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Bioengineering, Volume 8, Issue 11 (November 2021) – 43 articles

Cover Story (view full-size image): Cheese whey (CW) can be used as an excellent carbon source for polyhydroxyalkanoate (PHA) production, but it has not been explored, especially CW, which contains a low amount of lactose. Therefore, in the present study, PHA production was evaluated using low-lactose-content CW in a two-stage process using Acetobacter pasteurianus C1 and Bacillus sp. CYR1. Results discovered that, instead of the hydrolysis of lactose (which enhances the production cost), PHA production from CW with a two-stage process is economically feasible.View this paper
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17 pages, 5240 KiB  
Article
Engineering Breast Cancer Cells and hUMSCs Microenvironment in 2D and 3D Scaffolds: A Mechanical Study Approach of Stem Cells in Anticancer Therapy
by Despoina Nektaria Metsiou, Foteini K. Kozaniti and Despina D. Deligianni
Bioengineering 2021, 8(11), 189; https://doi.org/10.3390/bioengineering8110189 - 20 Nov 2021
Cited by 2 | Viewed by 2895
Abstract
Cell biomechanics plays a major role as a promising biomarker for early cancer diagnosis and prognosis. In the present study, alterations in modulus of elasticity, cell membrane roughness, and migratory potential of MCF-7 (ER+) and SKBR-3 (HER2+) cancer cells were elucidated prior to [...] Read more.
Cell biomechanics plays a major role as a promising biomarker for early cancer diagnosis and prognosis. In the present study, alterations in modulus of elasticity, cell membrane roughness, and migratory potential of MCF-7 (ER+) and SKBR-3 (HER2+) cancer cells were elucidated prior to and post treatment with conditioned medium from human umbilical mesenchymal stem cells (hUMSCs-CM) during static and dynamic cell culture. Moreover, the therapeutic potency of hUMSCs-CM on cancer cell’s viability, migratory potential, and F-actin quantified intensity was addressed in 2D surfaces and 3D scaffolds. Interestingly, alterations in ER+ cancer cells showed a positive effect of treatment upon limiting cell viability, motility, and potential for migration. Moreover, increased post treatment cell stiffness indicated rigid cancer cells with confined cell movement and cytoskeletal alterations with restricted lamellipodia formation, which enhanced these results. On the contrary, the cell viability and the migratory potential were not confined post treatment with hUMSCs-CM on HER2+ cells, possibly due to their intrinsic aggressiveness. The increased post treatment cell viability and the decreased cell stiffness indicated an increased potency for cell movement. Hence, the therapy had no efficacy on HER2+ cells. Full article
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11 pages, 2445 KiB  
Article
In Silico Analysis of Glucose Oxidase from Aspergillus niger: Potential Cysteine Mutation Sites for Enhancing Protein Stability
by Sirawit Ittisoponpisan and Itthipon Jeerapan
Bioengineering 2021, 8(11), 188; https://doi.org/10.3390/bioengineering8110188 - 19 Nov 2021
Cited by 5 | Viewed by 3963
Abstract
Glucose oxidase (GOx) holds considerable advantages for various applications. Nevertheless, the thermal instability of the enzyme remains a grand challenge, impeding the success in applications outside the well-controlled laboratories, particularly in practical bioelectronics. Many strategies to modify GOx to achieve better thermal stability [...] Read more.
Glucose oxidase (GOx) holds considerable advantages for various applications. Nevertheless, the thermal instability of the enzyme remains a grand challenge, impeding the success in applications outside the well-controlled laboratories, particularly in practical bioelectronics. Many strategies to modify GOx to achieve better thermal stability have been proposed. However, modification of this enzyme by adding extra disulfide bonds is yet to be explored. This work describes the in silico bioengineering of GOx from Aspergillus niger by judiciously analyzing characteristics of disulfide bonds found in the Top8000 protein database, then scanning for amino acid residue pairs that are suitable to be replaced with cysteines in order to establish disulfide bonds. Next, we predicted and assessed the mutant GOx models in terms of disulfide bond quality (bond length and α angles), functional impact by means of residue conservation, and structural impact as indicated by Gibbs free energy. We found eight putative residue pairs that can be engineered to form disulfide bonds. Five of these are located in less conserved regions and, therefore, are unlikely to have a deleterious impact on functionality. Finally, two mutations, Pro149Cys and His158Cys, showed potential for stabilizing the protein structure as confirmed by a structure-based stability analysis tool. The findings in this study highlight the opportunity of using disulfide bond modification as a new alternative technique to enhance the thermal stability of GOx. Full article
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14 pages, 1961 KiB  
Article
Bioengineered System for High Throughput Screening of Kv1 Ion Channel Blockers
by George V. Sharonov, Oksana V. Nekrasova, Ksenia S. Kudryashova, Mikhail P. Kirpichnikov and Alexey V. Feofanov
Bioengineering 2021, 8(11), 187; https://doi.org/10.3390/bioengineering8110187 - 16 Nov 2021
Cited by 2 | Viewed by 3087
Abstract
Screening drug candidates for their affinity and selectivity for a certain binding site is a crucial step in developing targeted therapy. Here, we created a screening assay for receptor binding that can be easily scaled up and automated for the high throughput screening [...] Read more.
Screening drug candidates for their affinity and selectivity for a certain binding site is a crucial step in developing targeted therapy. Here, we created a screening assay for receptor binding that can be easily scaled up and automated for the high throughput screening of Kv channel blockers. It is based on the expression of the KcsA-Kv1 hybrid channel tagged with a fluorescent protein in the E. coli membrane. In order to make this channel accessible for the soluble compounds, E. coli were transformed into spheroplasts by disruption of the cellular peptidoglycan envelope. The assay was evaluated using a hybrid KcsA-Kv1.3 potassium channel tagged with a red fluorescent protein (TagRFP). The binding of Kv1.3 channel blockers was measured by flow cytometry either by using their fluorescent conjugates or by determining the ability of unconjugated compounds to displace fluorescently labeled blockers with a known affinity. A fraction of the occupied receptor was calculated with a dedicated pipeline available as a Jupyter notebook. Measured binding constants for agitoxin-2, charybdotoxin and kaliotoxin were in firm agreement with the earlier published data. By using a mid-range flow cytometer with manual sample handling, we measured and analyzed up to ten titration curves (eight data points each) in one day. Finally, we considered possibilities for multiplexing, scaling and automation of the assay. Full article
(This article belongs to the Special Issue Biomedical Applications of Ionic Liquids and Deep Eutectic Solvents)
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20 pages, 5134 KiB  
Article
Recapitulating the Angiogenic Switch in a Hydrogel-Based 3D In Vitro Tumor-Stroma Model
by Claudia Kuehlbach, Sabine Hensler and Margareta M. Mueller
Bioengineering 2021, 8(11), 186; https://doi.org/10.3390/bioengineering8110186 - 15 Nov 2021
Cited by 6 | Viewed by 3323
Abstract
To ensure nutrient and oxygen supply, tumors beyond a size of 1–2 mm3 need a connection to the vascular system. Thus, tumor cells modify physiological tissue homeostasis by secreting inflammatory and angiogenic cytokines. This leads to the activation of the tumor microenvironment [...] Read more.
To ensure nutrient and oxygen supply, tumors beyond a size of 1–2 mm3 need a connection to the vascular system. Thus, tumor cells modify physiological tissue homeostasis by secreting inflammatory and angiogenic cytokines. This leads to the activation of the tumor microenvironment and the turning of the angiogenic switch, resulting in tumor vascularization and growth. To inhibit tumor growth by developing efficient anti-angiogenic therapies, an in depth understanding of the molecular mechanism initiating angiogenesis is essential. Yet so far, predominantly 2D cell cultures or animal models have been used to clarify the interactions within the tumor stroma, resulting in poor transferability of the data obtained to the in vivo situation. Consequently, there is an abundant need for complex, humanized, 3D models in vitro. We established a dextran-hydrogel-based 3D organotypic in vitro model containing microtumor spheroids, macrophages, neutrophils, fibroblasts and endothelial cells, allowing for the analysis of tumor–stroma interactions in a controlled and modifiable environment. During the cultivation period of 21 days, the microtumor spheroids in the model grew in size and endothelial cells formed elongated tubular structures resembling capillary vessels, that appeared to extend towards the tumor spheroids. The tubular structures exhibited complex bifurcations and expanded without adding external angiogenic factors such as VEGF to the culture. To allow high-throughput screening of therapeutic candidates, the 3D cell culture model was successfully miniaturized to a 96-well format, while still maintaining the same level of tumor spheroid growth and vascular sprouting. The quantification of VEGF in the conditioned medium of these cultures showed a continuous increase during the cultivation period, suggesting the contribution of endogenous VEGF to the induction of the angiogenic switch and vascular sprouting. Thus, this model is highly suitable as a testing platform for novel anticancer therapeutics targeting the tumor as well as the vascular compartment. Full article
(This article belongs to the Special Issue Advances in 3D Printing Tissue Engineering)
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16 pages, 1152 KiB  
Review
3D Hepatic Organoid-Based Advancements in LIVER Tissue Engineering
by Amit Panwar, Prativa Das and Lay Poh Tan
Bioengineering 2021, 8(11), 185; https://doi.org/10.3390/bioengineering8110185 - 14 Nov 2021
Cited by 18 | Viewed by 9949
Abstract
Liver-associated diseases and tissue engineering approaches based on in vitro culture of functional Primary human hepatocytes (PHH) had been restricted by the rapid de-differentiation in 2D culture conditions which restricted their usability. It was proven that cells growing in 3D format can better [...] Read more.
Liver-associated diseases and tissue engineering approaches based on in vitro culture of functional Primary human hepatocytes (PHH) had been restricted by the rapid de-differentiation in 2D culture conditions which restricted their usability. It was proven that cells growing in 3D format can better mimic the in vivo microenvironment, and thus help in maintaining metabolic activity, phenotypic properties, and longevity of the in vitro cultures. Again, the culture method and type of cell population are also recognized as important parameters for functional maintenance of primary hepatocytes. Hepatic organoids formed by self-assembly of hepatic cells are microtissues, and were able to show long-term in vitro maintenance of hepato-specific characteristics. Thus, hepatic organoids were recognized as an effective tool for screening potential cures and modeling liver diseases effectively. The current review summarizes the importance of 3D hepatic organoid culture over other conventional 2D and 3D culture models and its applicability in Liver tissue engineering. Full article
(This article belongs to the Special Issue Material and Engineering-Based Approaches for Organoids)
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15 pages, 3232 KiB  
Article
Heat-Inactivation of Fetal and Newborn Sera Did Not Impair the Expansion and Scaffold Engineering Potentials of Fibroblasts
by Félix-Antoine Pellerin, Christophe Caneparo, Ève Pellerin, Stéphane Chabaud, Martin Pelletier and Stéphane Bolduc
Bioengineering 2021, 8(11), 184; https://doi.org/10.3390/bioengineering8110184 - 13 Nov 2021
Cited by 6 | Viewed by 3277
Abstract
Heat inactivation of bovine sera is routinely performed in cell culture laboratories. Nevertheless, it remains debatable whether it is still necessary due to the improvement of the production process of bovine sera. Do the benefits balance the loss of many proteins, such as [...] Read more.
Heat inactivation of bovine sera is routinely performed in cell culture laboratories. Nevertheless, it remains debatable whether it is still necessary due to the improvement of the production process of bovine sera. Do the benefits balance the loss of many proteins, such as hormones and growth factors, that are very useful for cell culture? This is even truer in the case of tissue engineering, the processes of which is often very demanding. This balance is examined here, from nine populations of fibroblasts originating from three different organs, by comparing the capacity of adhesion and proliferation of cells, their metabolism, and the capacity to produce the stroma; their histological appearance, thickness, and mechanical properties were also evaluated. Overall, serum inactivation does not appear to provide a significant benefit. Full article
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16 pages, 4081 KiB  
Article
Design and Analysis of a Biodegradable Polycaprolactone Flow Diverting Stent for Brain Aneurysms
by Kaitlyn Tidwell, Seth Harriet, Vishal Barot, Andrew Bauer, Melville B. Vaughan and Mohammad R. Hossan
Bioengineering 2021, 8(11), 183; https://doi.org/10.3390/bioengineering8110183 - 12 Nov 2021
Cited by 4 | Viewed by 4691
Abstract
The flow diverting stent (FDS) has become a promising endovascular device for the treatment of aneurysms. This research presents a novel biodegradable and non-braided Polycaprolactone (PCL) FDS. The PCL FDS was designed and developed using an in-house fabrication unit and coated on two [...] Read more.
The flow diverting stent (FDS) has become a promising endovascular device for the treatment of aneurysms. This research presents a novel biodegradable and non-braided Polycaprolactone (PCL) FDS. The PCL FDS was designed and developed using an in-house fabrication unit and coated on two ends with BaSO4 for angiographic visibility. The mechanical flexibility and quality of FDS surfaces were examined with the UniVert testing machine, scanning electron microscope (SEM), and 3D profilometer. Human umbilical vein endothelial cell (HUVEC) adhesion, proliferation, and cell morphology studies on PCL FDS were performed. The cytotoxicity and NO production by HUVECs with PCL FDS were also conducted. The longitudinal tensile, radial, and bending flexibility were found to be 1.20 ± 0.19 N/mm, 0.56 ± 0.11 N/mm, and 0.34 ± 0.03 N/mm, respectively. The FDS was returned to the original shape and diameter after repeated compression and bending without compromising mechanical integrity. Results also showed that the proliferation and adhesion of HUVECs on the FDS surface increased over time compared to control without FDS. Lactate dehydrogenase (LDH) release and NO production showed that PCL FDS were non-toxic and satisfactory. Cell morphology studies showed that HUVECs were elongated to cover the FD surface and developed an endothelial monolayer. This study is a step forward toward the development and clinical use of biodegradable flow diverting stents for endovascular treatment of the aneurysm. Full article
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13 pages, 1838 KiB  
Review
High-Resolution Imaging for the Analysis and Reconstruction of 3D Microenvironments for Regenerative Medicine: An Application-Focused Review
by Michail E. Klontzas and Alexandros Protonotarios
Bioengineering 2021, 8(11), 182; https://doi.org/10.3390/bioengineering8110182 - 10 Nov 2021
Cited by 11 | Viewed by 2789
Abstract
The rapid evolution of regenerative medicine and its associated scientific fields, such as tissue engineering, has provided great promise for multiple applications where replacement and regeneration of damaged or lost tissue is required. In order to evaluate and optimise the tissue engineering techniques, [...] Read more.
The rapid evolution of regenerative medicine and its associated scientific fields, such as tissue engineering, has provided great promise for multiple applications where replacement and regeneration of damaged or lost tissue is required. In order to evaluate and optimise the tissue engineering techniques, visualisation of the material of interest is crucial. This includes monitoring of the cellular behaviour, extracellular matrix composition, scaffold structure, and other crucial elements of biomaterials. Non-invasive visualisation of artificial tissues is important at all stages of development and clinical translation. A variety of preclinical and clinical imaging methods—including confocal multiphoton microscopy, optical coherence tomography, magnetic resonance imaging (MRI), and computed tomography (CT)—have been used for the evaluation of artificial tissues. This review attempts to present the imaging methods available to assess the composition and quality of 3D microenvironments, as well as their integration with human tissues once implanted in the human body. The review provides tissue-specific application examples to demonstrate the applicability of such methods on cardiovascular, musculoskeletal, and neural tissue engineering. Full article
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15 pages, 542 KiB  
Article
Automatic Electrodiagnosis of Carpal Tunnel Syndrome Using Machine Learning
by Konstantinos I. Tsamis, Prokopis Kontogiannis, Ioannis Gourgiotis, Stefanos Ntabos, Ioannis Sarmas and George Manis
Bioengineering 2021, 8(11), 181; https://doi.org/10.3390/bioengineering8110181 - 10 Nov 2021
Cited by 9 | Viewed by 3346
Abstract
Recent literature has revealed a long discussion about the importance and necessity of nerve conduction studies in carpal tunnel syndrome management. The purpose of this study was to investigate the possibility of automatic detection, based on electrodiagnostic features, for the median nerve mononeuropathy [...] Read more.
Recent literature has revealed a long discussion about the importance and necessity of nerve conduction studies in carpal tunnel syndrome management. The purpose of this study was to investigate the possibility of automatic detection, based on electrodiagnostic features, for the median nerve mononeuropathy and decision making about carpal tunnel syndrome. The study included 38 volunteers, examined prospectively. The purpose was to investigate the possibility of automatically detecting the median nerve mononeuropathy based on common electrodiagnostic criteria, used in everyday clinical practice, as well as new features selected based on physiology and mathematics. Machine learning techniques were used to combine the examined characteristics for a stable and accurate diagnosis. Automatic electrodiagnosis reached an accuracy of 95% compared to the standard neurophysiological diagnosis of the physicians with nerve conduction studies and 89% compared to the clinical diagnosis. The results show that the automatic detection of carpal tunnel syndrome is possible and can be employed in decision making, excluding human error. It is also shown that the novel features investigated can be used for the detection of the syndrome, complementary to the commonly used ones, increasing the accuracy of the method. Full article
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9 pages, 2538 KiB  
Technical Note
A New Orthodontic-Surgical Approach to Mandibular Retrognathia
by Francisco Vale, Joana Queiroga, Flávia Pereira, Madalena Ribeiro, Filipa Marques, Raquel Travassos, Catarina Nunes, Anabela Baptista Paula and Inês Francisco
Bioengineering 2021, 8(11), 180; https://doi.org/10.3390/bioengineering8110180 - 8 Nov 2021
Cited by 6 | Viewed by 4765
Abstract
(1) Background: Mandibular deficiency is one of the most common growth disorders of the facial skeleton. Recently, distraction osteogenesis has been suggested as the treatment of choice for overcoming the limitations of conventional orthognathic surgery; (2) Methods: A new custom-manufactured dental-anchored distractor was [...] Read more.
(1) Background: Mandibular deficiency is one of the most common growth disorders of the facial skeleton. Recently, distraction osteogenesis has been suggested as the treatment of choice for overcoming the limitations of conventional orthognathic surgery; (2) Methods: A new custom-manufactured dental-anchored distractor was built and anchored in the first molar and lower canine. It consists of a stainless-steel disjunction screw, adapted and welded to the orthodontic bands through two 1.2 mm diameter connector bars with a universal silver-based and cadmium-free solder; (3) Results: The distractor described can be a useful tool to correct mandibular retrognathia and is better tolerated by patients, especially in severe cases; (4) Conclusions: The dental-anchored distractor increases the anterior mandibular bone segment without affecting the gonial angle or transverse angulation of the segments and avoids posterior mandibular rotation, overcoming the limitations of conventional surgical treatment. Full article
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10 pages, 991 KiB  
Article
Biosynthesis of Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) from CO2 by a Recombinant Cupriavidusnecator
by Kenji Tanaka, Kazumasa Yoshida, Izumi Orita and Toshiaki Fukui
Bioengineering 2021, 8(11), 179; https://doi.org/10.3390/bioengineering8110179 - 7 Nov 2021
Cited by 30 | Viewed by 4639
Abstract
The copolyester of 3-hydroxybutyrate (3HB) and 3-hydoxyhexanoate (3HHx), PHBHHx, is one of the most practical kind of bacterial polyhydroxyalkanoates due to its high flexibility and marine biodegradability. PHBHHx is usually produced from vegetable oils or fatty acids through β-oxidation, whereas biosynthesis from sugars [...] Read more.
The copolyester of 3-hydroxybutyrate (3HB) and 3-hydoxyhexanoate (3HHx), PHBHHx, is one of the most practical kind of bacterial polyhydroxyalkanoates due to its high flexibility and marine biodegradability. PHBHHx is usually produced from vegetable oils or fatty acids through β-oxidation, whereas biosynthesis from sugars has been achieved by recombinant strains of hydrogen-oxidizing bacterium Cupriavidus necator. This study investigated the biosynthesis of PHBHHx from CO2 as the sole carbon source by engineered C. necator strains. The recombinant strains capable of synthesizing PHBHHx from fructose were cultivated in a flask using complete mineral medium and a substrate gas mixture (H2/O2/CO2 = 8:1:1). The results of GC and 1H NMR analyses indicated that the recombinants of C. necator synthesized PHBHHx from CO2 with high cellular content. When 1.0 g/L (NH4)2SO4 was used as a nitrogen source, the 3HHx composition of PHBHHx in the strain MF01∆B1/pBBP-ccrMeJ4a-emd was 47.7 ± 6.2 mol%. Further investigation demonstrated that the PHA composition can be regulated by using (R)-enoyl-CoA hydratase (PhaJ) with different substrate specificity. The composition of 3HHx in PHBHHx was controlled to about 11 mol%, suitable for practical applications, and high cellular content was kept in the strains transformed with pBPP-ccrMeJAc-emd harboring short-chain-length-specific PhaJ. Full article
(This article belongs to the Special Issue Advances in Polyhydroxyalkanoate (PHA) Production, Volume 3)
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30 pages, 3051 KiB  
Review
Bioprinting Scaffolds for Vascular Tissues and Tissue Vascularization
by Peter Viktor Hauser, Hsiao-Min Chang, Masaki Nishikawa, Hiroshi Kimura, Norimoto Yanagawa and Morgan Hamon
Bioengineering 2021, 8(11), 178; https://doi.org/10.3390/bioengineering8110178 - 6 Nov 2021
Cited by 24 | Viewed by 6222
Abstract
In recent years, tissue engineering has achieved significant advancements towards the repair of damaged tissues. Until this day, the vascularization of engineered tissues remains a challenge to the development of large-scale artificial tissue. Recent breakthroughs in biomaterials and three-dimensional (3D) printing have made [...] Read more.
In recent years, tissue engineering has achieved significant advancements towards the repair of damaged tissues. Until this day, the vascularization of engineered tissues remains a challenge to the development of large-scale artificial tissue. Recent breakthroughs in biomaterials and three-dimensional (3D) printing have made it possible to manipulate two or more biomaterials with complementary mechanical and/or biological properties to create hybrid scaffolds that imitate natural tissues. Hydrogels have become essential biomaterials due to their tissue-like physical properties and their ability to include living cells and/or biological molecules. Furthermore, 3D printing, such as dispensing-based bioprinting, has progressed to the point where it can now be utilized to construct hybrid scaffolds with intricate structures. Current bioprinting approaches are still challenged by the need for the necessary biomimetic nano-resolution in combination with bioactive spatiotemporal signals. Moreover, the intricacies of multi-material bioprinting and hydrogel synthesis also pose a challenge to the construction of hybrid scaffolds. This manuscript presents a brief review of scaffold bioprinting to create vascularized tissues, covering the key features of vascular systems, scaffold-based bioprinting methods, and the materials and cell sources used. We will also present examples and discuss current limitations and potential future directions of the technology. Full article
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24 pages, 1624 KiB  
Article
Predictive Monitoring of Shake Flask Cultures with Online Estimated Growth Models
by Barbara Pretzner, Rüdiger W. Maschke, Claudia Haiderer, Gernot T. John, Christoph Herwig and Peter Sykacek
Bioengineering 2021, 8(11), 177; https://doi.org/10.3390/bioengineering8110177 - 6 Nov 2021
Cited by 4 | Viewed by 3605
Abstract
Simplicity renders shake flasks ideal for strain selection and substrate optimization in biotechnology. Uncertainty during initial experiments may, however, cause adverse growth conditions and mislead conclusions. Using growth models for online predictions of future biomass (BM) and the arrival of critical events like [...] Read more.
Simplicity renders shake flasks ideal for strain selection and substrate optimization in biotechnology. Uncertainty during initial experiments may, however, cause adverse growth conditions and mislead conclusions. Using growth models for online predictions of future biomass (BM) and the arrival of critical events like low dissolved oxygen (DO) levels or when to harvest is hence important to optimize protocols. Established knowledge that unfavorable metabolites of growing microorganisms interfere with the substrate suggests that growth dynamics and, as a consequence, the growth model parameters may vary in the course of an experiment. Predictive monitoring of shake flask cultures will therefore benefit from estimating growth model parameters in an online and adaptive manner. This paper evaluates a newly developed particle filter (PF) which is specifically tailored to the requirements of biotechnological shake flask experiments. By combining stationary accuracy with fast adaptation to change the proposed PF estimates time-varying growth model parameters from iteratively measured BM and DO sensor signals in an optimal manner. Such proposition of inferring time varying parameters of Gompertz and Logistic growth models is to our best knowledge novel and here for the first time assessed for predictive monitoring of Escherichia coli (E. coli) shake flask experiments. Assessments that mimic real-time predictions of BM and DO levels under previously untested growth conditions demonstrate the efficacy of the approach. After allowing for an initialization phase where the PF learns appropriate model parameters, we obtain accurate predictions of future BM and DO levels and important temporal characteristics like when to harvest. Statically parameterized growth models that represent the dynamics of a specific setting will in general provide poor characterizations of the dynamics when we change strain or substrate. The proposed approach is thus an important innovation for scientists working on strain characterization and substrate optimization as providing accurate forecasts will improve reproducibility and efficiency in early-stage bioprocess development. Full article
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13 pages, 1623 KiB  
Article
Impact of Operational Factors, Inoculum Origin, and Feedstock Preservation on the Biochemical Methane Potential
by Audrey Lallement, Aline Siaud, Christine Peyrelasse, Prasad Kaparaju, Blandine Schraauwers, Samuel Maunas and Florian Monlau
Bioengineering 2021, 8(11), 176; https://doi.org/10.3390/bioengineering8110176 - 5 Nov 2021
Cited by 5 | Viewed by 2211
Abstract
Anaerobic digestion for the valorization of organic wastes into biogas is gaining worldwide interest. Nonetheless, the sizing of the biogas plant units require knowledge of the quantity of feedstock, and their associated methane potentials, estimated widely by Biochemical Methane Potential (BMP) tests. Discrepancies [...] Read more.
Anaerobic digestion for the valorization of organic wastes into biogas is gaining worldwide interest. Nonetheless, the sizing of the biogas plant units require knowledge of the quantity of feedstock, and their associated methane potentials, estimated widely by Biochemical Methane Potential (BMP) tests. Discrepancies exist among laboratories due to variability of protocols adopted and operational factors used. The aim of this study is to verify the influence of some operational factors (e.g., analysis frequency, trace elements and vitamins solution addition and flushing gas), feedstock conservation and the source of inoculum on BMP. Among the operational parameters tested on cellulose degradation, only the type of gas used for flushing headspace of BMP assays had shown a significant influence on methane yields from cellulose. Methane yields of 344 ± 6 NL CH4 kg−1 VS and 321 ± 10 NL CH4 kg−1 VS obtained from assays flushed with pure N2 and N2/CO2 (60/40 v/v). The origin of inoculum (fed in co-digestion) only significantly affected the methane yields for straw, 253 ± 3 and 333 ± 3 NL CH4 kg−1 VS. Finally, freezing/thawing cycle effect depended of the substrate (tested on biowaste, manure, straw and WWTP sludge) with a possible effect of water content substrate. Full article
(This article belongs to the Topic Bioreactors: Control, Optimization and Applications)
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14 pages, 3753 KiB  
Article
Patient-Specific Analysis of Ascending Thoracic Aortic Aneurysm with the Living Heart Human Model
by Salvatore Cutugno, Valentina Agnese, Giovanni Gentile, Giuseppe M. Raffa, Andrew D. Wisneski, Julius M. Guccione, Michele Pilato and Salvatore Pasta
Bioengineering 2021, 8(11), 175; https://doi.org/10.3390/bioengineering8110175 - 4 Nov 2021
Cited by 9 | Viewed by 3884
Abstract
In ascending thoracic aortic aneurysms (ATAAs), aneurysm kinematics are driven by ventricular traction occurring every heartbeat, increasing the stress level of dilated aortic wall. Aortic elongation due to heart motion and aortic length are emerging as potential indicators of adverse events in ATAAs; [...] Read more.
In ascending thoracic aortic aneurysms (ATAAs), aneurysm kinematics are driven by ventricular traction occurring every heartbeat, increasing the stress level of dilated aortic wall. Aortic elongation due to heart motion and aortic length are emerging as potential indicators of adverse events in ATAAs; however, simulation of ATAA that takes into account the cardiac mechanics is technically challenging. The objective of this study was to adapt the realistic Living Heart Human Model (LHHM) to the anatomy and physiology of a patient with ATAA to assess the role of cardiac motion on aortic wall stress distribution. Patient-specific segmentation and material parameter estimation were done using preoperative computed tomography angiography (CTA) and ex vivo biaxial testing of the harvested tissue collected during surgery. The lumped-parameter model of systemic circulation implemented in the LHHM was refined using clinical and echocardiographic data. The results showed that the longitudinal stress was highest in the major curvature of the aneurysm, with specific aortic quadrants having stress levels change from tensile to compressive in a transmural direction. This study revealed the key role of heart motion that stretches the aortic root and increases ATAA wall tension. The ATAA LHHM is a realistic cardiovascular platform where patient-specific information can be easily integrated to assess the aneurysm biomechanics and potentially support the clinical management of patients with ATAAs. Full article
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4 pages, 198 KiB  
Editorial
Modern Approaches in Cardiovascular Disease Therapeutics: From Molecular Genetics to Tissue Engineering
by Panagiotis Mallis, Efstathios Michalopoulos and Catherine Stavropoulos-Giokas
Bioengineering 2021, 8(11), 174; https://doi.org/10.3390/bioengineering8110174 - 4 Nov 2021
Cited by 4 | Viewed by 2116
Abstract
Cardiovascular disease (CVD) currently represents one of the leading causes of death worldwide. It is estimated that more than 17.9 million people die each year due to CVD manifestations. Often, occlusion or stenosis of the vascular network occurs, either in large- or small-diameter [...] Read more.
Cardiovascular disease (CVD) currently represents one of the leading causes of death worldwide. It is estimated that more than 17.9 million people die each year due to CVD manifestations. Often, occlusion or stenosis of the vascular network occurs, either in large- or small-diameter blood vessels. Moreover, the obstruction of small vessels such as the coronary arteries may be related to more pronounced events, which can be life-threatening. The gold standard procedure utilizes the transplantation of secondary vessels or the use of synthetic vascular grafts. However, significant adverse reactions have accompanied the use of the above grafts. Therefore, modern therapeutic strategies must be evaluated for better disease administration. In the context of alternative therapies, advanced tissue-engineering approaches including the decellularization procedure and the 3D additive bioprinting methods, have been proposed. In this way the availability of bioengineered vascular grafts will be increased, covering the great demand that exists globally. In this Special Issue of Bioengineering, we tried to highlight the modern approaches which are focused on CVD therapeutics. This issue includes articles related to the efficient development of vascular grafts, 3D printing approaches and suitable atherosclerosis models. Full article
14 pages, 4021 KiB  
Article
A Biomechanical Evaluation of a Novel Airbag Bicycle Helmet Concept for Traumatic Brain Injury Mitigation
by Kwong Ming Tse and Daniel Holder
Bioengineering 2021, 8(11), 173; https://doi.org/10.3390/bioengineering8110173 - 3 Nov 2021
Cited by 11 | Viewed by 4710
Abstract
In this study, a novel expandable bicycle helmet, which integrates an airbag system into the conventional helmet design, was proposed to explore the potential synergetic effect of an expandable airbag and a standard commuter-type EPS helmet. The traumatic brain injury mitigation performance of [...] Read more.
In this study, a novel expandable bicycle helmet, which integrates an airbag system into the conventional helmet design, was proposed to explore the potential synergetic effect of an expandable airbag and a standard commuter-type EPS helmet. The traumatic brain injury mitigation performance of the proposed expandable helmet was evaluated against that of a typical traditional bicycle helmet. A series of dynamic impact simulations on both a helmeted headform and a representative human head with different configurations were carried out in accordance with the widely recognised international bicycle helmet test standards. The impact simulations were initially performed on a ballast headform for validation and benchmarking purposes, while the subsequent ones on a biofidelic human head model were used for assessing any potential intracranial injury. It was found that the proposed expandable helmet performed admirably better when compared to a conventional helmet design—showing improvements in impact energy attenuation, as well as kinematic and biometric injury risk reduction. More importantly, this expandable helmet concept, integrating the airbag system in the conventional design, offers adequate protection to the cyclist in the unlikely case of airbag deployment failure. Full article
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17 pages, 4551 KiB  
Article
Characterization, Cytotoxicity and Anti-Inflammatory Effect Evaluation of Nanocapsules Containing Nicotine
by Carolina Landau Albrecht, Laura Elena Sperling, Daikelly Iglesias Braghirolli and Patricia Pranke
Bioengineering 2021, 8(11), 172; https://doi.org/10.3390/bioengineering8110172 - 3 Nov 2021
Cited by 2 | Viewed by 2766
Abstract
(1) Background: Nanotechnology is an emerging field that can be applied in the biomedical area. In this study, Eudragit nanocapsules (NCs) containing nicotine were produced. Nicotine is the main alkaloid found in tobacco and has anti-inflammatory properties. NCs containing nicotine may be used [...] Read more.
(1) Background: Nanotechnology is an emerging field that can be applied in the biomedical area. In this study, Eudragit nanocapsules (NCs) containing nicotine were produced. Nicotine is the main alkaloid found in tobacco and has anti-inflammatory properties. NCs containing nicotine may be used as an adjuvant therapy in the treatment of inflammation in the central nervous system. (2) Methods: Nanocapsules were prepared by the interfacial deposition of the pre-formed polymer method and characterized in terms of zeta potential, diameter, polydispersity index, pH, encapsulation efficiency (EE), stability and sustained release profile. In vitro tests with the PC12 cell line were performed, such as MTT, LIVE/DEAD and ELISA assays, to verify their cytotoxic and anti-inflammatory effects. (3) Results: The nanocapsules presented satisfactory values of the characterization parameters; however, poor encapsulation was obtained for nicotine (8.17% ± 0.47). The in vitro tests showed that the treatment with nanocapsules reduced cell viability, which suggests that the Eudragit or the amount of polymer on top of the cells may be detrimental to them, as the cells were able to survive when treated with bulk nicotine. ELISA showed an increment in the expression of IL-6 and IL-1β, corroborating the hypothesis that NCs were toxic to the cells because of the increase in the levels of these pro-inflammatory cytokines. (4) Conclusions: This study demonstrates that NCs of Eudragit present toxicity. It is therefore necessary to improve NC formulation to obtain better values for the encapsulation efficiency and reduce toxicity of these nanodevices. Full article
(This article belongs to the Collection Nanoparticles for Therapeutic and Diagnostic Applications)
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15 pages, 2149 KiB  
Article
Interferences of Waxes on Enzymatic Saccharification and Ethanol Production from Lignocellulose Biomass
by Marttin Paulraj Gundupalli, Santi Chuetor, Kraipat Cheenkachorn, Kittipong Rattanaporn, Pau-Loke Show, Yu-Shen Cheng and Malinee Sriariyanun
Bioengineering 2021, 8(11), 171; https://doi.org/10.3390/bioengineering8110171 - 2 Nov 2021
Cited by 15 | Viewed by 2904
Abstract
Wax is an organic compound found on the surface of lignocellulose biomass to protect plants from physical and biological stresses in nature. With its small mass fraction in biomass, wax has been neglected from inclusion in the design of the biorefinery process. This [...] Read more.
Wax is an organic compound found on the surface of lignocellulose biomass to protect plants from physical and biological stresses in nature. With its small mass fraction in biomass, wax has been neglected from inclusion in the design of the biorefinery process. This study investigated the interfering effect of wax in three types of lignocellulosic biomass, including rice straw (RS), Napier grass (NG), and sugarcane bagasse (SB). In this study, although small fractions of wax were extracted from RS, NG, and SB at 0.57%, 0.61%, and 1.69%, respectively, dewaxing causes changes in the plant compositions and their functional groups and promotes dissociations of lignocellulose fibrils. Additionally, dewaxing of biomass samples increased reducing sugar by 1.17-, 1.04-, and 1.35-fold in RS, NG, and SB, respectively. The ethanol yield increased by 1.11-, 1.05-, and 1.23-fold after wax removal from RS, NG, and SB, respectively. The chemical composition profiles of the waxes obtained from RS, NG, and SB showed FAME, alcohol, and alkane as the major groups. According to the conversion rate of the dewaxing process and ethanol fermentation, the wax outputs of RS, NG, and SB are 5.64, 17.00, and 6.00 kg/ton, respectively. The current gasoline price is around USD 0.903 per liter, making ethanol more expensive than gasoline. Therefore, in order to reduce the cost of ethanol in the biorefinery industry, other valuable products (such as wax) should be considered for commercialization. The cost of natural wax ranges from USD 2 to 22 per kilogram, depending on the source of the extracted wax. The wax yields obtained from RS, SB, and NG have the potential to increase profits in the biorefining process and could provide an opportunity for application in a wider range of downstream industries than just biofuels. Full article
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22 pages, 3242 KiB  
Review
Innovative High-Pressure Fabrication Processes for Porous Biomaterials—A Review
by Mythili Prakasam, Jean-François Silvain and Alain Largeteau
Bioengineering 2021, 8(11), 170; https://doi.org/10.3390/bioengineering8110170 - 1 Nov 2021
Cited by 6 | Viewed by 2975
Abstract
Biomaterials and their clinical application have become well known in recent years and progress in their manufacturing processes are essential steps in their technological advancement. Great advances have been made in the field of biomaterials, including ceramics, glasses, polymers, composites, glass-ceramics and metal [...] Read more.
Biomaterials and their clinical application have become well known in recent years and progress in their manufacturing processes are essential steps in their technological advancement. Great advances have been made in the field of biomaterials, including ceramics, glasses, polymers, composites, glass-ceramics and metal alloys. Dense and porous ceramics have been widely used for various biomedical applications. Current applications of bioceramics include bone grafts, spinal fusion, bone repairs, bone fillers, maxillofacial reconstruction, etc. One of the common impediments in the bioceramics and metallic porous implants for biomedical applications are their lack of mechanical strength. High-pressure processing can be a viable solution in obtaining porous biomaterials. Many properties such as mechanical properties, non-toxicity, surface modification, degradation rate, biocompatibility, corrosion rate and scaffold design are taken into consideration. The current review focuses on different manufacturing processes used for bioceramics, polymers and metals and their alloys in porous forms. Recent advances in the manufacturing technologies of porous ceramics by freeze isostatic pressure and hydrothermal processing are discussed in detail. Pressure as a parameter can be helpful in obtaining porous forms for biomaterials with increased mechanical strength. Full article
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18 pages, 2585 KiB  
Communication
Composite Scaffolds from Gelatin and Bone Meal Powder for Tissue Engineering
by Darlin Lantigua, Xinchen Wu, Sanika Suvarnapathaki, Michelle A. Nguyen and Gulden Camci-Unal
Bioengineering 2021, 8(11), 169; https://doi.org/10.3390/bioengineering8110169 - 1 Nov 2021
Cited by 23 | Viewed by 3024
Abstract
Bone tissue engineering offers versatile solutions to broaden clinical options for treating skeletal injuries. However, the variety of robust bone implants and substitutes remains largely uninvestigated. The advancements in hydrogel scaffolds composed of natural polymeric materials and osteoinductive microparticles have shown to be [...] Read more.
Bone tissue engineering offers versatile solutions to broaden clinical options for treating skeletal injuries. However, the variety of robust bone implants and substitutes remains largely uninvestigated. The advancements in hydrogel scaffolds composed of natural polymeric materials and osteoinductive microparticles have shown to be promising solutions in this field. In this study, gelatin methacrylate (GelMA) hydrogels containing bone meal powder (BP) particles were investigated for their osteoinductive capacity. As natural source of the bone mineral, we expect that BP improves the scaffold’s ability to induce mineralization. We characterized the physical properties of GelMA hydrogels containing various BP concentrations (0, 0.5, 5, and 50 mg/mL). The in vitro cellular studies revealed enhanced mechanical performance and the potential to promote the differentiation of pre-osteoblast cells. The in vivo studies demonstrated both promising biocompatibility and biodegradation properties. Overall, the biological and physical properties of this biomaterial is tunable based on BP concentration in GelMA scaffolds. The findings of this study offer a new composite scaffold for bone tissue engineering. Full article
(This article belongs to the Special Issue Cell-Biomaterial Interactions: Vol II)
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14 pages, 1162 KiB  
Review
Advanced Glycation End-Products in Skeletal Muscle Aging
by Lucas C. Olson, James T. Redden, Zvi Schwartz, David J. Cohen and Michael J. McClure
Bioengineering 2021, 8(11), 168; https://doi.org/10.3390/bioengineering8110168 - 1 Nov 2021
Cited by 26 | Viewed by 5848
Abstract
Advanced age causes skeletal muscle to undergo deleterious changes including muscle atrophy, fast-to-slow muscle fiber transition, and an increase in collagenous material that culminates in the age-dependent muscle wasting disease known as sarcopenia. Advanced glycation end-products (AGEs) non-enzymatically accumulate on the muscular collagens [...] Read more.
Advanced age causes skeletal muscle to undergo deleterious changes including muscle atrophy, fast-to-slow muscle fiber transition, and an increase in collagenous material that culminates in the age-dependent muscle wasting disease known as sarcopenia. Advanced glycation end-products (AGEs) non-enzymatically accumulate on the muscular collagens in old age via the Maillard reaction, potentiating the accumulation of intramuscular collagen and stiffening the microenvironment through collagen cross-linking. This review contextualizes known aspects of skeletal muscle extracellular matrix (ECM) aging, especially the role of collagens and AGE cross-linking, and underpins the motor nerve’s role in this aging process. Specific directions for future research are also discussed, with the understudied role of AGEs in skeletal muscle aging highlighted. Despite more than a half century of research, the role that intramuscular collagen aggregation and cross-linking plays in sarcopenia is well accepted yet not well integrated with current knowledge of AGE’s effects on muscle physiology. Furthermore, the possible impact that motor nerve aging has on intramuscular cross-linking and muscular AGE levels is posited. Full article
(This article belongs to the Special Issue Extracellular Matrix in Musculoskeletal Regeneration)
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16 pages, 2177 KiB  
Article
Modulation of Macrophage Activity by Pulsed Electromagnetic Fields in the Context of Fracture Healing
by Yangmengfan Chen, Maximilian M. Menger, Benedikt J. Braun, Sara Schweizer, Caren Linnemann, Karsten Falldorf, Michael Ronniger, Hongbo Wang, Tina Histing, Andreas K. Nussler and Sabrina Ehnert
Bioengineering 2021, 8(11), 167; https://doi.org/10.3390/bioengineering8110167 - 29 Oct 2021
Cited by 17 | Viewed by 3008
Abstract
Delayed fracture healing and fracture non-unions impose an enormous burden on individuals and society. Successful healing requires tight communication between immune cells and bone cells. Macrophages can be found in all healing phases. Due to their high plasticity and long life span, they [...] Read more.
Delayed fracture healing and fracture non-unions impose an enormous burden on individuals and society. Successful healing requires tight communication between immune cells and bone cells. Macrophages can be found in all healing phases. Due to their high plasticity and long life span, they represent good target cells for modulation. In the past, extremely low frequency pulsed electromagnet fields (ELF-PEMFs) have been shown to exert cell-specific effects depending on the field conditions. Thus, the aim was to identify the specific ELF-PEMFs able to modulate macrophage activity to indirectly promote mesenchymal stem/stromal cell (SCP-1 cells) function. After a blinded screening of 22 different ELF-PEMF, two fields (termed A and B) were further characterized as they diversely affected macrophage function. These two fields have similar fundamental frequencies (51.8 Hz and 52.3 Hz) but are emitted in different groups of pulses or rather send–pause intervals. Macrophages exposed to field A showed a pro-inflammatory function, represented by increased levels of phospho-Stat1 and CD86, the accumulation of ROS, and increased secretion of pro-inflammatory cytokines. In contrast, macrophages exposed to field B showed anti-inflammatory and pro-healing functions, represented by increased levels of Arginase I, increased secretion of anti-inflammatory cytokines, and growth factors are known to induce healing processes. The conditioned medium from macrophages exposed to both ELF-PEMFs favored the migration of SCP-1 cells, but the effect was stronger for field B. Furthermore, the conditioned medium from macrophages exposed to field B, but not to field A, stimulated the expression of extracellular matrix genes in SCP-1 cells, i.e., COL1A1, FN1, and BGN. In summary, our data show that specific ELF-PEMFs may affect immune cell function. Thus, knowing the specific ELF-PEMFs conditions and the underlying mechanisms bears great potential as an adjuvant treatment to modulate immune responses during pathologies, e.g., fracture healing. Full article
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19 pages, 3120 KiB  
Article
Single-Cell RNA-Sequencing Identifies Infrapatellar Fat Pad Macrophage Polarization in Acute Synovitis/Fat Pad Fibrosis and Cell Therapy
by Dimitrios Kouroupis, Thomas M. Best, Lee D. Kaplan, Diego Correa and Anthony J. Griswold
Bioengineering 2021, 8(11), 166; https://doi.org/10.3390/bioengineering8110166 - 29 Oct 2021
Cited by 8 | Viewed by 3291
Abstract
The pathogenesis and progression of knee inflammatory pathologies is modulated partly by residing macrophages in the infrapatellar fat pad (IFP), thus, macrophage polarization towards pro-inflammatory (M1) or anti-inflammatory (M2) phenotypes is important in joint disease pathologies. Alteration of M1/M2 balance contributes to the [...] Read more.
The pathogenesis and progression of knee inflammatory pathologies is modulated partly by residing macrophages in the infrapatellar fat pad (IFP), thus, macrophage polarization towards pro-inflammatory (M1) or anti-inflammatory (M2) phenotypes is important in joint disease pathologies. Alteration of M1/M2 balance contributes to the initiation and progression of joint inflammation and can be potentially altered with mesenchymal stem cell (MSC) therapy. In an acute synovial/IFP inflammation rat model a single intra-articular injection of IFP-MSC was performed, having as controls (1) diseased rats not receiving IFP-MSC and (2) non-diseased rats. After 4 days, cell specific transcriptional profiling via single-cell RNA-sequencing was performed on isolated IFP tissue from each group. Eight transcriptomically distinct cell populations were identified within the IFP across all three treatment groups with a noted difference in the proportion of myeloid cells across the groups. Largely myeloid cells consisted of macrophages (>90%); one M1 sub-cluster highly expressing pro-inflammatory markers and two M2 sub-clusters with one of them expressing higher levels of canonical M2 markers. Notably, the diseased samples (11.9%) had the lowest proportion of cells expressing M2 markers relative to healthy (14.8%) and MSC treated (19.4%) samples. These results suggest a phenotypic polarization of IFP macrophages towards the pro-inflammatory M1 phenotype in an acute model of inflammation, which are alleviated by IFP-MSC therapy inducing a switch towards an alternate M2 status. Understanding the IFP cellular heterogeneity and associated transcriptional programs may offer insights into novel therapeutic strategies for disabling joint disease pathologies. Full article
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13 pages, 2359 KiB  
Article
Effect on Osteogenic Differentiation of Genetically Modified IL4 or PDGF-BB Over-Expressing and IL4-PDGF-BB Co-Over-Expressing Bone Marrow-Derived Mesenchymal Stromal Cells In Vitro
by Masanori Tsubosaka, Masahiro Maruyama, Elijah Ejun Huang, Ning Zhang, Takeshi Utsunomiya, Qi Gao, Huaishuang Shen, Xueping Li, Junichi Kushioka, Hirohito Hirata, Zhenyu Yao, Yunzhi Peter Yang and Stuart B. Goodman
Bioengineering 2021, 8(11), 165; https://doi.org/10.3390/bioengineering8110165 - 29 Oct 2021
Cited by 4 | Viewed by 2744
Abstract
The use of genetically modified (GM) mesenchymal stromal cells (MSCs) and preconditioned MSCs (pMSCs) may provide further opportunities to improve the outcome of core decompression (CD) for the treatment of early-stage osteonecrosis of the femoral head (ONFH). GM interleukin-4 (IL4) over-expressing MSCs (IL4-MSCs), [...] Read more.
The use of genetically modified (GM) mesenchymal stromal cells (MSCs) and preconditioned MSCs (pMSCs) may provide further opportunities to improve the outcome of core decompression (CD) for the treatment of early-stage osteonecrosis of the femoral head (ONFH). GM interleukin-4 (IL4) over-expressing MSCs (IL4-MSCs), platelet-derived growth factor (PDGF)-BB over-expressing MSCs (PDGF-BB-MSCs), and IL4-PDGF-BB co-over-expressing MSCs (IL4-PDGF-BB-MSCs) and their respective pMSCs were used in this in vitro study and compared with respect to cell proliferation and osteogenic differentiation. IL4-MSCs, PDGF-BB-MSCs, IL4-PDGF-BB-MSCs, and each pMSC treatment significantly increased cell proliferation compared to the MSC group alone. The percentage of Alizarin red-stained area in the IL4-MSC and IL4-pMSC groups was significantly lower than in the MSC group. However, the percentage of Alizarin red-stained area in the PDGF-BB-MSC group was significantly higher than in the MSC and PDGF-BB-pMSC groups. The percentage of Alizarin red-stained area in the IL4-PDGF-BB-pMSC was significantly higher than in the IL4-PDGF-BB-MSC group. There were no significant differences in the percentage of Alizarin red-stained area between the MSC and IL4-PDGF-BB-pMSC groups. The use of PDGF-BB-MSCs or IL4-PDGF-BB-pMSCs increased cell proliferation. Furthermore, PDGF-BB-MSCs promoted osteogenic differentiation. The addition of GM MSCs may provide a useful supplementary cell-based therapy to CD for treatment of ONFH. Full article
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14 pages, 3387 KiB  
Article
Facile Fabrication of Three-Dimensional Hydrogel Film with Complex Tissue Morphology
by Young-Hyeon An and Su-Hwan Kim
Bioengineering 2021, 8(11), 164; https://doi.org/10.3390/bioengineering8110164 - 27 Oct 2021
Cited by 2 | Viewed by 2782
Abstract
In this study, we proposed a simple and easy method for fabricating a three-dimensional (3D) structure that can recapitulate the morphology of a tissue surface and deliver biological molecules into complex-shaped target tissues. To fabricate the 3D hydrogel film structure, we utilized a [...] Read more.
In this study, we proposed a simple and easy method for fabricating a three-dimensional (3D) structure that can recapitulate the morphology of a tissue surface and deliver biological molecules into complex-shaped target tissues. To fabricate the 3D hydrogel film structure, we utilized a direct tissue casting method that can recapitulate tissue structure in micro-/macroscale using polydimethylsiloxane (PDMS). A replica 3D negative mold was manufactured by a polyurethane acrylate (PUA)-based master mold. Then, we poured the catechol-conjugated alginate (ALG-C) solution into the mold and evaporated it to form a dried film, followed by crosslinking the film using calcium chloride. The ALG-C hydrogel film had a tensile modulus of 725.2 ± 123.4 kPa and maintained over 95% of initial weight after 1 week without significant degradation. The ALG-C film captured over 4.5 times as much macromolecule (FITC-dextran) compared to alginate film (ALG). The cardiomyoblast cells exhibited high cell viability over 95% on ALG-C film. Moreover, the ALG-C film had about 70% of surface-bound lentivirus (1% in ALG film), which finally exhibited much higher viral transfection efficiency of GFP protein to C2C12 cells on the film than ALG film. In conclusion, we demonstrated a 3D film structure of biofunctionalized hydrogel for substrate-mediated drug delivery, and this approach could be utilized to recapitulate the complex-shaped tissues. Full article
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18 pages, 1950 KiB  
Review
Understanding the Role of Fibroblasts following a 3D Tumoroid Implantation for Breast Tumor Formation
by Girdhari Rijal
Bioengineering 2021, 8(11), 163; https://doi.org/10.3390/bioengineering8110163 - 27 Oct 2021
Cited by 2 | Viewed by 3719
Abstract
An understanding of the participation and modulation of fibroblasts during tumor formation and growth is still unclear. Among many speculates, one might be the technical challenge to reveal the versatile function of fibroblasts in tissue complexity, and another is the dynamics in tissue [...] Read more.
An understanding of the participation and modulation of fibroblasts during tumor formation and growth is still unclear. Among many speculates, one might be the technical challenge to reveal the versatile function of fibroblasts in tissue complexity, and another is the dynamics in tissue physiology and cell activity. The histology of most solid tumors shows a predominant presence of fibroblasts, suggesting that tumor cells recruit fibroblasts for breast tumor growth. In this review paper, therefore, the migration, activation, differentiation, secretion, and signaling systems that are associated with fibroblasts and cancer-associated fibroblasts (CAFs) after implantation of a breast tumoroid, i.e., a lab-generated tumor tissue into an animal, are discussed. Full article
(This article belongs to the Topic Human Anatomy and Pathophysiology)
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14 pages, 34598 KiB  
Article
Liquid Helium Enhanced Vitrification Efficiency of Human Bone-Derived Mesenchymal Stem Cells and Human Embryonic Stem Cells
by Mengjia Dou, Chennan Lu, Jing Liu and Wei Rao
Bioengineering 2021, 8(11), 162; https://doi.org/10.3390/bioengineering8110162 - 26 Oct 2021
Cited by 6 | Viewed by 2521
Abstract
Stem cells have the capacity to self-renew and differentiate to specialized cells, which are usually sensitive to cryopreservation. Therefore, the cell survival rate of stem cells using common cryopreservation protocol is generally not ideal. High cooling rates are crucial for decreasing the usage [...] Read more.
Stem cells have the capacity to self-renew and differentiate to specialized cells, which are usually sensitive to cryopreservation. Therefore, the cell survival rate of stem cells using common cryopreservation protocol is generally not ideal. High cooling rates are crucial for decreasing the usage of cryoprotectants (CPAs) and promoting the successful vitrification of stem cells. In this study, we adopted liquid helium (LHe) instead of liquid nitrogen (LN2) as the cryogen to achieve high cooling rates for vitrifying stem cells with high viability and complete functions. A numerical model was established to simulate the cooling processes of vitrifying specimens by immersing them in LHe and LN2. The calculated results revealed higher cooling rates when plunging specimens into LHe than into LN2. The high viability of human bone-derived mesenchymal stem cells (hBMSCs) and human embryonic stem cells (hESCs) after vitrifying into LHe also shows the superiority of LHe as the cryogen. Furthermore, considerable cell viability was achieved by vitrification in LHe, even when decreasing the concentrations of CPAs. Additionally, post-vitrification, the cells still maintained high attachment and proliferation efficiency, normal stemness, and multipotential differentiation both for hBMSCs and hESCs. LHe is prospective to be employed as a universal cryogen for vitrification which has a great potential for widespread applications, including bioengineering and clinical medicine. Full article
(This article belongs to the Special Issue Multiscale Thermal Engineering for Biomedical Applications)
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28 pages, 2487 KiB  
Review
Recent Advances in Natural Materials for Corneal Tissue Engineering
by Julie F. Jameson, Marisa O. Pacheco, Henry H. Nguyen, Edward A. Phelps and Whitney L. Stoppel
Bioengineering 2021, 8(11), 161; https://doi.org/10.3390/bioengineering8110161 - 26 Oct 2021
Cited by 18 | Viewed by 7864
Abstract
Given the incidence of corneal dysfunctions and diseases worldwide and the limited availability of healthy, human donors, investigators are working to generate engineered cellular and acellular therapeutic approaches as alternatives to corneal transplants from human cadavers. These engineered strategies aim to address existing [...] Read more.
Given the incidence of corneal dysfunctions and diseases worldwide and the limited availability of healthy, human donors, investigators are working to generate engineered cellular and acellular therapeutic approaches as alternatives to corneal transplants from human cadavers. These engineered strategies aim to address existing complications with human corneal transplants, including graft rejection, infection, and complications resulting from surgical methodologies. The main goals of these research endeavors are to (1) determine ideal mechanical properties, (2) devise methodologies to improve the efficacy of engineered corneal grafts and cell-based therapies, and (3) optimize transplantation of engineered tissue structures in the eye. Thus, recent innovations have sought to address these challenges through both in vitro and in vivo studies. This review covers recent work aimed at evaluating engineered materials, potential therapeutic cells, and the resulting cell-material interactions that lead to optimal corneal graft properties. Furthermore, we discuss promising strategies in corneal tissue engineering techniques and in vivo studies in animal models. Full article
(This article belongs to the Special Issue Advances in the Fabrication of Tissue Mimetics)
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17 pages, 1109 KiB  
Article
Monte Carlo-Based Error Propagation for a More Reliable Regression Analysis across Specific Rates in Bioprocesses
by Julian Kager and Christoph Herwig
Bioengineering 2021, 8(11), 160; https://doi.org/10.3390/bioengineering8110160 - 26 Oct 2021
Cited by 6 | Viewed by 2957
Abstract
During process development, bioprocess data need to be converted into applicable knowledge. Therefore, it is crucial to evaluate the obtained data under the usage of transparent and reliable data reduction and correlation techniques. Within this contribution, we show a generic Monte Carlo error [...] Read more.
During process development, bioprocess data need to be converted into applicable knowledge. Therefore, it is crucial to evaluate the obtained data under the usage of transparent and reliable data reduction and correlation techniques. Within this contribution, we show a generic Monte Carlo error propagation and regression approach applied to two different, industrially relevant cultivation processes. Based on measurement uncertainties, errors for cell-specific growth, uptake, and production rates were determined across an evaluation chain, with interlinked inputs and outputs. These uncertainties were subsequently included in regression analysis to derive the covariance of the regression coefficients and the confidence bounds for prediction. The usefulness of the approach is shown within two case studies, based on the relations across biomass-specific rate control limits to guarantee high productivities in E. coli, and low lactate formation in a CHO cell fed-batch could be established. Besides the possibility to determine realistic errors on the evaluated process data, the presented approach helps to differentiate between reliable and unreliable correlations and prevents the wrong interpretations of relations based on uncertain data. Full article
(This article belongs to the Topic Bioreactors: Control, Optimization and Applications)
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