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A novel decellularized matrix of Wnt signaling-activated osteocytes accelerates the repair of critical-sized parietal bone defects with osteoclastogenesis, angiogenesis, and neurogenesis

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Bioactive Materials 2023年第3期21卷 110-128页

作者： Xiaofang Wang Yufei Ma Jie Chen Yujiao Liu Guangliang Liu Pengtao Wang Bo Wang Makoto M.Taketo Teresita Bellido Xiaolin Tu Laboratory of Skeletal Development and Regeneration Institute of Life SciencesChongqing Medical UniversityChongqing400016China Department of Pharmacology Graduate School of MedicineKyoto UniversityKyoto606-8501Japan Department of Physiology and Cell Biology University of Arkansas for Medical SciencesLittle RockAR72223USA Department of Anatomy and Cell Biology Indiana University School of MedicineIndianapolisIN46202USA

Cell source is the key to decellularized matrix(DM)strategy.This study compared 3 cell types,osteocytes with/without dominant active Wnt/β-catenin signaling(daCO and WTO)and bone marrow stromal cells(BMSCs)for their DMs in bone repair.Decellularization removes all organelles and>95%DNA,and retained>74%collagen and>71%GAG,maintains the integrity of cell basement membrane with dense boundaries showing oval and honeycomb structure in osteocytic DM and smooth but irregular shape in the BMSC-DM.DM produced higher cell survival rate(90%)and higher proliferative activity.In vitro,daCO-DM induces more and longer stress fibers in BMSCs,conducive to cell adhesion,spreading,and osteogenic differentiation.8-wk after implantation of the critical-sized parietal bone defect model,daCO-DM formed tight structures,composed of a large number of densely-arranged type-I collagen under polarized light microscope,which is similar to and integrated with host bone.BV/TV(>54%)was 1.5,2.9,and 3.5 times of WTO-DM,BMSC-DM,and none-DM groups,and N.Ob/T.Ar(3.2×10^(2)/mm^(2))was 1.7,2.9,and 3.3 times.At 4-wk,daCO-DM induced osteoclastogenesis,2.3 times higher than WTO-DM;but BMSC-DM or none-DM didn't.daCO-DM increased the expression of RANKL and MCSF,Vegfa and Angpt1,and Ngf in BMSCs,which contributes to osteoclastogenesis,angiogenesis,and neurogenesis,respectively.daCO-DM promoted H-type vessel formation and nerve markersβ3-tubulin and NeuN expression.Conclusion:daCO-DM produces metabolic and neurovascularized organoid bone to accelerate the repair of bone defects.These features are expected to achieve the effect of autologous bone transplantation,suitable for transformation application.

关键词： decellularized matrix Osteocyte Wnt signaling 3D printing Regenerative repair Metabolic and neurovascular organoid bone

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TGF-β1-supplemented decellularized annulus fibrosus matrix hydrogels promote annulus fibrosus repair

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Bioactive Materials 2023年第1期19卷 581-593页

作者： Qiang Wei Dachuan Liu Genglei Chu Qifan Yu Zhao Liu Jiaying Li Qingchen Meng Weishan Wang Fengxuan Han Bin Li Department of Orthopaedic Surgery Orthopaedic InstituteThe First Affiliated HospitalSuzhou Medical CollegeSoochow UniversitySuzhouJiangsuChina Department of Orthopaedic Surgery The First Affiliated HospitalShihezi University School of MedicineShiheziXinjiangChina Collaborative Innovation Center of Hematology Soochow UniversitySuzhouJiangsuChina

Annulus fibrosus(AF)repair remains a challenge because of its limited self-healing ability.Endogenous repair strategies combining scaffolds and growth factors show great promise in AF repair.Although the unique and beneficial characteristics of decellularized extracellular matrix(ECM)in tissue repair have been demonstrated,the poor mechanical property of ECM hydrogels largely hinders their applications in tissue regeneration.In the present study,we combined polyethylene glycol diacrylate(PEGDA)and decellularized annulus fibrosus matrix(DAFM)to develop an injectable,photocurable hydrogel for AF repair.We found that the addition of PEGDA markedly improved the mechanical strength of DAFM hydrogels while maintaining their porous structure.Transforming growth factor-β1(TGF-β1)was further incorporated into PEGDA/DAFM hydrogels,and it could be continuously released from the hydrogel.The in vitro experiments showed that TGF-β1 facilitated the migration of AF cells.Furthermore,PEGDA/DAFM/TGF-β1 hydrogels supported the adhesion,proliferation,and increased ECM production of AF cells.In vivo repair performance of the hydrogels was assessed using a rat AF defect model.The results showed that the implantation of PEGDA/DAFM/TGF-β1 hydrogels effectively sealed the AF defect,prevented nucleus pulposus atrophy,retained disc height,and partially restored the biomechanical properties of disc.In addition,the implanted hydrogel was infiltrated by cells resembling AF cells and well integrated with adjacent AF tissue.In summary,findings from this study indicate that TGF-β1-supplemented DAFM hydrogels hold promise for AF repair.

关键词： decellularized matrix Annulus fibrosus Hydrogel TGF-β1 Tissue repair

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Wnt3a-induced ST2 decellularized matrix ornamented PCL scaffold for bone tissue engineering

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BIOCELL 2022年第9期46卷 2089-2099页

作者： XIAOFANG WANG XIAOLIN TU YUFEI MA JIE CHEN YANG SONG GUANGLIANG LIU Laboratory of Skeletal Development and Regeneration The Institute of Life ScienceChongqing Medical UniversityChongqing400016China Department of Anatomy and Cell Biology Indiana University School of MedicineIndianapolisIN46202USA Bioengineering Collage Chongqing UniversityChongqing400044China

The limited bioactivity of scaffold materials is an important factor that restricts the development of bone tissue engineering.Wnt3a activates the classicWnt/β-catenin signaling pathway which effects bone growth and development by the accumulation ofβ-catenin in the nucleus.In this study,we fabricated 3D printed PCL scaffold with Wnt3a-induced murine bone marrow-derived stromal cell line ST2 decellularized matrix(Wnt3a-ST2-dCM-PCL)and ST2 decellularized matrix(ST2-dCM-PCL)by freeze-thaw cycle and DNase decellularization treatment which efficiently decellularized>90%DNA while preserved most protein.Compared to ST2-dCM-PCL,Wnt3a-ST2-dCM-PCL significantly enhanced newly-seeded ST2 proliferation,osteogenic differentiation and upregulated osteogenic marker genes alkaline phosphatase(Alp),Runx2,type I collagen(Col 1)and osteocalcin(Ocn)mRNA expression.After 14 days of osteogenic induction,Wnt3a-ST2-dCM-PCL promoted ST2 mineralization.These results demonstrated that Wnt3a-induced ST2 decellularized matrix improve scaffold materials’osteoinductivity and osteoconductivity.

关键词： Wnt3a decellularized matrix Bone marrow stromal cells Osteogenic differentiation Bone tissue engineering

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Sterilization and disinfection methods for decellularized matrix materials:Review,consideration and proposal

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Bioactive Materials 2021年第9期6卷 2927-2945页

作者： Meihan Tao Tianrang Ao Xiaoyan Mao Xinzhu Yan Rabia Javed Weijian Hou Yang Wang Cong Sun Shuang Lin Tianhao Yu Qiang Ao Department of Tissue Engineering China Medical UniversityShenyangChina Chinese Academy of Medical Sciences&Peking Union Medical College BeijingChina The VIP Department School and Hospital of StomatologyChina Medical UniversityLiaoning Provincial Key Laboratory of Oral DiseasesShenyangChina Department of Developmental Cell Biology Key Laboratory of Medical Cell BiologyMinistry of EducationChina Medical UniversityShenyangChina Institute of Regulatory Science for Medical Device National Engineering Research Center for BiomaterialsSichuan UniversityChengduChina

Sterilization is the process of killing all microorganisms,while disinfection is the process of killing or removing all kinds of pathogenic microorganisms except bacterial spores.Biomaterials involved in cell experiments,animal experiments,and clinical applications need to be in the aseptic state,but their physical and chemical properties as well as biological activities can be affected by sterilization or disinfection.decellularized matrix(dECM)is the low immunogenicity material obtained by removing cells from tissues,which retains many inherent components in tissues such as proteins and proteoglycans.But there are few studies concerning the effects of sterilization or disinfection on dECM,and the systematic introduction of sterilization or disinfection for dECM is even less.Therefore,this review systematically introduces and analyzes the mechanism,advantages,disadvantages,and applications of various sterilization and disinfection methods,discusses the factors influencing the selection of sterilization and disinfection methods,summarizes the sterilization and disinfection methods for various common dECM,and finally proposes a graphical route for selecting an appropriate sterilization or disinfection method for dECM and a technical route for validating the selected method,so as to provide the reference and basis for choosing more appropriate sterilization or disinfection methods of various dECM.

关键词： decellularized matrix Sterilization Disinfection Irradiation Ethylene oxide Supercritical carbon dioxide Peracetic acid Hydrogen peroxide Alcohol Antibiotic

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Ureteral reconstruction with decellularized small intestinal submucosa matrix for ureteral stricture: A preliminary report of two cases

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Asian Journal of Urology 2020年第1期7卷 51-55页

作者： Qingkang Xu Chen Chen Zhefeng Xu Feng Chen Yongtao Yu Xiang Hong Shengli Xu Jiajia Chen Qin Ding Hong Chen Department of Urology Zhejiang Provincial Corps Hospital of Chinese People’s Armed ForcesJiaxingChina Department of Urology First Affiliated HospitalSchool of MedicineZhejiang UniversityHangzhouChina

Objective:To determine the feasibility of decellularized small intestinal submu-cosa(5IS)matrix in repairing ureteral strictures.Methods:Two patients with ureteral stenoses underwent ureteral reconstruction with SIS ma-trix at the Zhejiang Provincial Corps Hospital of Chinese People's Armed Forces between June 2014 and June 2016.The ureteral stenoses were repaired with a semi-tubular SIS matrix and the postoperative recoveries were observed.Results:Both operations were successfully completed.The average operative time was 90 min and the average length of hospital stay was 15 days.No fevers,incision infections,intestinal obstruction,graft rejection,or other serious complications were noted.After 2 months,ure-teroscopic examinations showed that the surfaces of the original patches were covered by mu-Cosa and there were no apparent stenoses in the lumens.The ureteral stents were replaced every 2 months postoperatively and removed 12 months postoperatively.No infections or uri-nary leakage occurred after removal of the stents.Intravenous urography was performed 6 and 12 months postoperatively.The results showed that the ureters were not obstructed and there was no apparent stenosis at the anastomosis sites.The average follow-up time was>12 months.Long-term follow-up is still ongoing,and computed tomography examin ations of the urinary tract have been conducted in the outpatient department of our hospital 1,3,and 6 months after removal of the double-J stents,suggesting the absence of hydronephrosis.The serum creatinine levels remained stable during the follow-up.Conclusion:SIS matrix reconstruction is a feasible method to repair ureters stenosis.

关键词： Ureteral stenosis decellularized matrix Ureteroplasty Ureteral reconstruction Ureteral obstruction

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Construction of tissue-engineered heart valves by using decellularized scaffolds and endothelial progenitor cells

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Chinese Medical Journal 2007年第8期120卷 696-702页

作者： FANG Ning-tao XIE Shang-zhe WANG Song-mei GAO Hong-yang WU Chun-gen PAN Luan-feng Laboratory of Molecular Biology Shanghai Medical College Fudan University Shanghai 200032 China

Background Tissue-engineered heart valves have the potential to overcome the limitations of present heart valve replacements. This study was designed to develop a tissue engineering heart valve by using human umbilical cord blood-derived endothelial progenitor cells （EPCs） and decellularized valve scaffolds. Methods decellularized valve scaffolds were prepared from fresh porcine heart valves. EPCs were isolated from fresh human umbilical cord blood by density gradient centrifugation, cultured for 3 weeks in EGM-2-MV medium, by which time the resultant cell population became endothelial in nature, as assessed by immunofluorescent staining. EPC-derived endothelial cells were seeded onto the decellularized scaffold at 3 × 10^6 cells/cm^2 and cultured under static conditions for 7 days. Proliferation of the seeded cells on the scaffolds was detected using the MTT assay. Tissue-engineered heart valves were analyzed by HE staining, immunofluorescent staining and scanning electron microscopy. The anti-thrombogenic function of the endothelium on the engineered heart valves was evaluated by platelet adhesion experiments and reverse transcription-polymerase chain reaction （RT-PCR） analysis for the expression of endothelial nitric oxide synthase （eNOS） and tissue-type plasminogen activator （t-PA）.Results EPC-derived endothelial cells showed a histolytic cobblestone morphology, expressed specific markers of the endothelial cell lineage including von Willebrand factor （vWF） and CD31, bound a human endothelial cell-specific lectin, Ulex Europaeus agglutinin-1 （UEA-1）, and took up Dil-labeled low density lipoprotein （Dil-Ac-LDL）. After seeding on the decellularized scaffold, the cells showed excellent metabolic activity and proliferation. The cells formed confluent endothelial monolayers atop the decellularized matrix, as assessed by HE staining and immunostaining for vWF and CD31. Scanning electron microscopy demonstrated the occurrence of tight junctions between cells forming the confluent monolayer. Platelets adhesion ex

关键词： endothelial progenitor cell tissue-engineered heart valve decellularized matrix platelet adhesion

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