Immune Tolerance of Human Dental Pulp-Derived Mesenchymal Stem Cells Mediated by CD4⁺CD25⁺FoxP3⁺ Regulatory T-Cells and Induced by TGF-β1 and IL-10

PURPOSE: Most studies on immune tolerance of mesenchymal stem cells (MSCs) have been performed using MSCs derived from bone marrow, cord blood, or adipose tissue. MSCs also exist in the craniofacial area, specifically in teeth. The purpose of this study was to evaluate the mechanisms of immune tolerance of dental pulp-derived MSC (DP-MSC) in vitro and in vivo. MATERIALS AND METHODS: We isolated DP-MSCs from human dental pulp and co-cultured them with CD4⁺ T-cells. To evaluate the role of cytokines, we blocked TGF-β and IL-10, separately and together, in co-cultured DP-MSCs and CD4⁺ T-cells. We analyzed CD25 and FoxP3 to identify regulatory T-cells (Tregs) by fluorescence-activated cell sorting (FACS) and real-time PCR. We performed alloskin grafts with and without DP-MSC injection in mice. We performed mixed lymphocyte reactions (MLRs) to check immune tolerance. RESULTS: Co-culture of CD4⁺ T-cells with DP-MSCs increased the number of CD4⁺CD25⁺FoxP3⁺ Tregs (p<0.01). TGF-β or/and IL-10 blocking suppressed Treg induction in co-cultured cells (p<0.05). TGF-β1 mRNA levels were higher in co-cultured DP-MSCs and in co-cultured CD4⁺ T-cells than in the respective monocultured cells. However, IL-10 mRNA levels were not different. There was no difference in alloskin graft survival rate and area between the DP-MSC injection group and the non-injection group. Nonetheless, MLR was reduced in the DP-MSC injected group (p<0.05). CONCLUSION: DP-MSCs can modulate immune tolerance by increasing CD4⁺CD25⁺FoxP3⁺ Tregs. TGF-β1 and IL-10 are factors in the immune-tolerance mechanism. Pure DP-MSC therapy may not be an effective treatment for rejection, although it may module immune tolerance in vivo.

Diagnostic Performance of Diffusion-Weighted Steady-State Free Precession in Differential Diagnosis of Neoplastic and Benign Osteoporotic Vertebral Compression Fractures: Comparison to Diffusion-Weighted Echo-Planar Imaging

PURPOSE: To evaluate the diagnostic performance of diffusion-weighted steady-state free precession (DW-SSFP) in comparison to diffusion-weighted echo-planar imaging (DW-EPI) for differentiating the neoplastic and benign osteoporotic vertebral compression fractures. MATERIALS AND METHODS: The subjects were 40 patients with recent vertebral compression fractures but no history of vertebroplasty, spine operation, or chemotherapy. They had received 3-Tesla (T) spine magnetic resonance imaging (MRI), including both DW-SSFP and DW-EPI sequences. The 40 patients included 20 with neoplastic vertebral fracture and 20 with benign osteoporotic vertebral fracture. In each fracture lesion, we obtained the signal intensity normalized by the signal intensity of normal bone marrow (SI norm) on DW-SSFP and the apparent diffusion coefficient (ADC) on DW-EPI. The correlation between the SI norm and the ADC in each lesion was analyzed using linear regression. The optimal cut-off values for the diagnosis of neoplastic fracture were determined in each sequence using Youden's J statistics and receiver operating characteristic curve analyses. RESULTS: In the neoplastic fracture, the median SI norm on DW-SSFP was higher and the median ADC on DW-EPI was lower than the benign osteoporotic fracture (5.24 vs. 1.30, P = 0.032, and 0.86 vs. 1.48, P = 0.041, respectively). Inverse linear correlations were evident between SI norm and ADC in both neoplastic and benign osteoporotic fractures (r = −0.45 and −0.61, respectively). The optimal cut-off values for diagnosis of neoplastic fracture were SI norm of 3.0 in DW-SSFP with the sensitivity and specificity of 90.4% (95% confidence interval [CI]: 81.0–99.0) and 95.3% (95% CI: 90.0–100.0), respectively, and ADC of 1.3 in DW-EPI with the sensitivity and specificity of 90.5% (95% CI: 80.0–100.0) and 70.4% (95% CI: 60.0–80.0), respectively. CONCLUSION: In 3-T MRI, DW-SSFP has comparable sensitivity and specificity to DW-EPI in differentiating the neoplastic vertebral fracture from the benign osteoporotic vertebral fracture.

Clinical Nutrition Service at Medical Centers in Seoul

The purpose of this study was to investigate the status of clinical nutrition services at various medical centers in Seoul, Korea. A questionnaire was distributed to the departments of nutrition at 44 hospitals in Seoul on July 2009. Nutritional screening carried out at a rate of 59.1% at the medical centers, and a significant difference was found according to the type of center, from 100% in tertiary hospitals to 18.8% in normal hospitals. On annual average, the numbers of inpatients, inpatients for malnutritional screening, inpatients with malnutrition, and inpatients for malnutrition management were 15,169.5, 10,870.9, 2,224.8, and 1,546.2, respectively. On average the group nutrition education was done 36.1 times/year for diabetes, 8.2 times/year for cancer, and 1.9 times/year for renal disease, and the numbers of participants 423.1, 95.1, and 31.5, respectively. On average the individual nutrition education of inpatients with diabetes was done 135.4 times/year for ordered-type, and 119.3 times/year for unordered-type, 106.2 times/year for paid-type, and 148.5 times/year for unpaid-type. The mean fee for education and counseling was the highest for peritoneal dialysis (73,090.9 won) but the lowest for heart disease (23,609.1 won). On average the individual nutrition education of outpatients with diabetes was done 234.6 times/year for ordered-type, and 2.5 times/year for unordered-type, 204.4 times/year for paid-type, and 32.7 times/year for unpaid-type. The mean fee for education and counseling was also the highest for peritoneal dialysis (63,500.0 won) but the lowest for heart disease (21,336.4 won). To implement more effective clinical nutrition service, a national medical insurance imbursement policy should be urgently instituted such that diseases left as unpaid are covered by health insurance, including all nutrition-related disease.

Analysis of Hospital Foodservice Management and Health Insurance Coverage of Inpatient Meals in Seoul

The objectives of this study were to explore hospital foodservice management and to investigate conditions related to health insurance coverage of inpatient meals. A questionnaire was distributed to the nutrition departments of 44 hospitals in Seoul on July 2009. The average kitchen area was 0.5 m2, and centralized distribution systems were in place. Partition walls from contamination zones, separate work tables to prevent cross-contamination, exclusive areas for preparing tube feeding, and split carts with refrigerated and convection heat settings were largely used in tertiary hospitals. Most dietitians did meal rounds (93.2%) and surveyed for patient satisfaction (86.4%). The major theme of QI (Quality Improvement) was menu management (31.8%). The health insurance fees for meals were 4,938.9 won for a general diet, 5,199.8 won for a therapeutic diet, 4,067.0 won for tube feeding, 9,950.0 won for sterilized diet, and 18,383.4 won for diets not covered by health insurance. The prices for general and therapeutic diets were significantly lower in hospitals compared to tertiary or general hospitals (P<0.001). The cost composed of 48.3% food, 44.0% labor and 7.7% overhead for general diets and 47.9%, 44.5% and 7.6% for therapeutic diets. In the case of health insurance coverage for patient meals, the number of items applied to general diets averaged 2.8 out of 4 and for therapeutic diets it averaged 1.9 out of 3. To reform the health insurance coverage system for patient meals, it is urgent that the qualified level of patient meals is presented from a national viewpoint, and monitoring should be performed consistently by developing the evaluation tools.

Increased Immunoendocrine Cells in Intestinal Mucosa of Postinfectious Irritable Bowel Syndrome Patients 3 Years after Acute Shigella Infection: An Observation in a Small Case Control Study

PURPOSE: Postinfectiously irritable bowel syndrome (PI-IBS) develops in 3-30% of individuals with bacterial gastroenteritis. Recent studies demonstrated increases in inflammatory components in gut mucosa of PI-IBS patients even after complete resolution of infection. We aimed to investigate histological changes in colon and rectum of PI-IBS subjects after long term period of infection. MATERIALS AND METHODS: We recruited PI-IBS subjects who had been diagnosed IBS after complete resolution of enteritis caused by shigellosis outbreak 3 years earlier. We compared unmatched four groups, PI-IBS (n = 4), non PI-IBS (n = 7), D-IBS (n = 7, diarrhea predominant type) and healthy controls (n = 10). All of them underwent colonoscopic biopsy at three areas, including descending colon (DC), sigmoid colon (SC) and rectum, which were assessed for 5-hydroxytryptamine (5-HT)/peptide YY (PYY)-containing enterochromaffin (EC) cell, intraepithelial (IEL) and lamina propria T lymphocyte (CD3), CD8 lymphocytes, mast cells and CD68/calprotectin+ macrophages. RESULTS: All subjects had no structural or gross abnormalities at colonoscopy. In PI-IBS, 5-HT containing EC cells, PYY containing EC cells, IELs, CD3 lymphocytes, CD8 lymphocytes, mast cells, and CD68 + macrophages were increased compared to control (p < 0.05). In D-IBS, PYY containing EC cells, IELs, and CD3 lymphocytes were increased compared to control (p < 0.05). In PI-IBS, 5-HT containing EC cells tended to increase and PYY containing EC cells, CD8 lymphocytes, mast cells, and CD68+ macrophages were increased compared to non PI-IBS (p < 0.05). Calprotectin + marcrophages were decreased in PI-IBS, non PI-IBS and IBS compared to control. CONCLUSION: The immunoendocrine cells were sporadically increased in PI-IBS, non PI-IBS and D-IBS compared with control. Our findings in a very small number of patients suggest that mucosal inflammation may play a role in long-term PI-IBS, and that other sub-groups of IBS and larger scale studies are needed to confirm this observation.

Effect of Itopride Hydrochloride on the Ileal and Colonic Motility in Guinea Pig In Vitro

PURPOSE: Itopride hydrochloride (itopride) inhibits acetylcholinesterase (AChE) and antagonizes dopamine D(2) receptor, and has been used as a gastroprokinetic agent. However, its prokinetic effect on the small bowel or colon has not yet been thoroughly investigated. The aim of this study was to investigate the effects of itopride on motor functions of the ileum and colon in guinea pigs. MATERIALS AND METHODS: The distal ileum was excised and the activity of peristaltic contraction was determined by measuring the amplitude and propagation velocity of peristaltic contraction. The distal colon was removed and connected to the chamber containing Krebs-Henseleit solution (K-H solution). Artificial fecal matter was inserted into the oral side of the lumen, and moved toward the anal side by intraluminal perfusion via peristaltic pump. Colonic transit times were measured by the time required for the artificial feces to move a total length of 10cm with 2-cm intervals. RESULTS: In the ileum, itopride accelerated peristaltic velocity at higher dosage (10(-10)-10(-6)M) whereas neostigmine accelerated it only with a lower dosage (10(-10)-10(-9)M). Dopamine (10(-8)M) decelerated the velocity that was recovered by itopride infusion. Itopride and neostigmine significantly shortened colonic transit at a higher dosage (10(-10)-10(-6)M). Dopamine (10(-8)M) delayed colonic transit time that was also recovered after infusion of itopride. CONCLUSION: Itopride has prokinetic effects on both the ileum and colon, which are regulated through inhibitory effects on AChE and antagonistic effects on dopamine D(2) receptor.

Access Type as A Predictor of Dialysis Dose and Efficiency in Hemodialysis Patients / 대한신장학회잡지

BACKGROUND: Use of venous catheter is associated with increased mortality in hemodialysis patients, but little data are available on impact of vascular access type on the discrepancy between dialysis prescription and dose. Dialysis prescription commonly exceeds the delivered dialysis dose. The purpose of this study are to evaluate the dialysis dose and efficiency between a tunneled cuffed catheters (TCCs) and arteriovenous fistulas (AVF) to the same patients on different dialysis days. METHODS: This study enrolled 35 hemodialysis patients. All patients started dialysis via tunneled cuffed catheters (Permcath(R)) in the internal jugular vein and a vascular surgeon created AVF later. We investigated prescribed spKT/V (pKt/V) and delivered spKt/V (dKt/V) within 2 months after the TCCs placement and after the AVF cannulation. Dialysis efficiencies (dKt/V/pKt/V) were compared over this two distinct times. RESULTS: Patient demographics were similar between two distinct times. The mean age of patients was 52.7 years (18 men, 17 women) and mean BMI was 21.9. 17 patients (48.6%) were diabetics. The mean pKt/V was not different (TCCs 1.63+/-0.30; AVF 1.62+/-0.29) but mean dKt/V was significantly different (TCCs 1.38+/-0.31; AVF 1.46+/-0.34) (p<0.05). The dialysis efficiencies were 85.14% for TCCs and 90.74% for AVF (p<0.05). There were no differences in sex, or diabetics. Underdialysis (delivered spKt/V <1.3) was greater in TCCs (17/35; 48.6 %), compared to AVF (10/35; 28.6%). Underdialysis was more common in larger patients. CONCLUSION: Our data suggest that AVF tend to receive more dialysis dose than tunneled cuffed catheters and underdialysis exposure are more common with catheter use.

Effect of Recombinant Fragment of Fibronectin on the Cellular Functions of Human Bone Marrow-Derived Mesenchymal Stem Cells / 대한정형외과연구학회지

PURPOSE: We investigated the effects of recombinant 9-10th type III repeat of fibronectin (rhFNIII9-10) on the adhesion, proliferation, and the osteogenic differentiation of human bone marrow-derived mesenchymal stem cells(hMSCs). MATERIALS AND METHODS: Adhesion and blocking assay for hMSCs were performed on the plates which had been coated with 100 microgram/ml rhFNIII9-10 or fibronectin. hMSCs seeded on the precoated plates were cultured in the osteogenic media for 3 weeks. MTS(Dimethylthiazole carboxymethoxyphenyl sulfophenyl tetrazolium compound) assay for the cell number, [Methyl-3H] thymidine incorporation study, alkaline phosphatase activity assay, calcium content assay and RT-PCR for alkaline phosphatase, osteopontin, cbfa-1, and type I collagen were performed during the osteogenic differentiation. RESULTS: hMSCs showed significantly increased adhesion to rhFNIII9-10-coated plates and fibronectin-coated plates. A monoclonal antibody to the integrin alpha 5 beta 1 inhibited adhesion to rhFNIII9-10-coated plates and fibronectin-coated plates in dose-dependent manner. hMSCs seeded on the rhFNIII9-10-coated plates showed increased proliferation during the osteogenic differentiation. However, there was no significant difference in the alkaline phosphatase activity, calcium content and expression levels of mRNAs for alkaline phosphatase, osteopontin, cbfa-1, and type I collagen of hMSCs seeded on the rhFNIII9-10-coated plates. CONCLUSION: rhFNIII9-10 stimulates hMSCs adhesion and increases hMSCs proliferation during the osteogenic differentiation. Although osteogenic differentiation is not promoted, adsorption of rhFNIII9-10 onto appropriate biomaterials can enhance integrin-mediated hMSCs adhesion and proliferation. This biomolecular engineering strategy represents a robust approach to increase biofunctional activity and integrin specificity of hMSCs.

Investigation into the Effects of Mosapride on Motility of Guinea Pig Stomach, Ileum, and Colon

Mosapride citrate (Mosapride) is a new prokinetic agent that enhances the gastrointestinal (GI) motility by stimulation of 5-HT4 receptors. This agent stimulates acetylcholine release from enteric cholinergic neurons in the GI wall. It was reported in several studies that mosapride selectively enhanced the upper, but not lower, GI motor activity. However, in these studies other 5-HT4 receptor agonists exerted stimulating effects on the motility of the colon. Moreover, it is well known that the receptors of 5-HT4 are also located in the colon. The purpose of this study was to estimate the effect of mosapride on the motility of the stomach, ileum and colon in the guinea pig and to investigate whether or not mosapride influenced the colonic motility. Mosapride significantly increased the amplitude of the contraction waves in the guinea pig stomach by electrical stimulation. In addition, it significantly increased the number of peaks, the area under the curve and the propagation velocity of the peristaltic contraction of the guinea pig ileum in a concentration dependent fashion. Mosapride also significantly shortened the transit time of the guinea pig colon. Accordingly, we concluded that mosapride exerted prokinetic effect on the entire GI tract of the guinea pig. Based on the possibility of similar results in humans, we suggest the potential use of mosapride for lower GI motor disorders such as constipation and upper GI motor disorders such as gastroesophageal reflex disease or gastroparesis.

Altered Colonic Motor Functions in Experimental Colitis of Guinea Pigs / 대한소화관운동학회지

BACKGROUND/AIMS: Trinitrobenzene sulphonic acid (TNBS)-induced colonic inflammation in rat alters colonic motor function. The aims of this study were to investigate the effect of TNBS-induced colitis on the colonic motor function of a guinea pig along the course of colitis and to document persistently altered colonic motor function after resolution of inflammation. METHODS: Colitis was induced in about 300 g male guinea pigs (Hartley) by intrarectal administration of 0.3 mL TNBS in 50% ethanol, while controls received 0.3 mL of 50% ethanol or not. After 24 hours, 48 hours, 72 hours, 1 week, and 2 weeks, the distal colon was taken for the investigation of gross and microscopic findings, muscle tension, and colonic transit. RESULTS: Maximal mucosal injury and inflammation were evident from the 2nd day following the induction of colitis. Seven days after the induction of colitis, some portions of the damaged mucosa began to recover. Development of tension in response to carbachol was not altered significantly along the course of colitis. Colonic transit was delayed significantly at 3, 7, and 14 days after administration of TNBS. CONCLUSIONS: Colonic transit in TNBS-induced colitis of guinea pigs is delayed, but not linearly related to the degree of inflammation. Delayed colonic transit is not related to the muscarinic receptor-mediated contractions of circular muscle from the inflamed colon. Further studies are required to determine the mechanism(s) involved in this motor change.

Small Circumscribed Aortic Dissection Complicating Annuloaortic Ectasia in a Non-Marfanoid Patient

Annuloaortic ectasia, cystic medial degeneration of the afflicted aortic wall leading to progressive dilatation, is often accompanied by Marfan's syndrome. Some portions of intimal flap is commonly demonstrated along the aorta in the noninvasive diagnosis of aortic dissection. We report the first case of circumscribed aortic dissection developed in a 28 year old obese non-Marfanoid patient. He was transferred after thrombolytic therapy at a community hospital because of severe chest pain and ST segment elevation. Transthoracic echocardiography showed markedly dilated aortic root, moderate amount of pericardial effusion, mild aortic regurgitation in spite of normal regional wall motion of left ventricle. Intimal flap, characteristic of aortic dissection, was not seen with computed tomography. Intimal tear was demonstrated just above aortic valve only by transesophageal echocardiography. Two parallel intimal tear and small circumscribed dissection was demonstrated by autopsy.