Tissue engineering of functional intestine using a decellularized scaffold
Professor Paolo de Coppi
Description of Project/Background:
In the last two decades, regenerative medicine has shown the potential for ‘‘bench-to-bedside’’ translational research in specific clinical settings. Progress made in cell and stem cell biology, materials science, and tissue engineering enabled researchers to develop cutting-edge technology that has lead to the creation of non-modular tissue constructs such as skin, bladder, vessels and upper airway. In all cases, autologous cells were seeded on either artificial or natural supporting scaffolds. Engineering of modular organs (namely, organs organized in functioning units referred to as modules and requiring the reconstruction of the vascular supply) is more complex and challenging. Models of functioning hearts and livers have been engineered using ‘‘natural tissue’’ scaffolds and efforts are underway to produce kidneys, pancreata and small intestines.
There are several gastrointestinal conditions characterized by functional or anatomical loss of intestine. Examples include congenital or acquired absence of the oesophagus (e.g. long-gap oesophageal atresia, oesophageal cancer), massive small bowel resection (neonatal necrotizing enterocolitis, congenital intestinal atresia, midgut volvulus, inflammatory bowel disease, trauma and mesenteric vascular disease), fistulating diseases of the intestine, long segment Hirschsprungs’s disease and other intestinal neuropathies and myopathies (e.g. hollow visceral myopathy). Such conditions render the gut of many children and adults incapable of sustaining its nutritional function (i.e. intestinal failure), and such patients become transiently or permanently dependent on artificial feeding and parenteral nutrition.
Children and adults with intestinal failure often have a poor quality of life and suffer significant morbidity and mortality, generally related to complications of parenteral nutrition such as intravenous line sepsis and liver disease. Several operative interventions have been described to promote enteral feeding, but unfortunately none seem to have contributed significantly to improved survival. Alternative strategies such as small bowel transplantation are limited by the scarcity of donor organs, the need for aggressive immunosuppression, and the low survival rates. Thus the development of new treatment strategies for intestinal failure is an area requiring further investigation.
One option is the use of tissue engineering to generate functional gut tissue. This is now a realistic possibility given recent and significant advances in biomaterial science, bioreactor technology, and molecular biology including cell characterization, isolation and expansion. These methodologies are relevant not only to develop engineered intestine that could be used in patients but they could also become powerful tools for studying tissue and disease physiology in a controlled environment as well as being used for basic studies on tissue development and cell functions in response to genetic alterations, drugs and physiological stimuli.
Recent studies suggest that primary human cell systems can be designed to model many aspects of disease biology and that robust and automated assays can be engineered to detect and discriminate disease-relevant mechanisms and potential therapies. Human cell-based testing has proven to be a valuable tool to quickly explore toxic and non-functional compounds. The low-cost and high-speed testing of compounds in cell culture, and the obvious advantages of using intact cells as a primary representation of living patients, makes cell-based testing an appealing key component of therapy discovery programs. Studies have validated the potential of harvesting plugs of intestine, termed organoid units, seeding them on biodegradable scaffolds and implanting the engineered intestine into small animal models in order to substitute intestine.
However, newer approaches have emerged that utilize harvested stem cells to facilitate a more rapid generation of engineered tissues. We have applied this approach focusing specifically on individual cell components of the gut. Having investigated the role of various polymers we are now in a unique position to further explore the integration of the different components of the gut within three-dimensional constructs to generate functional intestine. We believe these models will help prevent the need for animal models as a necessary prerequisite to clinical trials on human patients since some of the limitations of the current animal models may be overcome by the use of human-derived cells and engineered tissues. Such substitutes will potentially give more reliable outcomes when responding to drug therapies and serve as promising prospects for increasing viable gut tissue, thus diminishing the need for parenteral nutrition or intestinal transplantation for patients with intestinal failure.
1. Cells derived from the amnion. Maghsoudlou P, De Coppi P. In Stem Cell Transplantation, 2012.
2. Tissue engineering and stem cell research. Maghsoudlou P, Loukogeorgakis SP, De Coppi P. In Seminars in Paediatric Surgery, 2013.
3. Tissue engineering for oesophageal diseases: a new option for oesophageal replacement. Totonelli G, Maghsoudlou P, De Coppi P. In Regenerative Medicine Applications in Organ Transplantation, 2013.
1. Amniotic Fluid Stem Cells Are Cardioprotective Following Acute Myocardial Infarction. Bollini S, Cheung KK, Riegler J, Dong X, Smart N, Ghionzoli M, Loukogeorgakis SP, Maghsoudlou P, Dubé KN, Riley PR, Lythgoe MF, De Coppi P. Stem Cells and Development. 2011 Nov; 20(11): 1985-94.
2. A rat decellularized small bowel scaffold that preserves villus-crypt architecture for intestinal regeneration. Totonelli G, Maghsoudlou P, Garriboli M, Riegler J, Orlando G, Burns AJ, Sebire NJ, Smith VV, Fishman JM, Ghionzoli M, Turmaine M, Birchall MA, Atala A, Soker S, Lythgoe MF, Seifalian A, Pierro A, Eaton S, De Coppi P. Biomaterials. 2012 Apr; 33(12): 3401-10.
3. Detergent enzymatic treatment for the development of a natural acellular matrix for oesophageal regeneration. Totonelli G, Maghsoudlou P, Georgiades F, Garriboli M, Koshy K, Turmaine M, Ashworth M, Sebire NJ, Pierro A, Eaton S, De Coppi P. Pediatric Surgery International. 2013 Jan; 29(1): 87-95.
4. Esophageal tissue engineering; a new approach for esophageal replacement. Totonelli G, Maghsoudlou P, Fishman JM, Orlando G, Ansari T, Sibbons P, Birchall MA, Pierro A, Eaton S, De Coppi P. World Journal of Gastroenterology. 2012 Dec; 18(47): 6900-7.
5. Skeletal muscle tissue engineering: which cell to use? Fishman JM*, Tyraskis A*, Maghsoudlou P, Urbani L, Totonelli G, Birchall MA, De Coppi P. Tissue Engineering Part B. 2013 Dec; 19(6): 503-15. (*These authors contributed equally)
6. Discarded human kidneys as a source of ECM scaffold for kidney regeneration technologies. Orlando G, Booth C, Wang Z, Totonelli G, Ross CL, Moran E, Salvatori M, Maghsoudlou P, Turmaine M, Delario G, Al-Shraideh Y, Farooq U, Farney AC, Rogers J, Iskandar SS, Burns A, Marini FC, De Coppi P, Stratta RJ, Soker S. Biomaterials. 2013 Aug; 34(24): 5915-25.
7. Preservation of micro-architecture and angiogenic potential in a pulmonary acellular matrix obtained using intermittent intra-tracheal flow of detergent enzymatic treatment. Maghsoudlou P, Georgiades F, Tyraskis A, Totonelli G, Loukogeorgakis SP, Orlando G, Shangaris P, Lange P, Delalande JM, Burns AJ, Cenedese A, Sebire NJ, Turmaine M, Guest BN, Alcorn JF, Atala A, Birchall MA, Elliott MJ, Eaton S, Pierro A, Gilbert TW, De Coppi P. Biomaterials. 2013 Sep; 34(28): 6638-48.
8. Recent developments in therapies with stem cells from amniotic fluid and placenta. Loukogeorgakis SP, Maghsoudlou P, De Coppi P. Fetal and Maternal Medicine. 2013 Aug; 24(03): 148-68.
9. A decellularization methodology for the production of a natural acellular intestinal matrix. Maghsoudlou P, Totonelli G, Loukogeorgakis SP, Eaton S, De Coppi P. Journal of Visualized Experiments. 2013 Oct; (80).
10. Person-specific health promotion in the health-care setting: an exploratory study. Razzaki S*, Theodorou A*, Maghsoudlou P, Protopapa E, Beynon H. The Lancet. 2013 Nov; 382: S83. (*These authors contributed equally)
11. Tissue engineering of the oesophagus. Maghsoudlou P, De Coppi P. Seminars in Pediatric Surgery. 2014 Jun; 23(3): 127-34.
12. Sheep CD34+ amniotic fluid cells have haematopoietic potential and engraft after autologous in utero transplantation. Shaw SW, Blundell MP, Pipino C, Shangaris P, Maghsoudlou P, Ramachandra DL, Georgiades F, Boyd M, Thrasher AJ, Porada CD, Almeida-Porada G, Cheng PJ, David AL, De Coppi P. Stem Cells. 2015 Jan; 33(1): 122-32.
13. Isolation of esophageal stem cells with potential for therapy. Maghsoudlou P, Ditchfield D, Klepacka DH, Shangaris P, Urbani L, Loukogeorgakis SP, Eaton S, De Coppi P. Pediatric Surgery International. 2014 Dec; 30(12): 1249-56.
14. Organ bioengineering for the newborn. Maghsoudlou P, Urbani L, De Coppi P. Seminars in Pediatric Surgery. 2014 Oct; 23(5): 314-23.
15. A potential platform for developing 3D tubular scaffolds for paediatric organ development. De Mel A, Yap T, Cittadella G, Hale LR, Maghsoudlou P, de Coppi P, Birchall MA, Seifalian AM. Journal of Materials Science. 2015 Mar; 26(3): 141.
16. Decellularized human liver as a natural scaffold for liver bioengineering and transplantation. Mazza G, Rombouts K, Hall A, Urbani L, Luong TV, Al-Akkad W, Longato, L, Brown D, Maghsoudlou P, Dhillon AP, Fuller B, Davidson B, Moore K, Dhar D, De Coppi P, Malago M, Pinzani M. Nature Scientific Reports. 2015 Aug; 5: 13079.
17. OP-1 artificial oesophagus engineering in a 3D dynamic culture. Camilli C, Urbani L, Scottoni F, Crowley C, Wong RR, Maghsoudlou P, Fallas ME, Lowdell M, Birchall M, Cossu G, De Coppi P. Journal of Pediatric Gastroenterology and Nutrition. 2015 Oct; 61(4): 509.
18. The human pancreas as a source of pro-tolerogenic extracellular matrix scaffold for a new generation bioartificial endocrine pancreas. Peloso A, Urbani L, Katari R, Maghsoudlou P, Alvarez M, Purroy C, Mcquilling J, Sittadjody S, Farney AC, Iskandar SS, Rogers J, Stratta RJ, Opara EC, Piemonti L, Soker S, De Coppi P. Annals of Surgery 2015 (In print).
19. X-ray phase contrast tomography for microstructural assessment of acellular organ scaffolds in regenerative medicine. Hagen CK, Maghsoudlou P, Totonelli G, Diemoz PC, Endrizzi M, Rigon L, Menk RH, Arfelli F, Dreossi D, Longo R, Brun E, Coan P, Bravin A, De Coppi P, Olivo A. Nature Scientific Reports 2015 (In print).
Awards & Prizes:
- Young Investigator Award - EUPSA conference 2013 - ‘Intestinal tissue engineering in vivo through the combination of mouse crypt organoid units and decellularised matrices’ (£2,000)
- AG Leventis Award for academic excellence (£15,000)
- Graduate School Travel Award (£1,500)
- ICH Dean Travel Award (£600)
- Medical Research Council Centenary Award - ‘Amniotic fluid stem cell transdifferentiation using extracellular matrix components from decellularised matrices’ (£45,398)
- ICH Poster Competition Award, 2nd Prize - ‘Three-dimensional scanning electron microscopy for fine tuning of pulmonary decellularisation protocols’ (£200)
- UCL Charlotte and Yule Bogue Research Fellowship - 'Intestinal tissue engineering in vivo through the combination of mouse crypt organoid units and decellularised matrices” (£3,336)
- Medical Research Council MB PhD Studentship - “Intestinal tissue engineering using natural acellular matrices” (£49,000)
- Merit in Clinical Sciences, MBBS Year 2
- Merit in Clinical Sciences, MBBS Year 1
- Republic of Cyprus Scholarship for academic excellence (£15,000)