Lena Rössler, Ramin Nasehi, Nadja Hansen, Sanja Aveic, Horst Fischer
Despite recent advances in the field of tissue engineering, the development of complex tissue-like structures in vitro is compromised by the lack of integration of a functioning vasculature. In this study, we propose a mesoscale three-dimensional (3D) in vitro vascularized connective tissue model and demonstrate its feasibility to prompt the self-assembly of endothelial cells into vessel-like structures. Moreover, we investigate the effect of perfusion on the organization of the cells. For this purpose, primary endothelial cells (HUVECs) and a cell line of human foreskin fibroblasts are cultivated in ECM-like matrices made up of freeze-dried collagen scaffolds permeated with collagen type I hydrogel. A tailored bioreactor is designed to investigate the effect of perfusion on self-organization of HUVECs. Immunofluorescent staining, two-photon microscopy, second-harmonic generation imaging, and scanning electron microscopy are applied to visualize the spatial arrangement of the cells. The analyses reveal the formation of hollow, vessel-like structures of HUVECs in hydrogel-permeated collagen scaffolds under both static and dynamic conditions. In conclusion, we demonstrate the feasibility of a 3D porous collagen scaffolding system that enables and maintains the self-organization of HUVECs into vessel-like structures independent of a dynamic flow.
尽管组织工程领域取得了最新进展,但体外复杂组织样结构的发展因缺乏功能性血管的整合而受到影响。在这项研究中,我们提出了一种中尺度三维体外血管化结缔组织模型,并证明了它在促使内皮细胞自组装成血管样结构方面的可行性。此外,我们还研究了灌注对细胞组织的影响。为此,我们将原代内皮细胞(HUVECs)和人包皮成纤维细胞系培养在类似 ECM 的基质中,该基质由冷冻干燥的胶原支架和 I 型胶原水凝胶组成。为研究灌注对 HUVECs 自组织的影响,设计了一种量身定制的生物反应器。应用免疫荧光染色、双光子显微镜、二次谐波发生成像和扫描电子显微镜观察细胞的空间排列。分析结果显示,在静态和动态条件下,HUVEC 在水凝胶渗透的胶原支架中形成了中空的血管样结构。总之,我们证明了三维多孔胶原支架系统的可行性,该系统能使 HUVEC 自组织成血管样结构,且不受动态流动的影响。
{"title":"Porous collagen scaffolds enable endothelial lumen formation in vitro under both static and dynamic growth conditions","authors":"Lena Rössler, Ramin Nasehi, Nadja Hansen, Sanja Aveic, Horst Fischer","doi":"10.1002/jbm.b.35444","DOIUrl":"10.1002/jbm.b.35444","url":null,"abstract":"<p>Despite recent advances in the field of tissue engineering, the development of complex tissue-like structures in vitro is compromised by the lack of integration of a functioning vasculature. In this study, we propose a mesoscale three-dimensional (3D) in vitro vascularized connective tissue model and demonstrate its feasibility to prompt the self-assembly of endothelial cells into vessel-like structures. Moreover, we investigate the effect of perfusion on the organization of the cells. For this purpose, primary endothelial cells (HUVECs) and a cell line of human foreskin fibroblasts are cultivated in ECM-like matrices made up of freeze-dried collagen scaffolds permeated with collagen type I hydrogel. A tailored bioreactor is designed to investigate the effect of perfusion on self-organization of HUVECs. Immunofluorescent staining, two-photon microscopy, second-harmonic generation imaging, and scanning electron microscopy are applied to visualize the spatial arrangement of the cells. The analyses reveal the formation of hollow, vessel-like structures of HUVECs in hydrogel-permeated collagen scaffolds under both static and dynamic conditions. In conclusion, we demonstrate the feasibility of a 3D porous collagen scaffolding system that enables and maintains the self-organization of HUVECs into vessel-like structures independent of a dynamic flow.</p>","PeriodicalId":15269,"journal":{"name":"Journal of biomedical materials research. Part B, Applied biomaterials","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2024-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jbm.b.35444","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141457105","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
David Johnson, Connor Tobo, Jeffrey Au, Aakash Nagarapu, Natalia Ziemkiewicz, Hannah Chauvin, Jessica Robinson, Saloni Shringarpure, Jamshid Tadiwala, Julia Brockhouse, Colin A. Flaveny, Koyal Garg
Volumetric muscle loss (VML) injury causes irreversible deficits in muscle mass and function, often resulting in permanent disability. The current standard of care is physical therapy, but it is limited in mitigating functional deficits. We have previously optimized a rehabilitation technique using electrically stimulated eccentric contraction training (EST) that improved muscle mass, strength, and size in VML-injured rats. A biosponge scaffold composed of extracellular matrix proteins has previously enhanced muscle function postVML. This study aimed to determine whether combining a regenerative therapy (i.e., biosponge) with a novel rehabilitation technique (i.e., EST) could enhance recovery in a rat model of VML. A VML defect was created by removing ~20% of muscle mass from the tibialis anterior muscle in adult male Lewis rats. Experimental groups included VML-injured rats treated with biosponge with EST or biosponge alone (n = 6/group). EST was implemented 2 weeks postinjury at 150 Hz and was continued for 4 weeks. A linear increase in eccentric torque over 4 weeks showed the adaptability of the VML-injured muscle to EST. Combining biosponge with EST improved peak isometric torque by ~52% compared with biosponge treatment alone at 6 weeks postinjury. Application of EST increased MyoD gene expression and the percentage of large (>2000 μm2) type 2B myofibers but reduced fibrotic tissue deposition in VML-injured muscles. Together, these changes may provide the basis for improved torque production. This study demonstrates the potential for combined regenerative and rehabilitative therapy to improve muscle recovery following VML.
{"title":"Combined regenerative rehabilitation improves recovery following volumetric muscle loss injury in a rat model","authors":"David Johnson, Connor Tobo, Jeffrey Au, Aakash Nagarapu, Natalia Ziemkiewicz, Hannah Chauvin, Jessica Robinson, Saloni Shringarpure, Jamshid Tadiwala, Julia Brockhouse, Colin A. Flaveny, Koyal Garg","doi":"10.1002/jbm.b.35438","DOIUrl":"10.1002/jbm.b.35438","url":null,"abstract":"<p>Volumetric muscle loss (VML) injury causes irreversible deficits in muscle mass and function, often resulting in permanent disability. The current standard of care is physical therapy, but it is limited in mitigating functional deficits. We have previously optimized a rehabilitation technique using electrically stimulated eccentric contraction training (EST) that improved muscle mass, strength, and size in VML-injured rats. A biosponge scaffold composed of extracellular matrix proteins has previously enhanced muscle function postVML. This study aimed to determine whether combining a regenerative therapy (i.e., biosponge) with a novel rehabilitation technique (i.e., EST) could enhance recovery in a rat model of VML. A VML defect was created by removing ~20% of muscle mass from the tibialis anterior muscle in adult male Lewis rats. Experimental groups included VML-injured rats treated with biosponge with EST or biosponge alone (<i>n</i> = 6/group). EST was implemented 2 weeks postinjury at 150 Hz and was continued for 4 weeks. A linear increase in eccentric torque over 4 weeks showed the adaptability of the VML-injured muscle to EST. Combining biosponge with EST improved peak isometric torque by ~52% compared with biosponge treatment alone at 6 weeks postinjury. Application of EST increased MyoD gene expression and the percentage of large (>2000 μm<sup>2</sup>) type 2B myofibers but reduced fibrotic tissue deposition in VML-injured muscles. Together, these changes may provide the basis for improved torque production. This study demonstrates the potential for combined regenerative and rehabilitative therapy to improve muscle recovery following VML.</p>","PeriodicalId":15269,"journal":{"name":"Journal of biomedical materials research. Part B, Applied biomaterials","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2024-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141457165","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Nik S. A. N. Sharifulden, Lady V. Barrios Silva, Nandin-Erdene Mandakhbayar, Seong-Jin Shin, Hae-Won Kim, Jonathan C. Knowles, Linh T. B. Nguyen, David Y. S. Chau
An ideal wound dressing should create a healing environment that relieves pain, protects against infections, maintains moisture, removes debris, and speeds up wound closure and repair. However, conventional options like gauze often fall short in fulfilling these requirements, especially for chronic or nonhealing wounds. Hence there is a critical need for inventive formulations that offer efficient, cost-effective, and eco-friendly alternatives. This study focuses on assessing the innovative formulation based on a microbial-derived copolymer known as poly(3-hydroxybutyrate-co-4-hydroxybutyrate), P(3HB-co-4HB) bioactive glass and graphene particles, and exploring their biological response in vitro and in vivo—to find the best combination that promotes cell adhesion and enhances wound healing. The formulation optimized at concentration of bioactive glass (1 w/w%) and graphene (0.01 w/w%) showed accelerated degradation and enhanced blood vessel formation. Meanwhile biocompatibility was evaluated using murine osteoblasts, human dermal fibroblasts, and standard cell culture assays, demonstrating no adverse effects after 7 days of culture and well-regulated inflammatory kinetics. Whole thickness skin defect using mice indicated the feasibility of the biocomposites for a faster wound closure and reduced inflammation. Overall, this biocomposite appears promising as an ideal wound dressing material and positively influencing wound healing rates.
{"title":"The biological and therapeutic assessment of a P(3HB-co-4HB)-bioactive glass-graphene composite biomaterial for tissue regeneration","authors":"Nik S. A. N. Sharifulden, Lady V. Barrios Silva, Nandin-Erdene Mandakhbayar, Seong-Jin Shin, Hae-Won Kim, Jonathan C. Knowles, Linh T. B. Nguyen, David Y. S. Chau","doi":"10.1002/jbm.b.35441","DOIUrl":"10.1002/jbm.b.35441","url":null,"abstract":"<p>An ideal wound dressing should create a healing environment that relieves pain, protects against infections, maintains moisture, removes debris, and speeds up wound closure and repair. However, conventional options like gauze often fall short in fulfilling these requirements, especially for chronic or nonhealing wounds. Hence there is a critical need for inventive formulations that offer efficient, cost-effective, and eco-friendly alternatives. This study focuses on assessing the innovative formulation based on a microbial-derived copolymer known as poly(3-hydroxybutyrate-co-4-hydroxybutyrate), P(3HB-co-4HB) bioactive glass and graphene particles, and exploring their biological response in vitro and in vivo—to find the best combination that promotes cell adhesion and enhances wound healing. The formulation optimized at concentration of bioactive glass (1 w/w%) and graphene (0.01 w/w%) showed accelerated degradation and enhanced blood vessel formation. Meanwhile biocompatibility was evaluated using murine osteoblasts, human dermal fibroblasts, and standard cell culture assays, demonstrating no adverse effects after 7 days of culture and well-regulated inflammatory kinetics. Whole thickness skin defect using mice indicated the feasibility of the biocomposites for a faster wound closure and reduced inflammation. Overall, this biocomposite appears promising as an ideal wound dressing material and positively influencing wound healing rates.</p>","PeriodicalId":15269,"journal":{"name":"Journal of biomedical materials research. Part B, Applied biomaterials","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2024-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jbm.b.35441","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141457106","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sensorineural hearing loss (SNHL) is mainly caused by injury or loss of hair cells (HCs) and associated spiral ganglion neurons (SGNs) in the inner ear. At present, there is still no effective treatment for SNHL in clinic. Recently, advances in organoid bring a promising prospect for research and treatment of SNHL. Meanwhile, three-dimensional (3D) printing provides a tremendous opportunity to construct versatile organoids for tissue engineering and regenerative medicine. In this study, gelatin (Gel), sodium alginate (SA), and polyvinyl alcohol (PVA) were used to fabricate biomimetic scaffold through 3D printing. The organ of Corti derived from neonatal mice inner ear was seeded on the PVA/Gel/SA scaffold to construct organ of Corti organoid. Then, the organ of Corti organoid was used to study the potential protective effects of berberine sulfate on neomycin-juried auditory HCs and SGNs. The results showed that the PVA/Gel/SA biomimetic 3D scaffolds had good cytocompatibilities and mechanical properties. The constructed organoid could maintain organ of Corti activity well in vitro. In addition, the injury intervention results showed that berberine sulfate could significantly inhibit neomycin-induced HC and SGN damage. This study suggests that the fabricated organoid is highly biomimetic to the organ of Corti, which may provide an effective model for drug development, cell and gene therapy for SNHL.
{"title":"Construction of organ of Corti organoid to study the effects of berberine sulfate on damaged auditory cells","authors":"Junming Zhang, Li Liu, Rong Shen, Xiangxin Lou","doi":"10.1002/jbm.b.35439","DOIUrl":"10.1002/jbm.b.35439","url":null,"abstract":"<p>Sensorineural hearing loss (SNHL) is mainly caused by injury or loss of hair cells (HCs) and associated spiral ganglion neurons (SGNs) in the inner ear. At present, there is still no effective treatment for SNHL in clinic. Recently, advances in organoid bring a promising prospect for research and treatment of SNHL. Meanwhile, three-dimensional (3D) printing provides a tremendous opportunity to construct versatile organoids for tissue engineering and regenerative medicine. In this study, gelatin (Gel), sodium alginate (SA), and polyvinyl alcohol (PVA) were used to fabricate biomimetic scaffold through 3D printing. The organ of Corti derived from neonatal mice inner ear was seeded on the PVA/Gel/SA scaffold to construct organ of Corti organoid. Then, the organ of Corti organoid was used to study the potential protective effects of berberine sulfate on neomycin-juried auditory HCs and SGNs. The results showed that the PVA/Gel/SA biomimetic 3D scaffolds had good cytocompatibilities and mechanical properties. The constructed organoid could maintain organ of Corti activity well in vitro. In addition, the injury intervention results showed that berberine sulfate could significantly inhibit neomycin-induced HC and SGN damage. This study suggests that the fabricated organoid is highly biomimetic to the organ of Corti, which may provide an effective model for drug development, cell and gene therapy for SNHL.</p>","PeriodicalId":15269,"journal":{"name":"Journal of biomedical materials research. Part B, Applied biomaterials","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2024-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141457166","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Lori M. Estes Bright, Manjyot Kaur Chug, Stephen Thompson, Megan Brooks, Elizabeth J. Brisbois, Hitesh Handa
The development of drug-resistant microorganisms is taking a heavy toll on the biomedical world. Clinical infections are costly and becoming increasingly dangerous as bacteria that once responded to standard antibiotic treatment are developing resistance mechanisms that require innovative treatment strategies. Nitric oxide (NO) is a gaseous molecule produced endogenously that has shown potent antibacterial capabilities in numerous research studies. Its multimechanistic antibacterial methods prevent the development of resistance and have shown potential as an alternative to antibiotics. However, there has yet to be a direct comparison study evaluating the antibacterial properties of NO against antibiotic susceptible and antibiotic-resistant clinically isolated bacterial strains. Herein, standardized lab and clinically isolated drug-resistant bacterial strains are compared side-by-side for growth and viability following treatment with NO released from S-nitrosoglutathione (GSNO), an NO donor molecule. Evaluation of growth kinetics revealed complete killing of E. coli lab and clinical strains at 17.5 mM GSNO, though 15 mM displayed >50% killing and significantly reduced metabolic activity, with greater dose dependence for membrane permeability. Clinical P. aeruginosa showed greater susceptibility to GSNO during growth curve studies, but metabolic activity and membrane permeability demonstrated similar effects for 12.5 mM GSNO treatment of lab and clinical strains. MRSA lab and clinical strains exhibited total killing at 17.5 mM treatment, though metabolic activity was decreased, and membrane permeation began at 12.5 mM for both strains. Lastly, both S. epidermidis strains were killed by 15 mM GSNO, with sensitivities in metabolic activity and membrane permeability at 12.5 mM GSNO. The mirrored antibacterial effects seen by the lab and clinical strains of two Gram-negative and two Gram-positive bacteria reveal the translational success of NO as an antibacterial therapy and potential alternative to standard antibiotic treatment.
{"title":"Analysis of the broad-spectrum potential of nitric oxide for antibacterial activity against clinically isolated drug-resistant bacteria","authors":"Lori M. Estes Bright, Manjyot Kaur Chug, Stephen Thompson, Megan Brooks, Elizabeth J. Brisbois, Hitesh Handa","doi":"10.1002/jbm.b.35442","DOIUrl":"10.1002/jbm.b.35442","url":null,"abstract":"<p>The development of drug-resistant microorganisms is taking a heavy toll on the biomedical world. Clinical infections are costly and becoming increasingly dangerous as bacteria that once responded to standard antibiotic treatment are developing resistance mechanisms that require innovative treatment strategies. Nitric oxide (NO) is a gaseous molecule produced endogenously that has shown potent antibacterial capabilities in numerous research studies. Its multimechanistic antibacterial methods prevent the development of resistance and have shown potential as an alternative to antibiotics. However, there has yet to be a direct comparison study evaluating the antibacterial properties of NO against antibiotic susceptible and antibiotic-resistant clinically isolated bacterial strains. Herein, standardized lab and clinically isolated drug-resistant bacterial strains are compared side-by-side for growth and viability following treatment with NO released from <i>S</i>-nitrosoglutathione (GSNO), an NO donor molecule. Evaluation of growth kinetics revealed complete killing of <i>E. coli</i> lab and clinical strains at 17.5 mM GSNO, though 15 mM displayed >50% killing and significantly reduced metabolic activity, with greater dose dependence for membrane permeability. Clinical <i>P. aeruginosa</i> showed greater susceptibility to GSNO during growth curve studies, but metabolic activity and membrane permeability demonstrated similar effects for 12.5 mM GSNO treatment of lab and clinical strains. MRSA lab and clinical strains exhibited total killing at 17.5 mM treatment, though metabolic activity was decreased, and membrane permeation began at 12.5 mM for both strains. Lastly, both <i>S. epidermidis</i> strains were killed by 15 mM GSNO, with sensitivities in metabolic activity and membrane permeability at 12.5 mM GSNO. The mirrored antibacterial effects seen by the lab and clinical strains of two Gram-negative and two Gram-positive bacteria reveal the translational success of NO as an antibacterial therapy and potential alternative to standard antibiotic treatment.</p>","PeriodicalId":15269,"journal":{"name":"Journal of biomedical materials research. Part B, Applied biomaterials","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2024-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jbm.b.35442","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141457164","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Promise Chinonso Okpe, Oladipo Folorunso, Victor Sunday Aigbodion, Camillus Obayi
Hydroxyapatites (HAps) synthesized from waste animal bones have recently gained attention due to their outstanding properties. This is because there is a need to fabricate scaffolds with desirable mechanical strength, ability to withstand high temperatures, and insoluble in solvents such as water, acetone, ethanol, and isopropyl alcohol. This study is an extensive summary of many articles on the routes of synthesis/preparation of HAp, and the optimum processing parameter, and the biomedical application areas, such as: drug administration, dental implants, bone tissue engineering, orthopedic implant coatings, and tissue regeneration/wound healing. A broad catalog of the synthesis methods (and combination methods), temperature/time, shape/size, and the calcium-to-phosphorous (Ca/P) value of diverse waste animal bone sources were reported. The alkaline hydrolysis method is proposed to be suitable for synthesizing HAp from natural sources due to the technique's ability to produce intrinsic HAp. The method is also preferred to the calcination method owing to the phase transformation that takes place at high temperatures during calcinations. However, calcinations aid in removing impurities and germs during heating at high temperatures. When compared to calcination technique, alkaline hydrolysis method results in crystalline HAp; the higher degree of crystallinity is disadvantageous to HAp bioactivity. In addition, the standardization and removal of impurities and contaminants, thorough biocompatibility to ensure clinical safety of the HAp to the human body, and improvement of the mechanical strength and toughness to match specific requirements for the various biomedical applications are the important areas for future studies.
{"title":"Hydroxyapatite synthesis and characterization from waste animal bones and natural sources for biomedical applications","authors":"Promise Chinonso Okpe, Oladipo Folorunso, Victor Sunday Aigbodion, Camillus Obayi","doi":"10.1002/jbm.b.35440","DOIUrl":"10.1002/jbm.b.35440","url":null,"abstract":"<p>Hydroxyapatites (HAps) synthesized from waste animal bones have recently gained attention due to their outstanding properties. This is because there is a need to fabricate scaffolds with desirable mechanical strength, ability to withstand high temperatures, and insoluble in solvents such as water, acetone, ethanol, and isopropyl alcohol. This study is an extensive summary of many articles on the routes of synthesis/preparation of HAp, and the optimum processing parameter, and the biomedical application areas, such as: drug administration, dental implants, bone tissue engineering, orthopedic implant coatings, and tissue regeneration/wound healing. A broad catalog of the synthesis methods (and combination methods), temperature/time, shape/size, and the calcium-to-phosphorous (Ca/P) value of diverse waste animal bone sources were reported. The alkaline hydrolysis method is proposed to be suitable for synthesizing HAp from natural sources due to the technique's ability to produce intrinsic HAp. The method is also preferred to the calcination method owing to the phase transformation that takes place at high temperatures during calcinations. However, calcinations aid in removing impurities and germs during heating at high temperatures. When compared to calcination technique, alkaline hydrolysis method results in crystalline HAp; the higher degree of crystallinity is disadvantageous to HAp bioactivity. In addition, the standardization and removal of impurities and contaminants, thorough biocompatibility to ensure clinical safety of the HAp to the human body, and improvement of the mechanical strength and toughness to match specific requirements for the various biomedical applications are the important areas for future studies.</p>","PeriodicalId":15269,"journal":{"name":"Journal of biomedical materials research. Part B, Applied biomaterials","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2024-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141457104","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}