioengineering9050216 - 17 May 2022
Cited by 18 | Viewed by 2228
Abstract
The objective of this paper is to define normal values of a novel 3D cephalometric analysis and to define the links through an artificial neural network (ANN). Methods: One hundred and fifteen CBCTs of Class I young patients, distributed among gender-adjusted developmental groups, [...] Read more.
The objective of this paper is to define normal values of a novel 3D cephalometric analysis and to define the links through an artificial neural network (ANN). Methods: One hundred and fifteen CBCTs of Class I young patients, distributed among gender-adjusted developmental groups, were selected. Three operators identified 18 cephalometric landmarks from which 36 measurements were obtained. The repeatability was assessed through the ICC. Two-dimensional values were extracted by an automatic function, and the mean value and standard deviation were compared by paired Student’s t-tests. Correlation coefficient gave the relationships between 2D and 3D measurements for each group. The values were computed with the ANN to evaluate the parameters normality link and displayed by Pajek software. Results: The ICC assessed an excellent (≥0.9) repeatability. Normal values were extracted, and compared with 2D measurements, they showed a high correlation on the mid-sagittal plane, reaching 1.00, with the lowest 0.71 on the lateral plane. The ANN showed strong links between the values with the centrality of the go-sagittal plane compared to the rest. Conclusions: The study provides a set of 3D cephalometric values obtained by the upper and lower 95% CI for the mean divided into the developmental stage subgroups. The two-dimensional measurements showed variable concordance, while the ANN showed a centrality between the parameters. Full article
(This article belongs to the Section Regenerative Engineering)
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13 pages, 3182 KiB  
Article
Self-Assembled Hydrogel Microparticle-Based Tooth-Germ Organoids
by Cemile Kilic Bektas, Weibo Zhang, Yong Mao, Xiaohuan Wu, Joachim Kohn and Pamela C. Yelick
Bioengineering 2022, 9(5), 215; https://doi.org/10.3390/bioengineering9050215 - 17 May 2022
Cited by 10 | Viewed by 4051
Abstract
Here, we describe the characterization of tooth-germ organoids, three-dimensional (3D) constructs cultured in vitro with the potential to develop into living teeth. To date, the methods used to successfully create tooth organoids capable of forming functional teeth have been quite limited. Recently, hydrogel [...] Read more.
Here, we describe the characterization of tooth-germ organoids, three-dimensional (3D) constructs cultured in vitro with the potential to develop into living teeth. To date, the methods used to successfully create tooth organoids capable of forming functional teeth have been quite limited. Recently, hydrogel microparticles (HMP) have demonstrated utility in tissue repair and regeneration based on their useful characteristics, including their scaffolding ability, effective cell and drug delivery, their ability to mimic the natural tissue extracellular matrix, and their injectability. These outstanding properties led us to investigate the utility of using HMPs (average diameter: 158 ± 32 µm) derived from methacrylated gelatin (GelMA) (degree of substitution: 100%) to create tooth organoids. The tooth organoids were created by seeding human dental pulp stem cells (hDPSCs) and porcine dental epithelial cells (pDE) onto the HMPs, which provided an extensive surface area for the cells to effectively attach and proliferate. Interestingly, the cell-seeded HMPs cultured on low-attachment tissue culture plates with gentle rocking self-assembled into organoids, within which the cells maintained their viability and morphology throughout the incubation period. The self-assembled organoids reached a volume of ~50 mm3 within two weeks of the in vitro tissue culture. The co-cultured hDPSC-HMP and pDE-HMP structures effectively attached to each other without any externally applied forces. The presence of polarized, differentiated dental cells in these composite tooth-bud organoids demonstrated the potential of self-assembled dental cell HMPs to form tooth-bud organoid-like structures for potential applications in tooth regeneration strategies. Full article
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21 pages, 1401 KiB  
Article
Identification of Radiation-Induced miRNA Biomarkers Using the CGL1 Cell Model System
by Jayden Peterson, Christopher D. McTiernan, Christopher Thome, Neelam Khaper, Simon J. Lees, Douglas R. Boreham, Tze Chun Tai and Sujeenthar Tharmalingam
Bioengineering 2022, 9(5), 214; https://doi.org/10.3390/bioengineering9050214 - 16 May 2022
Cited by 2 | Viewed by 2199
Abstract
MicroRNAs (miRNAs) have emerged as a potential class of biomolecules for diagnostic biomarker applications. miRNAs are small non-coding RNA molecules, produced and released by cells in response to various stimuli, that demonstrate remarkable stability in a wide range of biological fluids, in extreme [...] Read more.
MicroRNAs (miRNAs) have emerged as a potential class of biomolecules for diagnostic biomarker applications. miRNAs are small non-coding RNA molecules, produced and released by cells in response to various stimuli, that demonstrate remarkable stability in a wide range of biological fluids, in extreme pH fluctuations, and after multiple freeze–thaw cycles. Given these advantages, identification of miRNA-based biomarkers for radiation exposures can contribute to the development of reliable biological dosimetry methods, especially for low-dose radiation (LDR) exposures. In this study, an miRNAome next-generation sequencing (NGS) approach was utilized to identify novel radiation-induced miRNA gene changes within the CGL1 human cell line. Here, irradiations of 10, 100, and 1000 mGy were performed and the samples were collected 1, 6, and 24 h post-irradiation. Corroboration of the miRNAome results with RT-qPCR verification confirmed the identification of numerous radiation-induced miRNA expression changes at all doses assessed. Further evaluation of select radiation-induced miRNAs, including miR-1228-3p and miR-758-5p, as well as their downstream mRNA targets, Ube2d2, Ppp2r2d, and Id2, demonstrated significantly dysregulated reciprocal expression patterns. Further evaluation is needed to determine whether the candidate miRNA biomarkers identified in this study can serve as suitable targets for radiation biodosimetry applications. Full article
(This article belongs to the Special Issue Molecular Diagnostics in Postgenomic Era)
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11 pages, 564 KiB  
Article
Cardiac Function after Modern Radiation Therapy with Volumetric Modulated Arc Therapy or Helical Tomotherapy for Advanced Left-Breast Cancer Receiving Regional Nodal Irradiation
by Pei-Yu Hou, Chen-Hsi Hsieh, Le-Jung Wu, Chen-Xiong Hsu, Deng-Yu Kuo, Yueh-Feng Lu, Yen-Wen Wu, Hui-Ju Tien, Shih-Ming Hsu and Pei-Wei Shueng
Bioengineering 2022, 9(5), 213; https://doi.org/10.3390/bioengineering9050213 - 16 May 2022
Cited by 3 | Viewed by 1723
Abstract
Background: Protecting cardiac function in patients with advanced left-breast cancer receiving radiation therapy (RT) with regional nodal irradiation (RNI) is an important issue. Modern RT techniques can limit cardiac exposure. The aim of this study was to explore the association be-tween cardiac dose [...] Read more.
Background: Protecting cardiac function in patients with advanced left-breast cancer receiving radiation therapy (RT) with regional nodal irradiation (RNI) is an important issue. Modern RT techniques can limit cardiac exposure. The aim of this study was to explore the association be-tween cardiac dose and cardiac function. Methods: Between 2017 and 2020, we retrospectively reviewed left-breast cancer patients who received adjuvant RT, including RNI with either volumetric-modulated arc therapy (VMAT) or helical tomotherapy (HT). Left ventricular ejection fraction (LVEF) was assessed by echocardiography before RT and 1 year after RT to detect any early deterioration in cardiac systolic function. Results: A total of 30 eligible patients were enrolled. The median follow-up time from the initiation of RT was 3.9 years (range 0.6–5 years). Seventeen patients received VMAT, and the other 13 patients received HT. The median RT dose was 55 Gray (Gy), and the mean heart dose was 3.73 Gy (range 1.95–9.36 Gy). The median LVEF before and after RT was 68% and 68.5%, respectively. No obvious deterioration was found. There was no association between cardiac dose (mean heart dose, V5–V30) and LVEF (change in values or post-RT). Conclusions: For left-breast cancer patients undergoing RT with RNI, VMAT, or HT can be used to limit cardiac exposure. Cardiac function as evaluated by LVEF revealed no obvious deterioration after RT in our patients, and no association was found between cardiac dose and LVEF in those treated with either VMAT or HT in early cardiac surveillance. Full article
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12 pages, 794 KiB  
Article
3D Modeling of the Crystalline Lens Complex under Pseudoexfoliation
by Leonor Jud, André P. G. Castro, Rui B. Ruben, Bernardo Feijóo, Filomena J. Ribeiro and Paulo R. Fernandes
Bioengineering 2022, 9(5), 212; https://doi.org/10.3390/bioengineering9050212 - 13 May 2022
Viewed by 1853
Abstract
Pseudoexfoliation, one of the most frequent crystalline lens complex disorders, is prevalent in up to 30% of individuals older than 60 years old. This disease can lead to severe conditions, such as subluxation or dislocation of the lens, due to the weakening of [...] Read more.
Pseudoexfoliation, one of the most frequent crystalline lens complex disorders, is prevalent in up to 30% of individuals older than 60 years old. This disease can lead to severe conditions, such as subluxation or dislocation of the lens, due to the weakening of the zonules. The goal for the present study was to understand the relevant biomechanical features that can lead to the worsening of an individual’s visual capacity under pseudoexfoliation. To this end, finite element models based on a 62-year-old lens complex were developed, composed by the capsular bag, cortex, nucleus, anterior, equatorial, and posterior zonular fibers. Healthy and pseudoexfoliative conditions were simulated, varying the location of the zonulopathy (inferior/superior) and the degenerated layer. The accommodative capacity of the models with inferior dialysis of the zonular fibers was, on average, 4.7% greater than for the cases with superior dialysis. If the three sets of zonules were disrupted, this discrepancy increased to 14.9%. The present work provides relevant data to be further analyzed in clinical scenarios, as these models (and their future extension to a wider age range) can help in identifying the most influential regions for the reduction of the visual capacity of the lens. Full article
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6 pages, 1270 KiB  
Article
Telehealth and Burn Care: From Faxes to Augmented Reality
by Caroline Park, Youngwoo Cho, Jalen Harvey, Brett Arnoldo and Benjamin Levi
Bioengineering 2022, 9(5), 211; https://doi.org/10.3390/bioengineering9050211 - 13 May 2022
Cited by 1 | Viewed by 2024
Abstract
Despite advances in telemedicine, practices remain diverse, ranging from telephonic to still images and video-based conferencing. We review the various modes of telemedicine in burn care and summarize relevant studies, including their contributions and limitations. We also review the role of a more [...] Read more.
Despite advances in telemedicine, practices remain diverse, ranging from telephonic to still images and video-based conferencing. We review the various modes of telemedicine in burn care and summarize relevant studies, including their contributions and limitations. We also review the role of a more recent technology, augmented reality, and its role in the triage and management of burn patients. Telemedicine in burn care remains diverse, with varied outcomes in accuracy and efficiency. Newer technologies such as augmented reality have not been extensively studied or implemented but show promise in immersive, real-time triage. Full article
(This article belongs to the Special Issue Bioengineered Strategies for Surgical Innovation)
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21 pages, 7783 KiB  
Article
Design, Spectral Characteristics, Photostability, and Possibilities for Practical Application of BODIPY FL-Labeled Thioterpenoid
by Galina B. Guseva, Elena V. Antina, Mikhail B. Berezin, Anastassia S. Smirnova, Roman S. Pavelyev, Ilmir R. Gilfanov, Oksana G. Shevchenko, Svetlana V. Pestova, Evgeny S. Izmest’ev, Svetlana A. Rubtsova, Olga V. Ostolopovskaya, Sergey V. Efimov, Vladimir V. Klochkov, Ilfat Z. Rakhmatullin, Ayzira F. Timerova, Ilya A. Khodov, Olga A. Lodochnikova, Daut R. Islamov, Pavel V. Dorovatovskii, Liliya E. Nikitina and Sergei V. Boichukadd Show full author list remove Hide full author list
Bioengineering 2022, 9(5), 210; https://doi.org/10.3390/bioengineering9050210 - 12 May 2022
Cited by 3 | Viewed by 2222
Abstract
This paper presents the design and a comparative analysis of the structural and solvation factors on the spectral and biological properties of the BODIPY biomarker with a thioterpene fragment. Covalent binding of the thioterpene moiety to the butanoic acid residue of meso-substituted [...] Read more.
This paper presents the design and a comparative analysis of the structural and solvation factors on the spectral and biological properties of the BODIPY biomarker with a thioterpene fragment. Covalent binding of the thioterpene moiety to the butanoic acid residue of meso-substituted BODIPY was carried out to find out the membranotropic effect of conjugate to erythrocytes, and to assess the possibilities of its practical application in bioimaging. The molecular structure of the conjugate was confirmed via X-ray, UV/vis-, NMR-, and MS-spectra. It was found that dye demonstrates high photostability and high fluorescence quantum yield (to ~100%) at 514–519 nm. In addition, the marker was shown to effectively penetrate the erythrocytes membrane in the absence of erythrotoxicity. The conjugation of BODIPY with thioterpenoid is an excellent way to increase affinity dyes to biostructures, including blood components. Full article
(This article belongs to the Special Issue Molecular Diagnostics in Postgenomic Era)
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10 pages, 3073 KiB  
Article
Soluble Papain to Digest Monoclonal Antibodies; Time and Cost-Effective Method to Obtain Fab Fragment
by Matthew Collins and Hanieh Khalili
Bioengineering 2022, 9(5), 209; https://doi.org/10.3390/bioengineering9050209 - 12 May 2022
Cited by 4 | Viewed by 7836
Abstract
Antigen binding fragments (Fabs) used in research (e.g., antibody mimetics, antibody-drug conjugate, bispecific antibodies) are frequently obtained by enzymatic digestion of monoclonal antibodies using immobilised papain. Despite obtaining pure Fab, using immobilised papain to digest IgG has limitations, most notably slow digestion time [...] Read more.
Antigen binding fragments (Fabs) used in research (e.g., antibody mimetics, antibody-drug conjugate, bispecific antibodies) are frequently obtained by enzymatic digestion of monoclonal antibodies using immobilised papain. Despite obtaining pure Fab, using immobilised papain to digest IgG has limitations, most notably slow digestion time (more than 8 h), high cost and limited scalability. Here we report a time and cost-effective method to produce pure, active and stable Fab using soluble papain. Large laboratory scale digestion of an antibody (100 mg) was achieved using soluble papain with a digestion time of 30 min and isolated yields of 55–60%. The obtained Fabs displayed similar binding activity as Fabs prepared via immobilised papain digestion. Site-specific conjugation between Fabs and polyethylene glycol (PEG) was carried out to obtain antibody mimetics FpF (Fab-PEG-Fab) indicating that the native disulphide bond had been preserved. Surface-plasmon resonance (SPR) of prepared FpFs showed that binding activity towards the intended antigen was maintained. We anticipate that this work will provide a fast and less costly method for researchers to produce antibody fragments at large scale from whole IgG suitable for use in research. Full article
(This article belongs to the Special Issue Bioengineering of Biotherapeutics)
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15 pages, 2329 KiB  
Article
Hybrid Zero Dynamics Control for Gait Guidance of a Novel Adjustable Pediatric Lower-Limb Exoskeleton
by Anthony Goo, Curt A. Laubscher, Jason J. Wiebrecht, Ryan J. Farris and Jerzy T. Sawicki
Bioengineering 2022, 9(5), 208; https://doi.org/10.3390/bioengineering9050208 - 12 May 2022
Cited by 4 | Viewed by 2465
Abstract
Exoskeleton technology has undergone significant developments for the adult population but is still lacking for the pediatric population. This paper presents the design of a hip–knee exoskeleton for children 6 to 11 years old with gait abnormalities. The actuators are housed in an [...] Read more.
Exoskeleton technology has undergone significant developments for the adult population but is still lacking for the pediatric population. This paper presents the design of a hip–knee exoskeleton for children 6 to 11 years old with gait abnormalities. The actuators are housed in an adjustable exoskeleton frame where the thigh part can adjust in length and the hip cradle can adjust in the medial-lateral and posterior-anterior directions concurrently. Proper control of exoskeletons to follow nominal healthy gait patterns in a time-invariant manner is important for ease of use and user acceptance. In this paper, a hybrid zero dynamics (HZD) controller was designed for gait guidance by defining the zero dynamics manifold to resemble healthy gait patterns. HZD control utilizes a time-invariant feedback controller to create dynamically stable gaits in robotic systems with hybrid models containing both discrete and continuous dynamics. The effectiveness of the controller on the novel pediatric exoskeleton was demonstrated via simulation. The presented preliminary results suggest that HZD control provides a viable method to control the pediatric exoskeleton for gait guidance. Full article
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19 pages, 333 KiB  
Review
Research Progress on the Synthetic Biology of Botanical Biopesticides
by Jianbo Zhao, Dongmei Liang, Weiguo Li, Xiaoguang Yan, Jianjun Qiao and Qinggele Caiyin
Bioengineering 2022, 9(5), 207; https://doi.org/10.3390/bioengineering9050207 - 12 May 2022
Cited by 9 | Viewed by 3353
Abstract
The production and large-scale application of traditional chemical pesticides will bring environmental pollution and food safety problems. With the advantages of high safety and environmental friendliness, botanical biopesticides are in line with the development trend of modern agriculture and have gradually become the [...] Read more.
The production and large-scale application of traditional chemical pesticides will bring environmental pollution and food safety problems. With the advantages of high safety and environmental friendliness, botanical biopesticides are in line with the development trend of modern agriculture and have gradually become the mainstream of modern pesticide development. However, the traditional production of botanical biopesticides has long been faced with prominent problems, such as limited source and supply, complicated production processes, and excessive consumption of resources. In recent years, the rapid development of synthetic biology will break through these bottlenecks, and many botanical biopesticides are produced using synthetic biology, such as emodin, celangulin, etc. This paper reviews the latest progress and application prospect of synthetic biology in the development of botanical pesticides so as to provide new ideas for the analysis of synthetic pathways and heterologous and efficient production of botanical biopesticides and accelerate the research process of synthetic biology of natural products. Full article
20 pages, 3956 KiB  
Article
Numerical and Experimental Investigation of the Hydrodynamics in the Single-Use Bioreactor Mobius® CellReady 3 L
by Diana Kreitmayer, Srikanth R. Gopireddy, Tomomi Matsuura, Yuichi Aki, Yuta Katayama, Taihei Sawada, Hirofumi Kakihara, Koichi Nonaka, Thomas Profitlich, Nora A. Urbanetz and Eva Gutheil
Bioengineering 2022, 9(5), 206; https://doi.org/10.3390/bioengineering9050206 - 11 May 2022
Cited by 4 | Viewed by 5417
Abstract
Two-way Euler-Lagrange simulations are performed to characterize the hydrodynamics in the single-use bioreactor Mobius® CellReady 3 L. The hydrodynamics in stirred tank bioreactors are frequently modeled with the Euler–Euler approach, which cannot capture the trajectories of single bubbles. The present study employs [...] Read more.
Two-way Euler-Lagrange simulations are performed to characterize the hydrodynamics in the single-use bioreactor Mobius® CellReady 3 L. The hydrodynamics in stirred tank bioreactors are frequently modeled with the Euler–Euler approach, which cannot capture the trajectories of single bubbles. The present study employs the two-way coupled Euler–Lagrange approach, which accounts for the individual bubble trajectories through Langrangian equations and considers their impact on the Eulerian liquid phase equations. Hydrodynamic process characteristics that are relevant for cell cultivation including the oxygen mass transfer coefficient, the mixing time, and the hydrodynamic stress are evaluated for different working volumes, sparger types, impeller speeds, and sparging rates. A microporous sparger and an open pipe sparger are considered where bubbles of different sizes are generated, which has a pronounced impact on the bubble dispersion and the volumetric oxygen mass transfer coefficient. It is found that only the microporous sparger provides sufficiently high oxygen transfer to support typical suspended mammalian cell lines. The simulated mixing time and the volumetric oxygen mass transfer coefficient are successfully validated with experimental results. Due to the small reactor size, mixing times are below 25 s across all tested conditions. For the highest sparging rate of 100 mL min1, the mixing time is found to be two seconds shorter than for a sparging rate of 50 mL min1, which again, is 0.1 s longer than for a sparging rate of 10 mL min1 at the same impeller speed of 100 rpm and the working volume of 1.7 L. The hydrodynamic stress in this bioreactor is found to be below critical levels for all investigated impeller speeds of up to 150 rpm, where the maximum levels are found in the region where the bubbles pass behind the impeller blades. Full article
(This article belongs to the Section Biochemical Engineering)
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13 pages, 22329 KiB  
Article
Epidermal Electrodes with Ferrimagnetic/Conductive Properties for Biopotential Recordings
by Andrea Spanu, Mohamad Taki, Giulia Baldazzi, Antonello Mascia, Piero Cosseddu, Danilo Pani and Annalisa Bonfiglio
Bioengineering 2022, 9(5), 205; https://doi.org/10.3390/bioengineering9050205 - 11 May 2022
Cited by 3 | Viewed by 2237
Abstract
Interfacing ultrathin functional films for epidermal applications with external recording instruments or readout electronics still represents one of the biggest challenges in the field of tattoo electronics. With the aim of providing a convenient solution to this ever-present limitation, in this work we [...] Read more.
Interfacing ultrathin functional films for epidermal applications with external recording instruments or readout electronics still represents one of the biggest challenges in the field of tattoo electronics. With the aim of providing a convenient solution to this ever-present limitation, in this work we propose an innovative free-standing electrode made of a composite thin film based on the combination of the conductive polymer PEDOT:PSS and ferrimagnetic powder. The proposed epidermal electrode can be directly transferred onto the skin and is structured in two parts, namely a conformal conductive part with a thickness of 3 μm and a ferrimagnetic-conductive part that can be conveniently connected using magnetic connections. The films were characterized for ECG recordings, revealing a performance comparable to that of commercial pre-gelled electrodes in terms of cross-spectral coherence, signal-to-noise ratio, and baseline wandering. These new, conductive, magnetically interfaceable, and free-standing conformal films introduce a novel concept in the domain of tattoo electronics and can set the basis for the development of a future family of epidermal devices and electrodes. Full article
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18 pages, 1157 KiB  
Article
Lab-Scale Cultivation of Cupriavidus necator on Explosive Gas Mixtures: Carbon Dioxide Fixation into Polyhydroxybutyrate
by Vera Lambauer and Regina Kratzer
Bioengineering 2022, 9(5), 204; https://doi.org/10.3390/bioengineering9050204 - 10 May 2022
Cited by 18 | Viewed by 4433
Abstract
Aerobic, hydrogen oxidizing bacteria are capable of efficient, non-phototrophic CO2 assimilation, using H2 as a reducing agent. The presence of explosive gas mixtures requires strict safety measures for bioreactor and process design. Here, we report a simplified, reproducible, and safe cultivation [...] Read more.
Aerobic, hydrogen oxidizing bacteria are capable of efficient, non-phototrophic CO2 assimilation, using H2 as a reducing agent. The presence of explosive gas mixtures requires strict safety measures for bioreactor and process design. Here, we report a simplified, reproducible, and safe cultivation method to produce Cupriavidus necator H16 on a gram scale. Conditions for long-term strain maintenance and mineral media composition were optimized. Cultivations on the gaseous substrates H2, O2, and CO2 were accomplished in an explosion-proof bioreactor situated in a strong, grounded fume hood. Cells grew under O2 control and H2 and CO2 excess. The starting gas mixture was H2:CO2:O2 in a ratio of 85:10:2 (partial pressure of O2 0.02 atm). Dissolved oxygen was measured online and was kept below 1.6 mg/L by a stepwise increase of the O2 supply. Use of gas compositions within the explosion limits of oxyhydrogen facilitated production of 13.1 ± 0.4 g/L total biomass (gram cell dry mass) with a content of 79 ± 2% poly-(R)-3-hydroxybutyrate in a simple cultivation set-up with dissolved oxygen as the single controlled parameter. Approximately 98% of the obtained PHB was formed from CO2. Full article
(This article belongs to the Special Issue Advances in Polyhydroxyalkanoate (PHA) Production, Volume 3)
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11 pages, 2654 KiB  
Article
Decellularised Cartilage ECM Culture Coatings Drive Rapid and Robust Chondrogenic Differentiation of Human Periosteal Cells
by Wollis J. Vas, Mittal Shah, Helen C. Roberts and Scott J. Roberts
Bioengineering 2022, 9(5), 203; https://doi.org/10.3390/bioengineering9050203 - 10 May 2022
Cited by 2 | Viewed by 2327
Abstract
The control of cell behaviour in an effort to create highly homogeneous cultures is becoming an area of intense research, both to elucidate fundamental biology and for regenerative applications. The extracellular matrix (ECM) controls many cellular processes in vivo, and as such is [...] Read more.
The control of cell behaviour in an effort to create highly homogeneous cultures is becoming an area of intense research, both to elucidate fundamental biology and for regenerative applications. The extracellular matrix (ECM) controls many cellular processes in vivo, and as such is a rich source of cues that may be translated in vitro. Herein, we describe the creation of cell culture coatings from porcine decellularised hyaline cartilage through enzymatic digestion. Surprisingly, heat-mediated sterilisation created a coating with the capacity to rapidly and robustly induce chondrogenic differentiation of human periosteal cells. This differentiation was validated through the alteration of cell phenotype from a fibroblastic to a cuboidal/cobblestone chondrocyte-like appearance. Moreover, chondrogenic gene expression further supported this observation, where cells cultured on heat sterilised ECM-coated plastic displayed higher expression of COL2A1, ACAN and PRG4 (p < 0.05) compared to non-coated plastic cultures. Interestingly, COL2A1 and ACAN expression in this context were sensitive to initial cell density; however, SOX9 expression appeared to be mainly driven by the coating independent of seeding density. The creation of a highly chondrogenic coating may provide a cost-effective solution for the differentiation and/or expansion of human chondrocytes aimed towards cartilage repair strategies. Full article
(This article belongs to the Special Issue Extracellular Matrix in Musculoskeletal Regeneration)
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17 pages, 1842 KiB  
Review
Engineering Extracellular Microenvironment for Tissue Regeneration
by Dake Hao, Juan-Maria Lopez, Jianing Chen, Alexandra Maria Iavorovschi, Nora Marlene Lelivelt and Aijun Wang
Bioengineering 2022, 9(5), 202; https://doi.org/10.3390/bioengineering9050202 - 08 May 2022
Cited by 9 | Viewed by 3132
Abstract
The extracellular microenvironment is a highly dynamic network of biophysical and biochemical elements, which surrounds cells and transmits molecular signals. Extracellular microenvironment controls are of crucial importance for the ability to direct cell behavior and tissue regeneration. In this review, we focus on [...] Read more.
The extracellular microenvironment is a highly dynamic network of biophysical and biochemical elements, which surrounds cells and transmits molecular signals. Extracellular microenvironment controls are of crucial importance for the ability to direct cell behavior and tissue regeneration. In this review, we focus on the different components of the extracellular microenvironment, such as extracellular matrix (ECM), extracellular vesicles (EVs) and growth factors (GFs), and introduce engineering approaches for these components, which can be used to achieve a higher degree of control over cellular activities and behaviors for tissue regeneration. Furthermore, we review the technologies established to engineer native-mimicking artificial components of the extracellular microenvironment for improved regenerative applications. This review presents a thorough analysis of the current research in extracellular microenvironment engineering and monitoring, which will facilitate the development of innovative tissue engineering strategies by utilizing different components of the extracellular microenvironment for regenerative medicine in the future. Full article
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12 pages, 2715 KiB  
Article
Concept, Design, and Early Prototyping of a Low-Cost, Minimally Invasive, Fully Implantable Left Ventricular Assist Device
by Florin Alexandru Pleșoianu, Carmen Elena Pleșoianu, Iris Bararu Bojan, Andrei Bojan, Andrei Țăruș and Grigore Tinică
Bioengineering 2022, 9(5), 201; https://doi.org/10.3390/bioengineering9050201 - 06 May 2022
Cited by 3 | Viewed by 2331
Abstract
Despite evidence associating the use of mechanical circulatory support (MCS) devices with increased survival and quality of life in patients with advanced heart failure (HF), significant complications and high costs limit their clinical use. We aimed to design an innovative MCS device to [...] Read more.
Despite evidence associating the use of mechanical circulatory support (MCS) devices with increased survival and quality of life in patients with advanced heart failure (HF), significant complications and high costs limit their clinical use. We aimed to design an innovative MCS device to address three important needs: low cost, minimally invasive implantation techniques, and low risk of infection. We used mathematical modeling to calculate the pump characteristics to deliver variable flows at different pump diameters, turbomachinery design software CFturbo (2020 R2.4 CFturbo GmbH, Dresden, Germany) to create the conceptual design of the pump, computational fluid dynamics analysis with Solidworks Flow Simulation to in silico test pump performance, Solidworks (Dassault Systèmes SolidWorks Corporation, Waltham, MA, USA) to further refine the design, 3D printing with polycarbonate filament for the initial prototype, and a stereolithography printer (Form 2, Formlabs, Somerville, MA, USA) for the second variant materialization. We present the concept, design, and early prototyping of a low-cost, minimally invasive, fully implantable in a subcutaneous pocket MCS device for long-term use and partial support in patients with advanced HF which unloads the left heart into the arterial system containing a rim-driven, hubless axial-flow pump and the wireless transmission of energy. We describe a low-cost, fully implantable, low-invasive, wireless power transmission left ventricular assist device that has the potential to address patients with advanced HF with higher impact, especially in developing countries. In vitro testing will provide input for further optimization of the device before proceeding to a completely functional prototype that can be implanted in animals. Full article
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24 pages, 4956 KiB  
Review
Gold Nanorod-Assisted Photothermal Therapy and Improvement Strategies
by Mitchell Lee Taylor, Raymond Edward Wilson, Jr., Kristopher Daniel Amrhein and Xiaohua Huang
Bioengineering 2022, 9(5), 200; https://doi.org/10.3390/bioengineering9050200 - 05 May 2022
Cited by 32 | Viewed by 4276
Abstract
Noble metal nanoparticles have been sought after in cancer nanomedicine during the past two decades, owing to the unique localized surface plasmon resonance that induces strong absorption and scattering properties of the nanoparticles. A popular application of noble metal nanoparticles is photothermal therapy, [...] Read more.
Noble metal nanoparticles have been sought after in cancer nanomedicine during the past two decades, owing to the unique localized surface plasmon resonance that induces strong absorption and scattering properties of the nanoparticles. A popular application of noble metal nanoparticles is photothermal therapy, which destroys cancer cells by heat generated by laser irradiation of the nanoparticles. Gold nanorods have stood out as one of the major types of noble metal nanoparticles for photothermal therapy due to the facile tuning of their optical properties in the tissue penetrative near infrared region, strong photothermal conversion efficiency, and long blood circulation half-life after surface modification with stealthy polymers. In this review, we will summarize the optical properties of gold nanorods and their applications in photothermal therapy. We will also discuss the recent strategies to improve gold nanorod-assisted photothermal therapy through combination with chemotherapy and photodynamic therapy. Full article
(This article belongs to the Special Issue Multiscale Thermal Engineering for Biomedical Applications)
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