Abstract
101 Our laboratory is investigating the tissue-engineering of small intestine using intestinal epithelial organoid units seeded on biodegradable polymer tubes. Previous studies have demonstrated the formation of cysts with tissue morphology resembling small intestine. This study investigates methods of stimulation for optimizing neointestinal regeneration. Intestinal epithelial organoid units (4-6 × 104 units) harvested from neonatal Lewis rats were seeded onto synthetic biodegradable polymer tubes (n=34) created from nonwoven sheets of PGA fibers sprayed on the outer surface with 5% PLA. These organoid unit/polymer constructs were implanted into the omentum of adult male Lewis rats (175-250 grams) in the following conditions: 1). C: control, implantation alone (n=9); 2). SBR: after 75% small bowel resection (n=9); 3). PCS: after portacaval shunt (n=8); and 4). PH: after 75% partial hepatectomy (n=8). Ultrasonography was performed at 3, 7, and 10 weeks to document neointestinal cyst size and growth. The constructs were harvested at 10 weeks and examined by histology. Morphometric analysis of the neomucosa including villus number, villus height, mucosal surface area, crypt number, and crypt area was obtained using a video-microscope integrated computer program (NIH Image, version 1.59) by a blinded observer. Cyst development was noted in all animals. Ultrasonography demonstrated no significant differences in cyst size at 3 weeks. As shown in the table, at 7 and 10 weeks, cyst lengths and diameters were significantly greater in the SBR group compared to the other groups (*P<0.05, ANOVA, Fisher's PLSD). Histology revealed a neomucosa lining the lumen of the cyst with invaginations resembling crypt-villus structures. Morphometric analysis demonstrated a significantly greater villus number (18±2), villus height (133.9±22.6µm), and mucosal surface area (8533.3±977.6µm2) in the SBR group compared to the other groups (C: 11.4±3.6, 87.1±7.8µm, 5998.4±416.5 µm2; PCS: 12.3±1.8, 89.9±10.3µm, 6131.6±548.4 µm2; PH: 12±4, 97.3±9.6µm, 6000.1±777.1 µm2; P<0.01, ANOVA, Fisher's PLSD), and a significantly greater crypt number (23.3±2.2) and crypt area (68127.3±12431.5µm2) in the PCS group compared to control (14.6±5.7; 40150.8±16388.7µm2; P<0.01, ANOVA, Fisher's PLSD).TableIntestinal epithelial organoid units transplanted on biodegradable polymer tubes can successfully survive and regenerate into complex tissue resembling small intestine. Small bowel resection, and to a lesser extent, portacaval shunt, contribute significant regenerative stimuli for the morphogenesis and differentiation of tissue-engineered small intestine.
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