Korean J Urol.  2007 Apr;48(4):433-438. 10.4111/kju.2007.48.4.433.

Augmentation Cystoplasty using Hydroxapatite/chitosan Composite Sheet Seeded with Autologous Muscle-derived Stem Cells

• Byun SS
• Chung YS
• Lee SS
• Lee HN
• Lee JY
• Lee JY

• KMID: 1997112
• DOI: http://doi.org/10.4111/kju.2007.48.4.433

Abstract

PURPOSE
This study was designed to investigate the feasibility of a hydroxyapatite/chitosan (HAp/chitosan) composite, seeded with autologous muscle-derived stem cells, as a partial bladder substitute in rats.
MATERIALS AND METHODS
Muscle-derived stem cells were isolated from the gastrocnemius muscle of 6 female Sprague-Dawley rats, using the preplate technique, and cultured on HAp/chitosan composite sheets. Sheets with 10mm diameters were implanted into the urinary bladder of rats following a hemicystectomy in an autologous fashion. Three rats were sacrificed 4 and 8 weeks postoperatively, and the morphological changes subsequently assessed by H&E and immunofluorescence staining using DAPI, myogenin and alpha-smooth muscle actin (SMA).
RESULTS
All rats survived the scheduled duration. Adequate epithelialization was observed to be completed after postoperative week 4. Abundant muscle bundles, showing positive alpha-SMA staining, were observed after the 4th week. The bladder shape was well preserved after the 8th week. Ingrowing smooth muscles were observed on the periphery of the composite and muscular bundles, with positive myogenin immunostaining in the middle of the composite.
CONCLUSIONS
A HAp/chitosan composite sheet, seeded with autologous muscle-derived stem cells, showed a degree of skeletal muscle differentiation 8 weeks after augmentation cystoplasty, in an autologous fashion. This new material seeded with muscle-derived stem cells may, in the future, prove to be a viable option as a partial bladder substitute.

Keyword

Bladder; Atrophy; Regeneration; Stem cells

MeSH Terms

Actins
Animals
Atrophy
Female
Fluorescent Antibody Technique
Humans
Muscle, Skeletal
Muscle, Smooth
Myogenin
Rats
Rats, Sprague-Dawley
Regeneration
Stem Cells*
Urinary Bladder
Actins
Myogenin

Figure

• 

  Fig. 1. Postoperative morphological changes in a rat bladder implanted with a hydroxyapatite/chitosan (HAp/chitosan) composite, seeded with autologous muscle-derived stem cells, as a partial bladder substitute. (A) Scattered muscular bundles on the HAp/chitosan composite sheet are observed in the 4-week postoperative specimen. The black arrows indicate the boundary of the composite sheet seeded with muscle-derived stem cells. The broken arrow indicates adipose tissue surrounding the sheet (H&E, x40). (B) Ingrowing muscular bundles are observed at the periphery of the composite sheet; the black arrow indicates adipose tissue surrounding the sheet (H&E, x20).

• 

  Fig. 2. Immunostaining of the 4-week postoperative specimen. (A) Scattered, unorganized muscular bundles are observed (H&E, x100). (B) Muscular bundles show positive DAPI and α -SMA staining. (C) Very weak myogenin staining is observed in the sheet.

• 

  Fig. 3. Immunostaining of the 8-week postoperative specimen. (A) Complete luminal epithelialization is observed. The bladder shape is well preserved (H&E, x20). (B) Ingrowing muscular bundles are observed at the periphery of the sheet (H&E, x100). (C) Area showing positive DAPI and α -SMA staining. (D) Myogenin immunostaining reveals negative expression of the muscular bundles. (E) Connective tissue is predominant in this part (H&E, x100). (F) No α -SMA immunostaining is evident. (G) Weak myogenin immunostaining is observed in this area.

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