4.2 Article

Development of a porcine acellular bladder matrix for tissue-engineered bladder reconstruction

Journal

PEDIATRIC SURGERY INTERNATIONAL
Volume 38, Issue 5, Pages 665-677

Publisher

SPRINGER
DOI: 10.1007/s00383-022-05094-2

Keywords

Bladder augmentation; De-cellularisation; Extracellular matrix; Tissue engineering

Funding

  1. Fondazione Alberto Mascherpa Onlus
  2. Great Ormond Street Hospital Charity
  3. Fondazione Citta della Speranza
  4. MRC [MR/L013347/1, MR/N028414/1]
  5. NIHR Great Ormond Street Hospital Biomedical Research Centre
  6. Foundation Eugenio Litta
  7. MRC [MR/N028414/1, MR/L013347/1] Funding Source: UKRI

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A protocol for creating a natural-derived bladder extracellular matrix (BEM) was established for developing tissue-engineered bladder, and its structural and functional characteristics were investigated. The results showed that BEM retained structural characteristics, increased strength and stiffness, and had better compliance compared to native tissue, as well as angiogenic potential. These findings suggest the potential use of BEM for developing tissue-engineered bladder for patients requiring bladder augmentation.
Purpose Enterocystoplasty is adopted for patients requiring bladder augmentation, but significant long-term complications highlight need for alternatives. We established a protocol for creating a natural-derived bladder extracellular matrix (BEM) for developing tissue-engineered bladder, and investigated its structural and functional characteristics. Methods Porcine bladders were de-cellularised with a dynamic detergent-enzymatic treatment using peristaltic infusion. Samples and fresh controls were evaluated using histological staining, ultrastructure (electron microscopy), collagen, glycosaminoglycans and DNA quantification and biomechanical testing. Compliance and angiogenic properties (Chicken chorioallantoic membrane [CAM] assay) were evaluated. T test compared stiffness and glycosaminoglycans, collagen and DNA quantity. p value of < 0.05 was regarded as significant. Results Histological evaluation demonstrated absence of cells with preservation of tissue matrix architecture (collagen and elastin). DNA was 0.01 mu g/mg, significantly reduced compared to fresh tissue 0.13 mu g/mg (p < 0.01). BEM had increased tensile strength (0.259 +/- 0.022 vs 0.116 +/- 0.006, respectively, p < 0.0001) and stiffness (0.00075 +/- 0.00016 vs 0.00726 +/- 0.00216, p = 0.011). CAM assay showed significantly increased number of convergent allantoic vessels after 6 days compared to day 1 (p < 0.01). Urodynamic studies showed that BEM maintains or increases capacity and compliance. Conclusion Dynamic detergent-enzymatic treatment produces a BEM which retains structural characteristics, increases strength and stiffness and is more compliant than native tissue. Furthermore, BEM shows angiogenic potential. These data suggest the use of BEM for development of tissue-engineered bladder for patients requiring bladder augmentation.

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