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A functional tacrolimus-releasing nerve wrap for enhancing nerve regeneration following surgical nerve repair

Neural regeneration Research 2025年第1期20卷 291-304页

作者： Simeon C.Daeschler Katelyn J.W.So Konstantin Feinberg Marina Manoraj Jenny Cheung Jennifer Zhang Kaveh Mirmoeini JPaul Santerre Tessa Gordon Gregory HBorschel SickKids Research Institute Neuroscience and Mental Health ProgramTorontoONCanada Institute of Biomaterials and Biomedical Engineering University of TorontoTorontoONCanada Indiana University School of Medicine IndianapolisINUSA Division of Plastic and Reconstructive Surgery the Hospital for Sick ChildrenTorontoONCanada Translational Biology and Engineering Program Ted Rogers Centre for Heart ResearchInstitute of Biomedical EngineeringFaculty of DentistryUniversity of TorontoTorontoONCanada

Axonal regeneration following surgical nerve repair is slow and often incomplete,resulting in poor functional recovery which sometimes contributes to lifelong disability.Currently,there are no FDA-approved therapies available to promote nerve regeneration.Tacrolimus accelerates axonal regeneration,but systemic side effects presently outweigh its potential benefits for peripheral nerve surgery.The authors describe herein a biodegradable polyurethane-based drug delivery system for the sustained local release of tacrolimus at the nerve repair site,with suitable properties for scalable production and clinical application,aiming to promote nerve regeneration and functional recovery with minimal systemic drug exposure.Tacrolimus is encapsulated into co-axially electrospun polycarbonate-urethane nanofibers to generate an implantable nerve wrap that releases therapeutic doses of bioactive tacrolimus over 31 days.Size and drug loading are adjustable for applications in small and large caliber nerves,and the wrap degrades within 120 days into biocompatible byproducts.Tacrolimus released from the nerve wrap promotes axon elongation in vitro and accelerates nerve regeneration and functional recovery in preclinical nerve repair models while off-target systemic drug exposure is reduced by 80%compared with systemic delivery.Given its surgical suitability and preclinical efficacy and safety,this system may provide a readily translatable approach to support axonal regeneration and recovery in patients undergoing nerve surgery.

关键词： biodegradable local drug delivery nerve injury nerve regeneration nerve wrap tacrolimus

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Sustained delivery of vascular endothelial growth factor mediated by bioactive methacrylic anhydride hydrogel accelerates peripheral nerve regeneration after crush injury

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Neural regeneration Research 2022年第9期17卷 2064-2071页

作者： Wanlin Xu Yifan Wu Hao Lu Yun Zhu Jinhai Ye Wenjun Yang Department of Oral and Maxillofacial-Head and Neck Oncology Shanghai Ninth People’s HospitalShanghai Jiao Tong University School of MedicineCollege of StomatologyShanghai Jiao Tong UniversityNational Center for StomatologyNational Clinical Research Center for Oral DiseasesShanghai Key Laboratory of StomatologyShanghaiChina Jiangsu Key Laboratory of Oral Diseases Department of Oral and Maxillofacial SurgeryThe Affiliated Stomatological Hospital of Nanjing Medical UniversityNanjingJiangsu ProvinceChina

Neurotrophic factors,currently administered orally or by intravenous drip or intramuscular injection,are the main method for the treatment of peripheral nerve crush injury.However,the low effective drug concentration arriving at the injury site results in unsatisfactory outcomes.Therefore,there is an urgent need for a treatment method that can increase the effective drug concentration in the injured area.In this study,we first fabricated a gelatin modified by methacrylic anhydride hydrogel and loaded it with vascular endothelial growth factor that allowed the controlled release of the neurotrophic factor.This modified gelatin exhibited good physical and chemical properties,biocompatibility and supported the adhesion and proliferation of RSC96 cells and human umbilical vein endothelial cells.When injected into the epineurium of crushed nerves,the composite hydrogel in the rat sciatic nerve crush injury model promoted nerve regeneration,functional recovery and vascularization.The results showed that the modified gelatin gave sustained delivery of vascular endothelial growth factors and accelerated the repair of crushed peripheral nerves.

关键词： bioactive material controlled release crush injury hydrogel muscle function nerve regeneration peripheral nerve sciatic function index vascular endothelial growth factor vascularization

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Optical tissue clearing enables rapid,precise and comprehensive assessment of three-dimensional morphology in experimental nerve regeneration research

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Neural regeneration Research 2022年第6期17卷 1348-1356页

作者： Simeon C.Daeschler Jennifer Zhang Tessa Gordon Gregory H.Borschel SickKids Research Institute Neuroscience and Mental Health ProgramTorontoONCanada Institute of Biomaterials and Biomedical Engineering University of TorontoTorontoONCanada Division of Plastic and Reconstructive Surgery the Hospital for Sick ChildrenTorontoONCanada Indiana University School of Medicine IndianapolisINUSA

Morphological analyses are key outcome assessments for nerve regeneration studies but are historically limited to tissue sections.Novel optical tissue clearing techniques enabling three-dimensional imaging of entire organs at a subcellular resolution have revolutionized morphological studies of the brain.To extend their applicability to experimental nerve repair studies we adapted these techniques to nerves and their motor and sensory targets in rats.The solvent-based protocols rendered harvested peripheral nerves and their target organs transparent within 24 hours while preserving tissue architecture and fluorescence.The optical clearing was compatible with conventional laboratory techniques,including retrograde labeling studies,and computational image segmentation,providing fast and precise cell quantitation.Further,optically cleared organs enabled three-dimensional morphometry at an unprecedented scale including dermatome-wide innervation studies,tracing of intramuscular nerve branches or mapping of neurovascular networks.Given their wide-ranging applicability,rapid processing times,and low costs,tissue clearing techniques are likely to be a key technology for next-generation nerve repair studies.All procedures were approved by the Hospital for Sick Children’s Laboratory Animal Services Committee(49871/9)on November 9,2019.

关键词： histology image segmentation morphometry nerve regeneration outcome assessment peripheral nerve three-dimensional imaging tissue clearing

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Inhibition of LncRNA Vof-16 expression promotes nerve regeneration and functional recovery after spinal cord injury

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Neural regeneration Research 2022年第1期17卷 217-227页

作者： Xiao-Min Zhang Li-Ni Zeng Wan-Yong Yang Lu Ding Kang-Zhen Chen Wen-Jin Fu Si-Quan Zeng Yin-Ru Liang Gan-Hai Chen Hong-Fu Wu Key Laboratory of Stem Cell and Regenerative Tissue Engineering Guangdong Medical UniversityDongguanGuangdong ProvinceChina Biology Research Group Guangzheng Experimental SchoolHuizhouGuangdong ProvinceChina Geriatric Medicine Center Dongguan Waterfront Zone Central HospitalDongguanGuangdong ProvinceChina Scientific Research Center the Seventh Affiliated HospitalSun Yat-sen UniversityShenzhenGuangdong ProvinceChina Clinical Laboratory Affiliated Houjie HospitalGuangdong Medical UniversityDongguanGuangdong ProvinceChina Department of Intensive Care Unit Affiliated Houjie HospitalGuangdong Medical UniversityDongguanGuangdong ProvinceChina

Our previous RNA sequencing study showed that the long non-coding RNA ischemia-related factor Vof-16(lncRNA Vof-16)was upregulated after spinal cord injury,but its precise role in spinal cord injury remains unclear.Bioinformatics predictions have indicated that lncRNA Vof-16 may participate in the pathophysiological processes of inflammation and apoptosis.PC12 cells were transfected with a pHBLV-U6-MCS-CMV-ZsGreen-PGK-PURO vector to express an lncRNA Vof-16 knockdown lentivirus and a pHLV-CMVIE-ZsGree-Puro vector to express an lncRNA Vof-16 overexpression lentivirus.The overexpression of lncRNA Vof-16 inhibited PC12 cell survival,proliferation,migration,and neurite extension,whereas lncRNA Vof-16 knockdown lentiviral vector resulted in the opposite effects in PC12 cells.Western blot assay results showed that the overexpression of lncRNA Vof-16 increased the protein expression levels of interleukin 6,tumor necrosis factor-α,and Caspase-3 and decreased Bcl-2 expression levels in PC12 cells.Furthermore,we established rat models of spinal cord injury using the complete transection at T10.Spinal cord injury model rats were injected with the lncRNA Vof-16 knockdown or overexpression lentiviral vectors immediately after injury.At 7 days after spinal cord injury,rats treated with lncRNA Vof-16 knockdown displayed increased neuronal survival and enhanced axonal extension.At 8 weeks after spinal cord injury,rats treated with the lncRNA Vof-16 knockdown lentiviral vector displayed improved neurological function in the hind limb.Notably,lncRNA Vof-16 knockdown injection increased Bcl-2 expression and decreased tumor necrosis factor-αand Caspase-3 expression in treated animals.Rats treated with the lncRNA Vof-16 overexpression lentiviral vector displayed opposite trends.These findings suggested that lncRNA Vof-16 is associated with the regulation of inflammation and apoptosis.The inhibition of lncRNA Vof-16 may be useful for promoting nerve regeneration and functional recovery after spinal cord injury.The experiments were app

关键词： apoptosis functional recovery inflammation long non-coding RNA ischemia related factor Vof-16 nerve regeneration nerve repair neurite extension neuronal survival proliferation spinal cord injury

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Sequential expression of miR-221-3p and miR-338-3p in Schwann cells as a therapeutic strategy to promote nerve regeneration and functional recovery

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Neural regeneration Research 2023年第3期18卷 671-682页

作者： Li-Li Wen Tian-Hao Yu Yi-Zhan Ma Xiao-Yan Mao Tian-Rang Ao Rabia Javed Hirotomo Ten Akira Matsuno Qiang Ao Department of Tissue Engineering School of Intelligent MedicineChina Medical UniversityShenyangLiaoning ProvinceChina The VIP Department School and Hospital of StomatologyLiaoning Provincial Key Laboratory of Oral DiseasesChina Medical UniversityShenyangLiaoning ProvinceChina Department of Oncology Cancer HospitalChinese Academy of Medical Sciences&Peking Union Medical CollegeBeijingChina Department of Neurosurgery Teikyo University School of MedicineTokyoJapan NMPA Key Laboratory for Quality Research and Control of Tissue Regenerative Biomaterial&Institute of Regulatory Science for Medical Device&National Engineering Research Center for Biomaterials Sichuan UniversityChengduSichuan ProvinceChina

The functional properties of endogenous Schwann cells(SCs)during nerve repair are dynamic.Optimizing the functional properties of SCs at different stages of nerve repair may have therapeutic benefit in improving the repair of damaged nerves.Previous studies showed that miR-221-3p promotes the proliferation and migration of SCs,and miR-338-3p promotes the myelination of SCs.In this study,we established rat models of sciatic nerve injury by bridging the transected sciatic nerve with a silicone tube.We injected a miR-221 lentiviral vector system together with a doxycycline-inducible Tet-On miR-338 lentiviral vector system into the cavity of nerve conduits of nerve stumps to sequentially regulate the biological function of endogenous SCs at different stages of nerve regeneration.We found that the biological function of SCs was sequentially regulated,the diameter and density of myelinated axons were increased,the expression levels of NF200 and myelin basic protein were increased,and the function of injured peripheral nerve was improved using this system.miRNA Target Prediction Database prediction,Nanopore whole transcriptome sequencing,quantitative PCR,and dual luciferase reporter gene assay results predicted and verified Cdkn1b and Nrp1 as target genes of miR-221-3p and miR-338-3p,respectively,and their regulatory effects on SCs were confirmed in vitro.In conclusion,here we established a new method to enhance nerve regeneration through sequential regulation of biological functions of endogenous SCs,which establishes a new concept and model for the treatment of peripheral nerve injury.The findings from this study will provide direct guiding significance for clinical treatment of sciatic nerve injury.

关键词： cdkn1b miR-221 miR-338 miRNA nerve regeneration Nrp1 peripheral nerve injury regulation Schwann cells sequential expression

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Exosomes-loaded electroconductive nerve dressing for nerve regeneration and pain relief against diabetic peripheral nerve injury

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Bioactive Materials 2023年第8期26卷 194-215页

作者： Qinfeng Yang Shenghui Su Shencai Liu Sheng Yang Jing Xu Yixiu Zhong Yusheng Yang Liangjie Tian Zilin Tan Jian Wang Zhiqiang Yu Zhanjun Shi Fangguo Liang Division of Orthopaedic Surgery Department of OrthopaedicsNanfang HospitalSouthern Medical UniversityGuangzhouGuangdong510515China Department of Orthopaedics Ningde Municipal Hospital of Ningde Normal UniversityNingdeFujian352100China Department of Ophthalmology Nanfang HospitalSouthern Medical UniversityGuangzhouGuangdong510515China Department of Dermatology Shenzhen People’s Hospital(The Second Clinical Medical CollegeJinan UniversityThe First Affiliated HospitalSouthern University of Science and Technology)ShenzhenGuangdong518020China Division of Orthopaedics and Traumatology Department of OrthopaedicsNanfang HospitalSouthern Medical UniversityGuangzhouGuangdong510515China Department of General Surgery Nanfang HospitalSouthern Medical UniversityGuangzhouGuangdong510515China Cancer Center Integrated Hospital of Traditional Chinese MedicineSouthern Medical UniversityGuangzhouGuangdong510515China Guangdong Provincial Key Laboratory of New Drug Screening School of Pharmaceutical SciencesSouthern Medical UniversityGuangzhouGuangdong510515China Department of Plastic and Cosmetic Surgery Nanfang HospitalSouthern Medical UniversityGuangzhouGuangdong510515China

Over the years,electroconductive hydrogels(ECHs)have been extensively applied for stimulating nerve regeneration and restoring locomotor function after peripheral nerve injury(PNI)with diabetes,given their favorable mechanical and electrical properties identical to endogenous nerve tissue.Nevertheless,PNI causes the loss of locomotor function and inflammatory pain,especially in diabetic patients.It has been established that bone marrow stem cells-derived exosomes(BMSCs-Exos)have analgesic,anti-inflammatory and tissue regeneration properties.Herein,we designed an ECH loaded with BMSCs-Exos(ECH-Exos)electroconductive nerve dressing to treat diabetic PNI to achieve functional recovery and pain relief.Given its potent adhesive and self-healing properties,this laminar dressing is convenient for the treatment of damaged nerve fibers by automatically wrapping around them to form a size-matched tube-like structure,avoiding the cumbersome implantation process.Our in vitro studies showed that ECH-Exos could facilitate the attachment and migration of Schwann cells.Meanwhile,Exos in this system could modulate M2 macrophage polarization via the NF-κB pathway,thereby attenuating inflammatory pain in diabetic PNI.Additionally,ECH-Exos enhanced myelinated axonal regeneration via the MEK/ERK pathway in vitro and in vivo,consequently ameliorating muscle denervation atrophy and further promoting functional restoration.Our findings suggest that the ECH-Exos system has huge prospects for nerve regeneration,functional restoration and pain relief in patients with diabetic PNI.

关键词： Diabetic peripheral nerve injury Exosomes Electroconductive hydrogel nerve regeneration Pain relief

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Microtubes with gradient decellularized porcine sciatic nerve matrix from microfluidics for sciatic nerve regeneration

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Bioactive Materials 2023年第3期21卷 511-519页

作者： Binghui Jin Yunru Yu Xiangxiang Chen Yanhong Yang Yushan Xiong Young Jun Im Yuanjin Zhao Jian Xiao Oujiang Laboratory(Zhejiang Lab for Regenerative Medicine Vision and Brain Health)School of Pharmaceutical SciencesWenzhou Medical UniversityWenzhouZhejiang Province325035China Department of Rheumatology and Immunology Nanjing Drum Tower HospitalSchool of Biological Science and Medical EngineeringSoutheast UniversityNanjing210096China College of Pharmacy Chonnam National UniversityGwangju61186Republic of Korea Wenzhou Institute University of Chinese Academy of SciencesWenzhouZhejiang325001China

Long-range peripheral nerve defect is a severe and worldwide disease.With the increasing development of tissue engineering,the excellent ability of nerve extracellular matrix(ECM)in peripheral nerve injury(PNI)has been widely studied and verified.Here,we present a novel microtube that contains gradient decellularized porcine sciatic nerve ECM hydrogel(pDScNM-gel)from microfluidics for sciatic nerve regeneration.The pDScNM is confirmed to enhance cell proliferation and migration,and improve the axon growth of primary dorsal root ganglions(DRGs)in a concentration-related manner.These behaviors were also achieved when cells were co-cultured in a gradient pDScNM microtube.The in vivo sciatic nerve regeneration and functional recovery were also demonstrated by assembling the gradient pDScNM microtubes with a medical silicon tube.These results indicated that the microtubes with gradient pDScNM could act as a promising alternative for repairing peripheral nerve defects and showed great potential in clinical use.

关键词： Microfiber Peripheral nerve injury Extracellular matrix Microfluidics nerve regeneration

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Platelet-rich plasma promotes peripheral nerve regeneration after sciatic nerve injury

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Neural regeneration Research 2023年第2期18卷 375-381页

作者： Su-Long Wang Xi-Lin Liu Zhi-Chen Kang Yue-Shu Wang Department of Hand Surgery China-Japan Union Hospital of Jilin UniversityChangchunJilin ProvinceChina Department of Rehabilitation The Second Hospital of Jilin UniversityChangchunJilin ProvinceChina

The effect of platelet-rich plasma on nerve regeneration remains controversial.In this study,we established a rabbit model of sciatic nerve small-gap defects with preserved epineurium and then filled the gaps with platelet-rich plasma.Twenty-eight rabbits were divided into the following groups(7 rabbits/group):model,low-concentrati on PRP(2.5-3.5-fold concentration of whole blood platelets),medium-concentration PRP(4.5-6.5-fold concentration of whole blood platelets),and high-concentration PRP(7.5-8.5-fold concentration of whole blood platelets).Electrophysiological and histomorphometrical assessments and proteomics analysis we re used to evaluate regeneration of the sciatic nerve.Our results showed that platelet-rich plasma containing 4.5-6.5-and 7.5-8.5-fold concentrations of whole blood platelets promoted repair of sciatic nerve injury.Proteomics analysis was performed to investigate the possible mechanism by which platelet-rich plasma promoted nerve regeneration.Proteomics analysis showed that after sciatic nerve injury,platelet-rich plasma increased the expression of integrin subunitβ-8(ITGB8),which participates in angiogenesis,and differentially expressed proteins were mainly enriched in focal adhesion pathways.Additionally,two key proteins,ribosomal protein S27 a(RSP27 a)and ubiquilin 1(UBQLN1),which were selected after protein-protein interaction analysis,are involved in the regulation of ubiquitin levels in vivo.These data suggest that platelet-rich plasma promotes peripheral nerve regeneration after sciatic nerve injury by affecting angiogenesis and intracellular ubiquitin levels.

关键词： bioinformatic analysis ITGB8 leukocyte-platelet rich plasma nerve regeneration peripheral nerve injury platelet-rich plasma proteomic analysis sciatic nerve injury

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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

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Decellularized Extracellular Matrix Containing Electrospun Fibers for nerve regeneration:A Comparison Between Core–Shell Structured and Preblended Composites

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Advanced Fiber Materials 2022年第3期4卷 503-519页

作者： Rongli Deng Ziling Luo Zilong Rao Zudong Lin Shihao Chen Jing Zhou Qingtang Zhu Xiaolin Liu Ying Bai Daping Quan PCFM Lab GD HPPC LabSchool of ChemistrySun YatSen UniversityGuangzhou 510006China Guangdong Engineering Technology Research Centre for Functional Biomaterials School of Materials Science and EngineeringSun Yat-Sen UniversityGuangzhou 510006China Guangdong Peripheral nerve Tissue Engineering and Technology Research Center Department of Orthopedic and MicrosurgeryThe First Afliated Hospital of Sun Yat-Sen UniversityGuangzhou 510080China Guangdong Provincial Soft Tissue Biofabrication Engineering Laboratory GuangdongGuangzhou 510080China

Advanced biomaterial-based strategies for treatment of peripheral nerve injury require precise control over both topological and biological cues for facilitating rapid and directed nerve regeneration.As a highly bioactive and tissue-specifc natural material,decellularized extracellular matrix(dECM)derived from peripheral nerves(decellularized nerve matrix,DNM)has drawn increasing attention in the feld of regenerative medicine,due to its outstanding capabilities in facilitating neurite outgrowth and remyelination.To induce and maintain sufcient topological guidance,electrospinning was conducted for fabrication of axially aligned nanofbers consisting of DNM and poly(ε-caprolactone)(PCL).Core–shell structured fbers were prepared by coaxial electrospinning using DNM as the shell and PCL as the core.Compared to the aligned electrospun fbers using preblended DNM/PCL,the core–shell structured fbers exhibited lower tensile strength,faster degradation,but considerable toughness for nerve guidance conduit preparation and relatively intact fbrous structure after long-term degradation.More importantly,the full DNM surface coverage of the aligned core–shell fbers efectively promoted axonal extension and Schwann cells migration.The DNM contents further triggered neurite bundling and myelin formation toward nerve fber maturation and functionalization.Herein,we not only pursue a multi-functional scafold design for nerve regeneration,a detailed comparison between core–shell structured and preblended electrospinning of DNM/PCL composites was also provided as an applicable paradigm for advanced tissue-engineered strategies using dECM-based biomaterials.

关键词： Decellularized extracellular matrix Coaxial electrospinning Core–shell structure Preblended composites nerve regeneration

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