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Biomaterial engineering strategies for modeling the Bruch's membrane in age-related macular degeneration

Neural Regeneration Research 2024年第12期19卷 2626-2636页

作者： Blanca Molins Andrea Rodríguez Víctor Llorenç Alfredo Adán Group of Ocular Inflammation:Clinical and Experimental Studies Institut d’Investigacions Biomèdiques AgustíPi I Sunyer(IDIBAPS)BarcelonaSpain Institut Clínic d’Oftalmologia(ICOF) Hospital Clínic BarcelonaSpain

Age-related macular degeneration,a multifactorial inflammatory degenerative retinal disease,ranks as the leading cause of blindness in the elderly.Strikingly,there is a scarcity of curative therapies,especially for the atrophic advanced form of age-related macular degeneration,likely due to the lack of models able to fully recapitulate the native structure of the outer blood retinal barrier,the prime to rget tissue of age-related macular degeneration.Standard in vitro systems rely on 2D monocultures unable to adequately reproduce the structure and function of the outer blood retinal barrier,integrated by the dynamic interaction of the retinal pigment epithelium,the Bruch's membrane,and the underlying choriocapillaris.The Bruch's membrane provides structu ral and mechanical support and regulates the molecular trafficking in the outer blood retinal barrier,and therefo re adequate Bruch's membrane-mimics are key for the development of physiologically relevant models of the outer blood retinal barrie r.In the last years,advances in the field of biomaterial engineering have provided novel approaches to mimic the Bruch's membrane from a variety of materials.This review provides a discussion of the integrated properties and function of outer blood retinal barrier components in healt hy and age-related macular degeneration status to understand the requirements to adequately fabricate Bruch's membrane biomimetic systems.Then,we discuss novel materials and techniques to fabricate Bruch's membrane-like scaffolds for age-related macular degeneration in vitro modeling,discussing their advantages and challenges with a special focus on the potential of Bruch's membrane-like mimics based on decellularized tissue.

关键词： age-related macular degeneration Bruch's membrane decellularization retinal pigment epithelium scaffold

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High quality repair of osteochondral defects in rats using the extracellular matrix of antler stem cells

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World Journal of Stem Cells 2024年第2期16卷 176-190页

作者： Yu-Su Wang Wen-Hui Chu Jing-Jie Zhai Wen-Ying Wang Zhong-Mei He Quan-Min Zhao Chun-Yi Li Institute of Antler Science and Product Technology Changchun Sci-Tech UniversityChangchun 130000Jilin ProvinceChina College of Chinese Medicinal Materials Jilin Agricultural UniversityChangchun 130118Jilin ProvinceChina School of Life Science Taizhou UniversityTaizhou 318000Zhejiang ProvinceChina Department of Oral Implantology Hospital of StomatologyJilin UniversityChangchun 130000Jilin ProvinceChina

BACKGROUND Cartilage defects are some of the most common causes of arthritis.Cartilage lesions caused by inflammation,trauma or degenerative disease normally result in osteochondral defects.Previous studies have shown that decellularized extracellular matrix(ECM)derived from autologous,allogenic,or xenogeneic mesenchymal stromal cells(MSCs)can effectively restore osteochondral integrity.AIM To determine whether the decellularized ECM of antler reserve mesenchymal cells(RMCs),a xenogeneic material from antler stem cells,is superior to the currently available treatments for osteochondral defects.METHODS We isolated the RMCs from a 60-d-old sika deer antler and cultured them in vitro to 70%confluence;50 mg/mL L-ascorbic acid was then added to the medium to stimulate ECM deposition.Decellularized sheets of adipocyte-derived MSCs(aMSCs)and antlerogenic periosteal cells(another type of antler stem cells)were used as the controls.Three weeks after ascorbic acid stimulation,the ECM sheets were harvested and applied to the osteochondral defects in rat knee joints.RESULTS The defects were successfully repaired by applying the ECM-sheets.The highest quality of repair was achieved in the RMC-ECM group both in vitro(including cell attachment and proliferation),and in vivo(including the simultaneous regeneration of well-vascularized subchondral bone and avascular articular hyaline cartilage integrated with surrounding native tissues).Notably,the antler-stem-cell-derived ECM(xenogeneic)performed better than the aMSC-ECM(allogenic),while the ECM of the active antler stem cells was superior to that of the quiescent antler stem cells.CONCLUSION Decellularized xenogeneic ECM derived from the antler stem cell,particularly the active form(RMC-ECM),can achieve high quality repair/reconstruction of osteochondral defects,suggesting that selection of decellularized ECM for such repair should be focused more on bioactivity rather than kinship.

关键词： Osteochondral defect repair Mesenchymal stem cells Extracellular matrix decellularization Antler stem cells Reserve mesenchymal cells Xenogeneic

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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 immunosuppression.In recent years,with the development of regenerative medicine,the decellularization-recellularization strategy seems to be a promising and attractive method to resolve these difficulties.The 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 organs.This 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 limbs.Finally,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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Preconditioning process for dermal tissue decellularization using electroporation with sonication

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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 tissues.Effective decellularization methods depend on the characteristics of tissues,and in particular,tissues with dense,complex structure and abundant lipid content are difficult to completely decellularize.Our 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 decellularization.In 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,respectively.Conditions 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 tissue.The combinatorial decellularization treatment formed micropores in the lipid bilayers of the skin tissues while removing all cell and cellular residues without affecting the ECM properties.Therefore,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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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 cross-linking.In addition,how to promote wound healing and how to improve scar formation require further investigation.To ensure the safety and efficacy of animal-derived products,we designed bioartificial corneas(BACs)according to the criteria for Class III medical devices.Our 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 vitro.Preclinical animal studies showed that the BACs can rapidly regenerate the epithelium and restore vision within a month.After 3 months,the BACs were gradually filled with epithelial,stromal,and neuronal cells,and after 6 months,their transparency and histology were almost normal.In addition,side effects such as corneal neovascularization,conjunctival hyperemia,and ciliary body hyperemia rarely occur in vivo.Therefore,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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Polyethylene glycol crosslinked decellularized single liver lobe scaffolds with vascular endothelial growth factor promotes angiogenesis in vivo

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Hepatobiliary & Pancreatic Diseases International 2023年第6期22卷 622-631页

作者： Jian-Se Zhang Zhi-Bin Wang Zhi-Zhen Lai Jing-Wen Yang Wen-Jing Song Yu-Bing Wei Jin Mei Jian-Guang Wang Anatomy Department School of Basic Medical SciencesWenzhou Medical UniversityWenzhou 325000China Institute of Bioscaffold Transplantation and Immunology Wenzhou Medical UniversityWenzhou 325000China Institute of Hypoxic Medicine School of Basic Medical SciencesWenzhou Medical UniversityWenzhou 325000China Intensive Care Unit Tongde Hospital of Zhejiang ProvinceHangzhou 310012China Department of Geriatric Medicine The First Affiliated HospitalWenzhou Medical UniversityWenzhou 325000China Department of Microbiology and Immunology Wenzhou Medical UniversityWenzhou 325000China Medical Research Center Ningbo City First HospitalNingbo 315000China Department of Biochemistry School of Basic Medical SciencesWenzhou Medical UniversityWenzhou 325000China

Background: Improving the mechanical properties and angiogenesis of acellular scaffolds before transplantation is an important challenge facing the development of acellular liver grafts. The present study aimed to evaluate the cytotoxicity and angiogenesis of polyethylene glycol(PEG) crosslinked decellularized single liver lobe scaffolds(DLSs), and establish its suitability as a graft for long-term liver tissue engineering. Methods: Using mercaptoacrylate produced by the Michael addition reaction, DLSs were first modified using N-succinimidyl S-acetylthioacetate(SATA), followed by cross-linking with PEG as well as vascular endothelial growth factor(VEGF). The optimal concentration of agents and time of the individual steps were identified in this procedure through biomechanical testing and morphological analysis. Subsequently, human umbilical vein endothelial cells(HUVECs) were seeded on the PEG crosslinked scaffolds to detect the proliferation and viability of cells. The scaffolds were then transplanted into the subcutaneous tissue of Sprague-Dawley rats to evaluate angiogenesis. In addition, the average number of blood vessels was evaluated in the grafts with or without PEG at days 7, 14, and 21 after implantation. Results: The PEG crosslinked DLS maintained their three-dimensional structure and were more translucent after decellularization than native DLS, which presented a denser and more porous network structure. The results for Young’s modulus proved that the mechanical properties of 0.5 PEG crosslinked DLS were the best and close to that of native livers. The PEG-VEGF-DLS could better promote cell proliferation and differentiation of HUVECs compared with the groups without PEG cross-linking. Importantly, the average density of blood vessels was higher in the PEG-VEGF-DLS than that in other groups at days 7, 14, and 21 after implantation in vivo. Conclusions: The PEG crosslinked DLS with VEGF could improve the biomechanical properties of native DLS, and most importantly, their lack of cytotoxicity provides

关键词： decellularization Single liver lobe Polyethylene glycol Angiogenesis Liver tissue engineering

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Harnessing the synergy of perfusable muscle flap matrix and adipose-derived stem cells for prevascularization and macrophage polarization to reconstruct volumetric muscle loss

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Bioactive Materials 2023年第4期22卷 588-614页

作者： Qixu Zhang Yulun Chiu Youbai Chen Yewen Wu Lina W.Dunne Rene D.Largo Edward I.Chang David M.Adelman Mark V.Schaverien Charles E.Butler Department of Plastic Surgery The University of Texas MD Anderson Cancer CenterHoustonTX77030USA Department of Melanoma Medical Oncology The University of Texas MD Anderson Cancer CenterHoustonTX77030USA Department of Plastic Surgery Chinese PLA General HospitalBeijing100853China

Muscle flaps must have a strong vascular network to support a large tissue volume and ensure successful engraftment.We developed porcine stomach musculofascial flap matrix(PDSF)comprising extracellular matrix(ECM)and intact vasculature.PDSF had a dominant vascular pedicle,microcirculatory vessels,a nerve network,well-retained 3-dimensional(3D)nanofibrous ECM structures,and no allo-or xenoantigenicity.In-depth proteomic analysis demonstrated that PDSF was composed of core matrisome proteins(e.g.,collagens,glycoproteins,proteoglycans,and ECM regulators)that,as shown by Gene Ontology term enrichment analysis,are functionally related to musculofascial biological processes.Moreover,PDSF􀀀human adipose-derived stem cell(hASC)synergy not only induced monocytes towards IL-10􀀀producing M2 macrophage polarization through the enhancement of hASCs’paracrine effect but also promoted the proliferation and interconnection of both human skeletal muscle myoblasts(HSMMs)and human umbilical vein endothelial cells(HUVECs)in static triculture conditions.Furthermore,PDSF was successfully prevascularized through a dynamic perfusion coculture of hASCs and HUVECs,which integrated with PDSF and induced the maturation of vascular networks in vitro.In a xenotransplantation model,PDSF demonstrated myoconductive and immunomodulatory properties associated with the predominance of M2 macrophages and regulatory T cells.In a volumetric muscle loss(VML)model,prevascularized PDSF augmented neovascularization and constructive remodeling,which was characterized by the predominant infiltration of M2 macrophages and significant musculofascial tissue formation.These results indicate that hASCs’integration with PDSF enhances the cells’dual function in immunomodulation and angiogenesis.Owing in part to this PDSF-hASC synergy,our platform shows promise for vascularized muscle flap engineering for VML reconstruction.

关键词： Muscle flap fabrication Extracellular matrix decellularization Vascularization Macrophage polarization Volumetric muscle loss

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Decellularized extracellular matrix scaffolds:Recent trends and emerging strategies in tissue engineering

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Bioactive Materials 2022年第4期7卷 15-31页

作者： Xuewei Zhang Xi Chen Hua Hong Rubei Hu Jiashang Liu Changsheng Liu Key Laboratory for Ultrafine Materials of Ministry of Education Frontiers Science Center for Materiobiology and Dynamic ChemistryEngineering Research Center for Biomaterials of Ministry of EducationSchool of Materials Science and EngineeringEast China University of Science and TechnologyShanghai200237PR China

The application of scaffolding materials is believed to hold enormous potential for tissue regeneration.Despite the widespread application and rapid advance of several tissue-engineered scaffolds such as natural and synthetic polymer-based scaffolds,they have limited repair capacity due to the difficulties in overcoming the immunogenicity,simulating in-vivo microenvironment,and performing mechanical or biochemical properties similar to native organs/tissues.Fortunately,the emergence of decellularized extracellular matrix(dECM)scaffolds provides an attractive way to overcome these hurdles,which mimic an optimal non-immune environment with native three-dimensional structures and various bioactive components.The consequent cell-seeded construct based on dECM scaffolds,especially stem cell-recellularized construct,is considered an ideal choice for regenerating functional organs/tissues.Herein,we review recent developments in dECM scaffolds and put forward perspectives accordingly,with particular focus on the concept and fabrication of decellularized scaffolds,as well as the application of decellularized scaffolds and their combinations with stem cells(recellularized scaffolds)in tissue engineering,including skin,bone,nerve,heart,along with lung,liver and kidney.

关键词： decellularization ECM 3D scaffolds Tissue regeneration Recellularization

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Structural and biomechanical characterizations of acellular porcine mitral valve scaffolds:anterior leaflets,posterior leaflets,and chordae tendineae

Engineered Regeneration

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Engineered Regeneration 2022年第4期3卷 374-386页

作者： Bo Wang Leslie N.Sierad Jeremy J.Mercuri Agneta Simionescu Dan T.Simionescu Lakiesha N.Williams Ryan Vela Pietro Bajona Matthias Peltz Sharan Ramaswamy Yi Hong Jun Liao Department of Bioengineering University of Texas at ArlingtonArlingtonTX 76010United States Joint Department of Biomedical Engineering Medical College of Wisconsin and Marquette UniversityMilwaukeeWI 53226United States Department of Bioengineering Clemson UniversityClemsonSC 29634United States Department of Biomedical Engineering Florida International UniversityMiamiFL 33174United States Department of Biomedical Engineering University of FloridaGainesvilleFL 32611United States Department of Cardiovascular and Thoracic Surgery University of Texas Southwestern Medical CenterDallasTX 75390United States Allegheny Health Network-Drexel University College of Medicine PittsburghPA 15212United States

Mitral valve(MV)tissue engineering is still in its early stage,and one major challenge in MV tissue engineering is to identify appropriate scaffold materials.With the potential of acellular MV scaffolds being demonstrated recently,it is important to have a full understanding of the biomechanics of the native MV components and their acellular scaffolds.In this study,we have successfully characterized the structural and mechanical properties of porcine MV components,including anterior leaflet(AL),posterior leaflet(PL),strut chordae,and basal chordae,before and after decellularization.Quantitative DNA assay showed more than 90%reduction in DNA content,and Griffonia simplicifolia(GS)lectin immunohistochemistry confirmed the complete lack of porcine𝛼-Gal antigen in the acellular MV components.In the acellular AL and PL,the atrialis,spongiosa,and fibrosa trilayered structure,along with its ECM constitutes,i.e.,collagen fibers,elastin fibers,and portion of GAGs,were preserved.Never-theless,the ECM of both AL and PL experienced a certain degree of disruption,exhibiting a less dense,porous ECM morphology.The overall anatomical morphology of the strut and basal chordae were also maintained af-ter decellularization,with longitudinal morphology experiencing minimum disruption,but the cross-sectional morphology exhibiting evenly-distributed porous structure.In the acellular AL and PL,the nonlinear anisotropic biaxial mechanical behavior was overall preserved;however,uniaxial tensile tests showed that the removal of cellular content and the disruption of structural ECM did result in small decreases in maximum tensile modulus,tissue extensibility,failure stress,and failure strain for both MV leaflets and chordae.

关键词： Mitral valve tissue engineering Heart valve biomechanics Valve leaflets Chordae tendineae decellularization

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A Novel Low Air Pressure-Assisted Approach for the Construction of Cells-Decellularized Tendon Scaffold Complex

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Current Medical Science 2022年第3期42卷 569-576页

作者： Xin HUANG Zheng-tao LV Peng CHENG An-min CHEN Department of Orthopedics Tongji HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430030China

Objective The goal of this study was to develop a decellularized tendon scaffold(DTS)and repopulate it with adipose-derived stem cells(ADSCs)assisted by low air pressure(LP).Methods The porcine superficial flexor tendons were processed into the DTSs using a combination of physical,chemical,and enzymatic treatments.The effectiveness of decellularization was verified by histological analysis and DNA quantification.The properties of the DTSs were evaluated by quantitative analysis of biochemical characterization,porosimetry,in vitro biocompatibility assessment,and biomechanical testing.Subsequently,the ADSCs-DTS complexes were constructed via cell injection assisted by LP or under atmospheric pressure.The differences in cell distribution,biomechanical properties,and the total DNA content were compared by histological analysis,biomechanical testing,and DNA quantification,respectively.Results Histological analysis confirmed that no cells or condensed nuclear materials were retained within the DTSs with widened interfibrillar space.The decellularization treatment resulted in a significant decrease in the content of DNA and glycosaminoglycans,and a significant increase in the porosity.The DTSs were cytocompatible in vitro and did not show reduced collagen content and inferior biomechanical properties compared with the fresh-frozen tendons.The assistance of LP promoted the broader distribution of cells into the adjacent interfibrillar space and cell proliferation in DTSs.The biomechanical properties of the scaffolds were not significantly affected by the recellularization treatments.Conclusion A novel LP-assisted approach for the construction of cells-DTS complex was established,which could be a methodological foundation for further bioreactor and in vitro studies.

关键词： tendon decellularization adipose-derived stem cells recellularization low air pressure tissue engineering

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