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Adipose-derived stem cells modified by BDNF gene rescue erectile dysfunction after cavernous nerve injury

Neural Regeneration Research 2020年第1期15卷 120-127页

作者： Mei Yang Jiang-Yang Sun Cheng-Cheng Ying Yong Wang Yong-Lian Guo Department of Endocrinology Tongren Hospital of Wuhan University(Wuhan Third Hospital)WuhanHubei ProvinceChina Department of Hepatobiliary Surgery The Central Hospital of WuhanTongji Medical CollegeHuazhong University of Science and TechnologyWuhanHubei ProvinceChina Department of Urology The Central Hospital of WuhanTongji Medical CollegeHuazhong University of Science and TechnologyWuhanHubei ProvinceChina

Cavernous nerve injury is the main cause of erectile dysfunction following radical prostatectomy.The recovery of erectile function following radical prostatectomy remains challenging.Our previous studies found that injecting Adipose-derived stem cells(ADSCs)into the cavernosa could repair the damaged cavernous nerves,but the erectile function of the treated rats could not be restored to a normal level.We evaluated the efficacy of ADSCs infected with a lentiviral vector encoding rat brain-derived neurotrophic factor(lenti-rBDNF)in a rat model of cavernous nerve injury.The rats were equally and randomly divided into four groups.In the control group,bilateral cavernous nerves were isolated but not injured.In the bilateral cavernous nerve injury group,bilateral cavernous nerves were isolated and injured with a hemostat clamp for 2 minutes.In the ADSCGFP and ADSCrBDNF groups,after injury with a hemostat clamp for 2 minutes,rats were injected with ADSCs infected with lenti-GFP(1×106 in 20μL)and lenti-rBDNF(1×106 in 20μL),respectively.Erectile function was assessed 4 weeks after injury by measuring intracavernosal pressures.Then,penile tissues were collected for histological detection and western blot assay.Results demonstrated that compared with the bilateral cavernous nerve injury group,erectile function was significantly recovered in the ADSCGFP and ADSCrBDNF groups,and to a greater degree in the ADSCrBDNF group.Neuronal nitric oxide synthase content in the dorsal nerves and the ratio of smooth muscle/collagen were significantly higher in the ADSCrBDNF and ADSCGFP groups than in the bilateral cavernous nerve injury group.Neuronal nitric oxide synthase expression was obviously higher in the ADSCrBDNF group than in the ADSCGFP group.These findings confirm that intracavernous injection with ADSCs infected with lenti-rBDNF can effectively improve erectile dysfunction caused by cavernous nerve injury.This study was approved by the Medical Animal Care and Welfare Committee of Wuhan University,China(approval No.2017-1638)on

关键词： Adipose-derived stem cells brain-derived neurotrophic factor cavernous nerve injury erectile dysfunction infection intracavernous injection lentiviral vector neuronal nitric oxide synthase radical prostatectomy

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Neural-like cells from Adipose-derived stem cells for cavernous nerve injury in rats

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Neural Regeneration Research 2019年第6期14卷 1085-1090页

作者： Cheng-Cheng Ying Mei Yang Yong Wang Yong-Lian Guo Wan-Li Hu Xin-Min Zheng Department of Urology the Central Hospital of WuhanTongji Medical CollegeHuazhong University of Science and Technology Department of Endocrinology Tongren Hospital of Wuhan University (Wuhan Third Hospital) Department of Urology Zhongnan Hospital of Wuhan University

Although the remaining nerve tissue can regenerate and partly restore erectile function when the cavernous nerve is compressed/severed and function lost, the limited regenerative ability of these nerve tissues often fails to meet clinical needs. Adipose-derived stem cells are easy to obtain and culture, and can differentiate into neural cells. Their proliferation rate is easy to control and they may be used to help restore injured cavernous nerve function. Sprague-Dawley male rats(n = 45) were equally randomized into three groups: fifteen rats as a sham-operated group, fifteen rats as a bilateral nerve crush(BINC) group(with no further intervention), fifteen rats as a BINC with intracavernous injection of one million neural-like cells from Adipose-derived stem cells(NAS)(BINC + NAS) group. After 4 weeks, erectile function was assessed by stimulating the cavernous body. The number of myelinated axons in the dorsal cavernous nerve was determined by toluidine blue staining. The area of neuronal nitric oxide synthase-positive fibers in the dorsal penile nerve was measured by immunohistochemical staining. Masson staining was used to analyze the ratio of smooth muscle to collagen in penile tissue. The results demonstrate that maximal intracavernous pressure, the ratio of maximal intracavernous pressure to mean arterial pressure, the numbers of myelinated axons and neuronal nitric oxide synthase-positive fibers in the dorsal penile nerve, and the ratio of smooth muscle to collagen could be increased after cell transplantation. These findings indicate that neural-like cells from Adipose-derived stem cells can effectively alleviate cavernous nerve injury and improve erectile function. All animal experiments were approved by the Animal Ethics Committee of Huazhong University of Science and Technology, China(approval No. 2017-1925) on September 15, 2017.

关键词： nerve regeneration Adipose-derived neural stem cells corpus cavernosum cavernous nerve erectile dysfunction radical prostatectomy neurons cell differentiation neural regeneration

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Epithelial defect repair in the auricle and auditory meatus by grafting with cultured Adipose-derived mesenchymal stem cell aggregate-extracellular matrix

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Chinese Medical Journal 2019年第6期132卷 680-689页

作者： Wen-Jin Zhang Lei-Guo Ming Jian-Jun Sun Center of Otolaryngology Head and Neck Surgery Chinese PLA Navy General Hospital Beijing 100048 China School of Postgraduate Anhui Medical University Hefei Anhui 230032 China State Key Laboratory of Military Stomatology Center for Tissue Engineering School of Stomatology Fourth Military Medical University Xi'an Shaanxi 710032 China Research and Development Center for Tissue Engineering Fourth Military Medical University Xi'an Shaanxi 710032 China

Background: Several patients experience persistent otorrhea after a flawless surgical procedure because of insufficient epithelial healing. Several efforts, such as autologous tissue allograft and xenograft, have been made to halt otorrhea. However, a stable technology to induce temporal epithelial repair is yet to be established. Therefore, this study aims to investigate whether implantation of seeding Adipose-derived mesenchymal stem cell (ADMSC) aggregates on extracellular matrix (ECM;herein, ADMSC aggregate-ECM) into damaged skin wound promotes skin regeneration. Methods: ADMSC aggregate-ECM was prepared using a previously described procedure that isolated ADMSCs from rabbits and applied to the auricle and auditory meatus wound beds of New Zealand white rabbits. Wound healing was assessed by general observation and hematoxylin and eosin (H&E) staining. Secretion of growth factor of the tissue was evaluated by western blotting. Two other groups, namely, ECM and control, were used. Comparisons of three groups were conducted by one-way analysis of variance analysis. Results: ADMSCs adhered tightly to the ECM and quickly formed cell sheets. At 2 weeks, general observation and H&E staining indicated that the wound healing rates in the ADMSC aggregate-ECM (69.02±6.36%) and ECM (59.32 + 4.10%) groups were higher than that in the control group (43.74± 12.15%;P=0.005, P<0.001, respectively) in ear auricle excisional wounds. At 7 weeks, The scar elevation index was evidently reduced in the ADMSC aggregate-ECM (2.08±0.87) and ECM (2.31 ±0.33) groups compared with the control group (4.06 ±0.45;P < 0.001, P < 0.001, respectively). In addition, the scar elevation index of the ADMSC aggregate-ECM group reached the lowest rate 4 weeks in advance. In auditory meatus excisional wounds, the ADMSC aggregate-ECM group had the largest range of normal skin-like structure at 4 weeks. The ADMSC aggregate-ECM and ECM groups secreted increased amounts of growth factors that contributed to skin regeneration at weeks 1 and 2, respectivel

关键词： Adipose-derived mesenchymal stem cell Extracellular matrix Otorrhea Rabbit Skin regeneration Wound healing

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Ameliorating liver fibrosis in an animal model using the secretome released from miR-122-transfected Adipose-derived stem cells

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World Journal of Stem Cells 2019年第11期11卷 990-1004页

作者： Kee-Hwan Kim Jae Im Lee Ok-Hee Kim Ha-Eun Hong Bong Jun Kwak Ho Joong Choi Joseph Ahn Tae Yun Lee Sang Chul Lee Say-June Kim Department of Surgery Uijeongbu St.Mary’s HospitalCollege of Medicinethe Catholic University of KoreaSeoul 11765South Korea Catholic Central Laboratory of Surgery Institute of Biomedical IndustryCollege of Medicinethe Catholic University of KoreaSeoul 06591South Korea Department of Surgery Seoul St.Mary’s HospitalCollege of Medicinethe Catholic University of KoreaSeoul 06591South Korea Department of Surgery Daejeon St.Mary’s HospitalCollege of MedicineThe Catholic University of KoreaSeoul 34943South Korea

BACKGROUND Recently,the exclusive use of mesenchymal stem cell(MSC)-secreted molecules,called secretome,rather than cells,has been evaluated for overcoming the limitations of cell-based therapy,while maintaining its advantages.However,the use of na?ve secretome may not fully satisfy the specificity of each disease.Therefore,it appears to be more advantageous to use the functionally reinforced secretome through a series of processes involving physico-chemical adjustments or genetic manipulation rather than to the use na?ve secretome.AIM To determine the therapeutic potential of the secretome released from miR-122-transfected Adipose-derived stromal cells(ASCs).METHODS We collected secretory materials released from ASCs that had been transfected with antifibrotic miR-122(MCM)and compared their antifibrotic effects with those of the na?ve secretome(CM).MCM and CM were intravenously administered to the mouse model of thioacetamide-induced liver fibrosis,and their therapeutic potentials were compared.RESULTS MCM infusion provided higher therapeutic potential in terms of:(A)Reducing collagen content in the liver;(B)Inhibiting proinflammatory cytokines;and(C)Reducing abnormally elevated liver enzymes than the infusion of the na?ve secretome.The proteomic analysis of MCM also indicated that the contents of antifibrotic proteins were significantly elevated compared to those in the na?ve secretome.CONCLUSION We could,thus,conclude that the secretome released from miR-122-transfected ASCs has higher antifibrotic and anti-inflammatory properties than the na?ve secretome.Because miR-122 transfection into ASCs provides a specific way of potentiating the antifibrotic properties of ASC secretome,it could be considered as an enhanced method for reinforcing secretome effectiveness.

关键词： Adipose-derived stem cells Liver fibrosis MicroRNAs miR-122 Mesenchymal stem cells Secretome

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Effect of Adipose-derived mesenchymal stem cells on hepatocellular carcinoma: In vitro inhibition of carcinogenesis

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World Journal of Gastroenterology 2019年第5期25卷 567-583页

作者： Rim Serhal Nagib Saliba George Hilal Mayssam Moussa Ghada S Hassan Oula El Atat Nada Alaaeddine Regenerative Medicine Laboratory Faculty of Medicine Saint-Joseph University Surgery Department Faculty of Medicine Saint-Joseph University and Hotel-Dieu de France Cancer and Metabolism Laboratory Faculty of Medicine Campus of Medical Sciences Saint-Joseph University Laboratoire d’Immunologie Cellulaire et Moléculaire Centre Hospitalier de l’Université de Montréal

AIM To investigate the effect of Adipose-derived mesenchymal stem cells(ADMSCs)and their conditioned media(CM) on hepatocellular carcinoma(HCC) cell tumorigenesis.METHODS The proliferation rate of HepG2 and PLC-PRF-5 HCC cancer cells was measured using the trypan blue exclusion method and confirmed using the cell-counting kit8(commonly known as CCK-8) assay. Apoptosis was detected by flow cytometry using annexin V-FITC. Protein and mRNA expression was quantified by ELISA and real time PCR, respectively. Migration and invasion rates were performed by Transwell migration and invasion assays. Wound healing was examined to confirm the data obtained from the migration assays.RESULTS Our data demonstrated that when co-culturing HCC cell lines with ADMSCs or treating them with ADMSC CM, the HCC cell proliferation rate was significantly inhibited and the apoptosis rate increased. The decreased proliferation rate was accompanied by an upregulation of P53 and Retinoblastoma mRNA and a downregulation of c-Myc and hTERT mRNA levels. More notably, ADMSCs and their CM suppressed the expression of the two important markers of HCC carcinogenicity, alpha-fetoprotein and Des-gamma-carboxyprothrombin. In addition, the migration and invasion levels of HepG2 and PLC-PRF-5 cells significantly decreased, potentially through increased expression of the tissue inhibitor metalloproteinases TIMP-1, TIMP-2 and TIMP-3.CONCLUSION These findings shed new light on a protective and therapeutic role for ADMSCs and their CM in controlling HCC invasiveness and carcinogenesis.

关键词： Hepatocellular carcinoma Adipose-derived mesenchymal stem cells Adipose-derived mesenchymal stem cell conditioned media Proliferation Apoptosis Migration Invasion

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Adipose-derived stromal/stem cells from different adipose depots in obesity development

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World Journal of Stem Cells 2019年第3期11卷 147-166页

作者： Karina Ribeiro Silva Leandra Santos Baptista Laboratory of Tissue Bioengineering Directory of Metrology Applied to Life Sciences National Institute of Metrology Quality and Technology Post-Graduation Program of Biotechnology National Institute of Metrology Quality and Technology Multidisciplinary Center for Biological Research (Numpex-Bio) Federal University of Rio de Janeiro Campus Duque de Caxias

The increasing prevalence of obesity is alarming because it is a risk factor for cardiovascular and metabolic diseases(such as type 2 diabetes). The occurrence of these comorbidities in obese patients can arise from white adipose tissue(WAT) dysfunctions, which affect metabolism, insulin sensitivity and promote local and systemic inflammation. In mammals, WAT depots at different anatomical locations(subcutaneous, preperitoneal and visceral) are highly heterogeneous in their morpho-phenotypic profiles and contribute differently to homeostasis and obesity development, depending on their ability to trigger and modulate WAT inflammation. This heterogeneity is likely due to the differential behavior of cells from each depot. Numerous studies suggest that adiposederived stem/stromal cells(ASC; referred to as adipose progenitor cells, in vivo)with depot-specific gene expression profiles and adipogenic and immunomodulatory potentials are keys for the establishment of the morphofunctional heterogeneity between WAT depots, as well as for the development of depot-specific responses to metabolic challenges. In this review, we discuss depot-specific ASC properties and how they can contribute to the pathophysiology of obesity and metabolic disorders, to provide guidance for researchers and clinicians in the development of ASC-based therapeutic approaches.

关键词： White adipose tissue Metabolic diseases Obesity Adipose-derived

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Isolation and identification of Adipose-derived stromal/stem cells from breast cancer patients after exposure neoadjuvant chemotherapy

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World Journal of Experimental Medicine 2020年第3期10卷 26-40页

作者： Ashleigh Rapp Hagaman Ping Zhang Kiavash R Koko Ryan S Nolan Marc W Fromer John Gaughan Martha Matthews Department of Surgery Cooper University HospitalCamdenNJ 08103United States Cooper Medical School of Rowan University CamdenNJ 08103United States

BACKGROUND With recent research advances,Adipose-derived stromal/stem cells(ASCs)have been demonstrated to facilitate the survival of fat grafts and thus are increasingly used for reconstructive procedures following surgery for breast cancer.Unfortunately,in patients,following radiation and chemotherapy for breast cancer suggest that these cancer treatment therapies may limit stem cell cellular functions important for soft tissue wound healing.For clinical translation to patients that have undergone cancer treatment,it is necessary to understand the effects of these therapies on the ASC's ability to improve fat graft survival in clinical practice.AIM To investigate whether the impact on ASCs function capacity and recovery in cancer patients may be due to the chemotherapy.METHODS ASCs were isolated from the cancerous side and noncancerous side of the breast from the same patients with receiving neoadjuvant chemotherapy(NAC)or notreceiving NAC.ASCs were in vitro treated with 5-fluorouracil(5-FU),doxorubicin(DXR),and cyclophosphamide(Cytoxan)at various concentrations.The stem cells yield,cell viability,and proliferation rates were measured by growth curves and MTT assays.Differentiation capacity for adipogenesis was determined by qPCR analysis of the specific gene markers and histological staining.RESULTS No significant differences were observed between the yield of ASCs in patients receiving NAC treatment and not-receiving NAC.ASCs yield from the cancerous side of the breast showed lower than the noncancerous side of the breast in both patients receiving NAC and not-receiving NAC.The proliferation rates of ASCs from patients didn’t differ much before and after NAC upon in vitro culture,and these cells appeared to retain the capacity to acquire adipocyte traits simile to the ASCs from patients not-receiving NAC.After cessation and washout of the drugs for another a week of culturing,ASCs showed a slow recovery of cell growth capacity in 5-FU-treated groups but was not observed in ASCs treated with DXR groups.CONCLUS

关键词： Human Adipose-derived stromal/stem cells Breast cancer Neoadjuvant chemotherapy Chemotherapeutic agents Adipogenic differentiation Proliferation

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Platelet-rich plasma enhances Adipose-derived stem cell-mediated angiogenesis in a mouse ischemic hindlimb model

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World Journal of Stem Cells 2018年第12期10卷 212-227页

作者： Chia-Fang Chen Han-Tsung Liao Department of Plastic and Reconstructive Surgery Chang Gung Memorial Hospital Craniofacial Research Center Chang Gung Memorial Hospital College of Medicine Chang Gung University

AIM To evaluate the angiogenic effect of platelet-rich plasma(PRP)-preconditioned Adipose-derived stem cells(ADSCs) both in vitro and in a mouse ischemic hindlimb model.METHODS ADSCs were divided based on culture medium: 2.5% PRP, 5% PRP, 7.5% PRP, and 10% PRP. Cell proliferation rate was analyzed using the MTS assay. The gene expression of CD31, vascular endothelial growth factor, hypoxia-inducible factors, and endothelial cell nitric oxide synthase was analyzed using reverse transcription polymerase chain reaction. Cell markers and structural changes were assessed through immunofluorescence staining and the tube formation assay. Subsequently, we studied the in vivo angiogenic capabilities of ADSCs by a mouse ischemic hindlimb model.RESULTS The proliferation rate of ADSCs was higher in the 2.5%, 5%, and 7.5% PRP groups. The expression of hypoxia-inducible factor, CD31, vascular endothelial growth factor, and endothelial cell nitric oxide synthase in the 5% and 7.5% PRP groups increased. The 5%, 7.5%, and 10% PRP groups showed higher abilities to promote both CD31 and vascular endothelial growth factor production and tubular structure formation in ADSCs. According to laser Doppler perfusion scan, the perfusion ratios of ischemic limb to normal limb were significantly higher in 5% PRP, 7.5% PRP, and human umbilical vein endothelial cells groups compared with the negative control and fetal bovine serum(FBS) groups(0.88 ± 0.08, 0.85 ± 0.07 and 0.81 ± 0.06 for 5%, 7.5% PRP and human umbilical vein endothelial cells compared with 0.42 ± 0.17 and 0.54 ± 0.14 for the negative control and FBS, P < 0.01).CONCLUSION PRP-preconditioned ADSCs presented endothelial cell characteristics in vitro and significantly improved neovascularization in ischemic hindlimbs. The optimal angiogenic effect occurred in 5% PRP-and 7.5% PRPpreconditioned ADSCs.

关键词： Platelet-rich plasma Adipose-derived stem cells Mesenchymal stem cell Angiogenesis Endothelial differentiation Mouse ischemic hindlimb model

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Intra-articular transplantation of porcine Adipose-derived stem cells for the treatment of canine osteoarthritis: A pilot study

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World Journal of Transplantation 2014年第3期4卷 196-205页

作者： Shen-Yang Tsai Yun-Ching Huang Ling-Ling Chueh Lih-Seng Yeh Ching-Shwun Lin Institute of Veterinary Clinical Sciences School of Veterinary Medicine National Taiwan University Division of Urology Department of SurgeryChang Gung Memorial Hospital Graduate Institute of Veterinary Medicine School of Veterinary Medicine National Taiwan University Department of Urology School of MedicineUniversity of California San Francisco CA 94143-0738 United States

AIM: To test whether intra-articular injection of porcine Adipose-derived stem cells(ADSCs) can treat canine osteoarthritis(OA).METHODS: To enroll in this study dogs must have stifle joint OA that had lasted ≥ 3 mo and been treated with OA medication without significant improvement. Three dogs fulfilled these criteria and were thus subjects for ADSCs treatment. ADSCs were isolated from abdominal adipose tissue of a 2-mo-old female Yorkshire pig. Their stem cell marker expression was examined by immunofluorescence staining. For treatment, 5 million ADSCs were injected into the diseased joint of each dog. In the next 48 h, the patient was observed for signs of inflammatory and allergic reactions. Thepatient was then discharged to the owner and, at 2, 6, and 12 wk, followed up with orthopedic assessment, owner questionnaire, X-ray imaging, and force-plate gait analysis.RESULTS: Porcine ADSCs expressed mesenchymal stem cell markers CD90 and CD105. Injection of porcine ADSCs into canine stifle joints did not cause any inflammatory or allergic reactions. Orthopedic evaluation found improvements in two dogs, particularly at the longest time point. Owners' evaluation found increased capacity and decreased pain in all three dogs' activities such as walking and running. Radiographic evaluation did not find statistically significant differences before and after treatment. Force-plate analysis found significant improvements in all three dogs after treatment.CONCLUSION: Xenotransplantation of ADSCs for the treatment of OA is feasible. Further studies are needed to validate this novel treatment modality, which can then be implemented for the routine treatment of OA in veterinary medicine.

关键词： Canine osteoarthritis Adipose-derived stem cells Intra-articular transplantation Intra-articular injection Veterinary clinical study

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0Adipose-derived stem cells: Implications in tissue regeneration

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World Journal of Stem Cells 2014年第3期6卷 312-321页

作者： Wakako Tsuji J Peter Rubin Kacey G Marra adipose Stem Cell Center Department of Plastic Surgery University of Pitts-burgh McGowan Institute for Regenerative Medicine University of Pittsburgh Department of Bioengineering University of Pittsburgh

Adipose-derived stem cells(ASCs) are mesenchymal stem cells(MSCs) that are obtained from abundant adipose tissue, adherent on plastic culture flasks, can be expanded in vitro, and have the capacity to differ-entiate into multiple cell lineages. Unlike bone marrow-derived MSCs, ASCs can be obtained from abundant adipose tissue by a minimally invasive procedure, which results in a high number of cells. Therefore, ASCs are promising for regenerating tissues and organs dam-aged by injury and diseases. This article reviews the implications of ASCs in tissue regeneration.

关键词： Mesenchymal stem cells Adipose-derived stem cells Differentiation Growth factors Tissue engi-neering Clinical trials

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