Clinical trial NCT03698721: Urothelium Tissue Engineering Using Biopsies From Transurethral Resection of Prostate
Official title: Urothelium Tissue Engineering Using Bladder Mucosa From Transurethral Resection of Prostate
Study type: OBSERVATIONAL
Overall status: NOT_YET_RECRUITING
Conditions: Spina Bifida
Urothelial Neoplasm
Bladder, Neurogenic
Bladder Exstrophy
Hypospadias
Keywords: Tissue engineering
Collagen scaffold
Alginate scaffold
Urothelial cells
Lead sponsor: Central Hospital, Nancy, France
Sponsor class: OTHER
First posted: 2018-10-09
Enrollment: 365
Sex: MALE
Minimum age (years): 18
Standard ages: ADULT
OLDER_ADULT
Accepts healthy volunteers: no
Observational model: OTHER
Time perspective: PROSPECTIVE
Sampling method: NON_PROBABILITY_SAMPLE
Study population: Male patient requiring TURP for Benign Prostatic Hyperplasia with prostate weight evaluated preoperatively greater than or equal to 30 grams

Brief summary: Different clinical conditions can require urinary bladder augmentation or replacement. Tissue engineered bladder has been clinically evaluated but is not recommended due to diverse side effects. Thus, there is a real interest for the development of regenerative approach with innovative scaffolds and cell transplantation.

The investigators propose the use of urothelial cells obtained by Trans-Urethral Resection of Prostate or bladder (TURP) to obtain a tissue engineered urothelium in association with different scaffolds.

Detailed description: Bladder biopsies will be obtained during cystoscopy, conserved in culture medium (DMEM®), digested by dispase and sowed on collagen-coated culture support. Keratinocyte Serum Free Medium (KSFM) will be used for proliferation. Microscopy, immunohistochemistry, RNA extraction, Reverse Transcription and quantitative Polymerase Chain Reaction (RT-qPCR) will be performed during passages. Cell culture conditions will be optimized to improve proliferation and avoid loss of differentiation. The investigators will develop scaffolds based on sodium alginate hydrogels, followed by freeze-drying to generate porous sponges (at -20°C and -80°C). Cultured cells will be associated to these original scaffolds and to other scaffolds, for example alginate hydrogels or Collagen Cell Carrier (CCC), cultivated for 28 days and analyzed. Histological and immunohistological appearance of cellularized scaffolds will be compared to assess the effectiveness of each scaffold for tissue engineering in urothelium.

Cellularized scaffolds will be studied in vitro (Transepithelial Electrical Resistance, impermeability, ability to be stitched, resistance to urine) and in vivo in ectopic location (subcutaneous location in Nude mice) or in orthotopic location (bladder augmentation in small animal).

Eligibility criteria: Inclusion Criteria:

* Patient needing a Transurethral Resection of Prostate (TURP)
* Weight of prostate (evaluated by ultrasonography)greater than or equal to 30 grams
* Affiliation to a social security system
* Patient over the age of majority
* Patient receiving complete information on research organization without opposition to the use of biological specimen

Exclusion Criteria:

* Patient for whom no TURP is realized during endoscopic procedure
* Patient for whom no resection is realized on the bladder neck
* Patient with prostate weight estimated under 30 grams
* Patient who opposed to the realization of the study

Primary outcomes:
measure: Histological analysis of biopsy.
timeFrame: 6 month
Secondary outcomes:
measure: Immunohistological analysis of biopsy.
timeFrame: 6 month
measure: RT-qPCR analysis of biopsy.
timeFrame: 6 month
measure: Digestion of the biopsy and culture with adapted medium. Optimization of culture conditions.
timeFrame: 12 months
measure: Histological analysis of the cultured cells.
timeFrame: 12 months
measure: Immunocytological analysis of the cultured cells.
timeFrame: 12 months
measure: RT-qPCR analysis of the cultured cells.
timeFrame: 12 months
measure: Development of original alginate freezed-dried scaffold
timeFrame: 12 months
measure: Association of the cultured cells with different scaffolds. Culture of cellularized scaffolds.
timeFrame: 36 months
measure: Histological analysis of cellularized scaffolds
timeFrame: 36 months
measure: Immunohistological analysis of cellularized scaffolds
timeFrame: 36 months
measure: RT-qPCR analysis of cellularized scaffolds
timeFrame: 36 months
measure: Biophysical analysis of cellularized scaffolds
timeFrame: 36 months
measure: Implantation of the cellularized scaffold in Nude mice
timeFrame: 12 months
measure: Histological analysis of implanted scaffolds
timeFrame: 12 months
measure: Immunohistological analysis of implanted scaffolds
timeFrame: 12 months
measure: RT-qPCR analysis of implanted scaffolds
timeFrame: 12 months
Other outcomes:
Locations:
Study officials:
References:
citation: Adamowicz J, Kowalczyk T, Drewa T. Tissue engineering of urinary bladder - current state of art and future perspectives. Cent European J Urol. 2013;66(2):202-6. doi: 10.5173/ceju.2013.02.art23. Epub 2013 Aug 13.
citation: Lam Van Ba O, Aharony S, Loutochin O, Corcos J. Bladder tissue engineering: a literature review. Adv Drug Deliv Rev. 2015 Mar;82-83:31-7. doi: 10.1016/j.addr.2014.11.013. Epub 2014 Nov 14.
citation: Garthwaite M, Hinley J, Cross W, Warwick RM, Ambrose A, Hardaker H, Eardley I, Southgate J. Use of donor bladder tissues for in vitro research. BJU Int. 2014 Jan;113(1):160-6. doi: 10.1111/bju.12285.
citation: Baker SC, Shabir S, Southgate J. Biomimetic urothelial tissue models for the in vitro evaluation of barrier physiology and bladder drug efficacy. Mol Pharm. 2014 Jul 7;11(7):1964-70. doi: 10.1021/mp500065m. Epub 2014 Apr 17.