Ajalloueian, Fatemeh

Chamorro, C. I.

Chronakis, Ioannis S.

Fossum, M.

Hilborn, Jöns

Online Research Database In Technology

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Downloaded from orbit.dtu.dk on: Dec 18, 2017A full-layer bladder wall patch by mincing both porcine bladder mucosa and detrusorin a natural-synthetic scaffoldAjalloueian, Fatemeh; Chamorro, C. I.; Chronakis, Ioannis S.; Hilborn, Jöns; Fossum, M.Published in:European Cells & MaterialsPublication date:2016Document VersionPublisher's PDF, also known as Version of recordLink back to DTU OrbitCitation (APA):Ajalloueian, F., Chamorro, C. I., Chronakis, I. S., Hilborn, J., & Fossum, M. (2016). A full-layer bladder wall patchby mincing both porcine bladder mucosa and detrusor in a natural-synthetic scaffold. European Cells &Materials, 31(1), 84-84.European Cells and Materials Vol. 31. Suppl. 1, 2016 (page 84)                                                             ISSN 1473-2262                                                  http://www.ecmjournal.org  A full-layer bladder wall patch by mincing both porcine bladder mucosa and detrusor in a natural-synthetic scaffold  F Ajalloueian1,2, C I Chamorro 2, I S Chonakis 1, J Hilborn 3 , M Fossum 2,4 1 Nano-BioScience Research Group, DTU-Food, Technical University of Denmark, Denmark. 2Department of Women's and Children's Health, Center of Molecular Medicine, Karolinska Institutet, Sweden. 3Division of Polymer Chemistry, Ångström Laboratory, Uppsala University, Sweden. 4Department of Pediatric Surgery, Astrid Lindgren Children’s Hospital, Sweden  INTRODUCTION: Full layer bladder regeneration has remained a challenge in bladder tissue engineering due to lack of or incomplete regeneration of smooth muscle cells, especially in large area defects [1, 2]. To create a suitable tissue transplant for bladder augmentation, we developed a tissue-engineered construct that includes 1) autologous urothelial cells for good barrier function, 2) autologous smooth muscle cells to regenerate a muscular layer, and 3) a biodegradable support that allows take of the transplant, tissue ingrowth and meets the need of low-pressure high-compliance storage function. METHODS: An electrospun PLGA sheet with optimized porosity [3] was placed on a semi-gel collagen inside a mold and covered by a second collagen hydrogel followed by distribution of minced detrusor from porcine bladder on top. Thereafter, third collagen hydrogel layer was added and minced mucosa  of porcine bladder was seeded on top. The whole construct underwent plastic compression (PC), and was incubated for 1, 2, and 4 weeks in vitro for electron microscopy, histology and immunoassay studies. RESULTS: SEM imaging was performed on both surfaces seeded by minced tissue. Imaging the top collagen surface demonstrated a complete coverage with proliferated urothelial cells after two (confluent) and four (multi- layered) weeks. SEM imaging on the middle-seeded surface also showed that, spindle-shaped smooth muscle cells completely covered the surface after two weeks.  Histology and immunoassays demonstrated that successful proliferation of both urothelial and smooth muscle cells was achieved. It was seen that cells migrated from the minced tissue specimens (either on top or inside the construct), proliferated, reorganized and formed a multilayer epithelium (4 weeks) as well as muscle cells. Furthermore, immunostaining for MNF116 and SMA indicated epithelial cell origin and smooth muscle cells respectively.  Fig. 1: A) SEM images (a) confluent urothelium, (b) smooth muscle cells and B) Cross-section of PLGA-PC collagen construct seeded with minced mucosa and detrusor after 2 weeks (Trichrome x10 magnification, (arrow) shows urothelium, (*) denotes muscle cells. DISCUSSION & CONCLUSIONS: With our previous achievements, based on mincing bladder mucosa to regenerate urothelium [3], we continued to develop the technique to regenerate bladder muscle, a tissue that is considered more challenging to expand. We successfully regenerated both urothelium and muscle cells in a hybrid nano-micro, natural-synthetic scaffold of compressed collagen-electrospun PLGA.   In a clinical setting the construct is easy to make and all steps: harvesting, mincing and constructing the hybrid scaffold could be performed just before transplantation in a one-stage procedure. REFERENCES: 1 P. Caione et al (2012) Pediatr Surg Int 28:421–8. 2 G.S. Jack et al (2009) Biomaterials 30: 3259-70. 3 F. Ajalloueian, et al (2014) Biomaterials 35: 5741-8 ACKNOWLEDGEMENTS: This work was supported by Danish Research Council Foundation (DFF–4093-00282), the Freemason Foundation for Children’s Welfare, the Stockholm City Council and the Swedish Society of Medicine.  

A full-layer bladder wall patch by mincing both porcine bladder mucosa and detrusor in a natural-synthetic scaffold

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A full-layer bladder wall patch by mincing both porcine bladder mucosa and detrusor in a natural-synthetic scaffold

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