Wenhao Cheng, Sundol Kim, Sandra Zivkovic, Hoyong Chung, Yi Ren, J. Guan
Phagocytosis performed by a macrophage involves complex membrane trafficking and reorganization among various membranous cellular structures including phagosomes and vesicles derived from the phagosomes known as phagosome-derived vesicles. The present work reports on development of a technique that allows to specifically label the phagosome-derived vesicles in macrophages with a membrane dye. The technique is based on the use of microfabricated microparticles that are made of a thermosensitive nonbiodegradable polymer poly(N-isopropylacrylamide) (PNIPAM) or its derivative and contain a membrane dye 1,1'-dialkyl-3,3,3',3'-tetramethylindodicarbocyanine (DiI). The microparticles can be phagocytosed by RAW264.7 macrophages into their phagosomes, resulting in formation of intracellular DiI-positive vesicles derived from the phagosomes. The DiI-positive vesicles are motile and acidic; can be stained by fluorescently labelled dextran added in the culture medium; and can accumulate around new phagosomes, indicating that they possess properties of lysosomes. This technique is also applicable to another membrane dye 3,3'-dioctadecyloxacarbocyanine (DiO) and holds great potential to be useful for advancing our understanding of phagocytosis. STATEMENT OF SIGNIFICANCE: : Phagocytosis performed by macrophages is a cellular process of great importance to various applications of biomaterials such as drug delivery and medical implantation. This work reports on a technique for characterizing phagocytosis based on the use of poly(N-isopropylacrylamide), which is a major biomaterial with numerous applications. This technique is the first of its kind and has generated an original finding about phagocytosis. In addition to drug delivery and medical implantation, phagocytosis plays critical roles in diseases, injuries and vaccination. This work could thus attract immediate and widespread interests in the field of biomaterials science and engineering.
{"title":"Specific Labelling of Phagosome-Derived Vesicles in Macrophages with a Membrane Dye Delivered with Microfabricated Microparticles","authors":"Wenhao Cheng, Sundol Kim, Sandra Zivkovic, Hoyong Chung, Yi Ren, J. Guan","doi":"10.2139/ssrn.3937057","DOIUrl":"https://doi.org/10.2139/ssrn.3937057","url":null,"abstract":"Phagocytosis performed by a macrophage involves complex membrane trafficking and reorganization among various membranous cellular structures including phagosomes and vesicles derived from the phagosomes known as phagosome-derived vesicles. The present work reports on development of a technique that allows to specifically label the phagosome-derived vesicles in macrophages with a membrane dye. The technique is based on the use of microfabricated microparticles that are made of a thermosensitive nonbiodegradable polymer poly(N-isopropylacrylamide) (PNIPAM) or its derivative and contain a membrane dye 1,1'-dialkyl-3,3,3',3'-tetramethylindodicarbocyanine (DiI). The microparticles can be phagocytosed by RAW264.7 macrophages into their phagosomes, resulting in formation of intracellular DiI-positive vesicles derived from the phagosomes. The DiI-positive vesicles are motile and acidic; can be stained by fluorescently labelled dextran added in the culture medium; and can accumulate around new phagosomes, indicating that they possess properties of lysosomes. This technique is also applicable to another membrane dye 3,3'-dioctadecyloxacarbocyanine (DiO) and holds great potential to be useful for advancing our understanding of phagocytosis. STATEMENT OF SIGNIFICANCE: : Phagocytosis performed by macrophages is a cellular process of great importance to various applications of biomaterials such as drug delivery and medical implantation. This work reports on a technique for characterizing phagocytosis based on the use of poly(N-isopropylacrylamide), which is a major biomaterial with numerous applications. This technique is the first of its kind and has generated an original finding about phagocytosis. In addition to drug delivery and medical implantation, phagocytosis plays critical roles in diseases, injuries and vaccination. This work could thus attract immediate and widespread interests in the field of biomaterials science and engineering.","PeriodicalId":106645,"journal":{"name":"MatSciRN: Tissue Engineering (Topic)","volume":"10 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"123854083","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Y. Yasenchuk, E. Marchenko, S. Gunther, G. Baigonakova, O. Kokorev, A. Volinsky, E. Topolnitsky
The mechanical behavior of samples of skin, tendon, muscle, and knitted mesh made from TiNi wire was investigated employing uniaxial cyclic tension and tension to rupture. The cyclic tensile stress-strain curves of all biological tissues exhibited the Mullins softening effect, characteristic of cyclic tension of hyperelastic materials. The mechanical behavior of knitted TiNi mesh made of 40-100 μm diameter wire is similar to soft biological tissues. All samples under uniaxial cyclic tension have exhibited the softening and delay effects, and in each of the diagrams, one can distinguish low and high-modulus regions. The deformation of the wire in the loading-unloading cycle is characterized by a superelastic behavior, which did not manifest itself in knitted TiNi mesh made from it. The knitted mesh from TiNi 60 μm wire at a minimum pre-tension has shown a minimal softening effect and a minimal decrease in stress hysteresis after the first cycle of physiological deformation of 6%. The discovered effects of hyperelastic behavior will make it possible to develop criteria for the selection and evaluation of knitted materials made of titanium nickelide for soft tissue reconstructive surgery. In vivo studies have shown good integration of the knitted TiNi mesh into living biological tissues under normal physiological stress.
{"title":"Mullins Effect in Soft Biological Tissues and Knitted Titanium Nickelide Under Cyclic Loading","authors":"Y. Yasenchuk, E. Marchenko, S. Gunther, G. Baigonakova, O. Kokorev, A. Volinsky, E. Topolnitsky","doi":"10.2139/ssrn.3807767","DOIUrl":"https://doi.org/10.2139/ssrn.3807767","url":null,"abstract":"The mechanical behavior of samples of skin, tendon, muscle, and knitted mesh made from TiNi wire was investigated employing uniaxial cyclic tension and tension to rupture. The cyclic tensile stress-strain curves of all biological tissues exhibited the Mullins softening effect, characteristic of cyclic tension of hyperelastic materials. The mechanical behavior of knitted TiNi mesh made of 40-100 μm diameter wire is similar to soft biological tissues. All samples under uniaxial cyclic tension have exhibited the softening and delay effects, and in each of the diagrams, one can distinguish low and high-modulus regions. The deformation of the wire in the loading-unloading cycle is characterized by a superelastic behavior, which did not manifest itself in knitted TiNi mesh made from it. The knitted mesh from TiNi 60 μm wire at a minimum pre-tension has shown a minimal softening effect and a minimal decrease in stress hysteresis after the first cycle of physiological deformation of 6%. The discovered effects of hyperelastic behavior will make it possible to develop criteria for the selection and evaluation of knitted materials made of titanium nickelide for soft tissue reconstructive surgery. In vivo studies have shown good integration of the knitted TiNi mesh into living biological tissues under normal physiological stress.","PeriodicalId":106645,"journal":{"name":"MatSciRN: Tissue Engineering (Topic)","volume":"19 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2021-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"124145707","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
A. McMillan, M. K. Nguyen, C. T. Huynh, Samantha M. Sarett, Peilin Ge, Melanie Chetverikova, Kien Nguyen, D. Grosh, C. Duvall, E. Alsberg
Delivery systems for controlled release of RNA interference (RNAi) molecules, including small interfering (siRNA) and microRNA (miRNA), have the potential to direct stem cell differentiation for regenerative musculoskeletal applications. To date, localized RNA delivery platforms in this area have focused predominantly on bulk scaffold-based approaches, which can interfere with cell-cell interactions important for recapitulating some native musculoskeletal developmental and healing processes in tissue regeneration strategies. In contrast, scaffold-free, high density human mesenchymal stem cell (hMSC) aggregates may provide an avenue for creating a more biomimetic microenvironment. Here, photocrosslinkable dextran microspheres (MS) encapsulating siRNA-micelles were prepared via an aqueous emulsion method and incorporated within hMSC aggregates for localized and sustained delivery of bioactive siRNA. siRNA-micelles released from MS in a sustained fashion over the course of 28 days, and the released siRNA retained its ability to transfect cells for gene silencing. Incorporation of fluorescently labeled siRNA (siGLO)-laden MS within hMSC aggregates exhibited tunable siGLO delivery and uptake by stem cells. Incorporation of MS loaded with siRNA targeting green fluorescent protein (siGFP) within GFP-hMSC aggregates provided sustained presentation of siGFP within the constructs and prolonged GFP silencing for up to 15 days. This platform system enables sustained gene silencing within stem cell aggregates and thus shows great potential in tissue regeneration applications.
{"title":"Hydrogel Microspheres for Spatiotemporally Controlled Delivery of Rna and Silencing Gene Expression within Scaffold-Free Tissue Engineered Constructs","authors":"A. McMillan, M. K. Nguyen, C. T. Huynh, Samantha M. Sarett, Peilin Ge, Melanie Chetverikova, Kien Nguyen, D. Grosh, C. Duvall, E. Alsberg","doi":"10.2139/ssrn.3687962","DOIUrl":"https://doi.org/10.2139/ssrn.3687962","url":null,"abstract":"Delivery systems for controlled release of RNA interference (RNAi) molecules, including small interfering (siRNA) and microRNA (miRNA), have the potential to direct stem cell differentiation for regenerative musculoskeletal applications. To date, localized RNA delivery platforms in this area have focused predominantly on bulk scaffold-based approaches, which can interfere with cell-cell interactions important for recapitulating some native musculoskeletal developmental and healing processes in tissue regeneration strategies. In contrast, scaffold-free, high density human mesenchymal stem cell (hMSC) aggregates may provide an avenue for creating a more biomimetic microenvironment. Here, photocrosslinkable dextran microspheres (MS) encapsulating siRNA-micelles were prepared via an aqueous emulsion method and incorporated within hMSC aggregates for localized and sustained delivery of bioactive siRNA. siRNA-micelles released from MS in a sustained fashion over the course of 28 days, and the released siRNA retained its ability to transfect cells for gene silencing. Incorporation of fluorescently labeled siRNA (siGLO)-laden MS within hMSC aggregates exhibited tunable siGLO delivery and uptake by stem cells. Incorporation of MS loaded with siRNA targeting green fluorescent protein (siGFP) within GFP-hMSC aggregates provided sustained presentation of siGFP within the constructs and prolonged GFP silencing for up to 15 days. This platform system enables sustained gene silencing within stem cell aggregates and thus shows great potential in tissue regeneration applications.","PeriodicalId":106645,"journal":{"name":"MatSciRN: Tissue Engineering (Topic)","volume":"11 17 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2021-01-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"116823630","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
M. Klimak, R. Nims, Lara Pferdehirt, K. Collins, N. Harasymowicz, S. Oswald, L. Setton, F. Guilak
Immunoengineering continues to revolutionize healthcare, generating new approaches for treating previously intractable diseases, particularly in regard to cancer immunotherapy. In joint diseases, such as osteoarthritis (OA) and rheumatoid arthritis (RA), biomaterials and anti-cytokine treatments have previously been at that forefront of therapeutic innovation. However, while many of the existing anti-cytokine treatments are successful for a subset of patients, these treatments can also pose severe risks, adverse events and off-target effects due to continuous delivery at high dosages or a lack of disease-specific targets. The inadequacy of these current treatments has motivated the development of new immunoengineering strategies that offer a safer and more efficacious alternative therapies through the precise and controlled targeting of specific upstream immune responses, including direct and mechanistically-driven immunoengineering approaches. Advances in the understanding of the immunomodulatory pathways involved in musculoskeletal disease, in combination with the growing emphasis on personalized medicine, stress the need for carefully considering the delivery strategies and therapeutic targets when designing therapeutics to better treat RA and OA. Here, we focus on recent advances in biomaterial and cell-based immunomodulation, in combination with genetic engineering, for therapeutic applications in joint diseases. The application of immunoengineering principles to the study of joint disease will not only help to elucidate the mechanisms of disease pathogenesis but will also generate novel disease-specific therapeutics by harnessing cellular and biomaterial responses. STATEMENT OF SIGNIFICANCE: It is now apparent that joint diseases such as osteoarthritis and rheumatoid arthritis involve the immune system at both local (i.e., within the joint) and systemic levels. In this regard, targeting the immune system using both biomaterial-based or cellular approaches may generate new joint-specific treatment strategies that are well-controlled, safe, and efficacious. In this review, we focus on recent advances in immunoengineering that leverage biomaterials and/or genetically engineered cells for therapeutic applications in joint diseases. The application of such approaches, especially synergistic strategies that target multiple immunoregulatory pathways, has the potential to revolutionize our understanding, treatment, and prevention of joint diseases.
{"title":"Immunoengineering the Next Generation of Arthritis Therapies","authors":"M. Klimak, R. Nims, Lara Pferdehirt, K. Collins, N. Harasymowicz, S. Oswald, L. Setton, F. Guilak","doi":"10.2139/ssrn.3751569","DOIUrl":"https://doi.org/10.2139/ssrn.3751569","url":null,"abstract":"Immunoengineering continues to revolutionize healthcare, generating new approaches for treating previously intractable diseases, particularly in regard to cancer immunotherapy. In joint diseases, such as osteoarthritis (OA) and rheumatoid arthritis (RA), biomaterials and anti-cytokine treatments have previously been at that forefront of therapeutic innovation. However, while many of the existing anti-cytokine treatments are successful for a subset of patients, these treatments can also pose severe risks, adverse events and off-target effects due to continuous delivery at high dosages or a lack of disease-specific targets. The inadequacy of these current treatments has motivated the development of new immunoengineering strategies that offer a safer and more efficacious alternative therapies through the precise and controlled targeting of specific upstream immune responses, including direct and mechanistically-driven immunoengineering approaches. Advances in the understanding of the immunomodulatory pathways involved in musculoskeletal disease, in combination with the growing emphasis on personalized medicine, stress the need for carefully considering the delivery strategies and therapeutic targets when designing therapeutics to better treat RA and OA. Here, we focus on recent advances in biomaterial and cell-based immunomodulation, in combination with genetic engineering, for therapeutic applications in joint diseases. The application of immunoengineering principles to the study of joint disease will not only help to elucidate the mechanisms of disease pathogenesis but will also generate novel disease-specific therapeutics by harnessing cellular and biomaterial responses. STATEMENT OF SIGNIFICANCE: It is now apparent that joint diseases such as osteoarthritis and rheumatoid arthritis involve the immune system at both local (i.e., within the joint) and systemic levels. In this regard, targeting the immune system using both biomaterial-based or cellular approaches may generate new joint-specific treatment strategies that are well-controlled, safe, and efficacious. In this review, we focus on recent advances in immunoengineering that leverage biomaterials and/or genetically engineered cells for therapeutic applications in joint diseases. The application of such approaches, especially synergistic strategies that target multiple immunoregulatory pathways, has the potential to revolutionize our understanding, treatment, and prevention of joint diseases.","PeriodicalId":106645,"journal":{"name":"MatSciRN: Tissue Engineering (Topic)","volume":"11 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2020-12-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"123689160","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The diagnosis of multiple sclerosis disease (MSD) is crucial because it is a neurological disease leading to communication failure between brain tissues and other parts of the body. Effective classification and segmentation of brain tissues are necessary for early detection of multiple sclerosis disease. In this proposed work, an ensemble learning-based classification technique is proposed to identify multiple sclerosis diseases from a database of healthy and unhealthy brain magnetic resonance (MR) images. Feature extraction from brain MR images is performed using an eighteen different Gray Level Co-occurrence Matrix (GLCoM) based features. Then, decision tree-based ensemble learning is accomplished on these features using three different boosting techniques for classification of healthy brain MR image from a weak brain MR image. Performance metrics like sensitivity ( PR T ), specificity ( NRT ), accuracy, precision (PPV), and F-score are utilized for MSD identification. It has been verified that the ensemble learning technique yielded higher accuracy of 94.91% from other states of the art techniques on the e-health dataset.
{"title":"Multiple Sclerosis Identification Based on Ensemble Machine Learning Technique","authors":"Shikha Jain, N. Rajpal, Jyotsna Yadav","doi":"10.2139/ssrn.3734806","DOIUrl":"https://doi.org/10.2139/ssrn.3734806","url":null,"abstract":"The diagnosis of multiple sclerosis disease (MSD) is crucial because it is a neurological disease leading to communication failure between brain tissues and other parts of the body. Effective classification and segmentation of brain tissues are necessary for early detection of multiple sclerosis disease. In this proposed work, an ensemble learning-based classification technique is proposed to identify multiple sclerosis diseases from a database of healthy and unhealthy brain magnetic resonance (MR) images. Feature extraction from brain MR images is performed using an eighteen different Gray Level Co-occurrence Matrix (GLCoM) based features. Then, decision tree-based ensemble learning is accomplished on these features using three different boosting techniques for classification of healthy brain MR image from a weak brain MR image. Performance metrics like sensitivity ( PR T ), specificity ( NRT ), accuracy, precision (PPV), and F-score are utilized for MSD identification. It has been verified that the ensemble learning technique yielded higher accuracy of 94.91% from other states of the art techniques on the e-health dataset.","PeriodicalId":106645,"journal":{"name":"MatSciRN: Tissue Engineering (Topic)","volume":"122 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2020-11-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"114317881","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ze Gong, Katrina M. Wisdom, Eóin McEvoy, Julie Chang, K. Adebowale, Christopher C. Price, Ovijit Chaudhuri, V. Shenoy
Most extracellular matrices (ECMs) are known to be dissipative, exhibiting viscoelastic and often plastic behaviors. However, the influence of dissipation, in particular mechanical plasticity in 3D confining microenvironments, on cell motility is not clear. In this study, we develop a chemo-mechanical model for dynamics of invadopodia, the protrusive structures that cancer cells use to facilitate invasion, by considering myosin recruitment, actin polymerization, matrix deformation, and mechano-sensitive signaling pathways. We demonstrate that matrix dissipation facilitates invadopodia growth by softening ECMs over repeated cycles, during which plastic deformation accumulates via cyclic ratcheting. Our model reveals that distinct protrusion patterns, oscillatory or monotonic, emerge from the interplay of timescales for polymerization-associated extension and myosin recruitment dynamics. Our model predicts the changes in invadopodia dynamics upon inhibition of myosin, adhesions, and the Rho-Rho-associated kinase (ROCK) pathway. Altogether, our work highlights the role of matrix plasticity in invadopodia dynamics and can help design dissipative biomaterials to modulate cancer cell motility.
{"title":"Recursive Feedback between Matrix Dissipation and Chemo-Mechanical Signaling Drives Oscillatory Growth of Cancer Cell Invadopodia","authors":"Ze Gong, Katrina M. Wisdom, Eóin McEvoy, Julie Chang, K. Adebowale, Christopher C. Price, Ovijit Chaudhuri, V. Shenoy","doi":"10.2139/ssrn.3692663","DOIUrl":"https://doi.org/10.2139/ssrn.3692663","url":null,"abstract":"Most extracellular matrices (ECMs) are known to be dissipative, exhibiting viscoelastic and often plastic behaviors. However, the influence of dissipation, in particular mechanical plasticity in 3D confining microenvironments, on cell motility is not clear. In this study, we develop a chemo-mechanical model for dynamics of invadopodia, the protrusive structures that cancer cells use to facilitate invasion, by considering myosin recruitment, actin polymerization, matrix deformation, and mechano-sensitive signaling pathways. We demonstrate that matrix dissipation facilitates invadopodia growth by softening ECMs over repeated cycles, during which plastic deformation accumulates via cyclic ratcheting. Our model reveals that distinct protrusion patterns, oscillatory or monotonic, emerge from the interplay of timescales for polymerization-associated extension and myosin recruitment dynamics. Our model predicts the changes in invadopodia dynamics upon inhibition of myosin, adhesions, and the Rho-Rho-associated kinase (ROCK) pathway. Altogether, our work highlights the role of matrix plasticity in invadopodia dynamics and can help design dissipative biomaterials to modulate cancer cell motility.","PeriodicalId":106645,"journal":{"name":"MatSciRN: Tissue Engineering (Topic)","volume":"620 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2020-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"132720193","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Matrix dynamics influence how individual cells develop into complex multicellular tissues. Here, we develop hydrogels with identical polymer components but different crosslinking capacities to enable the investigation of mechanisms underlying vascular morphogenesis. We show that dynamic (D) hydrogels increase the contractility of human endothelial colony-forming cells (hECFCs), promote the clustering of integrin β1, and promote the recruitment of vinculin, leading to the activation of focal adhesion kinase (FAK) and metalloproteinase expression. This leads to the robust assembly of vasculature and the deposition of new basement membrane. We also show that non-dynamic (N) hydrogels do not promote FAK signaling and that stiff D- and N-hydrogels are constrained for vascular morphogenesis. Furthermore, D-hydrogels promote hECFC microvessel formation and angiogenesis in vivo. Our results indicate that cell contractility mediates integrin signaling via inside-out signaling and emphasizes the importance of matrix dynamics in vascular tissue formation, thus informing future studies of vascularization and tissue engineering applications.
{"title":"Hydrogel Network Dynamics Regulate Vascular Morphogenesis","authors":"Zhao Wei, Rahel Schnellmann, S. Gerecht","doi":"10.2139/ssrn.3553544","DOIUrl":"https://doi.org/10.2139/ssrn.3553544","url":null,"abstract":"Matrix dynamics influence how individual cells develop into complex multicellular tissues. Here, we develop hydrogels with identical polymer components but different crosslinking capacities to enable the investigation of mechanisms underlying vascular morphogenesis. We show that dynamic (D) hydrogels increase the contractility of human endothelial colony-forming cells (hECFCs), promote the clustering of integrin β1, and promote the recruitment of vinculin, leading to the activation of focal adhesion kinase (FAK) and metalloproteinase expression. This leads to the robust assembly of vasculature and the deposition of new basement membrane. We also show that non-dynamic (N) hydrogels do not promote FAK signaling and that stiff D- and N-hydrogels are constrained for vascular morphogenesis. Furthermore, D-hydrogels promote hECFC microvessel formation and angiogenesis in vivo. Our results indicate that cell contractility mediates integrin signaling via inside-out signaling and emphasizes the importance of matrix dynamics in vascular tissue formation, thus informing future studies of vascularization and tissue engineering applications.","PeriodicalId":106645,"journal":{"name":"MatSciRN: Tissue Engineering (Topic)","volume":"4 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2020-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"127385153","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Weiwei Huang, C. Shu, Liangqun Hua, Yilin Zhao, Hanghang Xie, Jia-long Qi, Fulan Gao, Ruiyu Gao, Yongjun Chen, Qishu Zhang, Weiran Li, Mingcui Yuan, Chao Ye, Yanbing Ma
Using monoclonal antibodies to block tumor angiogenesis has yielded effective antitumor effects. However, this treatment method has long cycles and is very expensive; therefore, its long-term and extensive application is limited. In this study, we developed a nanovaccine using bacterial biomembranes as carriers for antitumor therapy. The whole basic fibroblast growth factor (BFGF) molecule (154 amino acids (aa)) was loaded onto bacterial outer membrane vesicles (OMVs) using gene recombination technology. The strong adjuvant effect of OMVs was used to induce the host to produce anti-BFGF autoantibodies. We proved that persistent anti-BFGF autoantibodies can be induced in mice after only 3 immunizations to antagonize BFGF functions. The effects included multiple tumor suppression functions, including inhibition of tumor angiogenesis, induction of tumor cell apoptosis, reversal of tumor immune barriers, and promotion of tumor-specific cytotoxic T lymphocytes (CTLs), eventually causing tumor regression. We confirmed that bacterial biomembranes can be used as a vaccine delivery system to induce the production of antibodies against autoantigens, which may be used for tumor therapy. This study expands the application fields of bacterial biomembrane systems and provides insight for tumor immunotherapy other than monoclonal antibody technology.
{"title":"Modified Bacterial Outer Membrane Vesicles Induce Autoantibodies for Tumor Therapy","authors":"Weiwei Huang, C. Shu, Liangqun Hua, Yilin Zhao, Hanghang Xie, Jia-long Qi, Fulan Gao, Ruiyu Gao, Yongjun Chen, Qishu Zhang, Weiran Li, Mingcui Yuan, Chao Ye, Yanbing Ma","doi":"10.2139/ssrn.3518841","DOIUrl":"https://doi.org/10.2139/ssrn.3518841","url":null,"abstract":"Using monoclonal antibodies to block tumor angiogenesis has yielded effective antitumor effects. However, this treatment method has long cycles and is very expensive; therefore, its long-term and extensive application is limited. In this study, we developed a nanovaccine using bacterial biomembranes as carriers for antitumor therapy. The whole basic fibroblast growth factor (BFGF) molecule (154 amino acids (aa)) was loaded onto bacterial outer membrane vesicles (OMVs) using gene recombination technology. The strong adjuvant effect of OMVs was used to induce the host to produce anti-BFGF autoantibodies. We proved that persistent anti-BFGF autoantibodies can be induced in mice after only 3 immunizations to antagonize BFGF functions. The effects included multiple tumor suppression functions, including inhibition of tumor angiogenesis, induction of tumor cell apoptosis, reversal of tumor immune barriers, and promotion of tumor-specific cytotoxic T lymphocytes (CTLs), eventually causing tumor regression. We confirmed that bacterial biomembranes can be used as a vaccine delivery system to induce the production of antibodies against autoantigens, which may be used for tumor therapy. This study expands the application fields of bacterial biomembrane systems and provides insight for tumor immunotherapy other than monoclonal antibody technology.","PeriodicalId":106645,"journal":{"name":"MatSciRN: Tissue Engineering (Topic)","volume":"23 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2020-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"114311206","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Nandini A. Pattanashetti, Divya D. Achari, Anand I. Torvi, Radha V. Doddamani, M. Kariduraganavar
Development of polymeric nanofibrous scaffolds has achieved wide applications in the field of tissue engineering by electrospinning technique. In order to further improvise this technique for bone tissue regeneration, multilayered nanofibrous scaffolds were fabricated in the present study. The hybrid composite scaffolds comprised of hydrophobic polycaprolactone (PCL) and hydrophilic blend of poly(vinyl alcohol):sodium alginate (P:S) nanofibres in a layer-by-layer pattern in order to incorporate the selective properties of different polymers in a single scaffold. The stability of these multilayered scaffolds was further improved by crosslinking with 5% CaCl2. The properties of neat PCL scaffold was compared with the multilayered PCL/P:S, PCL/P:S/PCL and crosslinked PCL/P:S and PCL/P:S/PCL scaffolds. The physicochemical properties of the developed scaffolds were systematically studied. Morphological and structural characteristics measured by scanning electron microscopy revealed uniform nanofibres thereby forming a porous mesh like structure suitable for cell growth. The effect of multilayer deposition of nanofibres was observed in terms of increase in rate of water absorption and a slower rate degradation for PCL/P:S and PCL/P:S/PCL scaffolds respectively. Improved mechanical properties were also observed for triple layered PCL/P:S/PCL hybrid scaffold as obtained by mechanical testing. MG-63 bone osteosarcoma cells were employed to determine the biocompatibility of the multilayered scaffolds, wherein none of the scaffolds possessed any cytotoxic effect, and cell proliferation of >90% was clearly observed for multilayered scaffolds. Based on these results the developed multilayered scaffolds were proved to be suitable for bone tissue regeneration.
{"title":"Multilayer Electrospinning of PCL and PVA: NaAlg Nanofibres for Bone Tissue Engineering","authors":"Nandini A. Pattanashetti, Divya D. Achari, Anand I. Torvi, Radha V. Doddamani, M. Kariduraganavar","doi":"10.2139/ssrn.3484669","DOIUrl":"https://doi.org/10.2139/ssrn.3484669","url":null,"abstract":"Development of polymeric nanofibrous scaffolds has achieved wide applications in the field of tissue engineering by electrospinning technique. In order to further improvise this technique for bone tissue regeneration, multilayered nanofibrous scaffolds were fabricated in the present study. The hybrid composite scaffolds comprised of hydrophobic polycaprolactone (PCL) and hydrophilic blend of poly(vinyl alcohol):sodium alginate (P:S) nanofibres in a layer-by-layer pattern in order to incorporate the selective properties of different polymers in a single scaffold. The stability of these multilayered scaffolds was further improved by crosslinking with 5% CaCl<sub>2</sub>. The properties of neat PCL scaffold was compared with the multilayered PCL/P:S, PCL/P:S/PCL and crosslinked PCL/P:S and PCL/P:S/PCL scaffolds. The physicochemical properties of the developed scaffolds were systematically studied. Morphological and structural characteristics measured by scanning electron microscopy revealed uniform nanofibres thereby forming a porous mesh like structure suitable for cell growth. The effect of multilayer deposition of nanofibres was observed in terms of increase in rate of water absorption and a slower rate degradation for PCL/P:S and PCL/P:S/PCL scaffolds respectively. Improved mechanical properties were also observed for triple layered PCL/P:S/PCL hybrid scaffold as obtained by mechanical testing. MG-63 bone osteosarcoma cells were employed to determine the biocompatibility of the multilayered scaffolds, wherein none of the scaffolds possessed any cytotoxic effect, and cell proliferation of >90% was clearly observed for multilayered scaffolds. Based on these results the developed multilayered scaffolds were proved to be suitable for bone tissue regeneration.","PeriodicalId":106645,"journal":{"name":"MatSciRN: Tissue Engineering (Topic)","volume":"36 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2019-11-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"132484039","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sayali Dharmadhikari, Lumei Liu, K. Shontz, Matthew G. Wiet, A. White, A. Goins, Himani Akula, Jed Johnson, S. Reynolds, C. Breuer, Tendy Chiang
The ideal construct for tracheal replacement remains elusive in the management of long segment airway defects. Tissue engineered tracheal grafts (TETG) have been limited by the development of graft stenosis or collapse, infection, or lack of an epithelial lining. We applied a mouse model of orthotopic airway surgery to assess the impact of three critical barriers encountered in clinical applications: the scaffold, the extent of intervention, and the impact of cell seeding and characterized their impact on graft performance. First, synthetic tracheal scaffolds electrospun from polyethylene terephthalate / polyurethane (PET/PU) were orthotopically implanted in anterior tracheal defects of C57BL/6 mice. Scaffolds demonstrated complete coverage with ciliated respiratory epithelium by 2 weeks. Epithelial migration was accompanied by macrophage infiltration which persisted at long term (>6 weeks) time points. We then assessed the impact of segmental tracheal implantation using syngeneic trachea as a surrogate for the ideal tracheal replacement. Graft recovery involved local upregulation of epithelial progenitor populations and there was no evidence of graft stenosis or necrosis. Implantation of electrospun synthetic tracheal scaffold for segmental replacement resulted in respiratory distress and required euthanasia at an early time point. There was limited epithelial coverage of the scaffold with and without seeded bone marrow-derived mononuclear cells (BM-MNCs). We conclude that synthetic scaffolds support re-epithelialization in orthotopic patch implantation, syngeneic graft integration occurs with focal repair mechanisms, however epithelialization in segmental synthetic scaffolds is limited and is not influenced by cell seeding.
{"title":"Deconstructing Tissue Engineered Trachea: Assessing the Role of Synthetic Scaffolds, Segmental Replacement and Cell Seeding on Graft Performance","authors":"Sayali Dharmadhikari, Lumei Liu, K. Shontz, Matthew G. Wiet, A. White, A. Goins, Himani Akula, Jed Johnson, S. Reynolds, C. Breuer, Tendy Chiang","doi":"10.2139/ssrn.3428072","DOIUrl":"https://doi.org/10.2139/ssrn.3428072","url":null,"abstract":"The ideal construct for tracheal replacement remains elusive in the management of long segment airway defects. Tissue engineered tracheal grafts (TETG) have been limited by the development of graft stenosis or collapse, infection, or lack of an epithelial lining. We applied a mouse model of orthotopic airway surgery to assess the impact of three critical barriers encountered in clinical applications: the scaffold, the extent of intervention, and the impact of cell seeding and characterized their impact on graft performance. First, synthetic tracheal scaffolds electrospun from polyethylene terephthalate / polyurethane (PET/PU) were orthotopically implanted in anterior tracheal defects of C57BL/6 mice. Scaffolds demonstrated complete coverage with ciliated respiratory epithelium by 2 weeks. Epithelial migration was accompanied by macrophage infiltration which persisted at long term (>6 weeks) time points. We then assessed the impact of segmental tracheal implantation using syngeneic trachea as a surrogate for the ideal tracheal replacement. Graft recovery involved local upregulation of epithelial progenitor populations and there was no evidence of graft stenosis or necrosis. Implantation of electrospun synthetic tracheal scaffold for segmental replacement resulted in respiratory distress and required euthanasia at an early time point. There was limited epithelial coverage of the scaffold with and without seeded bone marrow-derived mononuclear cells (BM-MNCs). We conclude that synthetic scaffolds support re-epithelialization in orthotopic patch implantation, syngeneic graft integration occurs with focal repair mechanisms, however epithelialization in segmental synthetic scaffolds is limited and is not influenced by cell seeding.","PeriodicalId":106645,"journal":{"name":"MatSciRN: Tissue Engineering (Topic)","volume":"84 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2019-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"133370652","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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