Porcine Small Intestine Research Articles

Development and Characterization of an Acellular Porcine Small Intestine Submucosa Scaffold for Use in Corneal Epithelium Tissue Engineering

ABSTRACTPurpose: To produce an acellular small intestine submucosa (SIS) that would be a suitable scaffold for corneal epithelium tissue engineering.Methods: The SIS was decellularized by immersion in 0.1% (wt/vol) sodium dodecyl sulfate (SDS). The efficacy of acellularization was confirmed by histological observation and DNA quantification. The mechanical properties were evaluated by uniaxial tensile testing. ELISA was performed to assess the growth factor contents. The cytotoxicity of SIS scaffolds and extracts to rabbit corneal epithelial cells was determined by CCK-8 assay. We also investigated the inflammatory reaction of SIS implanted subcutaneously in a rat. The biocompatibility was studied by rabbit interlamellar corneal transplantation and reseeding assay with cornea-derived cells. Immunofluorescent staining was used to detect the expression of CK3, ZO-1 and K13.Results: Histological analyses showed that complete cell removal was achieved, and the DNA quantity, which reflects the presence of cellular materials, was significantly diminished in acellular SIS. Collagen fibers were properly preserved and appeared in an orderly fashion. The tissue structure, the mechanical properties and the growth factor contents within the acellular SIS were well retained. The CCK8 assay demonstrated that the acellular SIS scaffolds and extracts had no cytotoxicity to rabbit corneal epithelial cells. There was no sign that an immune reaction occurred with acellular SIS implanted subcutaneously in a rat. In fact, in vivo implantation to rabbit interlamellar stromal pockets showed good biocompatibility. We also observed that clusters of rabbit corneal epithelial cells were growing well on the surface of the SIS in vitro and the distinctive CK3, ZO-1 for corneal epithelial cells was detected.Conclusions: The decellularized SIS retained the major structural components. The matrix is biocompatible with cornea-derived cells and might be a suitable scaffold for corneal epithelium tissue engineering.

Ex vivo intestinal perfusion model for investigating mucoadhesion of microcontainers.

Micro fabricated delivery systems have shown promise in increasing oral bioavailability of drugs. Micrometer-sized polymeric devices (microcontainers) have the potential to facilitate unidirectional drug release directly into the intestinal mucosa whereby, drug absorption can be enhanced. The aim of this study was to develop an ex vivo model to investigate mucosal adhesion and orientation of microcontainers. Furthermore, to investigate how microcontainers with varying height, shape and material behave in regards to mucoadhesion and orientation. Microcontainers were placed at the top of an inclined piece of porcine small intestine. The tissue was perfused with biorelevant medium followed by microscopic examination to observe the orientation and amount of microcontainers on the tissue. The mucoadhesion of the microcontainers were evaluated based on the observed position on the tissue after being exposed to flow. When comparing the varying types of microcontainers, good adhesion was in general observed since most of the microcontainers were located in the beginning of the intestine. Microcontainers fabricated from the epoxy-based photoresist SU-8 had a slightly better adherence than those fabricated from poly-ɛ-caprolactone (PCL). The orientation of the microcontainers appeare to be dictated mainly by the height. In general, the model showed promising results in evaluating mucoadhesion and orientation.

Adipose tissue-derived stem cells upon decellularized ovine small intestine submucosa for tissue regeneration: An optimization and comparison method.

The extracellular matrix of different mammalian tissues is commonly used as scaffolds in the field of tissue engineering. One of these tissues, which has frequently been studied due to its structural and biological features, is the small intestine submucosal membrane. These research are mainly done on the porcine small intestine. However, a report has recently been published about a scaffold produced from the submucosal layer of the ovine small intestine. In the present study, ovine small intestine submucosal (OSIS) was decellularized in a modified manner and its histological, morphological, and biomechanical properties were studied. Decellularization was performed in two phases: physical and chemical. In this method, a chloroform-methanol mixture, enzymatic digestion, and a constant dose of sodium dodecyl sulfate (SDS) was used in the least agitation time and its histological property and biocompatibility were evaluated in the presence of adipose tissue-derived stem cells (ADSCs); furthermore, ADSCs were isolated with a simple method (modified physical washing non-enzymatic isolation). The results were showed that the use of OSIS could be effective and operative. Mechanical properties, histological structure and shape, and glycosaminoglycan content were preserved. In the SDS-treated group, more than 90% of the native cells of tissue were deleted, and also in this group, no toxicity was observed and cell proliferation was supported, compared to the untreated group. Therefore, our results indicate that ADSCs seeded on OSIS scaffold could be used as a new approach in regenerative medicine as hybrid or hydrogel application.

Evaluation of decellularization protocols for production of porcine small intestine submucosa for use in abdominal wall reconstruction.

Porcine-derived acellular biologic grafts are increasingly used in abdominal wall reconstruction and other soft tissue repairs. In a previous work, we have shown porcine small intestine submucosa (PSIS) exhibits clear advantages over porcine pericardium (PPC) and porcine acellular dermal matrix (PADM) in repairing full-thickness abdominal wall defects. In the present study, we aim to determine, quantify, and compare the effects of two most commonly used decellularization protocols on biomechanical and biocompatible properties of PSIS. After mechanical preparation, PSIS was treated with either alkaline and acid (AA) protocol or sodium dodecyl sulfate (SDS) protocol. Cellular content removal, preservation of matrix components, micro- and ultra- structures, and mechanical properties were compared. The host responses were evaluated using PSIS for repairing rat abdominal wall defects. With regard to the absence of cellular contents, neatly arranged collagen fiber structures, better retention of growth factors, better mechanical strength, lower degrees of local and systemic inflammatory responses, higher degree of vascularization and tissue ingrowth, alkaline and acid protocol exhibits clear advantages over SDS protocol for the preparation of PSIS extracellular matrix.

Hyperglycaemia-Induced Downregulation in Expression of nNOS Intramural Neurons of the Small Intestine in the Pig.

Diabetic autonomic peripheral neuropathy (PN) involves a broad spectrum of organs. One of them is the gastrointestinal (GI) tract. The molecular mechanisms underlying the pathogenesis of digestive complications are not yet fully understood. Digestion is controlled by the central nervous system (CNS) and the enteric nervous system (ENS) within the wall of the GI tract. Enteric neurons exert regulatory effects due to the many biologically active substances secreted and released by enteric nervous system (ENS) structures. These include nitric oxide (NO), produced by the neural nitric oxide synthase enzyme (nNOS). It is a very important inhibitory factor, necessary for smooth muscle relaxation. Moreover, it was noted that nitrergic innervation can undergo adaptive changes during pathological processes. Additionally, nitrergic neurons function may be regulated through the synthesis of other active neuropeptides. Therefore, in the present study, using the immunofluorescence technique, we first examined the influence of hyperglycemia on the NOS- containing neurons in the porcine small intestine and secondly the co-localization of nNOS with vasoactive intestinal polypeptide (VIP), galanin (GAL) and substance P (SP) in all plexuses studied. Following chronic hyperglycaemia, we observed a reduction in the number of the NOS-positive neurons in all intestinal segments studied, as well as an increased in investigated substances in nNOS positive neurons. This observation confirmed that diabetic hyperglycaemia can cause changes in the neurochemical characteristics of enteric neurons, which can lead to numerous disturbances in gastrointestinal tract functions. Moreover, can be the basis of an elaboration of these peptides analogues utilized as therapeutic agents in the treatment of GI complications.

Use of a Novel, Reinforced, Low Immunogenic, Porcine Small Intestine Submucosa Patch to Repair a Supraspinatus Tendon Defect in a Rabbit Model.

Background Repairs of large to massive rotator cuff tears have a high failure rate. We investigated the efficacy of a novel, reinforced, low immunogenic, porcine small intestine submucosa (SIS) patch to repair a supraspinatus tendon defect in a rabbit model. We hypothesized that the histological and biomechanical results of SIS patch repair would be comparable with those of autologous fascia lata (FL) repair. Methods The study mainly comprised two parts. First, the characteristics of the SIS patch were evaluated, including its micromorphology, mechanical properties, and immunogenic properties. Second, a supraspinatus tendon defect model was created in 36 rabbits (72 shoulders). The bilateral shoulders were randomly chosen to undergo repair using either a SIS patch (SIS group) or autologous FL (FL group). At 4, 8, and 12 weeks, histological analysis was performed using four shoulders from each group, and biomechanical tests were performed using eight shoulders from each group. Results The SIS patch was a three-dimensional construct mainly composed of collagen fibers. The mean single and double suture retention loads of the SIS patch were 48.6 ± 5.8 N and 117.9 ± 2.7 N, respectively. The DNA content in the SIS patch was 53.9 ± 10.9 ng/mg dry weight. Both the histological score and ultimate load to failure increased in a time-dependent manner in both groups, with no significant differences between the SIS and FL groups at 12 weeks. Conclusion Repair of a large supraspinatus tendon defect using a reinforced, low immunogenic, SIS patch achieves similar effects as autologous FL in a rabbit model. This novel patch might be useful to be employed as a structural tissue replacement in medical activities.

Pathogenic E. coli HPI upregulate the expression of inflammatory factors in porcine small intestinal epithelial cells by ubiquitin proteasome pathway

To investigate the effects of pathogenic Escherichia coli high pathogenicity island (HPI) on the expression of inflammatory factors via ubiquitin proteasome pathway. Firstly, the UBC-sus-263 shRNA plasmid was successfully established and transfected into porcine small intestine epithelial cells (IPEC-J2) by liposome to silence the ubiquitinntion gene. Then the IPEC-J2 was infected with E. coli HPI+ and HPI− strains, respectively. Finally, the mRNA of intracellular NF-κB and IκB-α,and the protein levels of NF-κB, IκB-α, TNF-α and IL-1 in IPEC-J2 cell line transfected with UBC-sus-263 shRNA (Ub-shRNA) were detected. The results showed that the Ub-shRNA was effectively inhibited ubiquitination pathway in the IPEC-J2 cell. After infected with HPI+, the mRNA and protein levels of NF-κB and IκB-α were dramatically decreased in Ub-hsRNA transfected IPEC-J2 cells compared to the control and HPI−-infected groups. Consistently, the production of downstream cytokines such as TNF-α and IL-1 were highly expressed after HPI+-infection than that of HPI−-infected groups. However, whether the HPI+ or HPI−, both could induce increasingly expression of NF-κB and IκB-α and its downstream cytokines in normal IPEC-J2 cells. Thus, the E. coli HPI can upregulate the expression of IκB-α to promote the releasing of TNF-α and IL-1 via the ubiquitination pathway.

Verification of small intestinal submucosa graft stent in vitro

Objective To verify the feasibility of using small intestine submucosa for graft stents. Methods A Z-type Nitinol wire stent was used as the metal stent material, and porcine small intestine submucosa was used as the biofilm material to prepare the bio-coated stent. In vitro implantation, extracorporeal pulsation and extracorporeal flexion of the stents were designed based on the requirements for the small intestine submucosa graft stent implantation during the endovascular aneurysm repair and of the mechanical properties of the graft stent during human body activity. The feasibility of using small intestine submucosa for the graft stents was evaluated according to the patency of the stent lumen after the experiment, the presence or absence of cracks in the membrane, and the presence or absence of fracture of the metal stent. Results In the in vitro experiments, all the samples were able to maintain complete tunica and luminal patency except the 8th sample that showed suture rupture. Besides, all the samples showed good rebounding and adhering properties in both in vitro implantations and extracorporeal pulsation pressure experiments, and no dangerous problems were found such as film rupture, suture loss and stent fracture. Conclusion Small intestine submucosa can meet the durability requirements of graft stents, adapt to the conditions of graft stents such as implantation, bending and blood pulsation, and can be used as the covering material of graft stents. Key words: Biological graft; Small intestine submucosa; Graft stent; In vitro test

Trypsin decorated self-emulsifying drug delivery systems (SEDDS): Key to enhanced mucus permeation

It was the aim of this study to prepare trypsin decorated mucus permeating self-emulsifying drug delivery systems (SEDDS). Lipophilicity of enzyme was increased by hydrophobic ion pairing (HIP) with the anionic surfactants sodium dodecyl sulfate (SDS), sodium taurocholate (ST) and sodium deoxycholate (SDO) to facilitate its incorporation in SEDDS. Blank SEDDS and trypsin decorated SEDDS were characterized regarding droplet size, polydispersity index (PI), zeta potential and proteolytic activity using Nα-benzoyl-l-arginine ethyl ester (BAEE) assay. Log DSEDDS/release medium of each complex was determined to assess its affinity towards SEDDS oily droplet upon emulsification. Ability of trypsin decorated SEDDS to enhance mucus permeation was studied on mucus gel from porcine small intestine for the period of 4 h at 37 °C. Degree of enzyme precipitation via HIP was 94.5%, 85.7% and 48.2% for SDS, ST and SDO complex, respectively. SEDDS composed of 50% (w/w) cremophor EL, 20% (w/w) captex 300, and 30% (w/w) propylene glycol with a complex payload of 1% (w/w) exhibited a droplet size in the range of 29.92 ± 0.09 nm to 39.15 ± 0.37 nm, a polydispersity index of 0.116–0.265 and zeta potential in the range of −2.36 mv to −4.25 mv. The enzymatic activity of trypsin complexed with SDO, SDS and ST in SEDDS was 51.9%, 44.8%, and 40.7% respectively, of the corresponding activity of free trypsin. Log D values of trypsin, SDS, ST and SDO complex were −2.73, 1.97, 1.89 and 1.68, respectively, suggesting higher lipophilicity of trypsin complexes as compare to free trypsin and ability to reside on SEDDS droplets. Enzyme decorated SEDDS improved mucus permeation 1.6- to 2.6-fold in comparison to blank SEDDS. Results demonstrated that decorating SEDDS with trypsin can be a promising technique to improve their mucus permeating properties.

Acrylamide-induced alterations in the cocaine- and amphetamine-regulated peptide transcript (CART)-like immunoreactivity within the enteric nervous system of the porcine small intestines

The main goal of the present study was to determine the influence of low and high doses of acrylamide on CART-like immunoreactivity within the ENS of the porcine small intestines. Investigation was performed on 15 sexually immature female pigs, divided into three groups: control group, where empty gelatin capsules were administered, and two experimental groups, where capsules containing low or high doses of acrylamide were given. After 28days of acrylamide exposure all animals were euthanized with a gradual overdose of anaesthetics. Immediately after euthanasia fragments of the duodenum, jejunum and ileum were collected and fixed. Then, 14μm-thick cryostat sections were processed for routine double-labelling immunofluorescence using primary antisera directed towards a pan neuronal marker — protein gene-product 9.5 (PGP 9.5) and cocaine- and amphetamine-regulated peptide transcript (CART). During the present investigation, CART-LI cell bodies were detected in all types of enteric plexuses of duodenum, jejunum and ileum. Acrylamide intoxication resulted in a significant increase in expression of CART in the intramural neurons of the porcine small intestines, especially in myenteric plexuses. It may suggest participation of CART in neuronal protection and recovery processes within the gut. Moreover, results of the present study suggest that even low doses of acrylamide cause a significant response of ENS neurons.

2036 Extracellular matrix as a novel approach to glioma therapy

2036 Extracellular matrix as a novel approach to glioma therapy

Porcine small intestine submucosal grafts improve remucosalization and progenitor cell recruitment to sites of upper airway tissue remodeling.

To better understand upper airway tissue regeneration, the exposed cartilage and bone at donor sites of tissue flaps may serve as in vivo "Petri dishes" for active wound healing. The pedicled nasoseptal flap (NSF) for skull-base reconstruction creates an exposed donor site within the nasal airway. The objective of this study is to evaluate whether grafting the donor site with a sinonasal repair cover graft is effective in promoting wound healing. In this multicenter, prospective trial, subjects were randomized to intervention (graft) or control (no graft) intraoperatively after NSF elevation. Individuals were evaluated at 2, 6, and 12 weeks postintervention with endoscopic recordings. Videos were graded (Likert scale) by 3 otolaryngologists blinded to intervention on remucosalization, crusting, and edema. Scores were analyzed for interrater reliability and cohorts compared. Biopsy and immunohistochemistry at the leading edge of wound healing was performed in select cases. Twenty-one patients were randomized to intervention and 26 to control. Subjects receiving the graft had significantly greater overall remucosalization (p = 0.01) than controls over 12 weeks. Although crusting was less in the small intestine submucosa (SIS) group, this was not statistically significant (p = 0.08). There was no overall effect on nasal edema (p = 0.2). Immunohistochemistry demonstrated abundant upper airway basal cell progenitors in 2 intervention samples, suggesting that covering grafts may facilitate tissue proliferation via progenitor cell expansion. This prospective, randomized, controlled trial indicates that a porcine SIS graft placed on exposed cartilage and bone within the upper airway confers improved remucosalization compared to current practice standards.

Is Decellularized Porcine Small Intestine Sub-mucosa Patch Suitable for Aortic Arch Repair?

Introduction: We reviewed our experience with decellularized porcine small intestine sub-mucosa (DPSIS) patch, recently introduced for congenital heart defects.Materials and Methods: Between 10/2011 and 04/2016 a DPSIS patch was used in 51 patients, median age 1.1 months (5 days to 14.5 years), for aortic arch reconstruction (45/51 = 88.2%) or aortic coarctation repair (6/51 = 11.8%). All medical records were retrospectively reviewed, with primary endpoints interventional procedure (balloon dilatation) or surgery (DPSIS patch replacement) due to patch-related complications.Results: In a median follow-up time of 1.5 ± 1.1 years (0.6–2.3years) in 13/51 patients (25.5%) a re-intervention, percutaneous interventional procedure (5/51 = 9.8%) or re-operation (8/51 = 15.7%) was required because of obstruction in the correspondence of the DPSIS patch used to enlarge the aortic arch/isthmus, with median max velocity flow at Doppler interrogation of 4.0 ± 0.51 m/s. Two patients required surgery after failed interventional cardiology. The mean interval between DPSIS patch implantation and re-intervention (percutaneous procedure or re-operation) was 6 months (1–17 months). While there were 3 hospital deaths (3/51 = 5.9%) not related to the patch implantation, no early or late mortality occurred for the subsequent procedure required for DPSIS patch interventional cardiology or surgery. The median max velocity flow at Doppler interrogation through the aortic arch/isthmus for the patients who did not require interventional procedure or surgery was 1.7 ± 0.57 m/s.Conclusions: High incidence of re-interventions with DPSIS patch for aortic arch and/or coarctation forced us to use alternative materials (homografts and decellularized gluteraldehyde preserved bovine pericardial matrix).

Comparison of Human Denuded Amniotic Membrane and Porcine Small Intestine Submucosa as Scaffolds for Limbal Mesenchymal Stem Cells.

Blinding corneal scarring is usually treated with allogeneic graft tissue. Nevertheless, the global shortage of donors leaves millions of patients in need of therapy. Traditional tissue engineering strategies involves the combination of cells, growth factors, and scaffolds that can supply cellular biological components allowing to restore the tissue function. The mesenchymal stem cells found in the limbal stroma (L-MSCs) have a self-renewal potential for multilineage differentiation. Thus, in this work we compared the potential of human amniotic membrane (hAM) and porcine small intestine submucosa (SIS) as scaffolds for L-MSCs, aiming at potential applications in corneal regeneration. For that, L-MSCs were seeded on hAM and SIS and we analyzed their viability, actin cytoskeleton, nuclei morphology, cell density, adhesion and surface markers. Our results showed that cells adhered and integrated into both membranes with a high cell density, an important characteristic for cell therapy. However, due to its transparency, the hAM allowed a better observation of L-MSCs. In addition, the analysis of surface markers expression on L-MSCs after two weeks showed a slight increase in the percentages of negative markers for MSCs grown on SIS membrane. Thus, considering a long-term culture, the hAM was considered better in maintaining the MSCs phenotype. Regarding the function as scaffolds, SIS was as efficient as the amniotic membrane, considering that these two types of biological matrices maintained the cell viability, actin cytoskeleton, nuclei morphology and mesenchymal phenotype, without causing cell death. Therefore, our data in vitro provides evidence for future pre-clinical studies were these membranes can be used as a support to transport mesenchymal stem cells to the injured area, creating a kind of temporary curative, allowing the release of bioactive molecules, such as cytokines and growth factors and then promoting the tissue regeneration, both in human and veterinary medicine.

Fabrication of Magnetically Driven Biopsy Mechanism Applicable to Capsule-Type Medical Device

This article proposes two kinds of magnetic rotary actuators applicable to a next-generation capsule endoscope that can conduct a small intestinal biopsy. One actuator is for an anchoring mechanism that can stop the capsule in a specific place against the peristaltic movement of the small intestine, and press a biopsy instrument on a lesion. The other actuator is for a biopsy mechanism that can project a circular blade while rotating, and obtain a tissue sample. Both actuators have the same basic structure, which is composed of a bolt with a permanent magnet and a nut, and can be driven by a rotating magnetic field. Because they are arranged orthogonally to each other in the capsule, they can be individually operated by the corresponding rotating magnetic field. Given the results of an operational test similar to an actual environment in a porcine small intestine, these actuators could successfully perform the desired operation.

The Regulatory Role of MeAIB in Protein Metabolism and the mTOR Signaling Pathway in Porcine Enterocytes.

Amino acid transporters play an important role in cell growth and metabolism. MeAIB, a transporter-selective substrate, often represses the adaptive regulation of sodium-coupled neutral amino acid transporter 2 (SNAT2), which may act as a receptor and regulate cellular amino acid contents, therefore modulating cellular downstream signaling. The aim of this study was to investigate the effects of MeAIB to SNAT2 on cell proliferation, protein turnover, and the mammalian target of rapamycin (mTOR) signaling pathway in porcine enterocytes. Intestinal porcine epithelial cells (IPEC)-J2 cells were cultured in a high-glucose Dulbecco’s modified Eagle’s (DMEM-H) medium with 0 or 5 mmoL/L System A amino acid analogue (MeAIB) for 48 h. Cells were collected for analysis of proliferation, cell cycle, protein synthesis and degradation, intracellular free amino acids, and the expression of key genes involved in the mTOR signaling pathway. The results showed that SNAT2 inhibition by MeAIB depleted intracellular concentrations of not only SNAT2 amino acid substrates but also of indispensable amino acids (methionine and leucine), and suppressed cell proliferation and impaired protein synthesis. MeAIB inhibited mTOR phosphorylation, which might be involved in three translation regulators, EIF4EBP1, IGFBP3, and DDIT4 from PCR array analysis of the 84 genes related to the mTOR signaling pathway. These results suggest that SNAT2 inhibition treated with MeAIB plays an important role in regulating protein synthesis and mTOR signaling, and provide some information to further clarify its roles in the absorption of amino acids and signal transduction in the porcine small intestine.

Optimized procedures for generating an enhanced, near physiological 2D culture system from porcine intestinal organoids

An important practical limitation of the three-dimensional geometry of stem-cell derived intestinal organoids is that it prevents easy access to the apical epithelium for testing food components, microorganisms, bioactive and toxic compounds. To this end, we here report on a new robust method for generating confluent intestinal cell monolayers from single-cell suspensions of enzymatically-dissociated porcine organoids using modified culture conditions. With this method, cell seeding densities can be standardised, overcoming problems with methods based on mechanical dissociation of organoids. Confluent monolayers formed tight junctions with high transepithelial electrical resistance in three days and could be used in experiments for up to two weeks. Multilineage differentiation of ileal stem cells was demonstrated by immunohistochemistry and RT-qPCR of cell-specific transcripts, also unequivocally confirming the controversial existence of Paneth-like cells in the porcine small intestine. The method described here is useful to standardize primary epithelial monolayer formation from intestinal organoids and allows rapid and robust studies of intestinal physiology.

53: Long-term subjective outcomes of porcine-derived small intestine submucosa graft-augmented versus native tissue rectocele repair

To evaluate long-term subjective outcomes of porcine small intestine submucosa (SIS) graft-augmented compared to native tissue rectocele repair. This is a follow up study of a previously published multi-center, randomized trial in which 159 women with stage 2 or greater posterior wall prolapse were randomized to SIS graft-augmented versus native tissue repair, completed in 2009. Exclusion criteria were concurrent sacral-colpopexy, porcine allergy, or past pelvic radiation. We mailed surveys to all participants at one institution (147) and conducted follow-up phone calls. Surveys assessed the Pelvic Floor Distress Inventory (PFDI) and interval care-seeking, treatment and surgery for bowel or prolapse symptoms. Failure was defined as presence of mild, moderate or severe bother (“bother”) associated with: 1) vaginal bulge 2) “bowel symptoms” defined as straining, splinting, sensation of incomplete evacuation on the PFDI; and/or 3) stool trapping. Post hoc power calculation estimated the detectable difference in failure by response rate to be 33-37% (response rate 40%, two-sided alpha 0.05, power 80%, graft:native tissue - 1:1). We used Students’ t-test and Wilcoxon rank sum test to compare groups. Of the 147 subjects, we mailed surveys to 122 (25 - not located). We received 57 completed surveys (25/81 graft, 32/78 native tissue); the overall response rate was 36%, with no difference between groups (p=0.2). The duration between surgery and follow-up was 8-13 years. Among responding subjects, there was one demographic difference between groups: a higher proportion of the graft group reported preoperative incomplete evacuation with bowel movement compared to native tissue (96% vs. 70%, respectively; p=0.03). At 8-13 years, 12% of responders reported bother with vaginal bulge, 63% with bowel symptoms, and 35% with stool trapping; these did not differ between groups. Twenty percent reported interval care-seeking for bowel or bulge symptoms, 5% reported non-surgical treatment and 5% reported surgical repair for any prolapse symptoms, with no differences between groups. Of the women who did not report bothersome symptoms at 12 months, by 8-13 years, 12% (6) developed new bothersome vaginal bulge, 54% (15) developed bowel symptoms, and 31% (14) developed stool trapping symptoms; there were no differences between groups. Of the women who reported bothersome symptoms at 12 months, at 8-13 years, no women reported worsening vaginal bulge symptoms, 50% (14) reported worsening bowel symptoms and 27% (3) reported worsening stool trapping. However, without any interval treatment, 75% (3) reported improvement/resolution of vaginal bulge, 25% (7) had improvement of bowel symptoms, and 45% (5) had improvement of stool trapping. There were no differences between groups. Among women randomized to graft-augmented versus native tissue rectocele repair, 30-50% of patients with no rectocele symptom bother at 12 months postoperative developed bothersome symptoms at long-term follow up. However, 25-75% of symptomatic women had spontaneous symptom improvement by 8-13 years postoperative.

Degree of SGLT1 phosphorylation is associated with but does not determine segment-specific glucose transport features in the porcine small intestines.

Glucose‐induced electrogenic ion transport is higher in the porcine ileum compared with the jejunum despite equal apical abundance of SGLT1. The objective of this study was a detailed determination of SGLT1 and GLUT2 expressions at mRNA and protein levels along the porcine small intestinal axis. Phosphorylation of SGLT1 at serine 418 was assessed as a potential modulator of activity. Porcine intestinal tissues taken along the intestinal axis 1 h or 3 h after feeding were analyzed for relative mRNA (RT‐PCR) and protein levels (immunoblot) of SGLT1, pSGLT1, GLUT2, (p)AMPK, β2‐receptor, and PKA substrates. Functional studies on electrogenic glucose transport were done (Ussing chambers: short circuit currents (I sc)). Additionally, effects of epinephrine (Epi) administration on segment‐specific glucose transport and pSGLT1 content were examined. SGLT1 and GLUT2 expression was similar throughout the small intestines but lower in the duodenum and distal ileum. pSGLT1 abundance was significantly lower in the ileum compared with the jejunum associated with significantly higher glucose‐induced I sc. SGLT1 phosphorylation was not inducible by Epi. Epi treatment decreased glucose‐induced I sc and glucose flux rates in the jejunum but increased basal I sc in the ileum. Epi‐induced PKA activation was detectable in jejunal tissue. These results may indicate that SGLT1 phosphorylation at Ser418 represents a structural change to compensate for certain conditions that may decrease glucose transport (unfavorable driving forces/changed apical membrane potential) rather than being the cause for the overall differences in glucose transport characteristics between the jejunum and ileum.

Use of Decellularised Porcine Intestinal Submucosa Extracellular Matrix in Airway Reconstruction to Enable Lung-sparing Oncological Surgery

Background: Reconstruction of the airways following tracheal or lung-sparing oncological surgery is challenging. A novel approach is the use of bioscaffolds, which allow regeneration of respiratory epithelium and eliminate the need for immunosuppression. Initial experiences with decellularised porcine small intestine submucosa extracellular matrix (CorMatrix) as a scaffold for vascular and cardiac surgery have been encouraging. Its properties are potentially suitable for airway reconstruction.