Implant of mesenchymal cells decreases acute cellular rejection in small bowel transplantation.

OBJECTIVE: The objective of the study was to show adipose tissue-derived mesenchymal stem cells (AD-MSCs) immunomodulatory effects in small bowel transplantation (SBTx). MATERIALS AND METHODS: Forty Wistar Han rats (age: 10-12 weeks): were allogenic receptor rats and were allotted in 2 groups. Control group: rats undergoing orthopic SBTx ; AD-MSCs group: rats undergoing orthotopic SBTx plus AD-MSCs. Male Lewis rats were allogeneic small bowel donors. Rejection was confirmed by histological study of the explanted intestine, enterocyte apoptosis was determined in crypts and the lamina propria of the small bowel. Cytokine concentration levels (enzyme-linked immunosorbent assay) (interleukin [IL]-4, IL-10, IL-12, IL-17, IL-21, IL-23, tumor necrosis factor-alpha, and transforming growth factor [TGF]-b1) and cell percentages (flow cytometry) (CD3+ CD4+, CD8+, CD4+/25+, CD8+/25+, CD4+/25+/Foxp3+, and CD8+/25+/Foxp3+) were assessed in peripheral blood preoperatively and after death. RESULTS: Treatment with AD-MSCs produced a significantly lower risk of rejection in the first 7 post-operative days (five rejection cases among 20 rats in the control group and only one case in the AD-MSCs group). Treg cells and TGFb1 levels showed a significant increase in the AD-MSCs group. CONCLUSIONS: The local implantation of AD-MSC in the anastomosis and the intestinal lumen can induce a regulatory immune response, by increasing the percentages of Treg cells and TGb-1 levels, leading to a lower risk of acute rejection by cell mediation, in the first 7 days of the intestinal transplant. We think that the implantation of AD-MSCs, in the anastomoses and in the lumen of the donor intestine, could give rise to a chimera of donor-recipient cells.

OBJETIVO: Mostrar el efecto inmunomodulador de las células madre mesenquimales (AD-MSCs) en el trasplante de intestino delgado (SBTx). MÉTODO: 40 ratas Wistar Han (edad: 10-12 semanas): grupo control (SBTx) y grupo AD-MSCs (SBTx + AD-MSCs implantadas en las anastomosis distal y proximal del intestino delgado y en la luz intestinal). El intestino delgado provino de ratas Lewis. El rechazo se confirmó histológicamente. Se estudió la apoptosis de los enterocitos en las criptas y en la lámina propia del intestino delgado. Se determinaron por ELISA las citocinas (IL-4, IL-10, IL-12, IL-17, IL-21, IL-23, TNF-α, TGF-b1) en sangre periférica y por citometría de flujo los porcentajes celulares (CD3+ CD4+, CD8+, CD4+/25+, CD8+/25+, CD4+/25+/Foxp3+, CD8+/25+/Foxp3+) en el preoperatorio y después de la muerte. RESULTADOS: El empleo de AD-MSCs se asoció a una disminución significativa del riesgo de rechazo en los primeros 7 días posoperatorios (cinco casos de rechazo de 20 ratas en el grupo control y un solo caso en el grupo AD-MSCs). Las células Treg y los valores de TGFb1 mostraron un incremento significativo en el grupo AD-MSCs. CONCLUSIONES: El implante local de AD-MSCs en las anastomosis del trasplante de intestino delgado podría disminuir el rechazo celular agudo. Pensamos que la implantación de AD-MSCs, en las anastomosis y en el lumen del intestino donante, podría dar lugar a un quimera de células donante-receptor.

Effect of age on cytokine response in an experimental model of osteomyelitis.

To study the effect of age on cytokine response in an experimental model of osteomyelitis. Forty adult male Wistar rats received a stainless steel needle, intramedullarly in the left tibia. Young rats (3 months old) and old rats (22 months old) were allotted in: Group A: Sterile implant. Group B: Sterile implant + slime producing S. aureus. Rats were sacrificed 9 weeks after surgery. Determinations: Cytokines (ELISA) in blood and in tibia extract and the number of bacteria in tibia and implant. The Wilcoxon, Mann-Whitney U tests were used (P < or = 0.01 significant). Infection was detected in every old rat receiving S. aureus, and in 7 of 10 young rats. In blood: prior to surgery, old rats presented higher IL-2 and lower IL-4 levels. Surgery alone did not induce significant changes in old rats; surgery + S. aureus induced significant increases of IL-2 and IL-10 in young rats, and of IL-6 in old rats. Tibia analysis S. aureus group showed increased levels of: IL-10 in young rats, and IL-1beta in old rats. In experimentally induced osteomyelitis, significant differences were observed in cytokine response with regard to age.

Immune cell variations in patients with hip fracture.

Hip fracture is an increasing pathology in the patients with increasing age. Immunological response differences may appear between different age groups. The purpose of this study was to investigate the immune response in patients with subcapital hip fracture and the relationship with age. Prospective study of 100 patients with displaced subcapital femoral fracture between 2000 and 2004, divided into three age groups: over 90 years (13), 80-90 (56) and under 80 years (27). The chi(2)-test, analysis of variance and Student's t-test were applied. Correlation coefficient and the Spearman test were used to study linear correlation. The T helper cells decreased with age, this inverse correlation was significant. There was a direct correlation between CD16% and age. IgA, IgG and IgM levels did not show any significant relationship with age in our study. Nevertheless, the IgE levels in peripheral blood showed a significant direct correlation with age. Basophils percentage presented an inverse correlation with age. Age is associated to some immune changes in patients suffering hip fracture.

Interleukin-1, interleukin-6, and interleukin-10 responses after antibiotic treatment in experimental chronic Staphylococcus aureus osteomyelitis.

BACKGROUND: Staphylococcal implant infections' response to treatment may be correlated with cytokine production. We investigated the effect of certain antibiotics on the cytokine response in experimental osteomyelitis. METHODS: A stainless steel needle with an adherent slime-producing Staphylococcus aureus was implanted intramedullarly in the left tibia of 40 adult male Wistar rats. At 42 days after implantation, cefuroxime, vancomycin, tobramycin, and ciprofloxacin were administered intramuscularly every 12 h for 21 days. The control group was given no antibiotic. At the end of the treatment, implants and tibias were retrieved, and the bacterial numbers were estimated. Cytokines [interleukin-1alpha (IL-1alpha), IL-6, and IL-10] were determined (ELISA) in the tibial extract. RESULTS: Vancomycin and cefuroxime inhibited bone colonization in all tibias, and tobramycin and ciprofloxacin inhibited it only partially. Cefuroxime reduced the number of bacteria that adhered to the implants more than the other antibiotics. IL-1alpha and IL-6 showed higher levels in the ciprofloxacin-treated group than in the cefuroxime-treated and control groups. IL-6 levels in rats treated with cefuroxime were lower than in rats treated with tobramycin or vancomycin and the control group. Cefuroxime decreased IL-10 levels more than ciprofloxacin or vancomycin or those seen in the control group. CONCLUSIONS: The cefuroxime group showed the greatest decrease of pro-inflammatory cytokines. Different antibiotics produce different cytokine reactions that should be studied to choose the best treatment.

Musculoskeletal hydatid disease: a report of 13 cases.

This is a retrospective study of 13 patients with muscular hydatidosis--i.e., 4% of the 309 cases of hydatid disease treated in our department during 1983-1999. The commonest clinical finding was an asymptomatic and slowly growing mass (7). Puncture or incision of the mass was followed by an infection of the cystic cavity with fistulization in 2 patients. The immunological findings were false negative in 4 patients. MR images were obtained in 4 patients before diagnosis, and were highly suggetive of hydatid disease. The cystic cavities in all 9 patients subjected to radical surgery healed without chemotherapy. Radical surgery was not possible in 4 cases, in 3 of whom the sacrum was involved. Medical treatment of these patients did not eliminate the disease and new operations were necessary.