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Delivery of coenzyme Q10 loaded micelle targets mitochondrial ROS and enhances efficiency of mesenchymal stem cell therapy in intervertebral disc degeneration

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Bioactive Materials 2023年第5期23卷 247-260页

作者： Junyuan Sun Fei Yang Lianlei Wang Haichao Yu Zhijie Yang Jingjing Wei Krasimir Vasilev Xuesong Zhang Xinyu Liu Yunpeng Zhao Department of Orthopaedics Qilu HospitalCheeloo College of MedicineShandong UniversityJinanShandong250012PR China School of Pharmaceutical Sciences Shandong UniversityJinanShandong250012PR China School of Medicine Nankai UniversityTianjin300071PR China Department of Orthopaedics The Fourth Medical CentreChinese People’s Liberation Army General HospitalBeijing100048PR China Key Laboratory of Colloid and Interface Chemistry Ministry of EducationSchool of Chemistry and Chemical EngineeringShandong UniversityJinanShandong250100PR China Academic Unit of STEM University of South AustraliaMawson LakesAdelaide5095South AustraliaAustralia College of Medicine and Public Health Flinders UniversityBedford Park5042South AustraliaAustralia

Stem cell transplantation has been proved a promising therapeutic instrument in intervertebral disc degeneration(IVDD).However,the elevation of oxidative stress in the degenerated region impairs the efficiency of mesenchymal stem cells(BMSCs)transplantation treatment via exaggeration of mitochondrial ROS and promotion of BMSCs apoptosis.Herein,we applied an emulsion-confined assembly method to encapsulate Coenzyme Q10(Co-Q10),a promising hydrophobic antioxidant which targets mitochondria ROS,into the lecithin micelles,which renders the insoluble Co-Q10 dispersible in water as stable colloids.These micelles are injectable,which displayed efficient ability to facilitate Co-Q10 to get into BMSCs in vitro,and exhibited prolonged release of Co-Q10 in intervertebral disc tissue of animal models.Compared to mere use of Co-Q10,the Co-Q10 loaded micelle possessed better bioactivities,which elevated the viability,restored mitochondrial structure as well as function,and enhanced production of ECM components in rat BMSCs.Moreover,it is demonstrated that the injection of this micelle with BMSCs retained disc height and alleviated IVDD in a rat needle puncture model.Therefore,these Co-Q10 loaded micelles play a protective role in cell survival and differentiation through antagonizing mitochondrial ROS,and might be a potential therapeutic agent for IVDD.

关键词： intervertebral disc degeneration Coenzyme Q10 Mesenchymal stem cell Reactive oxygen species Micelle

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Engineered high-strength biohydrogel as a multifunctional platform to deliver nucleic acid for ameliorating intervertebral disc degeneration

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Bioactive Materials 2023年第7期25卷 107-121页

作者： Tao Chen Qiuping Qian Pooyan Makvandi Ehsan Nazarzadeh Zare Qizhu Chen Linjie Chen Zhiguang Zhang Hao Zhou Wenxian Zhou Hui Wang Xiangyang Wang Yu Chen Yunlong Zhou Aimin Wu Department of Orthopaedics Key Laboratory of Orthopaedics of Zhejiang ProvinceThe Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical UniversityWenzhouZhejiang325000China Zhejiang Engineering Research Center for Tissue Repair Materials Wenzhou InstituteUniversity of Chinese Academy of SciencesWenzhouZhejiang325000China Institute for Bioengineering School of EngineeringThe University of EdinburghEdinburghEH93FBUK School of Chemistry Damghan UniversityDamghan36716-45667Iran Wenzhou Medical University WenzhouZhejiang325000China Materdicine Lab School of Life SciencesShanghai UniversityShanghai200444China

intervertebral disc degeneration(IVDD)is a leading cause of low back pain.The strategy of using functional materials to deliver nucleic acids provides a powerful tool for ameliorating IVDD.However,the immunogenicity of nucleic acid vectors and the poor mechanical properties of functional materials greatly limit their effects.Herein,antagomir-204-3p(AM)shows low immunogenicity and effectively inhibits the apoptosis of nucleus pulposus cells.Moreover,a high-strength biohydrogel based on zinc-oxidized sodium alginate-gelatin(ZOG)is designed as a multifunctional nucleic acid delivery platform.ZOG loaded with AM(ZOGA)exhibits great hygroscopicity,antibacterial activity,biocompatibility,and biodegradability.Moreover,ZOGA can be cross-linked with nucleus pulposus tissue to form a high-strength collagen network that improves the mechanical properties of the intervertebral disc(IVD).In addition,ZOGA provides an advantageous microenvironment for genetic expression in which AM can play an efficient role in maintaining the metabolic balance of the extracellular matrix.The results of the radiological and histological analyses demonstrate that ZOGA restores the height of the IVD,retains moisture in the IVD,and maintains the tissue structure.The ZOGA platform shows the sustained release of nucleic acids and has the potential for application to ameliorate IVDD,opening a path for future studies related to IVD.

关键词： intervertebral disc degeneration Hydrogel Nucleic acid MicroRNA Delivery Extracellular matrix

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An esterase-responsive ibuprofen nano-micelle pre-modified embryo derived nucleus pulposus progenitor cells promote the regeneration of intervertebral disc degeneration

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Bioactive Materials 2023年第3期21卷 69-85页

作者： Kai-shun Xia Dong-dong Li Cheng-gui Wang Li-wei Ying Jing-kai Wang Biao Yang Jia-wei Shu Xian-peng Huang Yu-ang Zhang Chao Yu Xiao-peng Zhou Fang-cai Li Nigel K.H.Slater Jian-bin Tang Qi-xin Chen Cheng-zhen Liang Department of Orthopedics 2nd Affiliated HospitalSchool of MedicineZhejiang UniversityHangzhou310009ZhejiangPR China Orthopedics Research Institute of Zhejiang University Zhejiang UniversityHangzhou310009ZhejiangPR China Key Laboratory of Motor System Disease Research and Precision Therapy of Zhejiang Province Hangzhou310009Zhejiang ProvincePR China Clinical Research Center of Motor System Disease of Zhejiang Province Hangzhou310009Zhejiang ProvincePR China Key Laboratory of Biomass Chemical Engineering of Ministry of Education Center for Bionanoengineering Collage of Chemical and Biological EngineeringZhejiang UniversityHangzhou310027ZhejiangPR China The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University Wenzhou325000ZhejiangPR China Orthopedic Department Taizhou Hospital Affiliated to Wenzhou Medical UniversityLinhai317099ZhejiangPR China Department of Chemical Engineering and Biotechnology University of CambridgeCambridgeUnited Kingdom

Stem cell-based transplantation is a promising therapeutic approach for intervertebral disc degeneration(IDD).Current limitations of stem cells include with their insufficient cell source,poor proliferation capacity,low nucleus pulposus(NP)-specific differentiation potential,and inability to avoid pyroptosis caused by the acidic IDD microenvironment after transplantation.To address these challenges,embryo-derived long-term expandable nucleus pulposus progenitor cells(NPPCs)and esterase-responsive ibuprofen nano-micelles(PEG-PIB)were prepared for synergistic transplantation.In this study,we propose a biomaterial pre-modification cell strategy;the PEG-PIB were endocytosed to pre-modify the NPPCs with adaptability in harsh IDD microenvironment through inhibiting pyroptosis.The results indicated that the PEG-PIB pre-modified NPPCs exhibited inhibition of pyroptosis in vitro;their further synergistic transplantation yielded effective functional recovery,histological regeneration,and inhibition of pyroptosis during IDD regeneration.Herein,we offer a novel biomaterial pre-modification cell strategy for synergistic transplantation with promising therapeutic effects in IDD regeneration.

关键词： intervertebral disc degeneration Nucleus pulposus progenitor cells Esterase-responsive nano micell Biomaterial pre-modification Synergistic transplantation therapy

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Multiple nano-drug delivery systems for intervertebral disc degeneration: Current status and future perspectives

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Bioactive Materials 2023年第5期23卷 274-299页

作者： Wenzhao Liu Zhanjun Ma Yonggang Wang Jingjing Yang The Second Clinical Medical College Lanzhou UniversityLanzhou730030GansuChina Louvain Drug Research Institute Advanced Drug Delivery and BiomaterialsUniversit´e Catholique de LouvainUCLouvain1200BrusselsBelgium Department of Orthopedics Lanzhou University Second HospitalLanzhou730030GansuChina

Low back pain(LBP)is a common disease that imposes a huge social and economic burden on people.intervertebral disc(IVD)degeneration(IVDD)is often considered to be the leading cause of LBP and further aggravate and cause serious spinal problems.The established treatment strategy for IVDD consists of physiotherapy,pain medication by drug therapy,and,if necessary,surgery,but none of them can be treated from the etiology;that is,it cannot fundamentally reverse IVD and reconstruct the mechanical function of the spine.With the development of nanotechnology and regenerative medicine,nano-drug delivery systems(NDDSs)have improved treatment results because of their good biodegradability,biocompatibility,precise targeted specific drug delivery,prolonged drug release time,and enhanced drug efficacy,and various new NDDSs for drugs,proteins,cells,and genes have brought light and hope for the treatment of IVDD.This review summarizes the research progress of NDDSs in the treatment of IVDD and provides prospects for using NDDSs to address the challenges of IVDD.We hope that the ideas generated in this review will provide insight into the precise treatment of IVDD.

关键词： intervertebral disc degeneration Nanotechnology Drug delivery system Nanocarriers Nanoparticles

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Nimbolide targeting SIRT1 mitigates intervertebral disc degeneration by reprogramming cholesterol metabolism and inhibiting inflammatory signaling

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Acta Pharmaceutica Sinica B 2023年第5期13卷 2269-2280页

作者： Yun Teng Yixue Huang Hao Yu Cenhao Wu Qi Yan Yingjie Wang Ming Yang Haifeng Xie Tianyi Wu Huilin Yang Jun Zou Department of Orthopaedic Surgery the First Affiliated Hospital of Soochow UniversitySuzhou 215006China

Inflammation,abnormal cholesterol metabolism,and macrophage infiltration are involved in the destruction of the extracellular matrix of the nucleus pulposus(NP),culminating in intervertebral disc degeneration(IDD).Whether nimbolide(Nim),a natural extract,can alleviate IDD is unclear.In this study,we demonstrated that Nim promotes cholesterol efflux and inhibits the activation of the nuclear factor kappa B(NF-κB)and mitogen-activated protein kinase(MAPK)signaling pathways by activating sirtuin 1(SIRT1)in nucleus pulposus cells(NPCs)during inflammation.Thus,Nim balanced matrix anabolism and catabolism of NPCs.However,the inhibition of SIRT1 significantly attenuated the effects of Nim.We also found that Nim promoted the expression of SIRT1 in RAW 264.7,which enhanced the proportion of M2 macrophages by facilitating cholesterol homeostasis reprogramming and impeded M1-like macrophages polarization by blocking the activation of inflammatory signaling.Based on these results,Nim can improve the microenvironment and facilitate matrix metabolism equilibrium in NPCs.Furthermore,in vivo treatment with Nim delayed IDD progression by boosting SIRT1 expression,modulating macrophage polarization and preserving the extracellular matrix.In conclusion,Nim may represent a novel therapeutic strategy for treating IDD.

关键词： Nimbolide SIRT1 intervertebral disc degeneration Nucleus pulposus Macrophage polarization Cholesterol

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How to enhance the ability of mesenchymal stem cells to alleviate intervertebral disc degeneration

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World Journal of Stem Cells 2023年第11期15卷 989-998页

作者： Qing-Xiang Zhang Min Cui Department of Critical Care Medicine Union HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430022Hubei ProvinceChina Department of Critical Care Medicine Wuhan Jinyintan HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430048Hubei ProvinceChina Department of Orthopedics Union HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430022Hubei ProvinceChina

intervertebral disc(ID)degeneration(IDD)is one of the main causes of chronic low back pain,and degenerative lesions are usually caused by an imbalance between catabolic and anabolic processes in the ID.The environment in which the ID is located is harsh,with almost no vascular distribution within the disc,and the nutrient supply relies mainly on the diffusion of oxygen and nutrients from the blood vessels located under the endplate.The stability of its internal environment also plays an important role in preventing IDD.The main feature of disc degeneration is a decrease in the number of cells.Mesenchymal stem cells have been used in the treatment of disc lesions due to their ability to differentiate into nucleus pulposus cells in a nonspecific anti-inflammatory manner.The main purpose is to promote their regeneration.The current aim of stem cell therapy is to replace the aged and metamorphosed cells in the ID and to increase the content of the extracellular matrix.The treatment of disc degeneration with stem cells has achieved good efficacy,and the current challenge is how to improve this efficacy.Here,we reviewed current treatments for disc degeneration and summarize studies on stem cell vesicles,enhancement of therapeutic effects when stem cells are mixed with related substances,and improvements in the efficacy of stem cell therapy by adjuvants under adverse conditions.We reviewed the new approaches and ideas for stem cell treatment of disc degeneration in order to contribute to the development of new therapeutic approaches to meet current challenges.

关键词： Mesenchymal stem cells intervertebral disc degeneration Extracellular vesicles Nucleus pulposus cells Tissue regeneration

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Injectable cartilage matrix hydrogel loaded with cartilage endplate stem cells engineered to release exosomes for non-invasive treatment of intervertebral disc degeneration

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Bioactive Materials 2022年第9期7卷 29-43页

作者： Liwen Luo Junfeng Gong Zhouguang Wang Yao Liu Jiaming Cao Jinghao Qin Rui Zuo Hongyu Zhang Shuai Wang Ping Zhao Di Yang Mengjie Zhang Yanqiu Wang Junfeng Zhang Yue Zhou Changqing Li Bing Ni Zhiqiang Tian MingHan Liu Department of Orthopaedics Xinqiao HospitalArmy Medical University(Third Military Medical University)ChongqingChina Institute of Immunology PLAArmy Medical University(Third Military Medical University)ChongqingChina Molecular Pharmacology Research Center School of Pharmaceutical ScienceWenzhou Medical UniversityWenzhou325000ZhejiangChina Department of Orthopaedics The Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical UniversityWenzhou325000ZhejiangChina Department of Pharmacy Daping HospitalArmy Medical University(Third Military Medical University)ChongqingChina Institute of Geophysics and Geomatics China University of GeosciencesWuhanChina Department of Emergency Second Affiliated Hospital of Chongqing Medical UniversityChongqingChina Institute of Pathology and Southwest Cancer Center Southwest HospitalArmy Medical University(Third Military Medical University)ChongqingChina State Key Laboratory of Silkworm Genome Biology Biological Science Research CenterSouthwest UniversityChongqingChina Department of Pathophysiology College of High Altitude Military MedicineArmy Medical UniversityChongqingChina Key Laboratory of Extreme Environmental Medicine Ministry of Education of ChinaArmy Medical University(Third Military Medical University)ChongqingChina Institute of Hepatopancreatobiliary Surgery Chongqing General HospitalUniversity of Chinese Academy of SciencesChongqingChina

Low back pain,mainly caused by intervertebral disc degeneration(IVDD),is a common health problem;however,current surgical treatments are less than satisfactory.Thus,it is essential to develop novel non-invasive surgical methods for IVDD treatment.Here,we describe a therapeutic strategy to inhibit IVDD by injecting hydrogels modified with the extracellular matrix of costal cartilage(ECM-Gels)that are loaded with cartilage endplate stem cells(CESCs).After loaded with CESCs overexpressing Sphk2(Lenti-Sphk2-CESCs)and injected near the cartilage endplate(CEP)of rats in vivo,ECM-Gels produced Sphk2-engineered exosomes(Lenti-Sphk2-Exos).These exosomes penetrated the annulus fibrosus(AF)and transported Sphk2 into the nucleus pulposus cells(NPCs).Sphk2 activated the phosphatidylinositol 3-kinase(PI3K)/p-AKT pathway as well as the intracellular autophagy of NPCs,ultimately ameliorating IVDD.This study provides a novel and efficient non-invasive combinational strategy for IVDD treatment using injectable ECM-Gels loaded with CESCs that express Sphk2 with sustained release of functional exosomes.

关键词： intervertebral disc degeneration Engineered exosomes Hydrogels Extracellular matrix of costal cartilage Sphingosine kinase 2

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Co-delivery of luteolin and TGF-β1 plasmids with ROS-responsive virus-inspired nanoparticles for microenvironment regulation and chemo-gene therapy of intervertebral disc degeneration

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Nano Research 2022年第9期15卷 8214-8227页

作者： Yifan Ding Huan Wang Yunyun Wang Long Li Jiahui Ding Caiyan Yuan Tao Xu Haoran Xu Hui Xie Ning Zhu Xin Hu Huang Fang Songwei Tan Department of Orthopedic Surgery Tongji HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430030China School of Pharmacy Tongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430030China Department of Cardiology the Fifth Hospital in WuhanWuhan 430030China Department of Radiology Tongji HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430030China College of Biotechnology and Pharmaceutical Engineering Nanjing Tech UniversityNanjing 211800China State Key Laboratory of Materials-Oriented Chemical Engineering Nanjing Tech UniversityNanjing 211800China College of Materials Science and Engineering Nanjing Tech UniversityNanjing 211800China

intervertebral disc degeneration(IDD)is closely related to inflammation and imbalance of synthesis/catabolism of extracellular matrix(ECM)in intervertebral disc(IVD).Considering this,luteolin(LUT),a kind of natural flavonoid with good anti-inflammatory effect and TGF-β1(a gene that promotes the regeneration of ECM)plasmid was co-loaded and co-delivered to nucleus pulposus cells(NPCs).Reactive oxygen species(ROS)responsive cationic copolymer,poly(β-amino ester)-poly(ε-caprolactone)(PBC),with high plasmid DNA(pDNA)compression affinity was synthesized.It can self-assemble into nano-sized polyplexes(pDNA@PBC)with virus-inspired structure and function through which it can transfect pDNA into NPCs with very high efficiency and negligible cytotoxicity.LUT was encapsulated in the hydrophobic core of pDNA@PBC.The co-delivery system,LUT-pTGFβ1@PBC,could enhance the cellular uptake of NPCs and manifest excellent sustained drug release in IVD.Real time quantitative polymerase chain reaction(RT-qPCR)and Western blot experiments reveal that the co-delivery system could inhibit inflammation in NPCs and restore the balance of anabolism and catabolism in vitro by activating TGF/SMAD3 and inhibiting NFkB/p65.Moreover,LUT-pTGF-β1@PBC retards IDD in vivo as detected by radiological and histological methods with good biosafety in rats.LUT-pTGF-β1@PBC may be a promising option for the treatment of IDD.

关键词： intervertebral disc degeneration poly(β-amino ester) luteolin TGF-β1 co-delivery

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Transcription regulators differentiate mesenchymal stem cells into chondroprogenitors,and their in vivo implantation regenerated the intervertebral disc degeneration

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World Journal of Stem Cells 2022年第2期14卷 163-182页

作者： Shumaila Khalid Sobia Ekram Asmat Salim G.Rasul Chaudhry Irfan Khan Dr.Panjwani Center for Molecular Medicine and Drug Research International Center for Chemical and Biological SciencesUniversity of KarachiKarachi 75270SindhPakistan Department of Biological Sciences Oakland UniversityRochesterMI 48309United States

BACKGROUND intervertebral disc degeneration(IVDD)is the leading cause of lower back pain.disc degeneration is characterized by reduced cellularity and decreased production of extracellular matrix(ECM).Mesenchymal stem cells(MSCs)have been envisioned as a promising treatment for degenerative illnesses.Cell-based therapy using ECM-producing chondrogenic derivatives of MSCs has the potential to restore the functionality of the intervertebral disc(IVD).AIM To investigate the potential of chondrogenic transcription factors to promote differentiation of human umbilical cord MSCs into chondrocytes,and to assess their therapeutic potential in IVD regeneration.METHODS MSCs were isolated and characterized morphologically and immunologically by the expression of specific markers.MSCs were then transfected with Sox-9 and Six-1 transcription factors to direct differentiation and were assessed for chondrogenic lineage based on the expression of specific markers.These differentiated MSCs were implanted in the rat model of IVDD.The regenerative potential of transplanted cells was investigated using histochemical and molecular analyses of IVDs.RESULTS Isolated cells showed fibroblast-like morphology and expressed CD105,CD90,CD73,CD29,and Vimentin but not CD45 antigens.Overexpression of Sox-9 and Six-1 greatly enhanced the gene expression of transforming growth factor beta-1 gene,BMP,Sox-9,Six-1,and Aggrecan,and protein expression of Sox-9 and Six-1.The implanted cells integrated,survived,and homed in the degenerated intervertebral disc.Histological grading showed that the transfected MSCs regenerated the IVD and restored normal architecture.CONCLUSION Genetically modified MSCs accelerate cartilage regeneration,providing a unique opportunity and impetus for stem cell-based therapeutic approach for degenerative disc diseases.

关键词： intervertebral disc degeneration Human umbilical cord Transcription factors Mesenchymal stem cells Gene expression Regeneration

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Effect of Tuina along“bladder meridian”alleviating intervertebral disc degeneration by regulating the transforming growth factor-β1/Smad signaling pathway in a rabbit model

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中医杂志:英文版 2023年第5期43卷 991-1000页

作者： SU Chengguo ZHAO Xiaoyan YE Jiangnan ZHANG Xin JIANG Yuqing GUO Junjie ZHANG Xiyuan QI Wenchuan ZHU Jun Acupuncture and Tuina School Chengdu University of Traditional Chinese MedicineChengdu 610075China Clinical Medical School Affiliated Hospital of Chengdu University of Traditional Chinese MedicineChengdu 610075China

OBJECTIVE:The aim of this study was to investigate the protective effects of Tuina(a traditional Chinese massage therapy)on intervertebral disc(IVD)degeneration and the regulatory mechanisms of the transforming growth factor-β1(TGF-β1)/small mothers against decapentaplegic(Smad)signaling pathway.METHODS:Thirty New Zealand white rabbits were randomized into five groups:the control group,model group,model+Tuina group(Tuina group),model+TGF-β1 group(TGF-β1 group),and model+TGF-β1 inhibitor SB431542 group(SB431542 group).The model was established by posterolateral annulus fibrosus puncturing(AFP).Recombinant TGF-β1 and inhibitor SB431542 was injected into the TGF-β1 group and SB431542 group with a microsyringe,respectively.The rabbits in the Tuina group received Tuina treatment along the bladder meridian for 4 weeks.Magnetic resonance imaging(MRI)was performed on rabbits before AFP and after 4 weeks of intervention.Lumbar IVDs(L2-L3 to L4-L5)were harvested after intervention.Histopathological changes in the IVDs were measured by hematoxylin and eosin(HE)staining.Type I collagen was analyzed by immunohistochemistry detection.The expression level of matrix metalloproteinase-3(MMP3)was determined by enzyme-linked immunosorbent assay.Cell apoptosis was evaluated by terminal deoxynucleotidyl transferasemediated nick end labeling and Western blotting.Realtime polymerase chain reaction and Western blotting were used to analyze the expression of TGF-β1 and Smad2/3/4 and a disintegrin and metalloproteinase with thrombospondin motifs 5.RESULTS:Posterolateral AFP induced IVD degeneration in rabbits with histopathological damage and noticeable changes in MRI images.Tuina alleviated histopathological changes and reversed the expression of extracellular matrix degeneration-related molecules and apoptosis-related proteins.Furthermore,AFP induced the activation of TGF-β1 and Smad2/3/4,whereas Tuina therapy markedly reduced the protein expression of Smad2/3 and the gene expression of TGF-β1 and Smad2/3/4.Additionally,the TGF-β1/Smad

关键词： intervertebral disc degeneration transforming growth factor beta1 Smad proteins Tuina apoptosis extracellular matrix degradation

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