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Self-confocal NIR-II fluorescence microscopy for multifunctional in vivo imaging

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Journal of Innovative Optical Health Sciences 2024年第1期17卷 105-119页

作者： Jing Zhou Tianxiang Wu Runze Chen liang Zhu Hequn Zhang Yifei li liying Chen Jun Qian State Key Laboratory of Modern Optical Instrumentations Centre for Optical and Electromagnetic ResearchCollege of Optical Science and EngineeringInternational Research Center for Advanced PhotonicsZhejiang UniversityHangzhou 310058P.R.China dr.li dak sum&yip yio chin center for stem cell and regenerative medicine Zhejiang UniversityHangzhou 310058P.R.China College of Biomedical Engineering and Instrument Science Interdisciplinary Institute of Neuroscience and Technology(ZIINT)Zhejiang UniversityHangzhou 310027P.R.China

Fluorescence imaging in the second near-infrared window(NIR-II,900–1880 nm)with less scattering background in biological tissues has been combined with the confocal microscopic system for achieving deep in vivo imaging with high spatial resolution.However,the traditional NIR-IIfluorescence confocal microscope with separate excitation focus and detection pinhole makes it possess low confocal e±ciency,as well as di±cultly to adjust.Two types of upgraded NIR-IIfluorescence confocal microscopes,sharing the same pinhole by excitation and emission focus,leading to higher confocal e±ciency,are built in this work.One type is-ber-pinhole-based confocal microscope applicable to CW laser excitation.It is constructed forfluorescence intensity imaging with large depth,high stabilization and low cost,which could replace multiphotonfluorescence microscopy in some applications(e.g.,cerebrovascular and hepatocellular imaging).The other type is air-pinhole-based confocal microscope applicable to femtosecond(fs)laser excitation.It can be employed not only for NIR-IIfluorescence intensity imaging,but also for multi-channelfluorescence lifetime imaging to recognize different structures with similarfluorescence spectrum.Moreover,it can be facilely combined with multiphotonfluorescence microscopy.A single fs pulsed laser is utilized to achieve up-conversion(visible multiphotonfluorescence)and down-conversion(NIR-II one-photonfluorescence)excitation simultaneously,extending imaging spectral channels,and thus facilitates multi-structure and multi-functional observation.

关键词： Self-confocal fiber-pinhole air-pinhole multi-channe fluorescence lifetime imaging multi-color imaging

Immunoregulatory and Anti-cancer Activities of Combination Treatment of Novel Four-Herb Formula and Doxorubicin in 4T1-Breast Cancer Bearing Mice

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chinese Journal of Integrative medicine 2024年第4期30卷 311-321页

作者： Lea ling-Yu Kan Ben Chung-Lap Chan Grace Gar-Lee Yue li Pei-ting Sharon Sze-Man Hon HUANG Dan-qi Miranda Sin-Man Tsang Clara Bik-San Lau Ping-Chung Leung Chun-Kwok Wong Institute of chinese medicine The Chinese University of Hong KongHong Kong SARChina State Key Laboratory of Research on Bioactivities and Clinical Applications of Medicinal Plants The Chinese University of Hong KongHong KongChina department of Chemical Pathology The Chinese University of Hong KongHong Kong SARChina china-Australia International Research Centre for chinese medicine School of Health and Biomedical SciencesSTEM CollegeRMIT UniversityBundooraVictoriaAustralia li dak sum yip yio chin R&D Centre for chinese medicine The Chinese University of Hong Kong SARHong KongChina

Objective To investigate the in vivo immunomodulatory and anti-tumor mechanisms of the combined treatment of novel Four-Herb formula(4HF)and doxorubicin in triple-negative breast cancer(TNBC).Methods Murine-derived triple-negative mammary carcinoma cell line,4T1 cells,was cultured and inoculated into mouse mammary glands.Sixty-six mice were randomly assigned into 6 groups(n=11 in ench):naive,control,LD 4HF(low dose 4HF),HD 4HF(high dose 4HF),LD 4HF+D(low dose and doxorubicin),and D(doxorubicin).Apart from the naive group,each mouse received subcutaneous inoculation with 5×10^(5)4T1 cells resuspended in 100µL of normal saline in the mammary fat pads.Starting from the day of tumor cell inoculation,tumors were grown for 6 days.The LD and HD groups received daily oral gavage of 658 and 2,630 mg/kg 4HF,respectively.The LD 4HF+D group received daily oral gavage of 658 mg/kg 4HF and weekly intraperitoneal injection of doxorubicin(5 mg/kg).The D group received weekly intraperitoneal injections of doxorubicin(5 mg/kg).The treatment naive mice received daily oral gavage of 0.2 mL double distilled water and 0.1 mL normal saline via intraperitoneal injection once a week.The control group received daily oral gavage of 0.2 mL double-distilled water.The treatment period was 30 days.At the end of treatment,mice organs were harvested to analyze immunological activities via immunophenotyping,gene and multiplex analysis,histological staining,and gut microbiota analysis.Results Mice treated with the combination of 4HF and doxorubicin resulted in significantly reduced tumor and spleen burdens(P<0.05),altered the hypoxia and overall immune lymphocyte landscape,and manipulated gut microbiota to favor the anti-tumor immunological activities.Moreover,immunosuppressive genes,cytokines,and chemokines such as C-C motif chemokine 2 and interleukin-10 of tumors were significantly downregulated(P<0.05).4HF-doxorubicin combination treatment demonstrated synergetic activities and was most effective in activating the anti-tumor immune response(P<

关键词： adjunctivetherapy breast cancer cytokines chinese medicine tumor microenvironment

New idea to promote the clinical applications of stem cells or their extracellular vesicles in central nervous system disorders:Combining with intranasal delivery

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Acta Pharmaceutica Sinica B 2022年第8期12卷 3215-3232页

作者： Yaosheng li Honghui Wu Xinchi Jiang Yunfei Dong Juanjuan Zheng Jianqing Gao Institute of Pharmaceutics College of Pharmaceutical SciencesZhejiang UniversityHangzhou 310058China dr.li dak sum&yip yio chin center for stem cell and regenerative medicine Zhejiang UniversityHangzhou 310058China Hangzhou Institute of Innovative medicine Zhejiang UniversityHangzhou 310058China

The clinical translation of stem cells and their extracellular vesicles(EVs)-based therapy for central nervous system(CNS) diseases is booming. Nevertheless, the insufficient CNS delivery and retention together with the invasiveness of current administration routes prevent stem cells or EVs from fully exerting their clinical therapeutic potential. Intranasal(IN) delivery is a possible strategy to solve problems as IN route could circumvent the brain-blood barrier non-invasively and fit repeated dosage regimens. Herein, we gave an overview of studies and clinical trials involved with IN route and discussed the possibility of employing IN delivery to solve problems in stem cells or EVs-based therapy. We reviewed relevant researches that combining stem cells or EVs-based therapy with IN administration and analyzed benefits brought by IN route. Finally, we proposed possible suggestions to facilitate the development of IN delivery of stem cells or EVs.

关键词： stem cells Extracellular vesicles Central nervous system disorders Intranasal Clinical translation Stroke Neurodegenerative disease Glioma

Hierarchical ultrastructure:An overview of what is known about tendons and future perspective for tendon engineering

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Bioactive Materials 2022年第2期7卷 124-139页

作者： Shichen Zhang Wei Ju Xiaoyi Chen Yanyan Zhao lingchong Feng Zi Yin Xiao Chen dr.li dak sum&yip yio chin center for stem cell and regenerative medicine and department of orthopedic surgery of the second affiliated hospital School of MedicineZhejiang UniversityHangzhou310052China Key Laboratory of Tissue Engineering and regenerative medicine of Zhejiang Province School of MedicineZhejiang UniversityHangzhou310058China Guangxi Key Laboratory of regenerative medicine Guangxi-ASEAN Collaborative Innovation Center for Major Disease Prevention and TreatmentGuangxi Medical UniversityGuangxi530021China State Key Laboratory of Clean Energy Utilization Zhejiang UniversityHangzhou310058ZhejiangChina dr.li dak sum&yip yio chin center for stem cell and regenerative medicine and regenerative medicine and department of orthopedic surgery of Sir Run Run Shaw hospital School of MedicineZhejiang UniversityHangzhou310058China department of Sports medicine School of MedicineZhejiang UniversityHangzhou310058China china orthopedic regenerative medicine Group(CORMed) Hangzhou310058China

Abnormal tendons are rarely ever repaired to the natural structure and morphology of normal tendons.To better guide the repair and regeneration of injured tendons through a tissue engineering method,it is necessary to have insights into the internal morphology,organization,and composition of natural tendons.This review summarized recent researches on the structure and function of the extracellular matrix(ECM)components of tendons and highlight the application of multiple detection methodologies concerning the structure of ECMs.In addition,we look forward to the future of multi-dimensional biomaterial design methods and the potential of structural repair for tendon ECM components.In addition,focus is placed on the macro to micro detection methods for tendons,and current techniques for evaluating the extracellular matrix of tendons at the micro level are introduced in detail.Finally,emphasis is given to future extracellular matrix detection methods,as well as to how future efforts could concentrate on fabricating the biomimetic tendons.

关键词： Tendon Collagen fibrillogenesis Extracellular matrix FIB-SEM Biomimetic

Micropatterns and peptide gradient on the inner surface of a guidance conduit synergistically promotes nerve regeneration in vivo

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Bioactive Materials 2022年第3期7卷 134-146页

作者： Deteng Zhang Ziming li Haifei Shi Yuejun Yao Wang Du Pan Lu Kejiong liang liangjie Hong Changyou Gao MOE Key Laboratory of Macromolecular Synthesis and Functionalization Department of Polymer Science and EngineeringZhejiang UniversityHangzhou310027China dr.li dak sum&yip yio chin center for stem cell and regenerative medicine Zhejiang UniversityHangzhou310058China department of Hand surgery First Affiliated Hospital of Zhejiang UniversitySchool of Medicine.Hangzhou310009China

Both of the surface topographical features and distribution of biochemical cues can influence the cell-substrate interactions and thereby tissue regeneration in vivo.However,they have not been combined simultaneously onto a biodegradable scaffold to demonstrate the synergistic role so far.In this study,a proof-of-concept study is performed to prepare micropatterns and peptide gradient on the inner wall of a poly(D,L-lactide-co-caprolactone)(PLCL)guidance conduit and its advantages in regeneration of peripheral nerve in vivo.After linear ridges/grooves of 20/40μm in width are created on the PLCL film,its surface is aminolyzed in a kinetically controlled manner to obtain the continuous gradient of amino groups,which are then transferred to CQAASIKVAV peptide density gradient via covalent coupling of glutaraldehyde.The Schwann cells are better aligned along with the stripes,and show a faster migration rate toward the region of higher peptide density.Implantation of the nerve guidance conduit made of the PLCL film having both the micropatterns and peptide gradient can significantly accelerate the regeneration of sciatic nerve in terms of rate,function recovery and microstructures,and reduction of fibrosis in muscle tissues.Moreover,this nerve conduit can also benefit the M2 polarization of macrophages and promote vascularization in vivo.

关键词： Peptides gradient Micropatterns Contact guidance effect Nerve guidance conduits Nerve regeneration

Dexamethasone-loaded ROS-responsive poly(thioketal)nanoparticles suppress inflammation and oxidative stress of acute lung injury

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Bioactive Materials 2022年第8期7卷 430-442页

作者： Zihe Zhai Wei Ouyang Yuejun Yao Yuqi Zhang Haolan Zhang Feng Xu Changyou Gao MOE Key Laboratory of Macromolecular Synthesis and Functionalization Department of Polymer Science and EngineeringZhejiang UniversityHangzhou310027China department of Infectious Diseases The Second Affiliated HospitalZhejiang University School of MedicineHangzhou310009China dr.li dak sum&yip yio chin center for stem cell and regenerative medicine Zhejiang UniversityHangzhou310058China department of orthopedics The Second Affiliated HospitalZhejiang University School of MedicineHangzhou310009China

Acute lung injury(Ali)is associated with excessive inflammatory response,leading to acute respiratory distress syndrome(ARDS)without timely treatment.A fewer effective drugs are available currently to treat the Ali/ARDS.Herein,a therapeutic nanoplatform with reactive oxygen species(ROS)-responsiveness was developed for the regulation of inflammation.Dexamethasone acetate(Dex)was encapsulated into poly(thioketal)polymers to form polymeric nanoparticles(NPs)(PTKNPs@Dex).The NPs were composed of poly(1,4-phenyleneacetonedimethylene thioketal)(PPADT)and polythioketal urethane(PTKU),in which the thioketal bonds could be cleaved by the high level of ROS at the Ali site.The PTKNPs@Dex could accumulate in the pulmonary inflammatory sites and release the encapsulated payloads rapidly,leading to the decreased ROS level,less generation of pro-inflammatory cytokines,and reduced lung injury and mortality of mice.RNA sequencing(RNA-seq)analysis showed that the therapeutic efficacy of the NPs was associated with the modulation of many immune and inflammation-linked pathways.These findings provide a newly developed nanoplatform for the efficient treatment of Ali/ARDS.

关键词： Acute lung injury Reactive oxygen species Inflammation Nanoparticles RNA-Seq

A reactive oxygen species-responsive hydrogel encapsulated with bone marrow derived stem cells promotes repair and regeneration of spinal cord injury

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Bioactive Materials 2023年第1期19卷 550-568页

作者： Ziming li Tengfei Zhao Jie Ding Haochen Gu Qiaoxuan Wang Yifan Wang Deteng Zhang Changyou Gao MOE Key Laboratory of Macromolecular Synthesis and Functionalization Department of Polymer Science and EngineeringZhejiang UniversityHangzhou 310027China department of orthopedics the Second Affiliated Hospital of Zhejiang UniversityHangzhou 310009China School of medicine Zhejiang UniversityHangzhou 310009China Institute of Neuroregeneration and Neurorehabilitation Qingdao UniversityQingdao 266071China dr.li dak sum&yip yio chin center for stem cell and regenerative medicine Zhejiang UniversityHangzhou 310058China

Spinal cord injury(SCI)is an overwhelming and incurable disabling event accompanied by complicated inflammation-related pathological processes,such as excessive reactive oxygen species(ROS)produced by the infiltrated inflammatory immune cells and released to the extracellular microenvironment,leading to the widespread apoptosis of the neuron cells,glial and oligodendroctyes.In this study,a thioketal-containing and ROS-scavenging hydrogel was prepared for encapsulation of the bone marrow derived mesenchymal stem cells(BMSCs),which promoted the neurogenesis and axon regeneration by scavenging the overproduced ROS and re-building a regenerative microenvironment.The hydrogel could effectively encapsulate BMSCs,and played a remarkable neuroprotective role in vivo by reducing the production of endogenous ROS,attenuating ROS-mediated oxidative damage and downregulating the inflammatory cytokines such as interleukin-1 beta(IL-1β),interleukin-6(IL-6)and tumor necrosis factor-alpha(TNF-α),resulting in a reduced cell apoptosis in the spinal cord tissue.The BMSCs-encapsulated ROS-scavenging hydrogel also reduced the scar formation,and improved the neurogenesis of the spinal cord tissue,and thus distinctly enhanced the motor functional recovery of SCI rats.Our work provides a combinational strategy against ROS-mediated oxidative stress,with potential applications not only in SCI,but also in other central nervous system diseases with similar pathological conditions.

关键词： Spinal cord injury(SCI) ROS scavenging BMSCs Axon regeneration Anti-oxidation

Single-cell genomics: the human biomolecular and cell atlases

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Signal Transduction and Targeted Therapy 2023年第12期8卷 5570-5571页

作者： Yuqing Mei Jingjing Wang Guoji Guo Bone Marrow Transplantation center of the First affiliated hospital and Center for Stem Cell and Regenerative MedicineZhejiang University School of MedicineHangzhouZhejiang 310000China liangzhu Laboratory Zhejiang UniversityHangzhouZhejiang 311121China Zhejiang Provincial Key Lab for Tissue Engineering and regenerative medicine Dr.Li Dak Sum&Yip Yio Chin Center for Stem Cell and Regenerative MedicineHangzhouZhejiang 310058China Institute of Hematology Zhejiang UniversityHangzhouZhejiang 310000China

Recently,in a series of three papers published in Nature,the Human Biomolecular Atlas Program(HuBMAP)consortium reported three multi-model single-cell and spatial atlases.1–3 Each work covered more than a million cells and mapped them to specific locations in the body,providing new standards to study human organs in both health and disease states.

关键词： states. molecular organs

Synergistically targeting synovium STING pathway for rheumatoid arthritis treatment

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Bioactive Materials 2023年第6期24卷 37-53页

作者： Haotian Shen Lulu Jin Qiangqiang Zheng Ziqiang Ye linxiang Cheng Yuxu Wu Honghao Wu Tae Gyong Jon Wenduo liu Zongyou Pan Zhengwei Mao Yue Wang department of orthopedic surgery The First Affiliated HospitalZhejiang University School of MedicineHangzhou310003China MOE Key Laboratory of Macromolecular Synthesis and Functionalization Department of Polymer Science and EngineeringZhejiang UniversityHangzhou310027China dr.li dak sum&yip yio chin center for stem cells and regenerative medicine Zhejiang University School of MedicineHangzhou310000China International Institutes of medicine The Fourth Affiliated HospitalZhejiang University School of MedicineN1 Shangchen RoadYiwuZhejiang322000China

Rheumatoid arthritis(RA)is a common autoimmune disease leading to pain,disability,and even death.Although studies have revealed that aberrant activation of STING was implicated in various autoimmune diseases,the role of STING in RA remains unclear.In the current study,we demonstrated that STING activation was pivotal in RA pathogenesis.As the accumulation of dsDNA,a specific stimulus for STING,is a feature of RA,we developed a spherical polyethyleneimine-coated mesoporous polydopamine nanoparticles loaded with STING antagonist C-176(PEI-PDA@C-176 NPs)for treating RA.The fabricated NPs with biocompatibility had high DNA adsorption ability and could effectively inhibit the STING pathway and inflammation in macrophages.Intra-articular administration of PEI-PDA@C-176 NPs could effectively reduce joint damage in mice models of dsDNA-induced arthritis and collagen-induced arthritis by inhibiting STING pathway.We concluded that materials with synergistic effects of STING inhibition might be an efficacious strategy to treat RA.

关键词： Mesoporous nanoparticles STING Inflammation drug delivery Rheumatoid arthritis