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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

Co-culture and characterization of rabbit urethral epithelial cells on dHAS. (A) dHAS seeded with rabbit urethral epithelial cells were observed under an inverted microscope 2 weeks after co-culture; (B) Cell culture with epithelium origin was confirmed by red fluorescence of cytokeratin 18 (blue: DAPI, magnification at ×400).
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f2-medscimonit-20-2430: Co-culture and characterization of rabbit urethral epithelial cells on dHAS. (A) dHAS seeded with rabbit urethral epithelial cells were observed under an inverted microscope 2 weeks after co-culture; (B) Cell culture with epithelium origin was confirmed by red fluorescence of cytokeratin 18 (blue: DAPI, magnification at ×400).

Mentions: Rabbit urethral epithelial cells were successfully collected after trypsinization. The immunostaining using cytokeratin 18 confirmed the epithelial origin of harvested cells (Figure 2A). In the first few days of co-culture with rabbit urethral epithelial cells and dHAS, slow growth of urethral epithelial cells was observed. By the seventh day, colonies of urethral epithelial cells began to form on the surface of dHAS. After 2 weeks of co-culture, a confluent layer of urethral epithelial cells was found on the entire surface of the dHAS sheet (Figure 2B).


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)

Co-culture and characterization of rabbit urethral epithelial cells on dHAS. (A) dHAS seeded with rabbit urethral epithelial cells were observed under an inverted microscope 2 weeks after co-culture; (B) Cell culture with epithelium origin was confirmed by red fluorescence of cytokeratin 18 (blue: DAPI, magnification at ×400).
© Copyright Policy
Related In: Results  -  Collection

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

f2-medscimonit-20-2430: Co-culture and characterization of rabbit urethral epithelial cells on dHAS. (A) dHAS seeded with rabbit urethral epithelial cells were observed under an inverted microscope 2 weeks after co-culture; (B) Cell culture with epithelium origin was confirmed by red fluorescence of cytokeratin 18 (blue: DAPI, magnification at ×400).
Mentions: Rabbit urethral epithelial cells were successfully collected after trypsinization. The immunostaining using cytokeratin 18 confirmed the epithelial origin of harvested cells (Figure 2A). In the first few days of co-culture with rabbit urethral epithelial cells and dHAS, slow growth of urethral epithelial cells was observed. By the seventh day, colonies of urethral epithelial cells began to form on the surface of dHAS. After 2 weeks of co-culture, a confluent layer of urethral epithelial cells was found on the entire surface of the dHAS sheet (Figure 2B).

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