Fabrication of Tissue-Engineered Bionic Urethra Using Cell Sheet Technology and Labeling By Ultrasmall Superparamagnetic Iron Oxide for Full-Thickness Urethral Reconstruction

Zhou, Shengyan; Yang, Ranxin; Zou, Qingsong; Zhang, Kaile; Yin, Ting; Zhao, Weixin; Shapter, Joseph G.; Gao, Guo; Fu, Qiang

doi:10.7150/thno.18833

Cited by 50 publications

(40 citation statements)

References 58 publications

(56 reference statements)

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“…Oral mucosal epithelial cells were harvested from canine oral mucosal epithelium and prepared as reported previously [19]. Oral mucosal epithelial cells were obtained from the buccal mucosa of beagle dogs.…”

Section: Methodsmentioning

confidence: 99%

The Fabrication and Evaluation of a Potential Biomaterial Produced with Stem Cell Sheet Technology for Future Regenerative Medicine

Zhou

Wang

Zhang

et al. 2020

Stem Cells International

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To date, the decellularized scaffold has been widely explored as a source of biological scaffolds for regenerative medicine. However, the acellular matrix derived from natural tissues and organs has a lot of defects, including the limited amount of autogenous tissue and surgical complication such as risk of blood loss, wound infection, pain, shock, and functional damage in the donor part of the body. In this study, we prepared acellular matrix using adipose-derived stem cell (ADSC) sheets and evaluate the cellular compatibility and immunoreactivity. The ADSC sheets were fabricated and subsequently decellularized using repeated freeze-thaw, Triton X-100 and SDS decellularization. Oral mucosal epithelial cells were seeded onto the decellularized ADSC sheets to evaluate the cell replantation ability, and silk fibroin was used as the control. Then, acellular matrix was transplanted onto subcutaneous tissue for 1 week or 3 weeks; H&E staining and immunohistochemical analysis of CD68 expression and quantitative real-time PCR (qPCR) were performed to evaluate the immunogenicity and biocompatibility. The ADSC sheet-derived ECM scaffolds preserved the three-dimensional architecture of ECM and retained the cytokines by Triton X-100 decellularization protocols. Compared with silk fibroin in vitro, the oral mucosal epithelial cells survived better on the decellularized ADSC sheets with an intact and consecutive epidermal cellular layer. Compared with porcine small intestinal submucosa (SIS) in vivo, the homogeneous decellularized ADSC sheets had less monocyte-macrophage infiltrating in vivo implantation. During 3 weeks after transplantation, the mRNA expression of cytokines, such as IL-4/IL-10, was obviously higher in decellularized ADSC sheets than that of porcine SIS. A Triton X-100 method can achieve effective cell removal, retain major ECM components, and preserve the ultrastructure of ADSC sheets. The decellularized ADSC sheets possess good recellularization capacity and excellent biocompatibility. This study demonstrated the potential suitability of utilizing acellular matrix from ADSC sheets for soft tissue regeneration and repair.

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Section: Methodsmentioning

confidence: 99%

The Fabrication and Evaluation of a Potential Biomaterial Produced with Stem Cell Sheet Technology for Future Regenerative Medicine

Zhou

Wang

Zhang

et al. 2020

Stem Cells International

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“…Some animal studies indicated that BM-EPC cell sheet technology increased vasculogenesis [12]. However, BM-EPCs cell sheet can not harvested with trypsin and cell-to-cell interactions and the integral structure of the BM-EPCs sheet were unknown [13,14].…”

Section: Introductionmentioning

confidence: 99%

Fabrication and Paracrine Effect Research of Bone Marrow-Derived Endothelial Progenitor Cell Sheet

Xue¹,

Yin²,

Bai³

et al. 2020

Preprint

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Background: The release of a wide array of endothelial progenitor cell (EPC) sheet-secreted paracrine factors is central to the mechanism by which these cells contribute to tissue repair. The purpose of this study was to fabricate BM-EPC sheet and conduct preliminary investigation on the paracrine effect of SDF-1ɑ about the role of the stromal cell-derived factor-1α (SDF-1α)/CXCR4 axis in the form of tubular structures from BM-EPCs sheet.Methods: EPCs derived from rat bone marrow (BM-EPC) were identified and isolated by the cell-surface markers CD34/CD133/VE-cadherin/KDR using flow cytometry, as well as by dual affinity for acLDL and UEA-1. Single-cell suspensions were seeded on temperature-responsive cell culture dishes. After 7 days of incubation, the BM-EPC cells were easily harvested as cell sheets, and a series of biochemical experiments were performed in vitro. The expression levels of SDF-1α/CXCR4 axis-associated genes and proteins were examined using RT‑qPCR and western blot analysis, and enzyme-linked immunosorbent assay (ELISA) was applied to determine the concentrations of VEGF, EGF and SDF-1α in the cell culture medium.Results: The BM-EPC cell sheets were successfully harvested. Moreover, BM-EPC cell sheets have superior proliferation and tube formation activity when compare with single cell suspension. When capillary-like tube were formed from EPCs sheets, the releasing of paracrine factors such as VEGF, EGF, and SDF-1ɑ were increasing. To drive the tube formation of BM-EPCs sheets, our mechanism research showed that the activation of PI3K/AKT/eNOS pathway since the phosphorylation of protein CXCR, PI3K, AKT and eNOS were increasing. Conclusion: BM-EPC cell sheets have superior proliferation and tube formation activity that can be used for tissue repair. The strong ability to secrete paracrine factors was be related to the SDF-1α/CXCR4 axis through PI3K/AKT/eNOS pathway.

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“…Cell sheets have been widely used to repair defects with the development of tissue engineering (Joseph et al, 2006). Compared to scaffold and traditional cells composition, cell sheets feature uniform distribution of cells and complete ECM 3-dimensional structure (Zhou et al, 2017). BMSCs are dominant cells in the process of osseointegration and have advantages of accessibility, multiple differentiation potential, and immune-evasion.…”

Section: Discussionmentioning

confidence: 99%

Activation of Nell-1 in BMSC Sheet Promotes Implant Osseointegration Through Regulating Runx2/Osterix Axis

Lai

et al. 2020

Front. Cell Dev. Biol.

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Neural epidermal growth factor-like 1 protein (Nell-1) is first studied because of its association with human craniosynostosis. Nell-1 has been used to accelerate the process of fracture healing because of the osteoinductive ability in recent years. However, the role of Nell-1 during the process of osteointegration is unknown. Here we show that activation of Nell-1 in the BMSC sheet promotes osseointegration in vivo and in vitro. We found that overexpression of Nell-1 improved osteogenic differentiation and enhanced matrix mineralization of BMSCs through increasing expression of Runx2 and Osterix. Activation of Nell-1 up-regulated the expression ratio of OPG/RANKL, which might have a negative influence on osteoclast differentiation. Furthermore, we obtained BMSC sheet-implant complexes transfected with lentivirus overexpressing and interfering Nell-1 in in vivo study, and confirmed that overexpression of Nell-1 promoted new bone formation around the implant and increased the bone-implant contacting area percentage. Our results demonstrate that activation of Nell-1 improves implant osteointegration by regulating Runx2/Osterix axis and shows the potential of BMSC sheet-implant complexes in gene therapy.

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Fabrication of Tissue-Engineered Bionic Urethra Using Cell Sheet Technology and Labeling By Ultrasmall Superparamagnetic Iron Oxide for Full-Thickness Urethral Reconstruction

Cited by 50 publications

References 58 publications

The Fabrication and Evaluation of a Potential Biomaterial Produced with Stem Cell Sheet Technology for Future Regenerative Medicine

The Fabrication and Evaluation of a Potential Biomaterial Produced with Stem Cell Sheet Technology for Future Regenerative Medicine

Fabrication and Paracrine Effect Research of Bone Marrow-Derived Endothelial Progenitor Cell Sheet

Activation of Nell-1 in BMSC Sheet Promotes Implant Osseointegration Through Regulating Runx2/Osterix Axis

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