Showing papers in "Journal of Biological Engineering in 2022"
TL;DR: Decellularized extracellular matrix (dECM) can be used as a powder, hydrogel and electrospun scaffolds as mentioned in this paper to provide the mechanical and physical microenvironment needed by cells to proliferate and survive.
Abstract: Abstract Autologous bone grafts are commonly used as the gold standard to repair and regenerate diseased bones. However, they are strongly associated with postoperative complications, especially at the donor site, and increased surgical costs. In an effort to overcome these limitations, tissue engineering (TE) has been proposed as an alternative to promote bone repair. The successful outcome of tissue engineering depends on the microstructure and composition of the materials used as scaffold. Decellularized bone matrix - based biomaterials have been applied as bioscaffolds in bone tissue engineering. These biomaterials play an important role in providing the mechanical and physical microenvironment needed by cells to proliferate and survive. Decellularized extracellular matrix (dECM) can be used as a powder, hydrogel and electrospun scaffolds. These bioscaffolds mimic the native microenvironment due to their structure similar to the original tissue. The aim of this review is to highlight the bone decellularization techniques. Herein we discuss: (1) bone structure; (2) properties of an ideal scaffold; (3) the potential of decellularized bone as bioscaffolds; (4) terminal sterilization of decellularized bone; (5) cell removing confirmation in decellularized tissues; and (6) post decellularization procedures. Finally, the improvement of bone formation by dECM and the immunogenicity aspect of using the decellularized bone matrix are presented, to illustrate how novel dECM-based materials can be used as bioscaffold in tissue engineering. A comprehensive understanding of tissue engineering may allow for better incorporation of therapeutic approaches in bone defects allowing for bone repair and regeneration.
37 citations
TL;DR: Decellularized extracellular matrix (dECM) can be used as a powder, hydrogel and electrospun scaffolds as mentioned in this paper to provide the mechanical and physical microenvironment needed by cells to proliferate and survive.
Abstract: Abstract Autologous bone grafts are commonly used as the gold standard to repair and regenerate diseased bones. However, they are strongly associated with postoperative complications, especially at the donor site, and increased surgical costs. In an effort to overcome these limitations, tissue engineering (TE) has been proposed as an alternative to promote bone repair. The successful outcome of tissue engineering depends on the microstructure and composition of the materials used as scaffold. Decellularized bone matrix - based biomaterials have been applied as bioscaffolds in bone tissue engineering. These biomaterials play an important role in providing the mechanical and physical microenvironment needed by cells to proliferate and survive. Decellularized extracellular matrix (dECM) can be used as a powder, hydrogel and electrospun scaffolds. These bioscaffolds mimic the native microenvironment due to their structure similar to the original tissue. The aim of this review is to highlight the bone decellularization techniques. Herein we discuss: (1) bone structure; (2) properties of an ideal scaffold; (3) the potential of decellularized bone as bioscaffolds; (4) terminal sterilization of decellularized bone; (5) cell removing confirmation in decellularized tissues; and (6) post decellularization procedures. Finally, the improvement of bone formation by dECM and the immunogenicity aspect of using the decellularized bone matrix are presented, to illustrate how novel dECM-based materials can be used as bioscaffold in tissue engineering. A comprehensive understanding of tissue engineering may allow for better incorporation of therapeutic approaches in bone defects allowing for bone repair and regeneration.
35 citations
TL;DR: In this article , the authors provide a brief overview of dendrimer's history and properties and the latest developments of Dendrimers as drug delivery systems, including active drug release strategies to dissociate drug/gene from Dendrer in response to stimuli, size-adaptive and charge reversal Dendimer delivery systems that can better take advantage of the size and surface properties of Dender, and bulk and micro/nano Dender gel delivery systems.
Abstract: Since the first dendrimer was reported in 1978 by Fritz Vögtle, dendrimer research has grown exponentially, from synthesis to application in the past four decades. The distinct structure characteristics of dendrimers include nanoscopic size, multi-functionalized surface, high branching, cavernous interior, and so on, making dendrimers themselves ideal drug delivery vehicles. This mini review article provides a brief overview of dendrimer's history and properties and the latest developments of dendrimers as drug delivery systems. This review focuses on the latest progress in the applications of dendrimers as drug and gene carriers, including 1) active drug release strategies to dissociate drug/gene from dendrimer in response to stimuli; 2) size-adaptive and charge reversal dendrimer delivery systems that can better take advantage of the size and surface properties of dendrimer; 3) bulk and micro/nano dendrimer gel delivery systems. The recent advances in dendrimer formulations may lead to the generation of new drug and gene products and enable the development of novel combination therapies.
23 citations
TL;DR: In this article , the authors provide a brief overview of dendrimer's history and properties and the latest developments of Dendrimers as drug delivery systems, including active drug release strategies to dissociate drug/gene from Dendrer in response to stimuli, size-adaptive and charge reversal Dendimer delivery systems that can better take advantage of the size and surface properties of Dender, and bulk and micro/nano Dender gel delivery systems.
Abstract: Since the first dendrimer was reported in 1978 by Fritz Vögtle, dendrimer research has grown exponentially, from synthesis to application in the past four decades. The distinct structure characteristics of dendrimers include nanoscopic size, multi-functionalized surface, high branching, cavernous interior, and so on, making dendrimers themselves ideal drug delivery vehicles. This mini review article provides a brief overview of dendrimer's history and properties and the latest developments of dendrimers as drug delivery systems. This review focuses on the latest progress in the applications of dendrimers as drug and gene carriers, including 1) active drug release strategies to dissociate drug/gene from dendrimer in response to stimuli; 2) size-adaptive and charge reversal dendrimer delivery systems that can better take advantage of the size and surface properties of dendrimer; 3) bulk and micro/nano dendrimer gel delivery systems. The recent advances in dendrimer formulations may lead to the generation of new drug and gene products and enable the development of novel combination therapies.
22 citations
TL;DR: In this article , the authors reviewed the recent advances in vascular tissue engineering, mainly polyurethane grafts, and summarized the application of preferred cell sources to vascular regeneration, physicochemical properties, and possible degradation mechanisms of PU to provide a more extensive perspective for future research.
Abstract: Certain polymeric materials such as polyurethanes (PUs) are the most prevalent class of used biomaterials in regenerative medicine and have been widely explored as vascular substitutes in several animal models. It is thought that PU-based biomaterials possess suitable hemo-compatibility with comparable performance related to the normal blood vessels. Despite these advantages, the possibility of thrombus formation and restenosis limits their application as artificial functional vessels. In this regard, various surface modification approaches have been developed to enhance both hemo-compatibility and prolong patency. While critically reviewing the recent advances in vascular tissue engineering, mainly PU grafts, this paper summarizes the application of preferred cell sources to vascular regeneration, physicochemical properties, and some possible degradation mechanisms of PU to provide a more extensive perspective for future research.
19 citations
TL;DR: In this paper , the authors reviewed the recent advances in vascular tissue engineering, mainly polyurethane grafts, and summarized the application of preferred cell sources to vascular regeneration, physicochemical properties, and possible degradation mechanisms of PU to provide a more extensive perspective for future research.
Abstract: Certain polymeric materials such as polyurethanes (PUs) are the most prevalent class of used biomaterials in regenerative medicine and have been widely explored as vascular substitutes in several animal models. It is thought that PU-based biomaterials possess suitable hemo-compatibility with comparable performance related to the normal blood vessels. Despite these advantages, the possibility of thrombus formation and restenosis limits their application as artificial functional vessels. In this regard, various surface modification approaches have been developed to enhance both hemo-compatibility and prolong patency. While critically reviewing the recent advances in vascular tissue engineering, mainly PU grafts, this paper summarizes the application of preferred cell sources to vascular regeneration, physicochemical properties, and some possible degradation mechanisms of PU to provide a more extensive perspective for future research.
18 citations
TL;DR: In this article , a review of the literature on the causes of mesangial expansion and its impacts on cell and tissue function is presented, highlighting the gaps that still remain and the potential areas where bioengineering studies can bring insight to mesangia expansion in diabetic nephropathy.
Abstract: Diabetic nephropathy, a kidney complication arising from diabetes, is the leading cause of death in diabetic patients. Unabated, the growing epidemic of diabetes is increasing instances of diabetic nephropathy. Although the main causes of diabetic nephropathy have been determined, the mechanisms of their combined effects on cellular and tissue function are not fully established. One of many damages of diabetic nephropathy is the development of fibrosis within the kidneys, termed mesangial expansion. Mesangial expansion is an important structural lesion that is characterized by the aberrant proliferation of mesangial cells and excess production of matrix proteins. Mesangial expansion is involved in the progression of kidney failure in diabetic nephropathy, yet its causes and mechanism of impact on kidney function are not well defined. Here, we review the literature on the causes of mesangial expansion and its impacts on cell and tissue function. We highlight the gaps that still remain and the potential areas where bioengineering studies can bring insight to mesangial expansion in diabetic nephropathy.
13 citations
TL;DR: In this article , a review of the literature on the causes of mesangial expansion and its impacts on cell and tissue function is presented, highlighting the gaps that still remain and the potential areas where bioengineering studies can bring insight to mesangia expansion in diabetic nephropathy.
Abstract: Diabetic nephropathy, a kidney complication arising from diabetes, is the leading cause of death in diabetic patients. Unabated, the growing epidemic of diabetes is increasing instances of diabetic nephropathy. Although the main causes of diabetic nephropathy have been determined, the mechanisms of their combined effects on cellular and tissue function are not fully established. One of many damages of diabetic nephropathy is the development of fibrosis within the kidneys, termed mesangial expansion. Mesangial expansion is an important structural lesion that is characterized by the aberrant proliferation of mesangial cells and excess production of matrix proteins. Mesangial expansion is involved in the progression of kidney failure in diabetic nephropathy, yet its causes and mechanism of impact on kidney function are not well defined. Here, we review the literature on the causes of mesangial expansion and its impacts on cell and tissue function. We highlight the gaps that still remain and the potential areas where bioengineering studies can bring insight to mesangial expansion in diabetic nephropathy.
12 citations
TL;DR: In this article , a review of the current capabilities for studying native ECM dynamics and delineates new research directions in discovering and implementing new methods for revealing ECM compositional dynamics to push the frontier forward.
Abstract: Abstract The extracellular matrix (ECM) constitutes the main acellular microenvironment of cells in almost all tissues and organs. The ECM not only provides mechanical support, but also mediates numerous biochemical interactions to guide cell survival, proliferation, differentiation, and migration. Thus, better understanding the everchanging temporal and spatial shifts in ECM composition and structure – the ECM dynamics – will provide fundamental insight regarding extracellular regulation of tissue homeostasis and how tissue states transition from one to another during diverse pathophysiological processes. This review outlines the mechanisms mediating ECM-cell interactions and highlights how changes in the ECM modulate tissue development and disease progression, using the lung as the primary model organ. We then discuss existing methodologies for revealing ECM compositional dynamics, with a particular focus on tracking newly synthesized ECM proteins. Finally, we discuss the ramifications ECM dynamics have on tissue engineering and how to implement spatial and temporal specific extracellular microenvironments into bioengineered tissues. Overall, this review communicates the current capabilities for studying native ECM dynamics and delineates new research directions in discovering and implementing ECM dynamics to push the frontier forward.
12 citations
TL;DR: In this paper , a review of the current capabilities for studying native ECM dynamics and delineates new research directions in discovering and implementing new methods for revealing ECM compositional dynamics to push the frontier forward.
Abstract: Abstract The extracellular matrix (ECM) constitutes the main acellular microenvironment of cells in almost all tissues and organs. The ECM not only provides mechanical support, but also mediates numerous biochemical interactions to guide cell survival, proliferation, differentiation, and migration. Thus, better understanding the everchanging temporal and spatial shifts in ECM composition and structure – the ECM dynamics – will provide fundamental insight regarding extracellular regulation of tissue homeostasis and how tissue states transition from one to another during diverse pathophysiological processes. This review outlines the mechanisms mediating ECM-cell interactions and highlights how changes in the ECM modulate tissue development and disease progression, using the lung as the primary model organ. We then discuss existing methodologies for revealing ECM compositional dynamics, with a particular focus on tracking newly synthesized ECM proteins. Finally, we discuss the ramifications ECM dynamics have on tissue engineering and how to implement spatial and temporal specific extracellular microenvironments into bioengineered tissues. Overall, this review communicates the current capabilities for studying native ECM dynamics and delineates new research directions in discovering and implementing ECM dynamics to push the frontier forward.
11 citations
TL;DR: In this article , the authors identified natural compounds capable of inhibiting non-structural protein 4 (Nsp4) of the virus, which is involved in their replication and pathogenesis.
Abstract: Porcine reproductive and respiratory syndrome respiratory sickness in weaned and growing pigs, as well as sow reproductive failure, and its infection is regarded as one of the most serious swine illnesses worldwide. Given the current lack of an effective treatment, in this study, we identified natural compounds capable of inhibiting non-structural protein 4 (Nsp4) of the virus, which is involved in their replication and pathogenesis.We screened natural compounds (n = 97,999) obtained from the ZINC database against Nsp4 and selected the top 10 compounds for analysing protein-ligand interactions and physicochemical properties. The five compounds demonstrating strong binding affinity were then subjected to molecular dynamics simulations (100 ns) and binding free energy calculations. Based on analysis, we identified four possible lead compounds that represent potentially effective drug-like inhibitors.These methods identified that these natural compounds are capable of inhibiting Nsp4 and possibly effective as antiviral therapeutics against PRRSV.
TL;DR: In this paper , the authors identified natural compounds capable of inhibiting non-structural protein 4 (Nsp4) of the virus, which is involved in their replication and pathogenesis.
Abstract: Porcine reproductive and respiratory syndrome respiratory sickness in weaned and growing pigs, as well as sow reproductive failure, and its infection is regarded as one of the most serious swine illnesses worldwide. Given the current lack of an effective treatment, in this study, we identified natural compounds capable of inhibiting non-structural protein 4 (Nsp4) of the virus, which is involved in their replication and pathogenesis.We screened natural compounds (n = 97,999) obtained from the ZINC database against Nsp4 and selected the top 10 compounds for analysing protein-ligand interactions and physicochemical properties. The five compounds demonstrating strong binding affinity were then subjected to molecular dynamics simulations (100 ns) and binding free energy calculations. Based on analysis, we identified four possible lead compounds that represent potentially effective drug-like inhibitors.These methods identified that these natural compounds are capable of inhibiting Nsp4 and possibly effective as antiviral therapeutics against PRRSV.
TL;DR: Wang et al. as discussed by the authors developed a bioactive 3D PLA scaffold using exosome-based strategy to improve its osteogenic and immunoregulatory potential, which showed that PLA-Exo scaffold could reduce the pro-inflammatory marker expression and reactive oxygen species (ROS) production.
Abstract: Polylactic acid (PLA) is a versatile and biodegradable scaffold widely used in biomedical fields to repair tissue defects. Exosomes derived from mesenchymal stem cells (MSCs) are nano-sized extracellular vesicles, which play an important role in tissue engineering in recent years. The primary focus of this study was to develop a bioactive 3D PLA scaffold using exosome-based strategy to improve its osteogenic and immunoregulatory potential. We firstly successfully isolated MSC-derived exosomes (MSC-Exo). Morphological analysis revealed that MSC-Exo exhibits a typical cup-shaped morphology with high expression of exosomal marker CD63. MSC-Exo internalization into recipient cells were also investigated using flow cytometry and confocal laser scanning microscopy. Porous 3D PLA scaffold coated MSC-Exo were used for immunoregulatory and osteogenic testing. Exosomes released from 3D PLA scaffold were validated in RAW264.7 and hBMSCs. The cell proliferation and live/dead assay indicated high biocompatibility for PLA-Exo scaffold. Additionally, PLA-Exo scaffold could reduce the pro-inflammatory marker expression and reactive oxygen species (ROS) production, indicating potential immunoregulatory potential. It is also confirmed that PLA-Exo scaffold could potentiate osteogenic differentiation in the osteogenesis assay. In conclusion, our results demonstrate this bioactive 3D-printed PLA scaffolds with MSC-Exo modification holds immunoregulatory potential and favor osteogenic differentiation, thus having potential applications in bone tissue regeneration.
TL;DR: In this paper , three-dimensional bioartificial tendons (BATs) seeded with murine fibroblasts (cell line C3H10T1/2) were subjected to uniaxial sinusoidal elongation at either overload conditions (0-16, 8%), or physiological load (0 -8, 4%).
Abstract: Tenocytes as specialised fibroblasts and inherent cells of tendons require mechanical load for their homeostasis. However, how mechanical overload compared to physiological load impacts on the tenogenic differentiation potential of fibroblasts is largely unknown.Three-dimensional bioartificial tendons (BATs) seeded with murine fibroblasts (cell line C3H10T1/2) were subjected to uniaxial sinusoidal elongation at either overload conditions (0-16%, Ø 8%) or physiological load (0-8%, Ø 4%). This regime was applied for 2 h a day at 0.1 Hz for 7 days. Controls were unloaded, but under static tension.Cell survival did not differ among overload, physiological load and control BATs. However, gene expression of tenogenic and extra-cellular matrix markers (Scx, Mkx, Tnmd, Col1a1 and Col3a1) was significantly decreased in overload versus physiological load and controls, respectively. In contrast, Mmp3 was significantly increased at overload compared to physiological load, and significantly decreased under physiological load compared to controls. Mkx and Tnmd were significantly increased in BATs subjected to physiological load compared to controls. Proinflammatory interleukin-6 showed increased protein levels comparing load (both over and physiological) versus unloaded controls. Alignment of the cytoskeleton in strain direction was decreased in overload compared to physiological load, while other parameters such as nuclear area, roundness or cell density were less affected.Mechanical overload decreases tenogenic differentiation and increases ECM remodelling/inflammation in 3D-stimulated fibroblasts, whereas physiological load may induce opposite effects.
TL;DR: Wang et al. as mentioned in this paper developed a bioactive 3D PLA scaffold using exosome-based strategy to improve its osteogenic and immunoregulatory potential, which showed that PLA-Exo scaffold could reduce the pro-inflammatory marker expression and reactive oxygen species (ROS) production.
Abstract: Polylactic acid (PLA) is a versatile and biodegradable scaffold widely used in biomedical fields to repair tissue defects. Exosomes derived from mesenchymal stem cells (MSCs) are nano-sized extracellular vesicles, which play an important role in tissue engineering in recent years. The primary focus of this study was to develop a bioactive 3D PLA scaffold using exosome-based strategy to improve its osteogenic and immunoregulatory potential. We firstly successfully isolated MSC-derived exosomes (MSC-Exo). Morphological analysis revealed that MSC-Exo exhibits a typical cup-shaped morphology with high expression of exosomal marker CD63. MSC-Exo internalization into recipient cells were also investigated using flow cytometry and confocal laser scanning microscopy. Porous 3D PLA scaffold coated MSC-Exo were used for immunoregulatory and osteogenic testing. Exosomes released from 3D PLA scaffold were validated in RAW264.7 and hBMSCs. The cell proliferation and live/dead assay indicated high biocompatibility for PLA-Exo scaffold. Additionally, PLA-Exo scaffold could reduce the pro-inflammatory marker expression and reactive oxygen species (ROS) production, indicating potential immunoregulatory potential. It is also confirmed that PLA-Exo scaffold could potentiate osteogenic differentiation in the osteogenesis assay. In conclusion, our results demonstrate this bioactive 3D-printed PLA scaffolds with MSC-Exo modification holds immunoregulatory potential and favor osteogenic differentiation, thus having potential applications in bone tissue regeneration.
TL;DR: In this article , heat shock pretreatment was used to up-regulate 70 kilodalton heat shock proteins (HSP70) expression in bone marrow mesenchymal stem cells (BMSCs)-derived exosomes (HS-BMSC-Exo) to alleviate cisplatin-induced ototoxicity.
Abstract: Abstract NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome contributes to the development of cisplatin-induced ototoxicity. Whether heat shock pretreatment could be utilized to up-regulate 70 kilodalton heat shock proteins (HSP70) expression in bone marrow mesenchymal stem cells (BMSCs)-derived exosomes (HS-BMSC-Exo) to alleviate cisplatin-induced ototoxicity is deciphered in this study. Heat shock pretreatment was performed on BMSCs to induce HS-BMSC-Exo, which were further trans-tympanically administrated into cisplatin intraperitoneally injected C57BL/6 mice. Auditory brainstem response (ABR) was assessed to indicate auditory sensitivity at 8, 16, 24, and 32 kHz. Myosin 7a staining was utilized to detect the mature hair cells. The relative expressions of the NLRP3 inflammasome complex were determined with Western blot in the cochlea. Diminished auditory sensitivity and increased hair cell loss could be observed in the cisplatin exposed mice with increased content of interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, NLRP3, ASC, cleaved caspase-1, and pro-caspase-1, and decreased content of IL-10, which could be reversed by HS-BMSC-Exo or BMSC-Exo administration. It was worth noting that HS-BMSC-Exo demonstrated more treatment benefits than BMSC-Exo in cisplatin-induced ototoxicity. Heat shock precondition may provide a new therapeutic option to produce exosomal HSP70, and HS-BMSC-Exo could be utilized to relieve cisplatin-induced ototoxicity.
TL;DR: In this paper , NCK1-AS1 was detected by qRT-PCR and the expression of inflammatory cytokines (IL-1β, IL-6, and TNK-α) was measured by ELISA assays.
Abstract: Although long non-coding RNA (lncRNA) NCK1-AS1 plays important roles in human cancer, its function in atherosclerosis (AS) remains unclear.The expression of NCK1-AS1 in AS blood samples was detected by qRT-PCR. Oxidized low-density lipoprotein (ox-LDL) was used to construct the AS cell model, and quantitative real-time polymerase chain reaction (qRT-PCR) assay was used to evaluate NCK1-AS1 level. Cell phenotypes including proliferation and apoptosis were assessed by Cell Counting Kit-8 (CCK-8) assay and flow cytometer, respectively. The malondialdehyde level was measured to evaluate oxidative stress. The expression of apoptosis-related proteins was evaluated by western blot. The expression of inflammatory cytokines (IL-1β, IL-6 and TNK-α) was measured by qRT-PCR and ELISA assays. The relationship among NCK1-AS1, miR-1197 and COX10 was determined by bioinformatic analysis and luciferase reporter assay.NCK1-AS1 was significantly upregulated in AS blood samples and ox-LDL stimulated vascular smooth muscle cells (VSMCs). Knockdown of NCK1-AS1 increased cell viability, reduced cell apoptosis and MDA level, and also inhibited the expression of inflammatory cytokines (IL-1β, IL-6 and TNK-α) in ox-LDL stimulated VSMCs. NCK1-AS1 could positively regulate COX10 expression by directly sponging miR-1197. Moreover, co-transfection of sh-NCK1-AS1 and miR-1197 inhibitor, or co-transfection of sh-NCK1-AS1 and pc-COX10 (COX10 overexpressing plasmid) obviously reduced cell viability, promoted cell apoptosis, and increased MDA level in VSMCs followed by ox-LDL treatment for 24 h compared to that in sh-NCK1-AS1 transfected VSMCs.Our study revealed that knockdown of NCK1-AS1 attenuated the development of AS by regulating miR-1197/COX10 axis, suggesting that this lncRNA might be a potential therapeutic target for AS.
TL;DR: In this paper , two different carbohydrate-binding modules (CBM3 and CBM9) have been successfully fused to an alcohol dehydrogenase from Saccharomyces cerevisiae, which has been produced in bench-scale reactor using an auxotrophic M15-derived E. coli strain, following a fed-batch strategy with antibiotic-free medium.
Abstract: The feasibility of biochemical transformation processes is usually greatly dependent on biocatalysts cost. Therefore, immobilizing and reusing biocatalysts is an approach to be considered to bring biotransformations closer to industrial feasibility, since it does not only allow to reuse enzymes but can also improve their stability towards several reaction conditions. Carbohydrate-Binding Modules (CBM) are well-described domains involved in substrate binding which have been already used as purification tags.In this work, two different Carbohydrate-Binding Modules (CBM3 and CBM9) have been successfully fused to an alcohol dehydrogenase from Saccharomyces cerevisiae, which has been produced in bench-scale reactor using an auxotrophic M15-derived E. coli strain, following a fed-batch strategy with antibiotic-free medium. Around 40 mg·g- 1 DCW of both fusion proteins were produced, with a specific activity of > 65 AU·mg- 1. Overexpressed proteins were bound to a low-cost and highly selective cellulosic support by one-step immobilization/purification process at > 98% yield, retaining about a 90% of initial activity. Finally, the same support was also used for protein purification, aiming to establish an alternative to metal affinity chromatography, by which CBM9 tag proved to be useful, with a recovery yield of > 97% and 5-fold increased purity grade.CBM domains were proved to be suitable for one-step immobilization/purification process, retaining almost total activity offered. However, purification process was only successful with CBM9.
TL;DR: In this article , two different carbohydrate-binding modules (CBM3 and CBM9) have been successfully fused to an alcohol dehydrogenase from Saccharomyces cerevisiae, which has been produced in bench-scale reactor using an auxotrophic M15-derived E. coli strain, following a fed-batch strategy with antibiotic-free medium.
Abstract: The feasibility of biochemical transformation processes is usually greatly dependent on biocatalysts cost. Therefore, immobilizing and reusing biocatalysts is an approach to be considered to bring biotransformations closer to industrial feasibility, since it does not only allow to reuse enzymes but can also improve their stability towards several reaction conditions. Carbohydrate-Binding Modules (CBM) are well-described domains involved in substrate binding which have been already used as purification tags.In this work, two different Carbohydrate-Binding Modules (CBM3 and CBM9) have been successfully fused to an alcohol dehydrogenase from Saccharomyces cerevisiae, which has been produced in bench-scale reactor using an auxotrophic M15-derived E. coli strain, following a fed-batch strategy with antibiotic-free medium. Around 40 mg·g- 1 DCW of both fusion proteins were produced, with a specific activity of > 65 AU·mg- 1. Overexpressed proteins were bound to a low-cost and highly selective cellulosic support by one-step immobilization/purification process at > 98% yield, retaining about a 90% of initial activity. Finally, the same support was also used for protein purification, aiming to establish an alternative to metal affinity chromatography, by which CBM9 tag proved to be useful, with a recovery yield of > 97% and 5-fold increased purity grade.CBM domains were proved to be suitable for one-step immobilization/purification process, retaining almost total activity offered. However, purification process was only successful with CBM9.
TL;DR: In this article , three-dimensional bioartificial tendons (BATs) seeded with murine fibroblasts (cell line C3H10T1/2) were subjected to uniaxial sinusoidal elongation at either overload conditions (0-16, 8%), or physiological load (0 -8, 4%).
Abstract: Tenocytes as specialised fibroblasts and inherent cells of tendons require mechanical load for their homeostasis. However, how mechanical overload compared to physiological load impacts on the tenogenic differentiation potential of fibroblasts is largely unknown.Three-dimensional bioartificial tendons (BATs) seeded with murine fibroblasts (cell line C3H10T1/2) were subjected to uniaxial sinusoidal elongation at either overload conditions (0-16%, Ø 8%) or physiological load (0-8%, Ø 4%). This regime was applied for 2 h a day at 0.1 Hz for 7 days. Controls were unloaded, but under static tension.Cell survival did not differ among overload, physiological load and control BATs. However, gene expression of tenogenic and extra-cellular matrix markers (Scx, Mkx, Tnmd, Col1a1 and Col3a1) was significantly decreased in overload versus physiological load and controls, respectively. In contrast, Mmp3 was significantly increased at overload compared to physiological load, and significantly decreased under physiological load compared to controls. Mkx and Tnmd were significantly increased in BATs subjected to physiological load compared to controls. Proinflammatory interleukin-6 showed increased protein levels comparing load (both over and physiological) versus unloaded controls. Alignment of the cytoskeleton in strain direction was decreased in overload compared to physiological load, while other parameters such as nuclear area, roundness or cell density were less affected.Mechanical overload decreases tenogenic differentiation and increases ECM remodelling/inflammation in 3D-stimulated fibroblasts, whereas physiological load may induce opposite effects.
TL;DR: In this article , a multi-channel alternating current biosusceptometry (ACB) system with nineteen pick-up coils was used to obtain 2D quantitative images of magnetic nanoparticle distributions by solving an inverse problem.
Abstract: Non-invasive magnetic imaging techniques are necessary to assist magnetic nanoparticles in biomedical applications, mainly detecting their distribution inside the body. In Alternating Current Biosusceptometry (ACB), the magnetic nanoparticle's magnetization response under an oscillating magnetic field, which is applied through an excitation coil, is detected with a balanced detection coil system.We built a Multi-Channel ACB system (MC-ACB) containing nineteen pick-up coils and obtained 2D quantitative images of magnetic nanoparticle distributions by solving an inverse problem. We reconstructed the magnetic nanoparticles spatial distributions in a field of view of 14 × 14 cm2 with a spatial resolution of 2.0 cm and sensitivity in the milligram scale. A correlation coefficient between quantitative reconstructed and nominal magnetic nanoparticle distributions above 0.6 was found for all measurements.Besides other interesting features such as sufficient large field of view dimension for mice and rat studies, portability, and the ability to assess the quantitative magnetic nanoparticles distributions in real-time, the MC-ACB system is a promising tool for quantitative imaging of magnetic nanoparticles distributions in real-time, offering an affordable setup for easy access in clinical or laboratory environments.
TL;DR: In this paper , VR videos of the laboratory component of a Biomolecular Engineering course were provided to students and a survey was distributed for students to self-report their experience with the videos.
Abstract: Abstract Background The Covid-19 pandemic caused a sudden shift towards remote learning, moving classes to online formats. Not exempt from this switch, laboratory courses traditionally taught in-person were also moved to remote methods, costing students the opportunity to learn these skills hands-on. In order for instructors to provide course materials effectively and engagingly, non-traditional methods should be explored. Virtual reality (VR) has become more accessible in recent years. VR simulations have been used for many years as educational tools in high-risk settings such as flight or medical simulations. Immersive VR videos implemented in a remote laboratory course could provide the students with an engaging and suitable learning experience. To test the effectiveness of VR videos as a tool for remote education, VR videos of the laboratory component of a Biomolecular Engineering course were provided to students. A survey was distributed for students to self-report their experience with the videos. The survey contained quantitative and qualitative ratings of VR as an educational tool. Results The survey showed that students (~ 89% strongly agree or agree) believed the videos provided the opportunity to work at their own pace and were an appropriate length. While ~ 74% of students said that the videos provided enough information to understand the tasks, a small percentage felt that the videos improved their retention (~ 16%) and understanding (~ 9%) of the course material. About 28% of the students responded positively when asked about how VR videos improved their engagement with the material. ~ 30% reported confidence in applying the skills learned in the videos in the future and ~ 43% believe the VR videos were an acceptable alternative to in-person labs. Two-thirds of students reported feeling some form of discomfort while viewing the VR videos and 54% reported not using the headset for the videos and using the 3D video feature instead. Conclusions As many students reported the videos containing appropriate information, the content of the videos was not an issue. A combination of improved camera quality with motion stability, more comfortable headsets, and a reduction in editing issues could greatly improve the quality and effectiveness of VR videos.
TL;DR: In this article , a novel antibacterial flexible polymeric hydrogel films were designed via crosslinking polymeric chitosan (CS) with folic acid-based carbon quantum dots (CQDs).
Abstract: Abstract Background To provide effective healing in the wound, various carbohydrate polymers are commonly utilized that are highly potent platforms as wound dressing films. In this work, novel antibacterial flexible polymeric hydrogel films were designed via crosslinking polymeric chitosan (CS) with folic acid-based carbon quantum dots (CQDs). To end this, folic acid as a bio-precursor is used to synthesize CQDs through the hydrothermal technique. The synthesized CQDs as a crosslinking agent was performed at different concentrations to construct nanocomposite hydrogel films via the casting technique. Also, gentamicin (GM), L -Arginine and glycerol were supplemented in the formulation of nanocomposite since their antibiotic, bioactivity and plasticizing ability, respectively. Results The successful construction of films were verified with different methods (FT-IR, UV-Vis, PL, SEM, and AFM analyses). The GM release profile displayed a controlled release manner over 48 h with a low initial burst release in the simulated wound media (PBS, pH 7.4). Antibacterial and in vitro cytotoxicity results showed a significant activity toward different gram-positive and negative bacterial strains (about 2.5 ± 0.1 cm inhibition zones) and a desired cytocompatibility against Human skin fibroblast (HFF-1) cells (over 80% cell viability), respectively. Conclusion The obtained results recommend CQDs-crosslinked CS (CS/CQD) nanocomposite as a potent antimicrobial wound dressing. Graphical Abstract
TL;DR: In this paper , VR videos of the laboratory component of a Biomolecular Engineering course were provided to students and a survey was distributed for students to self-report their experience with the videos.
Abstract: Abstract Background The Covid-19 pandemic caused a sudden shift towards remote learning, moving classes to online formats. Not exempt from this switch, laboratory courses traditionally taught in-person were also moved to remote methods, costing students the opportunity to learn these skills hands-on. In order for instructors to provide course materials effectively and engagingly, non-traditional methods should be explored. Virtual reality (VR) has become more accessible in recent years. VR simulations have been used for many years as educational tools in high-risk settings such as flight or medical simulations. Immersive VR videos implemented in a remote laboratory course could provide the students with an engaging and suitable learning experience. To test the effectiveness of VR videos as a tool for remote education, VR videos of the laboratory component of a Biomolecular Engineering course were provided to students. A survey was distributed for students to self-report their experience with the videos. The survey contained quantitative and qualitative ratings of VR as an educational tool. Results The survey showed that students (~ 89% strongly agree or agree) believed the videos provided the opportunity to work at their own pace and were an appropriate length. While ~ 74% of students said that the videos provided enough information to understand the tasks, a small percentage felt that the videos improved their retention (~ 16%) and understanding (~ 9%) of the course material. About 28% of the students responded positively when asked about how VR videos improved their engagement with the material. ~ 30% reported confidence in applying the skills learned in the videos in the future and ~ 43% believe the VR videos were an acceptable alternative to in-person labs. Two-thirds of students reported feeling some form of discomfort while viewing the VR videos and 54% reported not using the headset for the videos and using the 3D video feature instead. Conclusions As many students reported the videos containing appropriate information, the content of the videos was not an issue. A combination of improved camera quality with motion stability, more comfortable headsets, and a reduction in editing issues could greatly improve the quality and effectiveness of VR videos.
TL;DR: In this paper , the authors reviewed the research progress on microfluidic chip-based recombinase polymerase isothermal amplification technology and highlighted future prospects for preventing and controlling infectious diseases.
Abstract: Abstract The frequency of outbreaks of newly emerging infectious diseases has increased in recent years. The coronavirus disease 2019 (COVID-19) outbreak in late 2019 has caused a global pandemic, seriously endangering human health and social stability. Rapid detection of infectious disease pathogens is a key prerequisite for the early screening of cases and the reduction in transmission risk. Fluorescence quantitative polymerase chain reaction (qPCR) is currently the most commonly used pathogen detection method, but this method has high requirements in terms of operating staff, instrumentation, venues, and so forth. As a result, its application in the settings such as poorly conditioned communities and grassroots has been limited, and the detection needs of the first-line field cannot be met. The development of point-of-care testing (POCT) technology is of great practical significance for preventing and controlling infectious diseases. Isothermal amplification technology has advantages such as mild reaction conditions and low instrument dependence. It has a promising prospect in the development of POCT, combined with the advantages of high integration and portability of microfluidic chip technology. This study summarized the principles of several representative isothermal amplification techniques, as well as their advantages and disadvantages. Particularly, it reviewed the research progress on microfluidic chip–based recombinase polymerase isothermal amplification technology and highlighted future prospects.
TL;DR: In this paper , the authors identify ELF3 as a transcription factor that promotes mesenchymal-epithelial transition and stabilizes hybrid E/M phenotypes, and show that it is associated with epithelial phenotype and is inhibited during EMT.
Abstract: Epithelial-mesenchymal plasticity (EMP) involves bidirectional transitions between epithelial, mesenchymal and multiple intermediary hybrid epithelial/mesenchymal phenotypes. While the process of epithelial-mesenchymal transition (EMT) and its associated transcription factors are well-characterised, the transcription factors that promote mesenchymal-epithelial transition (MET) and stabilise hybrid E/M phenotypes are less well understood.Here, we analyse multiple publicly-available transcriptomic datasets at bulk and single-cell level and pinpoint ELF3 as a factor that is strongly associated with an epithelial phenotype and is inhibited during EMT. Using mechanism-based mathematical modelling, we also show that ELF3 inhibits the progression of EMT. This behaviour was also observed in the presence of an EMT inducing factor WT1. Our model predicts that the MET induction capacity of ELF3 is stronger than that of KLF4, but weaker than that of GRHL2. Finally, we show that ELF3 levels correlates with worse patient survival in a subset of solid tumour types.ELF3 is shown to be inhibited during EMT progression and is also found to inhibit the progression of complete EMT suggesting that ELF3 may be able to counteract EMT induction, including in the presence of EMT-inducing factors, such as WT1. The analysis of patient survival data indicates that the prognostic capacity of ELF3 is specific to cell-of-origin or lineage.
TL;DR: A review of the progress in the field of tissue engineering for the gastrointestinal tract can be found in this article , where the authors provide a perspective on the path to translation and highlight newer advances in bioengineering strategies.
Abstract: The gastrointestinal (GI) tract is imperative for multiple functions including digestion, nutrient absorption, and timely waste disposal. The central feature of the gut is peristalsis, intestinal motility, which facilitates all of its functions. Disruptions in GI motility lead to sub-optimal GI function, resulting in a lower quality of life in many functional GI disorders. Over the last two decades, tissue engineering research directed towards the intestine has progressed rapidly due to advances in cell and stem-cell biology, integrative physiology, bioengineering and biomaterials. Newer biomedical tools (including optical tools, machine learning, and nuanced regenerative engineering approaches) have expanded our understanding of the complex cellular communication within the GI tract that lead to its orchestrated physiological function. Bioengineering therefore can be utilized towards several translational aspects: (i) regenerative medicine to remedy/restore GI physiological function; (ii) in vitro model building to mimic the complex physiology for drug and pharmacology testing; (iii) tool development to continue to unravel multi-cell communication networks to integrate cell and organ-level physiology. Despite the significant strides made historically in GI tissue engineering, fundamental challenges remain including the quest for identifying autologous human cell sources, enhanced scaffolding biomaterials to increase biocompatibility while matching viscoelastic properties of the underlying tissue, and overall biomanufacturing. This review provides historic perspectives for how bioengineering has advanced over time, highlights newer advances in bioengineering strategies, and provides a realistic perspective on the path to translation.
TL;DR: The biological engineering profession must be involved in the research and implementation of these solutions-developing new tools to aid in decision-making, methods to optimize across different objectives, and new messaging frameworks to assist in prioritizing among different options as discussed by the authors .
Abstract: Nature-based Climate Solutions are landscape stewardship techniques to reduce greenhouse gas emissions and increase soil or biomass carbon sequestration. These mitigation approaches to climate change present an opportunity to supplement energy sector decarbonization and provide co-benefits in terms of ecosystem services and landscape productivity. The biological engineering profession must be involved in the research and implementation of these solutions-developing new tools to aid in decision-making, methods to optimize across different objectives, and new messaging frameworks to assist in prioritizing among different options. Furthermore, the biological engineering curriculum should be redesigned to reflect the needs of carbon-based landscape management. While doing so, the biological engineering community has an opportunity to embed justice, equity, diversity, and inclusion within both the classroom and the profession. Together these transformations will enhance our capacity to use sustainable landscape management as an active tool to mitigate the risks of climate change.
TL;DR: The biological engineering profession must be involved in the research and implementation of these solutions-developing new tools to aid in decision-making, methods to optimize across different objectives, and new messaging frameworks to assist in prioritizing among different options as mentioned in this paper .
Abstract: Nature-based Climate Solutions are landscape stewardship techniques to reduce greenhouse gas emissions and increase soil or biomass carbon sequestration. These mitigation approaches to climate change present an opportunity to supplement energy sector decarbonization and provide co-benefits in terms of ecosystem services and landscape productivity. The biological engineering profession must be involved in the research and implementation of these solutions-developing new tools to aid in decision-making, methods to optimize across different objectives, and new messaging frameworks to assist in prioritizing among different options. Furthermore, the biological engineering curriculum should be redesigned to reflect the needs of carbon-based landscape management. While doing so, the biological engineering community has an opportunity to embed justice, equity, diversity, and inclusion within both the classroom and the profession. Together these transformations will enhance our capacity to use sustainable landscape management as an active tool to mitigate the risks of climate change.
TL;DR: In this paper , Dihydroartemisinin (DHART)-loaded polycaprolactone collagen nanofibers (PCL/Col NFs) were constructed as effective biocompatible scaffolds through adjusting the proportions of hydrophobic/ hydrophilic polymers for enhanced osteoblastic differentiation of human adipose-derived stem cells (hADSCs).
Abstract: Adipose tissue-derived stem cells (ASCs) are promising candidate in stem cell therapies, and maintaining their stemness potential is vital to achieve effective treatment. Natural-based scaffolds have been recently attracted increasing attention in nanomedicine and drug delivery. In this study, Dihydroartemisinin (DHART)-loaded polycaprolactone collagen nanofibers (PCL/Col NFs) were constructed as effective biocompatible scaffolds through adjusting the proportions of hydrophobic/ hydrophilic polymers for enhanced osteoblastic differentiation of human adipose-derived stem cells (hADSCs).The designed NFs were characterized through FTIR, XRD, TGA, FE-SEM, and tensile testing. DHART-loaded PCL/Col electrospun NFs provide an ideal solution, with the potential of sustained drug release as well as inhibition of drug re-crystallization. Interestingly, inhibiting DHART re-crystallization can improve its bioavailability and provide a more effective therapeutic efficacy. Besides, the data set found through FE-SEM, MTT, PicoGreen, qPCR, and alkaline phosphatase (ALP) assays revealed the improved adhesion and proliferation rate of hADSCs cultured on PCL/Col/DHART (5%) NFs after 14 and 21 days of incubation.These findings confirmed the potential of the designed NF scaffolds for sustained/controlled release of DHART therapeutic molecules toward bone tissue regeneration and engineering.