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Urethral reconstruction with tissue-engineered human amniotic scaffold in rabbit urethral injury models.

Wang F, Liu T, Yang L, Zhang G, Liu H, Yi X, Yang X, Lin TY, Qin W, Yuan J - Med. Sci. Monit. (2014)

Bottom Line: After the successful acquisition of dHAS from AM, cell-seeded dHAS were prepared and characterized.Immune responses were compared by histological evaluation and CD4 cell and CD8 cell infiltrations.Histopathological analysis revealed mild immune response in cell-seeded dHAS group (P<0.05).

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, Xijing Hospital, Fourth Military Medical University, Xi'an, China (mainland).

ABSTRACT

Background: Mitigating urethral injury remains a great challenge for urologists due to lack of ideal biomaterials for urethroplasty. The application of amniotic membranes (AM) over other synthetic materials make it a better potential source for urethral reconstruction. We separated the basement layer of AM to obtain denuded human amniotic scaffold (dHAS) and then inoculated primary rabbit urethral epithelial cells on the surface of dHAS to define whether this strategy minimize potential rejection and maximize the biocompatibility of human AM.

Material/methods: After the successful acquisition of dHAS from AM, cell-seeded dHAS were prepared and characterized. Both cell-seeded dHAS and acellular dHAS were subcutaneously implanted. Immune responses were compared by histological evaluation and CD4 cell and CD8 cell infiltrations. Then they were applied as urethroplastic materials in the rabbit models of urethral injury to fully explore the feasibility and efficacy of tissue-engineered dHAS xenografts in urethral substitution application.

Results: Mild inflammatory infiltration was observed in cell-seeded dHAS grafts, as revealed by fewer accumulations of CD4 cells and CD8 cells (or neutrophils or other immune cells). Urethral defects of rabbits in the urethroplastic group with dHAS implantation (n=6) were completely resolved in one month, while there were one infection and one fistula in the control group with acellular dHAS patches (n=6). Histopathological analysis revealed mild immune response in cell-seeded dHAS group (P<0.05).

Conclusions: Tissue-engineered dHAS minimize potential rejection and maximize the biocompatibility of AM, which makes it a potential ideal xenograft for urethral reconstruction.

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Related in: MedlinePlus

Urethral reconstruction using cell-seeded dHAS in a rabbit urethral injury model. (A) Urethral reconstruction surgery utilizing a cell-seeded dHAS patch. (B) immunohistochemical evaluation of CD4+ and CD8+ infiltrations 2 weeks after urethroplasty. Significant accumulation of CD4+ cells and CD8+ cells in the dHAS group were found (* P<0.05); (C) HE staining of cell-seeded dHAS graft 2 weeks after surgery. The graft was intact without obvious infiltration of inflammatory cells. The continuity of epithelial cells was recovered (magnification at ×200); (D) HE staining of dHAS patch 2 weeks after surgery revealed serious inflammatory infiltration (magnification at ×200).
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f5-medscimonit-20-2430: Urethral reconstruction using cell-seeded dHAS in a rabbit urethral injury model. (A) Urethral reconstruction surgery utilizing a cell-seeded dHAS patch. (B) immunohistochemical evaluation of CD4+ and CD8+ infiltrations 2 weeks after urethroplasty. Significant accumulation of CD4+ cells and CD8+ cells in the dHAS group were found (* P<0.05); (C) HE staining of cell-seeded dHAS graft 2 weeks after surgery. The graft was intact without obvious infiltration of inflammatory cells. The continuity of epithelial cells was recovered (magnification at ×200); (D) HE staining of dHAS patch 2 weeks after surgery revealed serious inflammatory infiltration (magnification at ×200).

Mentions: Urethral injury was induced and then urethroplasty was performed successfully in all rabbits (Figure 5A). They exhibited normal behaviors during the 3-month follow-up examination. One rabbit developed serious infection and another was found with fistula in the group that received dHAS patches. Neither infection nor fistula was observed in the group with cell-seeded dHAS implantation. Two weeks after surgery, cell-seeded dHAS was intact without obvious inflammatory cell infiltration, as confirmed by immunohistochemical evaluation of CD4+ cell and CD8+ cell infiltrations (Figure 5B). The anastomosis repair between the defective and normal urethral tissue was revealed by HE staining in the cell-seeded dHAS group (Figure 5C), in contrast to few repairs in the control group using dHAS (Figure 5D). Three months after surgery, the urethral defect was completely repaired in the cell-seeded dHAS group. The formation of a smooth muscular layer and rich blood vessels was apparent.


Urethral reconstruction with tissue-engineered human amniotic scaffold in rabbit urethral injury models.

Wang F, Liu T, Yang L, Zhang G, Liu H, Yi X, Yang X, Lin TY, Qin W, Yuan J - Med. Sci. Monit. (2014)

Urethral reconstruction using cell-seeded dHAS in a rabbit urethral injury model. (A) Urethral reconstruction surgery utilizing a cell-seeded dHAS patch. (B) immunohistochemical evaluation of CD4+ and CD8+ infiltrations 2 weeks after urethroplasty. Significant accumulation of CD4+ cells and CD8+ cells in the dHAS group were found (* P<0.05); (C) HE staining of cell-seeded dHAS graft 2 weeks after surgery. The graft was intact without obvious infiltration of inflammatory cells. The continuity of epithelial cells was recovered (magnification at ×200); (D) HE staining of dHAS patch 2 weeks after surgery revealed serious inflammatory infiltration (magnification at ×200).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4257484&req=5

f5-medscimonit-20-2430: Urethral reconstruction using cell-seeded dHAS in a rabbit urethral injury model. (A) Urethral reconstruction surgery utilizing a cell-seeded dHAS patch. (B) immunohistochemical evaluation of CD4+ and CD8+ infiltrations 2 weeks after urethroplasty. Significant accumulation of CD4+ cells and CD8+ cells in the dHAS group were found (* P<0.05); (C) HE staining of cell-seeded dHAS graft 2 weeks after surgery. The graft was intact without obvious infiltration of inflammatory cells. The continuity of epithelial cells was recovered (magnification at ×200); (D) HE staining of dHAS patch 2 weeks after surgery revealed serious inflammatory infiltration (magnification at ×200).
Mentions: Urethral injury was induced and then urethroplasty was performed successfully in all rabbits (Figure 5A). They exhibited normal behaviors during the 3-month follow-up examination. One rabbit developed serious infection and another was found with fistula in the group that received dHAS patches. Neither infection nor fistula was observed in the group with cell-seeded dHAS implantation. Two weeks after surgery, cell-seeded dHAS was intact without obvious inflammatory cell infiltration, as confirmed by immunohistochemical evaluation of CD4+ cell and CD8+ cell infiltrations (Figure 5B). The anastomosis repair between the defective and normal urethral tissue was revealed by HE staining in the cell-seeded dHAS group (Figure 5C), in contrast to few repairs in the control group using dHAS (Figure 5D). Three months after surgery, the urethral defect was completely repaired in the cell-seeded dHAS group. The formation of a smooth muscular layer and rich blood vessels was apparent.

Bottom Line: After the successful acquisition of dHAS from AM, cell-seeded dHAS were prepared and characterized.Immune responses were compared by histological evaluation and CD4 cell and CD8 cell infiltrations.Histopathological analysis revealed mild immune response in cell-seeded dHAS group (P<0.05).

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, Xijing Hospital, Fourth Military Medical University, Xi'an, China (mainland).

ABSTRACT

Background: Mitigating urethral injury remains a great challenge for urologists due to lack of ideal biomaterials for urethroplasty. The application of amniotic membranes (AM) over other synthetic materials make it a better potential source for urethral reconstruction. We separated the basement layer of AM to obtain denuded human amniotic scaffold (dHAS) and then inoculated primary rabbit urethral epithelial cells on the surface of dHAS to define whether this strategy minimize potential rejection and maximize the biocompatibility of human AM.

Material/methods: After the successful acquisition of dHAS from AM, cell-seeded dHAS were prepared and characterized. Both cell-seeded dHAS and acellular dHAS were subcutaneously implanted. Immune responses were compared by histological evaluation and CD4 cell and CD8 cell infiltrations. Then they were applied as urethroplastic materials in the rabbit models of urethral injury to fully explore the feasibility and efficacy of tissue-engineered dHAS xenografts in urethral substitution application.

Results: Mild inflammatory infiltration was observed in cell-seeded dHAS grafts, as revealed by fewer accumulations of CD4 cells and CD8 cells (or neutrophils or other immune cells). Urethral defects of rabbits in the urethroplastic group with dHAS implantation (n=6) were completely resolved in one month, while there were one infection and one fistula in the control group with acellular dHAS patches (n=6). Histopathological analysis revealed mild immune response in cell-seeded dHAS group (P<0.05).

Conclusions: Tissue-engineered dHAS minimize potential rejection and maximize the biocompatibility of AM, which makes it a potential ideal xenograft for urethral reconstruction.

Show MeSH
Related in: MedlinePlus