Background
Short bowel syndrome (SBS) is a life-threatening condition with few solutions. Tissue engineered intestine (TEI) is a potential treatment, but donor intestine is a limiting factor. Expanded epithelial surrogates termed enteroids may serve as a potential donor source.
Materials and Methods
To produce TEI from enteroids, crypts were harvested from mice and enteroid cultures established. Enteroids were seeded onto polymer scaffolds using Matrigel or culture medium, and implanted in immunosuppressed mice for 4 weeks. Histology was analyzed using Periodic Acid Schiff (PAS) staining and immunofluorescence (IF). Neomucosa was quantified using ImageJ software. To determine if TEI could be produced from enteroids established from small intestinal biopsies, 2×2mm pieces of jejunum were processed for enteroid culture, enteroids were expanded and seeded onto scaffolds, and scaffolds implanted for 4 weeks.
Results
Enteroids in Matrigel produced TEI in 15/15 scaffolds, whereas enteroids in medium produced TEI in 9/15 scaffolds. Use of Matrigel led to more neomucosal surface area compared to media (10,520±2,905 μm vs. 450±127 μm, p<0.05). Histologic examination confirmed the presence of crypts and blunted villi, normal intestinal epithelial lineages, intestinal subepithelial myofibroblasts, and smooth muscle cells. Crypts obtained from biopsies produced an average of 192±71 enteroids. A single passage produced 685±58 enteroids, which was adequate for scaffold seeding. TEI was produced in 8/9 scaffolds seeded with expanded enteroids.
Conclusions
Enteroids can be obtained from minimal starting material, expanded ex vivo, and implanted to produce TEI. This method shows promise as a solution to the limited donor intestine available for TEI production in patients with SBS.
Preoperative assessment of tissue anatomy and accurate surgical planning is crucial in conjoined twin separation surgery. We developed a new method that combines three-dimensional (3D) printing, assembling, and casting to produce anatomic models of high fidelity for surgical planning. The related anatomic features of the conjoined twins were captured by computed tomography (CT), classified as five organ groups, and reconstructed as five computer models. Among these organ groups, the skeleton was produced by fused deposition modeling (FDM) using acrylonitrile-butadiene-styrene. For the other four organ groups, shell molds were prepared by FDM and cast with silica gel to simulate soft tissues, with contrast enhancement pigments added to simulate different CT and visual contrasts. The produced models were assembled, positioned firmly within a 3D printed shell mold simulating the skin boundary, and cast with transparent silica gel. The produced phantom was subject to further CT scan in comparison with that of the patient data for fidelity evaluation. Further data analysis showed that the produced model reassembled the geometric features of the original CT data with an overall mean deviation of less than 2 mm, indicating the clinical potential to use this method for surgical planning in conjoined twin separation surgery.
Wrapping of seeded scaffolds in vascularized membranes produced the largest quantity and highest quality of TESI. Attaching seeded scaffolds to the abdominal wall produced an intermediate quantity of TESI, but the quality was still comparable to TESI produced in vascularized membranes. Insertion of seeded scaffolds into the subcutaneous space produced the smallest quantity and lowest quality of TESI. In summary, wrapping seeded scaffolds with vascularized membranes is favorable for the production of TESI, and wrapping with omentum may produce TESI that is most easily anastomosed with host intestine.
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