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P388. Perifistular application of adipose-derived mesenchymal stem cells - experimental study

O. Ryska1, Z. Serclova1, O. Mestak2, E. Matouskova3, P. Vesely2, I. Mrazova4, O. Luksan4, 1Central Military Hospital, Department of Surgery, Prague, Czech Republic, 2Hospital Bulovka, Department of Plastic Surgery, Prague, Czech Republic, 33rd Medical Faculty Charles University, Department of Burns Medicine, Prague, Czech Republic, 4Institute for Clinical and Experimental Medicine, Centrum of Experimental Medicine, Prague, Czech Republic

Background

Local application of adipose-derived mesenchymal stem cells (ADSCs) is considered as a promising method for the treatment of perianal fistula in patients with Crohn's disease. More clinical and experimental studies are needed to evaluate this technique. In vivo bioluminescence imaging using firefly luciferase can clarify the distribution and viability of the cells after implantation.

The aim of the study was to evaluate the fistula healing after application of ADSCs in perifistular tissue.

Methods

Coecostomy was used as a fistula model in 11 Lewis rats. The inguinal adipose tissue was harvested from transgenic donor expressing firefly luciferase (LEW-Tg(Rosa-luc)11Jmsk; Jichi Medical School, Japan). The ADSCs were isolated using collagenase technique and injected (1–2×106 cells/ml) in perifistular tissue. D-luciferin (25 mg/kg) was applied subcutaneously and rats were imaged in IVIS Lumina XR camera on days 0, 2, 7, 14 and 30. The light emission from predefined regions of interest (ROIs) surrounding the fistula (circular field; 2.5 cm in diameter) was monitored. Fistulae drainage assessment (FDA) was used to evaluate the fistula healing.

Results

There were no complications during the follow-up. The fistula was identified as healed or improved according to FDA in 4/11 animals. Bioluminescence was strongest 2 days after application (4.46×105 (5.1×104–2.7×106). The radiance decreased as follows 1.15×105 (4.05×104–6.4×105) on day 7; 7.29×104 (9.36×103–2.88×105) on day 14 and 2.94×104 (4.93×103–1.70×105) on day 30 (values are medians in p/s/cm2/sr). There was a trend to higher luminescence in animals with healed fistula during the whole follow-up (30 days after injection - 8.23×104 (1.18×104–1.70×105) vs. 2.75×104 (4.93×103–6.88×104); p = 0.18).

Conclusion

Local application of ADSCs can lead to fistula healing. Bioluminescence imaging is feasible for in vivo monitoring of long term ADSCs viability in perifistular tissue. The higher concentration of viable cells confirmed by bioluminescence was associated with fistula healing. Repeated application of ADSCs may improve the outcome.

Supported by NT13708