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Preconditioning process for dermal tissue decellularization using electroporation with sonication

Regenerative Biomaterials 2022年第1期9卷 22-29页

作者： Mi Hee Leeand Jong-Chul Park Min-Ah Koo HaKyeong Jeong Seung Hee Hong Gyeung Mi Seon Mi Hee Lee Jong-Chul Park Cellbiocontrol Laboratory Yonsei University College of Medicine50-1 Yonsei-roSeodaemun-guSeoul 03722Republic of Korea Department of Medical Engineering Graduate School of Medical ScienceBrain Korea 21 ProjectYonsei University College of Medicine50-1 Yonsei-roSeodaemun-guSeoul 03722Republic of Korea

decellularization to produce bioscaffolds composed of the extracellular matrix(ECM)uses enzymatic,chemical and physical methods to remove antigens and cellular components from *** decellularization methods depend on the characteristics of tissues,and in particular,tissues with dense,complex structure and abundant lipid content are difficult to completely *** study enables future research on the development of methods and treatments for fabricating bioscaffolds via decellularization of complex and rigid skin tissues,which are not commonly considered for decellularization to date as their structural and functional characteristics could not be preserved after severe *** this study,decellularization of human dermal tissue was done by a combination of both chemical(0.05%trypsin-EDTA,2%SDS and 1%Triton X-100)and physical methods(electroporation and sonication).After decellularization,the content of DNA remaining in the tissue was quantitatively confirmed,and the structural change of the tissue and the retention and distribution of ECM components were evaluated through histological and histochemical analysis,*** of the chemical pretreatment that increase the efficiency of physical stimulation as well as decellularization,and conditions for electroporation and sonication without the use of detergents,unlike the methods performed in previous studies,were established to enable the complete decellularization of the skin *** combinatorial decellularization treatment formed micropores in the lipid bilayers of the skin tissues while removing all cell and cellular residues without affecting the ECM ***,this procedure can be widely used to fabricate bioscaffolds by decellularizing biological tissues with dense and complex structures.

关键词： decellularization electroporation sonication extracellular matrix

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A comparison of three methods of decellularization of pig corneas to reduce immunogenicity

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International Journal of Ophthalmology(English edition) 2014年第4期7卷 587-593页

作者： Whayoung Lee Yuko Miyagawa Cassandra Long David K.C.Cooper Hidetaka Hara Department of Surgery Thomas E. Starzl Transplantation Institute University of Pittsburgh

·AIM: To investigate whether decellularization using different techniques can reduce immunogenicity of the cornea, and to explore the decellularized cornea as a scaffold for cultured corneal endothelial cells(CECs).Transplantation of decellularized porcine corneas increases graft transparency and survival for longer periods compared with fresh grafts.·METHODS: Six-month-old wild-type pig corneas were cut into 100-200 μm thickness, and then decellularized by three different methods: 1) 0.1% sodium dodecyl sulfate(SDS); 2) hypoxic nitrogen(N2); and 3) hypertonic NaCl. Thickness and transparency were assessed visually. Fresh and decellularized corneas were stained with hematoxylin/eosin(H&E), and for the presence of galactose-α1,3-galactose(Gal) and N-glycolylneuraminic acid(NeuGc, a nonGal antigen). Also, a human IgM/IgG binding assay was performed. Cultured porcine CECs were seeded on the surface of the decellularized cornea and examined after H&E staining.· RESULTS: All three methods of decellularization reduced the number of keratocytes in the stromal tissue by >80% while the collagen structure remained preserved. No remaining nuclei stained positive for Gal or NeuGc, and expression of these oligosaccharides on collagen was also greatly decreased compared to expression on fresh corneas. Human IgM/IgG binding to decellularized corneal tissue was considerably reduced compared to fresh corneal tissue. The cultured CECs formed a confluent monolayer on the surface of decellularized tissue.· CONCLUSION: Though incomplete, the significant reduction in the cellular component of the decellularized cornea should be associated with a significantly reduced in vivo immune response compared to fresh corneas.

关键词： cornea decellularization immuneresponse pig xenotransplantation

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Donkey pericardium as a select sourcing to manufacture percutaneous heart valves:decellularization has not yet demonstrated any clear cut advantage to glutaraldehyde treatment

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Medicine in Novel Technology and Devices 2019年第4期4卷 38-46页

作者： Aisa Rassoli Yiming Li Xianhao Bao Fabien Kawecki Xinzhe Zhao Daniel Chappard Gaetan Le-Bel Jiaxuan Feng Benedikt Weber Nasser Fatouraee Ze Zhang Zaiping Jing Lucie Germain Lu Wang Robert Guidoin Department of Surgery Faculty of MedicineUniversite Laval and Regenerative Medicine AxisCHU de Quebec Research CenterQuebecCanada Department of Biomedical Engineering Amirkabir University of TechnologyTehranIran Departnent of Vascular Surgery Changhai Hospital of Naval Military Medical UniversityShanghaiChina Key Laboratory of Textile Sciences and Technology Ministry of EducationColle of TextilesDonghua UniversityShanghaiChina GEROM Institut de Biologie en SanteCHU D'AngersAngersFrance Department of Dermatology Medical University of ViennaViennaAustria

Background:The donkey pericardium is considered a good candidate to manufacture percutaneous heart valves based upon its thinness,low cellularity and undulating collagen bundles and *** represents an avenue worth exploring,should its superiority to glutaraldehyde-treated pericardium be *** and methods:Donkey pericardium was divided into two groups:regular glutaraldehyde fixation and mild *** treated pericardia were observed using scanning electron microscopy,histology and transmission electron *** tests were performed along the axial and perpendicular directions,with the data fitted into both the Gasser–Ogden–Holzapfel(GOH)material model and the Fung’s anisotropic ***:The microstructures of the pericardia processed by the two protocols were similar,showing collagen bundles and laminates free of *** decellularization eliminated most of the cells,however leaving the structure somehow *** collagen filaments in bundles were slightly *** anisotropy rates of the non-decellularized specimens were almost identical to the decellularized *** decellularized pericardium appeared ***:The decellularization proved to be ***,it makes the tissue stiffer,which may lead to higher shear concentration during cardiac cycles and reduce its wavy ***,it appears premature to select decellularized donkey pericardium to manufacture heart valves.

关键词： Donkey pericardium decellularization Valve leaflet Transcatheter aortic valve implantation Wavy microstructure

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Application of decellularization-recellularization technique in plastic and reconstructive surgery

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Chinese Medical Journal 2023年第17期136卷 2017-2027页

作者： Yujia Shang Guanhuier Wang Yonghuan Zhen Na Liu Fangfei Nie Zhenmin Zhao Hua Li Yang An Department of Plastic Surgery Peking University Third HospitalHaidian DistrictBeijing 100191China Wuya College of Innovation Key Laboratory of Structure-Based Drug Design&DiscoveryMinistry of EducationShenyang Pharmaceutical UniversityShenyangLiaoning 110016China

In the field of plastic and reconstructive surgery,the loss of organs or tissues caused by diseases or injuries has resulted in challenges,such as donor shortage and *** recent years,with the development of regenerative medicine,the decellularization-recellularization strategy seems to be a promising and attractive method to resolve these *** decellularized extracellular matrix contains no cells and genetic materials,while retaining the complex ultrastructure,and it can be used as a scaffold for cell seeding and subsequent transplantation,thereby promoting the regeneration of diseased or damaged tissues and *** review provided an overview of decellularization-recellularization technique,and mainly concentrated on the application of decellularization-recellularization technique in the field of plastic and reconstructive surgery,including the remodeling of skin,nose,ears,face,and ***,we proposed the challenges in and the direction of future development of decellularization-recellularization technique in plastic surgery.

关键词： decellularization Recellularization Extracellular matrix Plastic surgery Tissue regeneration

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Preclinical evaluation of the safety and effectiveness of a new bioartificial cornea

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Bioactive Materials 2023年第11期29卷 265-278页

作者： Yansha Hao Jingting Zhou Ju Tan Feng Xiang Zhongliang Qin Jun Yao Gang Li Mingcan Yang Lingqin Zeng Wen Zeng Chuhong Zhu Department of Anatomy Engineering Research Center for Organ Intelligent Biological Manufacturing of Chongqingkey Lab for Biomechanics and Tissue Engineering of ChongqingThird Military Medical UniversityChongqing400038China Engineering Research Center of Tissue and Organ Regeneration and Manufacturing Ministry of EducationChongqing400038China State Key Laboratory of Traum1 Burnand Combined InjuryChongqingChina Department of Cell Biology Third Military Army Medical UniversityChongqing400038China Hong Chang Biotechnology Co. LtdGuangzhou510700China Zhong Zhi Yi Gu Research Institute Chongqing Jiukang Medical Research Institute Co.Ltd..China

Cross-linking agents are frequently used to restore corneal properties after decellularization,and it is especially important to select an appropriate method to avoid excessive *** addition,how to promote wound healing and how to improve scar formation require further *** ensure the safety and efficacy of animal-derived products,we designed bioartificial corneas(BACs)according to the criteria for Class III medical *** BACs do not require cross-linking agents and increase mechanical strength via self-cross-linking of aldehyde-modified hyaluronic acid(AHA)and carboxymethyl chitosan(CMC)on the surface of decellularized porcine corneas(DPCs).The results showed that the BACs had good biocompatibility and transparency,and the modification enhanced their antibacterial and anti-inflammatory properties in *** animal studies showed that the BACs can rapidly regenerate the epithelium and restore vision within a *** 3 months,the BACs were gradually filled with epithelial,stromal,and neuronal cells,and after 6 months,their transparency and histology were almost *** addition,side effects such as corneal neovascularization,conjunctival hyperemia,and ciliary body hyperemia rarely occur in ***,these BACs show promise for clinical application for the treatment of infectious corneal ulcers and as a temporary covering for corneal perforations to achieve the more time.

关键词： decellularization Bioartificial cornea Cross-linking Biocompatibility Transparency

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Mechanisms underlying the cell-matrixed nerve grafts repairing peripheral nerve defects

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Bioactive Materials 2024年第1期31卷 563-577页

作者： Shanshan Wang Hongkui Wang Panjian Lu Leilei Gong Xiaosong Gu Meiyuan Li Key Laboratory of Neuroregeneration of Jiangsu and Ministry of Education Co-innovation Center of NeuroregenerationNMPA Key Laboratory for Research and Evaluation of Tissue Engineering Technology ProductsNantong UniversityNantongJiangsu226001PR China Department of Obstetrics and Gynecology Affiliated Hospital of Nantong UniversityNantongJiangsu226001PR China

Decellularized extracellular matrix(dECM),with its distinct biological properties,has gained significant attention as a natural *** its potentials,we successfully developed a three-dimensional matrix-based oriented nerve graft by encapsulating a fibrous scaffold with multilayered conformationally intact and biologically active human bone marrow mesenchymal stem cell-derived decellularized extracellular matrix(hBMSC-dECM).Convincingly,the hBMSC-dECM group exhibited comparable functional recoveries to the autograft group by postoperative week *** the comprehensive analysis,the molecular regulations in the hBMSC-dECM group were more intricate and nuanced compared to the autograft ***,both groups displayed similar molecular regulatory processes in terms of vascularization and extracellular ***,the hBMSC-dECM group demonstrated sustained high levels of regulation in axon and myelin regeneration at week 12,while the immunomodulation returned to the normal levels after peaking at week ***,our findings illustrated the satisfactory construction of a cell-matrixed nerve graft that established a microenvironment conducive to nerve regeneration,and elucidated the distinct molecular regulation patterns and characteristics associated with different repair modes.

关键词： decellularization Extracellular matrix hBMSC Peripheral nerve regeneration Tissue engineering

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Immunological and functional features of decellularized xenogeneic heart valves after transplantation into GGTA1-KO pigs

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Regenerative Biomaterials 2021年第5期8卷 25-36页

作者： Robert Ramm Tobias Goecke Peter Kohler Igor Tudorache Serghei Cebotari Anatol Ciubotaru Samir Sarikouch Klaus Hoffler Friederike Bothe Bjorn Petersen Axel Haverich Heiner Niemann Andres Hilfiker Leibniz Research Laboratories for Biotechnology and Artificial Organs(LEBAO) Hannover Medical SchoolCarl-Neuberg Str.1Hannover 30625Germany Department of Cardiac- Thoracic-Transplantation and Vascular SurgeryHannover Medical SchoolCarl-Neuberg Str.1Hannover 30625Germany Institute of Farm Animal Genetics Friedrich-Loeffler-InstitutHoeltystr.1031535 Mariensee/Neustadt am RuebenbergeGermany Medimplant GmbH Bischofsholer Damm 15Hannover 30173Germany

decellularization of xenogeneic heart valves might lead to excellent regenerative implants,from which many patients could ***,this material carries various xenogeneic epitopes and thus bears a considerable inherent immunological ***,we investigated the regenerative and immunogenic potential of xenogeneic decellularized heart valve implants using pigs deficient for the galactosyltransferase gene(GGTA1-KO)as novel large animal *** aortic and pulmonary heart valves obtained from sheep,wild-type pigs or GGTA1-KO pigs were implanted into GGTA1-KO pigs for 3,or 6 months,*** were analyzed histologically,immunhistologically(CD3,CD21 and CD172a)and anti-aGal antibody serum titers were determined by *** sheep derived implants exhibited a strong immune reaction upon implantation into GGTA1-KO pigs,characterized by massive inflammatory cells infiltrates,presence of foreign body giant cells,a dramatic increase of anti-aGal antibody titers and ultimately destruction of the graft,whereas wild-type porcine grafts induced only a mild reaction in GGTA1-KO *** implants,wild-type/wild-type and GGTA1-KO/GGTA1-KO valves did not induce a measurable immune ***,GGTA1-KO pigs developed a‘human-like’immune response toward decellularized xenogeneic implants showing that immunogenicity of xenogeneic implants is not sufficiently reduced by decellularization,which detracts from their regenerative potential.

关键词： aGal-KO pig decellularization heart valves xenoantibodies large animal model

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Characterization of decellularized scaffold derived from porcine meniscus for tissue engineering applications

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Frontiers of Materials Science 2016年第2期10卷 101-112页

作者： Shuang GAO Zhiguo YUAN Tingfei XI Xiaojuan WEI Quanyi GUO Center for Biomaterial and Tissue Engineering Academy for Advanced Interdisciplinary Studies Peking University Beijing 100871 China Key Laboratory of PLA Institute of Orthopaedics Chinese PLA General Hospital Beijing 100853 China Department of Materials Science and Engineering College of Engineering Peking University Beijing 100871 China

Menisci are fundamental fibrocartilaginous organs in knee joints. The injury in meniscus can impair normal knee function and predisposes patients to osteoarthritis. This study prepared decellularized meniscus scaffolds using a 1% （w/w） sodium dodecyl sulfate solution and sufficient rinsing steps. Complete cell removal was verified by hematoxylin and eosin staining and DNA content assay. Decellularized menisci had accordant tension properties to intact ones, but with declined compression properties. This occurred because the collagen fiber was not damaged but glycosami- noglycans was significantly lost during the decellularization process, which was confirmed by biochemical assay and histology staining. In vitro cytotoxicity assay demonstrated that decellularized meniscus scaffolds have no toxicity on L929 murine fibroblasts and porcine chondrocytes. Further experiment showed that porcine chondrocytes could adhere and proliferate on the scaffold surface, and some cells even could infiltrate into the scaffold. All results showed the potential of this decellularized meniscus to be the scaffolds in tissue engineering.

关键词： meniscus scaffold decellularization biomechanical cytotoxicity tissueengineering

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3D tissue engineering,an emerging technique for pharmaceutical research

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Acta Pharmaceutica Sinica B 2018年第5期8卷 756-766页

作者： Gregory Jensen Christian Morrill Yu Huang Department of Biological Engineering Utah State University

Tissue engineering and the tissue engineering model have shown promise in improving methods of drug delivery, drug action, and drug discovery in pharmaceutical research for the attenuation of the central nervous system inflammatory response. Such inflammation contributes to the lack of regenerative ability of neural cells, as well as the temporary and permanent loss of function associated with neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, and traumatic brain injury. This review is focused specifically on the recent advances in the tissue engineering model made by altering scaffold biophysical and biochemical properties for use in the treatment of neurodegenerative diseases. A portion of this article will also be spent on the review of recent progress made in extracellular matrix decellularization as a new and innovative scaffold for disease treatment.

关键词： Tissue engineering decellularization Neurodegenerative disease Drug screening Neuroinflammation 3D cell culture

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Immobilization of heparin on decellularized kidney scaffold to construct microenvironment for antithrombosis and inducing reendothelialization

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Science China(Life Sciences) 2018年第10期61卷 1168-1177页

作者： Miao Wang Lili Bao Xinyu Qiu Xiaoshan Yang Siying Liu Yuting Su Lulu Wang Bo Liu Qing He Shiyu Liu Yan Jin State Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi International Joint Research Center for Oral Diseases Center for Tissue Engineering School of Stomatology Fourth Military Medical UniversityXi'an 710032 China Shaanxi Institute of Medical Device Quality Supervision and Inspection Xi'an 721046 China Research and Development Center for Tissue Engineering Fourth Military Medical UniversityXi'an 710032 China Xi'an Institute of Tissue Engineering and Regenerative Medicine Xi'an 710032 China State Key Laboratory of Military Stomatology and National Clinical Research Center for Oral Diseases and Shaanxi Clinical Research Center for Oral Diseases Department of Orthodontics School of Stomatology Fourth Military Medical UniversityXi'an 710032 China Department of aerospace Fourth Military Medical UniversityXi'an 710032 China State Key Laboratory of Military Stomatology and National Clinical Research Center for Oral Diseases and Shaanxi Clinical Research Center for Oral Diseases Department of Pediatric Dentistry School of Stomatology Fourth Military Medical UniversitXi'an 710032 China

In recent years, rapid development of tissue engineering technology provides possibilities for the construction of artificial tissues or organs. In construction of engineered kidneys, researchers used native decellularized extracellular matrix(ECM) as the scaffolds to recellularization. However, thrombosis has been a great issue that hinders the progress of transplantation in vivo. In this study, heparin was immobilized to the collagen part of decellularized scaffold with collagen-binding peptide(CBP). Through the anticoagulant and endothelial cell reperfusion experiments, it can be demonstrated that the heparinized scaffolds absorbed less platelets and red blood cells which can effectively reduce the formation of thrombosis. Moreover, it is conducive to longterm adhesion of endothelial cells which is important for the formation of subsequent vascularization. Taken together, our results reveal that the whole kidney can be modified by CBP-heparin composite to reduce the thrombosis and provide the better conditions for neovascularization.

关键词： decellularization kidney scaffold antithrombosis reendothelialization

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