Rachel R. Besser, Annie C. Bowles, Ahmad Alassaf, Daniel Carbonero, Isabella Claure, Ellery Jones, Joseph Reda, Laura Wubker, W. Batchelor, N. Ziebarth, R. Silvera, Aisha Khan, Renata Maciel, Mario Saporta, A. Agarwal
We report a water-soluble and non-toxic method to incorporate additional extracellular matrix proteins into gelatin hydrogels, while obviating the use of chemical crosslinkers such as glutaraldehyde. Gelatin hydrogels were fabricated using a range of gelatin concentrations (4%-10%) that corresponded to elastic moduli of approximately 1 kPa-25 kPa, respectively, a substrate stiffness relevant for multiple cell types. Microbial transglutaminase was then used to enzymatically crosslink a layer of laminin on top of gelatin hydrogels, resulting in 2-component gelatin-laminin hydrogels. Human induced pluripotent stem cell derived spinal spheroids readily adhered and rapidly extended axons on GEL-LN hydrogels. Axons displayed a more mature morphology and superior electrophysiological properties on GEL-LN hydrogels compared to the controls. Schwann cells on GEL-LN hydrogels adhered and proliferated normally, displayed a healthy morphology, and maintained the expression of Schwann cell specific markers. Lastly, skeletal muscle cells on GEL-LN hydrogels achieved long-term culture for up to 28 days without delamination, while expressing higher levels of terminal genes including myosin heavy chain, MyoD, MuSK, and M-cadherin suggesting enhanced maturation potential and myotube formation compared to the controls. Future studies will employ the superior culture outcomes of this hybrid substrate for engineering functional neuromuscular junctions and related organ on a chip applications.
{"title":"Enzymatically Crosslinked Gelatin-Laminin Hydrogels for Applications in Neuromuscular Tissue Engineering","authors":"Rachel R. Besser, Annie C. Bowles, Ahmad Alassaf, Daniel Carbonero, Isabella Claure, Ellery Jones, Joseph Reda, Laura Wubker, W. Batchelor, N. Ziebarth, R. Silvera, Aisha Khan, Renata Maciel, Mario Saporta, A. Agarwal","doi":"10.2139/ssrn.3431850","DOIUrl":"https://doi.org/10.2139/ssrn.3431850","url":null,"abstract":"We report a water-soluble and non-toxic method to incorporate additional extracellular matrix proteins into gelatin hydrogels, while obviating the use of chemical crosslinkers such as glutaraldehyde. Gelatin hydrogels were fabricated using a range of gelatin concentrations (4%-10%) that corresponded to elastic moduli of approximately 1 kPa-25 kPa, respectively, a substrate stiffness relevant for multiple cell types. Microbial transglutaminase was then used to enzymatically crosslink a layer of laminin on top of gelatin hydrogels, resulting in 2-component gelatin-laminin hydrogels. Human induced pluripotent stem cell derived spinal spheroids readily adhered and rapidly extended axons on GEL-LN hydrogels. Axons displayed a more mature morphology and superior electrophysiological properties on GEL-LN hydrogels compared to the controls. Schwann cells on GEL-LN hydrogels adhered and proliferated normally, displayed a healthy morphology, and maintained the expression of Schwann cell specific markers. Lastly, skeletal muscle cells on GEL-LN hydrogels achieved long-term culture for up to 28 days without delamination, while expressing higher levels of terminal genes including myosin heavy chain, MyoD, MuSK, and M-cadherin suggesting enhanced maturation potential and myotube formation compared to the controls. Future studies will employ the superior culture outcomes of this hybrid substrate for engineering functional neuromuscular junctions and related organ on a chip applications.","PeriodicalId":106645,"journal":{"name":"MatSciRN: Tissue Engineering (Topic)","volume":"78 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2019-08-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"134170442","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}
Shuang-Fei Li, C. Song, Shengbing Yang, Weijun Yu, Weiqi Zhang, Guohua Zhang, Zhenhao Xi, Eryi Lu
Bone tissue engineering has substantial potential for the treatment of massive bone defects; however, efficient vascularization coupled with bone regeneration still remains a challenge in this field. In the current study,supercritical carbon dioxide (scCO2) foaming technique was adopted to fabricate mesoporous bioactive glasses (MBGs) particle-poly (lactic-co-glycolic acid) (PLGA) composite scaffolds with appropriate mechanical and degradation properties as well as in vitro bioactivity. The MBG-PLGA scaffolds incorporating the bioactive lipid FTY720 (designated as FTY/MBG-PLGA) exhibited simultaneously sustained release of thebioactive lipid and ions. In addition to providing a favorable microenvironment for cellular adhesion and proliferation, FTY/MBG-PLGA scaffolds significantly facilitated the in vitro osteogenic differentiation of rBMSCs and also markedly stimulated the up regulation of Hif-1α expression via the activation of the Erk1/2 pathway, which mediated the osteogenic and pro-angiogenic effects on rBMSCs. Furthermore, FTY/MBG-PLGA extracts induced superior in vitro angiogenic performance of HUVECs. In vivo evaluation of critical-sized rat calvarial bone defects indicated that FTY/MBG-PLGA scaffolds potently promoted vascularized bone regeneration. Notably, the significantly enhanced formation of type Hvessels (CD31hiEmcnhineo-vessels) was observed in newly formed bone tissue in FTY/MBG-PLGA group, strongly suggesting that FTY720 and therapeutic ions released from the scaffolds synergistically induced moretype H vessel formation, which indicated the coupling of angiogenesis and osteogenesis to achieve efficiently vascularized bone regeneration. Overall,the results indicated that the foamed porous MBG-PLGA scaffolds incorporating bioactive lipids achieved desirable vascularization-coupled bone formation and could be a promising strategy for bone regenerative medicine.
{"title":"scCO 2 Foamed Composite Scaffolds Incorporating Bioactive Lipids Promote Vascularized Bone Regeneration Via Hif-1α Upregulation and Enhanced Type H Vessel Formation","authors":"Shuang-Fei Li, C. Song, Shengbing Yang, Weijun Yu, Weiqi Zhang, Guohua Zhang, Zhenhao Xi, Eryi Lu","doi":"10.2139/ssrn.3387694","DOIUrl":"https://doi.org/10.2139/ssrn.3387694","url":null,"abstract":"Bone tissue engineering has substantial potential for the treatment of massive bone defects; however, efficient vascularization coupled with bone regeneration still remains a challenge in this field. In the current study,supercritical carbon dioxide (scCO2) foaming technique was adopted to fabricate mesoporous bioactive glasses (MBGs) particle-poly (lactic-co-glycolic acid) (PLGA) composite scaffolds with appropriate mechanical and degradation properties as well as in vitro bioactivity. The MBG-PLGA scaffolds incorporating the bioactive lipid FTY720 (designated as FTY/MBG-PLGA) exhibited simultaneously sustained release of thebioactive lipid and ions. In addition to providing a favorable microenvironment for cellular adhesion and proliferation, FTY/MBG-PLGA scaffolds significantly facilitated the in vitro osteogenic differentiation of rBMSCs and also markedly stimulated the up regulation of Hif-1α expression via the activation of the Erk1/2 pathway, which mediated the osteogenic and pro-angiogenic effects on rBMSCs. Furthermore, FTY/MBG-PLGA extracts induced superior in vitro angiogenic performance of HUVECs. In vivo evaluation of critical-sized rat calvarial bone defects indicated that FTY/MBG-PLGA scaffolds potently promoted vascularized bone regeneration. Notably, the significantly enhanced formation of type Hvessels (CD31hiEmcnhineo-vessels) was observed in newly formed bone tissue in FTY/MBG-PLGA group, strongly suggesting that FTY720 and therapeutic ions released from the scaffolds synergistically induced moretype H vessel formation, which indicated the coupling of angiogenesis and osteogenesis to achieve efficiently vascularized bone regeneration. Overall,the results indicated that the foamed porous MBG-PLGA scaffolds incorporating bioactive lipids achieved desirable vascularization-coupled bone formation and could be a promising strategy for bone regenerative medicine.","PeriodicalId":106645,"journal":{"name":"MatSciRN: Tissue Engineering (Topic)","volume":"16 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2019-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"125960645","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}
Purpose: Encapsulated microbubbles (MBs) have been reported asnew theranostic carriers for simultaneous imaging and ultrasound (US)-triggered therapy. Here, we designed a dual-modality US/NIRF contrastagent and extended its applications from image contrast enhancement tocombined diagnosis and therapy with US-directed and site-specifictargeting.
Methods: Gold nanorods (AuNRs) resonant at 880 nm together with theNIR797 dye were first encapsulated in lipid-shelled MBs to constructfluorescent gold microbubbles (NIR797/AuMBs) via thin film hydration andmechanical shaking in the presence of sulfur hexafluoride (SF6) gas.Then, polyethylenimine (PEI)-DNA complexes were electrostaticallyconjugated onto the surface of the NIR797/AuMBs, forming theranosticencapsulated MBs (PEI-DNA/NIR797/AuMBs). The potential of the PEIDNA/NIR797/AuMBs for use as a dual-modality contrast enhancement agentwas evaluated in vitro and in vivo. The antitumor effect of US/NIR laserirradiation mediating double-fusion suicide gene and photothermal therapywas also investigated using Bel-7402 cells and xenografts.
Results: The developed theranostic AuMB complexes could not only provideexcellent US and NIRF imaging to detect tumors but also serve as anefficient US-triggered carrier for gene delivery and photothermalablation of tumors in xenografted nude mice. And US + laser exposuregroup showed a much higher rate of cell inhibition, apoptosis andnecrosis as well as a higher Bel-7402 xenograft inhibition rate than thesingle gene therapy or single exposure (US or laser) group.
Conclusions: PEI-DNA/NIR797/AuMBs would be of great value for providingmore comprehensive diagnostic information and to guide more accurate andeffective synergistic cancer therapy.
{"title":"Multifunctional Theranostic Contrast Agent for US/NIRF-Based Tumor Diagnosis and US-Triggered Combined Photothermal and Gene Therapy","authors":"Ling Wang, Hangqing Lu, Q. Gao, Chenyan Yuan, Fengan Ding, Jia Li, Dongsheng Zhang, Xilong Ou","doi":"10.2139/ssrn.3385801","DOIUrl":"https://doi.org/10.2139/ssrn.3385801","url":null,"abstract":"Purpose: Encapsulated microbubbles (MBs) have been reported asnew theranostic carriers for simultaneous imaging and ultrasound (US)-triggered therapy. Here, we designed a dual-modality US/NIRF contrastagent and extended its applications from image contrast enhancement tocombined diagnosis and therapy with US-directed and site-specifictargeting.<br><br>Methods: Gold nanorods (AuNRs) resonant at 880 nm together with theNIR797 dye were first encapsulated in lipid-shelled MBs to constructfluorescent gold microbubbles (NIR797/AuMBs) via thin film hydration andmechanical shaking in the presence of sulfur hexafluoride (SF6) gas.Then, polyethylenimine (PEI)-DNA complexes were electrostaticallyconjugated onto the surface of the NIR797/AuMBs, forming theranosticencapsulated MBs (PEI-DNA/NIR797/AuMBs). The potential of the PEIDNA/NIR797/AuMBs for use as a dual-modality contrast enhancement agentwas evaluated in vitro and in vivo. The antitumor effect of US/NIR laserirradiation mediating double-fusion suicide gene and photothermal therapywas also investigated using Bel-7402 cells and xenografts.<br><br>Results: The developed theranostic AuMB complexes could not only provideexcellent US and NIRF imaging to detect tumors but also serve as anefficient US-triggered carrier for gene delivery and photothermalablation of tumors in xenografted nude mice. And US + laser exposuregroup showed a much higher rate of cell inhibition, apoptosis andnecrosis as well as a higher Bel-7402 xenograft inhibition rate than thesingle gene therapy or single exposure (US or laser) group.<br><br>Conclusions: PEI-DNA/NIR797/AuMBs would be of great value for providingmore comprehensive diagnostic information and to guide more accurate andeffective synergistic cancer therapy.","PeriodicalId":106645,"journal":{"name":"MatSciRN: Tissue Engineering (Topic)","volume":"1 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2019-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"123805949","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}
Even with tremendous advancement in medical technology tumor segmentation remain most tedious and complex work for doctors. Magnetic Resonance Imaging (MRI) is the most commonly used technique by the radiologist for inspection of internal human body parts without any dissection, but manual inspection consumes time and precious work hours. A possible solution for early detection of diseases such as a tumor, cancer can be computer-aided image analysis. Precise computerized classification of brain tumor form the MRI image is very important as it will lead to early detection, reduction in work hour and mistakes, propagation of automation in tumor removal and will also help to decide the course of treatment. Considering the difficulties, this work is aimed to highlight the techniques proposed in contemporary literature by summarizing the novel facts of research.
{"title":"A Survey on Brain Tumor Detection and Segmentation from Magnetic Resonance Image","authors":"Priyanka Arya, A. K. Malviya","doi":"10.2139/ssrn.3350289","DOIUrl":"https://doi.org/10.2139/ssrn.3350289","url":null,"abstract":"Even with tremendous advancement in medical technology tumor segmentation remain most tedious and complex work for doctors. Magnetic Resonance Imaging (MRI) is the most commonly used technique by the radiologist for inspection of internal human body parts without any dissection, but manual inspection consumes time and precious work hours. A possible solution for early detection of diseases such as a tumor, cancer can be computer-aided image analysis. Precise computerized classification of brain tumor form the MRI image is very important as it will lead to early detection, reduction in work hour and mistakes, propagation of automation in tumor removal and will also help to decide the course of treatment. Considering the difficulties, this work is aimed to highlight the techniques proposed in contemporary literature by summarizing the novel facts of research.","PeriodicalId":106645,"journal":{"name":"MatSciRN: Tissue Engineering (Topic)","volume":"30 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2019-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"129412408","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}
Background: This study attempts to discuss the efficacy and safety of transthoracic echocardiogram (TTE) -guided percutaneous atrial septal defect (ASD) closure for infants under 10 Kg. Methods: This is a retrospective study involving 37 infants who received TTE-guided percutaneous ASD closure at our hospital from September 2014 to September 2020. There were 22 girls and 15 boys. The age and weight were respectively between 3-18 m (9.05±3.67, 95% CI 7.83–10.28 m) and 3.50-9.80 Kg (7.58±1.43, 95% CI 7.11–8.06 Kg). Symptoms were recurrent pneumonia in 29 cases, evidence of dysplasia in 22 cases, and pulmonary arterial hypertension in 12 cases. Right atrial and ventricular dilation were present in all the 37 cases of varying degrees. Intubation was applied to all cases under general anesthesia, followed by TTE-guided percutaneous ASD closure via the femoral vein approach. Results: The overall success rate was 94.59% (35/37). Postoperative TTE showed that the size of the right atrium and right ventricle was significantly decreased than preoperative baseline data (p<0.05). There were no deaths or any serious complications during the perioperative period. At follow-up (6-72 m), there was no death and other complication. Nineteen cases with development dysplasia (19/22, 86.36%) had recovered height and weight equivalent to peer infants, 21 cases with respiratory system symptoms (21/29, 72.41%) got improved symptoms and all 12 cases with pulmonary arterial hypertension (100%) recovered to normal. Conclusions: This study applied TTE-guided percutaneous ASD closure for infants under 10 Kg with appreciating efficacy and a safety profile.
{"title":"Percutaneous Closure of Atrial Septal Defect Under the Guidance of Complete Transthoracic Ultrasound in Infants Under 10kg","authors":"Zhi Dou, Zhao Yu, Yuqing Gong, Q. Xie, Yonghuo Ye, Guoxing Weng","doi":"10.2139/ssrn.3937112","DOIUrl":"https://doi.org/10.2139/ssrn.3937112","url":null,"abstract":"Background: This study attempts to discuss the efficacy and safety of transthoracic echocardiogram (TTE) -guided percutaneous atrial septal defect (ASD) closure for infants under 10 Kg. Methods: This is a retrospective study involving 37 infants who received TTE-guided percutaneous ASD closure at our hospital from September 2014 to September 2020. There were 22 girls and 15 boys. The age and weight were respectively between 3-18 m (9.05±3.67, 95% CI 7.83–10.28 m) and 3.50-9.80 Kg (7.58±1.43, 95% CI 7.11–8.06 Kg). Symptoms were recurrent pneumonia in 29 cases, evidence of dysplasia in 22 cases, and pulmonary arterial hypertension in 12 cases. Right atrial and ventricular dilation were present in all the 37 cases of varying degrees. Intubation was applied to all cases under general anesthesia, followed by TTE-guided percutaneous ASD closure via the femoral vein approach. Results: The overall success rate was 94.59% (35/37). Postoperative TTE showed that the size of the right atrium and right ventricle was significantly decreased than preoperative baseline data (p<0.05). There were no deaths or any serious complications during the perioperative period. At follow-up (6-72 m), there was no death and other complication. Nineteen cases with development dysplasia (19/22, 86.36%) had recovered height and weight equivalent to peer infants, 21 cases with respiratory system symptoms (21/29, 72.41%) got improved symptoms and all 12 cases with pulmonary arterial hypertension (100%) recovered to normal. Conclusions: This study applied TTE-guided percutaneous ASD closure for infants under 10 Kg with appreciating efficacy and a safety profile.","PeriodicalId":106645,"journal":{"name":"MatSciRN: Tissue Engineering (Topic)","volume":"31 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"132174887","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}
K. Fujimoto, A. Yamawaki-Ogata, Y. Narita, A. Usui, K. Uto, M. Ebara
For decades, researchers have investigated the ideal material for clinical use in the cardiovascular field. Several substitute materials are used clinically, but each has drawbacks. Recently we developed poly(e-caprolactone-co-D,L-lactide) (P(CL-DLLA)) polymers with optimized biodegradation and elasticity by adjusting the CL/DLLA composition, and used these polymers in right ventricular outflow tract (RVOT) replacement to evaluate long-term efficacy and outcomes. This P(CL-DLLA) material was processed into a circular patch and used to replace a surgical defect in the RVOT of adult rats. Control rats were implanted with expanded polytetrafluoroethylene (ePTFE). Histologic evaluation was performed at 8, 24, and 48 weeks post-surgery. All animals survived the surgery with no aneurysm formation or thrombus. In all periods, ePTFE demonstrated fibrous tissue. In contrast, at 8 weeks P(CL-DLLA) showed infiltration of macrophages and fibroblast-like cells into the remaining material. At 24 weeks, P(CL-DLLA) was absorbed completely, and muscle-like tissue was present with positive staining for α-sarcomeric actinin and cTnT. At 48 weeks, the cTnT-positive area had increased. The P(CL-DLLA) with optimized elasticity and biodegradation induced cardiac regeneration throughout the 48-week study period. Future application of this material as a cardiovascular scaffold seems promising.
{"title":"Long Term Efficacy and Fate of a Right Ventricular Outflow Tract Replacement Using a Novel Developed Material with Optimized Biodegradation and Elasticity","authors":"K. Fujimoto, A. Yamawaki-Ogata, Y. Narita, A. Usui, K. Uto, M. Ebara","doi":"10.2139/ssrn.3640711","DOIUrl":"https://doi.org/10.2139/ssrn.3640711","url":null,"abstract":"For decades, researchers have investigated the ideal material for clinical use in the cardiovascular field. Several substitute materials are used clinically, but each has drawbacks. Recently we developed poly(e-caprolactone-co-D,L-lactide) (P(CL-DLLA)) polymers with optimized biodegradation and elasticity by adjusting the CL/DLLA composition, and used these polymers in right ventricular outflow tract (RVOT) replacement to evaluate long-term efficacy and outcomes. This P(CL-DLLA) material was processed into a circular patch and used to replace a surgical defect in the RVOT of adult rats. Control rats were implanted with expanded polytetrafluoroethylene (ePTFE). Histologic evaluation was performed at 8, 24, and 48 weeks post-surgery. All animals survived the surgery with no aneurysm formation or thrombus. In all periods, ePTFE demonstrated fibrous tissue. In contrast, at 8 weeks P(CL-DLLA) showed infiltration of macrophages and fibroblast-like cells into the remaining material. At 24 weeks, P(CL-DLLA) was absorbed completely, and muscle-like tissue was present with positive staining for α-sarcomeric actinin and cTnT. At 48 weeks, the cTnT-positive area had increased. The P(CL-DLLA) with optimized elasticity and biodegradation induced cardiac regeneration throughout the 48-week study period. Future application of this material as a cardiovascular scaffold seems promising.","PeriodicalId":106645,"journal":{"name":"MatSciRN: Tissue Engineering (Topic)","volume":"9 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"132456525","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}
Sarah Klein, Ilka I. Rose, S. Mráz, Jens Rubner, S. Prünte, Suzana Djeljadini, A. Thiebes, C. Cornelissen, S. Jockenhoevel, J. Schneider, Matthias Wessling
Lacking hemocompatibility of extracorporeal membrane oxygenation led to a biohybrid lung concept, in which gas exchange membranes are seeded with endothelial cells to form long-term stable surfaces. However, extensive surface modification of large oxygenator membranes is required for cell adhesion. In this study, polymethylpentene (PMP), polydimethylsiloxane (PDMS) and polyethersulfone/polyvinylpyrrolidone (PES/PVP) membranes were functionalized with thin films of titanium dioxide (TiO 2 ) or titanium-niobium (Ti45Nb), and characterized by field emission scanning electron microscopy, ellipsometry, X-ray photoelectron spectroscopy and gas permeability measurements. Endothelialized membranes were cultured under static conditions to evaluate cell coverage. Cell retention was investigated in a bioreactor system under flow conditions with a wall shear stress of 0.5 Pa, and cells were stained for endothelial markers CD31 and von Willebrand factor (vWf). Film thicknesses of 3.3±0.2 nm and 5.5±0.3 nm were determined for TiO2 and Ti45Nb coatings, respectively. With the exception of a slight decrease for PDMS+Ti45Nb, gas permeabilities of coated PMP and PDMS membranes did not deteriorate significantly compared to uncoated membranes. The microporous structure and the high hydrophilicity of PES/PVP prevented permeability measurements and subsequent dynamic culture due to membrane wetting. However, mean cell density increased substantially for all coated membranes compared to uncoated membranes. Although endothelialized PDMS+Ti45Nb showed minor cell layer defects, all coated PMP and PDMS membranes demonstrated integral cell layers after dynamic culture that stained positive for CD31 and vWf. This study has shown the suitability of thin TiO2 and Ti45Nb films for flow-stable endothelialization of gas exchange membranes for application in a biohybrid lung.
{"title":"Endothelialized TiO 2- and Ti45Nb-Coated PMP, PDMS and PES/PVP Membranes for Biohybrid Lung Support","authors":"Sarah Klein, Ilka I. Rose, S. Mráz, Jens Rubner, S. Prünte, Suzana Djeljadini, A. Thiebes, C. Cornelissen, S. Jockenhoevel, J. Schneider, Matthias Wessling","doi":"10.2139/ssrn.3687910","DOIUrl":"https://doi.org/10.2139/ssrn.3687910","url":null,"abstract":"Lacking hemocompatibility of extracorporeal membrane oxygenation led to a biohybrid lung concept, in which gas exchange membranes are seeded with endothelial cells to form long-term stable surfaces. However, extensive surface modification of large oxygenator membranes is required for cell adhesion. In this study, polymethylpentene (PMP), polydimethylsiloxane (PDMS) and polyethersulfone/polyvinylpyrrolidone (PES/PVP) membranes were functionalized with thin films of titanium dioxide (TiO 2 ) or titanium-niobium (Ti45Nb), and characterized by field emission scanning electron microscopy, ellipsometry, X-ray photoelectron spectroscopy and gas permeability measurements. Endothelialized membranes were cultured under static conditions to evaluate cell coverage. Cell retention was investigated in a bioreactor system under flow conditions with a wall shear stress of 0.5 Pa, and cells were stained for endothelial markers CD31 and von Willebrand factor (vWf). Film thicknesses of 3.3±0.2 nm and 5.5±0.3 nm were determined for TiO<i>2</i> and Ti45Nb coatings, respectively. With the exception of a slight decrease for PDMS+Ti45Nb, gas permeabilities of coated PMP and PDMS membranes did not deteriorate significantly compared to uncoated membranes. The microporous structure and the high hydrophilicity of PES/PVP prevented permeability measurements and subsequent dynamic culture due to membrane wetting. However, mean cell density increased substantially for all coated membranes compared to uncoated membranes. Although endothelialized PDMS+Ti45Nb showed minor cell layer defects, all coated PMP and PDMS membranes demonstrated integral cell layers after dynamic culture that stained positive for CD31 and vWf. This study has shown the suitability of thin TiO<sub>2</sub> and Ti45Nb films for flow-stable endothelialization of gas exchange membranes for application in a biohybrid lung.","PeriodicalId":106645,"journal":{"name":"MatSciRN: Tissue Engineering (Topic)","volume":"1 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"129756495","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}
W. Zeng, Qiu-Ping Yu, Duan Wang, Yipeng Zeng, Hao Yang, Juan Li, Chengguang Zhou, Qing Jun Yang, Z. Deng, Zong-ke Zhou
Intra-articular injection of mesenchymal stem cells (MSCs) is an attractive treatment for osteoarthritis (OA) due to their ease of use in clinical settings, excellent safety profile, and remarkable spontaneous cartilage repair. However, significant challenges arise in relation to the control and induction of stem cell differentiation into the desired cartilaginous lineage. Kartogenin (KGN), a hydrophobic small molecule drug, may significantly promote chondrogenic differentiation of MSCs and induce cartilage regeneration in OA. However, low water solubility and poor bioavailability limits its biological application. In this report, we propose a new nano-drug delivery system based on multifunctional nanographene oxide (NGO) to efficiently load KGN molecules noncovalently via π–π stacking and hydrophobic interactions (PPG-KGN), which could quickly enter MSCs. Before intra-articular injection, MSCs are simply mixed and co-incubated with PPG-KGN to acquire KGN enhanced MSCs. This allows the efficient intracellular delivery of KGN, thereby promoting the chondrogenic differentiation potency of the MSCs. We investigated the effect of KGN-enhanced MSCs in the treatment of knee osteoarthritis. An in vitro study showed the PPG could be rapidly uptaken in the first 4 h after incubation, reaching saturation at 12 h, and accumulating in the lysosome and cytoplasm of MSCs. Thus, PPG-KGN could enhance the efficiency of the intracellular delivery of KGN, which showed remarkably high chondrogenic differentiation capacity of the MSCs. When applied to an OA model of cartilage injury in rats, MSCs were preconditioned with PPG-KGN before being injected into the knee joint. It demonstrated the PPG-KGN preconditioned MSCs contribute to protection from joint space narrowing, pathologic mineralization, osteoarthritis development, behavioral assessment of OA-induced pain, and tissue regeneration, as evidenced by radiographic, weight bearing, and histological analysis. Based on these findings, we propose the use of PPG for delivery of KGN to achieve enhanced MSC chondrogenic potential in osteoarthritis treatment.
{"title":"Intra-Articular Injection of Kartogenin Enhanced Bone Marrow-Derived Mesenchymal Stem Cells in the Treatment of Knee Osteoarthritis","authors":"W. Zeng, Qiu-Ping Yu, Duan Wang, Yipeng Zeng, Hao Yang, Juan Li, Chengguang Zhou, Qing Jun Yang, Z. Deng, Zong-ke Zhou","doi":"10.2139/ssrn.3677351","DOIUrl":"https://doi.org/10.2139/ssrn.3677351","url":null,"abstract":"Intra-articular injection of mesenchymal stem cells (MSCs) is an attractive treatment for osteoarthritis (OA) due to their ease of use in clinical settings, excellent safety profile, and remarkable spontaneous cartilage repair. However, significant challenges arise in relation to the control and induction of stem cell differentiation into the desired cartilaginous lineage. Kartogenin (KGN), a hydrophobic small molecule drug, may significantly promote chondrogenic differentiation of MSCs and induce cartilage regeneration in OA. However, low water solubility and poor bioavailability limits its biological application. In this report, we propose a new nano-drug delivery system based on multifunctional nanographene oxide (NGO) to efficiently load KGN molecules noncovalently via π–π stacking and hydrophobic interactions (PPG-KGN), which could quickly enter MSCs. Before intra-articular injection, MSCs are simply mixed and co-incubated with PPG-KGN to acquire KGN enhanced MSCs. This allows the efficient intracellular delivery of KGN, thereby promoting the chondrogenic differentiation potency of the MSCs. We investigated the effect of KGN-enhanced MSCs in the treatment of knee osteoarthritis. An in vitro study showed the PPG could be rapidly uptaken in the first 4 h after incubation, reaching saturation at 12 h, and accumulating in the lysosome and cytoplasm of MSCs. Thus, PPG-KGN could enhance the efficiency of the intracellular delivery of KGN, which showed remarkably high chondrogenic differentiation capacity of the MSCs. When applied to an OA model of cartilage injury in rats, MSCs were preconditioned with PPG-KGN before being injected into the knee joint. It demonstrated the PPG-KGN preconditioned MSCs contribute to protection from joint space narrowing, pathologic mineralization, osteoarthritis development, behavioral assessment of OA-induced pain, and tissue regeneration, as evidenced by radiographic, weight bearing, and histological analysis. Based on these findings, we propose the use of PPG for delivery of KGN to achieve enhanced MSC chondrogenic potential in osteoarthritis treatment.","PeriodicalId":106645,"journal":{"name":"MatSciRN: Tissue Engineering (Topic)","volume":"36 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"122626331","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}
Yunying He, Lingjie Li, He Zhang, Yuzhou Li, Fengyi Liu, Yiru Fu, L. Mei, R. Cannon, Sheng Yang, P. Ji
Tunneling nanotubular expressways (TNTs), which allow direct cell-to-cell transfer of intracellular organelles, have been widely identified in various cell types. However, the precise functions of TNTs in intercellular communication and their practical application in tissue regeneration is still uncertain. Mesenchymal stem cells (MSCs) are commonly employed as seed cells in tissue engineering. The differentiation of MSCs requires sufficient energy, which can be regulated by the fusion and fission of mitochondria. The phenomenon of mitochondrial shuttle between cells has been observed, and has led to the hypothesis that applying TNTs to deliver mitochondria into MSCs might be a promising approach to stimulate osteogenic differentiation. In proliferating endothelial cells (ECs), cellular dynamics including the fusion and fission of mitochondria is increased, and thus ECs are considered as ideal candidates for mitochondria donors. In order to exploit the application of TNT-mediated mitochondria transfer, we employed mesenchymal stem cell/ human umbilical vein endothelial cell (MSC/HUVEC) spheroids as a research model, and investigated the transfer among them, as well as the underlying mechanism. Fluorescence staining showed that directional transfer of mitochondria between MSC-HUVEC pairs, especially from HUVEC to MSC. Through TNT-mediated mitochondrial transfer, osteogenesis markers were up-regulated, accompanied by an increased amount of β-catenin in MSCs. Moreover, the improved generation of pre-vascular network has also been observed in the spheroids, as a result of β-catenin translocation to the periphery of HUVECs. However, all of these effects are abolished by the destruction of TNTs. Collectively, our results indicate that the TNT strategy can be applied widely to various aspects of biological research, such as but not limited to tissue regeneration and targeted drug delivery.
{"title":"Effects of Tunneling Nanotubes on the Mitochondrial Regulation of the Amount and Subcellular Localization of Β-Catenin During Osteogenesis of MSC/HUVEC Spheroids","authors":"Yunying He, Lingjie Li, He Zhang, Yuzhou Li, Fengyi Liu, Yiru Fu, L. Mei, R. Cannon, Sheng Yang, P. Ji","doi":"10.2139/ssrn.3855764","DOIUrl":"https://doi.org/10.2139/ssrn.3855764","url":null,"abstract":"Tunneling nanotubular expressways (TNTs), which allow direct cell-to-cell transfer of intracellular organelles, have been widely identified in various cell types. However, the precise functions of TNTs in intercellular communication and their practical application in tissue regeneration is still uncertain. Mesenchymal stem cells (MSCs) are commonly employed as seed cells in tissue engineering. The differentiation of MSCs requires sufficient energy, which can be regulated by the fusion and fission of mitochondria. The phenomenon of mitochondrial shuttle between cells has been observed, and has led to the hypothesis that applying TNTs to deliver mitochondria into MSCs might be a promising approach to stimulate osteogenic differentiation. In proliferating endothelial cells (ECs), cellular dynamics including the fusion and fission of mitochondria is increased, and thus ECs are considered as ideal candidates for mitochondria donors. In order to exploit the application of TNT-mediated mitochondria transfer, we employed mesenchymal stem cell/ human umbilical vein endothelial cell (MSC/HUVEC) spheroids as a research model, and investigated the transfer among them, as well as the underlying mechanism. Fluorescence staining showed that directional transfer of mitochondria between MSC-HUVEC pairs, especially from HUVEC to MSC. Through TNT-mediated mitochondrial transfer, osteogenesis markers were up-regulated, accompanied by an increased amount of β-catenin in MSCs. Moreover, the improved generation of pre-vascular network has also been observed in the spheroids, as a result of β-catenin translocation to the periphery of HUVECs. However, all of these effects are abolished by the destruction of TNTs. Collectively, our results indicate that the TNT strategy can be applied widely to various aspects of biological research, such as but not limited to tissue regeneration and targeted drug delivery.","PeriodicalId":106645,"journal":{"name":"MatSciRN: Tissue Engineering (Topic)","volume":"2 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"130111836","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}
Maojiao Li, X. Xing, Hai-sen Huang, C. Liang, Xin Gao, Q. Tang, Xun Xu, Jian Yang, L. Liao, W. Tian
The transplantation of bone marrow mesenchymal stem cells (BMSCs) promotes bone repair and regeneration. However, it has been shown that the majority of BMSCs die within a short period after transplantation. During apoptosis, BMSCs generate a large number of apoptotic cell-derived extracellular vesicles (ApoEVs). This study aims to understand the potential role of ApoEVs in bone defect repair and regeneration. We confirm that BMSCs undergo apoptosis 2 days after transplantation into the defect of the cranium. In vitro, we find that abundant ApoEVs were generated by apoptotic BMSCs and can be engulfed by BMSCs and promote the proliferation, migration, and osteogenic differentiation of recipient cells. ApoEVs from cells in the middle stage of apoptosis were the most efficient at enhancing the regeneration capacity of BMSCs. In vivo, transplantation of ApoEVs in the calvarial defect region significantly promoted bone regeneration in both mouse and rat models. Mechanistically, ApoEVs promote new bone formation by upregulating the Reactive Oxygen Species (ROS) and activating the JNK signaling. This study reveals a previously unknown role of the dying transplanted BMSCs in promoting the viability of endogenous BMSCs and repairing the bone defects. Since it could avoid several adverse effects and limits of BMSCs cytotherapy, treatment of ApoEVs might be a promising strategy in bone repair and regeneration.
{"title":"Apoptotic BMSCs-Derived Extracellular Vesicles Promote Bone Repair Via Activating the ROS-Induced JNK Signal","authors":"Maojiao Li, X. Xing, Hai-sen Huang, C. Liang, Xin Gao, Q. Tang, Xun Xu, Jian Yang, L. Liao, W. Tian","doi":"10.2139/ssrn.3844704","DOIUrl":"https://doi.org/10.2139/ssrn.3844704","url":null,"abstract":"The transplantation of bone marrow mesenchymal stem cells (BMSCs) promotes bone repair and regeneration. However, it has been shown that the majority of BMSCs die within a short period after transplantation. During apoptosis, BMSCs generate a large number of apoptotic cell-derived extracellular vesicles (ApoEVs). This study aims to understand the potential role of ApoEVs in bone defect repair and regeneration. We confirm that BMSCs undergo apoptosis 2 days after transplantation into the defect of the cranium. In vitro, we find that abundant ApoEVs were generated by apoptotic BMSCs and can be engulfed by BMSCs and promote the proliferation, migration, and osteogenic differentiation of recipient cells. ApoEVs from cells in the middle stage of apoptosis were the most efficient at enhancing the regeneration capacity of BMSCs. In vivo, transplantation of ApoEVs in the calvarial defect region significantly promoted bone regeneration in both mouse and rat models. Mechanistically, ApoEVs promote new bone formation by upregulating the Reactive Oxygen Species (ROS) and activating the JNK signaling. This study reveals a previously unknown role of the dying transplanted BMSCs in promoting the viability of endogenous BMSCs and repairing the bone defects. Since it could avoid several adverse effects and limits of BMSCs cytotherapy, treatment of ApoEVs might be a promising strategy in bone repair and regeneration.","PeriodicalId":106645,"journal":{"name":"MatSciRN: Tissue Engineering (Topic)","volume":"28 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"1900-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"128573590","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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