Urothelium Tissue Engineering Using Biopsies From Transurethral Resection of Prostate

Официальное название

Urothelium Tissue Engineering Using Bladder Mucosa From Transurethral Resection of Prostate

Обзор

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.

Подробное описание

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).

Вмешательства

• Процедура Transurethral Resection of Prostate
  Transurethral resection of the prostate is a urological operation used to treat benign prostatic hyperplasia (BPH). It is performed by visualising the prostate through the urethra and removing tissue by electrocautery or sharp dissection with a resectoscope. This is considered the most effective treatment for BPH. This procedure is done with spinal or general anaesthetic. A triple lumen catheter is inserted through the urethra to irrigate and drain the bladder after the surgical procedure is com

Первичные конечные точки

• Histological analysis of biopsy. [Срок оценки: 6 month]

Вторичные конечные точки (12)

• Immunohistological analysis of biopsy. [Срок оценки: 6 month]
• RT-qPCR analysis of biopsy. [Срок оценки: 6 month]
• Digestion of the biopsy and culture with adapted medium. Optimization of culture conditions. [Срок оценки: 12 months]
• Histological analysis of the cultured cells. [Срок оценки: 12 months]
• Immunocytological analysis of the cultured cells. [Срок оценки: 12 months]
• RT-qPCR analysis of the cultured cells. [Срок оценки: 12 months]
• Development of original alginate freezed-dried scaffold [Срок оценки: 12 months]
• Association of the cultured cells with different scaffolds. Culture of cellularized scaffolds. [Срок оценки: 36 months]
• Histological analysis of cellularized scaffolds [Срок оценки: 36 months]
• Immunohistological analysis of cellularized scaffolds [Срок оценки: 36 months]
• RT-qPCR analysis of cellularized scaffolds [Срок оценки: 36 months]
• Biophysical analysis of cellularized scaffolds [Срок оценки: 36 months]

Критерии участия

Критерии включения

• 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

Критерии исключения

• 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

Критерии приведены из реестра в оригинале (на английском). Окончательную оценку соответствия проводит исследовательский центр.

Здоровые добровольцы: Нет

Дизайн исследования

Модель наблюдения

Другое

Центры проведения

Список центров уточняется — проверьте первичный протокол.

Публикации

• 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. PMID 24579029
• 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. PMID 25446136
• 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. PMID 24053725
• 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. PMID 24697150

Идентификаторы

NCT: NCT03698721 · PSS2017/IMOPU-BERTE/MS

Первоисточники (государственные реестры)

• ClinicalTrials.gov (NIH, США)

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