M.A.S.F. Baptista

Effect of Whole Bone Marrow Cell Infusion in the Progression of Experimental Chronic Renal Failure

INTRODUCTIONnThe therapeutic potential of adult stem cells for the treatment of chronic diseases is becoming increasingly evident over the last few years. In the present study, we sought to assess whether the infusion of bone marrow-derived mononuclear cells (MoSCs) and mesenchymal cells (MSCs) could reduce/stabilize the rate of progression of chronic renal failure (CRF) in rats.nnnMETHODSnWe used the 5/6 renal mass reduction model to induce chronic renal failure in male Wistar rats. Renal function was assessed by measurements of serum creatinine (sCr), creatinine clearance (Clcr), and 24-hour proteinuria at baseline as well as 60 and 120 days after surgery. MoSCs and MSCs obtained from bone marrow aspirates were separated by the Ficoll-Hypaque method. After a 12- to 14-day culture, 1.5 x 10(6) MSCs and the same number of MoSCs were injected into the renal parenchyma of the remanant kidney of rats with CRF on the day of surgery.nnnRESULTSnAmong the control group, at day 120, the results were sCr = 1.31 +/- 0.5 mg/dL, Clcr = 0.64 +/- 0.35 mL/min, and proteinuria = 140.0 +/- 57.7 mg/24 h. Rats treated with MoSCs at day 120 had sCr = 0.81 +/- 0.20 mg/dL, Clcr = 1.05 +/- 0.26 mL/min, and proteinuria = 61 +/- 46.5 mg/24 h, while rats injected with MSCs had sCr = 0.95 +/- 0.1 mg/dL, Clcr = 0.68 +/- 0.24 mL/min, and proteinuria = 119.2 +/- 50.0 mg/24 h. Analysis of the progression to CRF showed that the treatment significantly reduced the rate of decline in Clcr after treatment with MoSc: control: -0.0049 +/- 0.0024 mL/min/d versus MSC: - 0.0013 +/- 0.0017 mL/min/d versus MoSC: +0.0002 +/- 0.0016 mL/min/d (P = .017). Proteinuria tended to be lower among the treated groups. Histological scores of chronic damage were not different, but distinct patterns of chronic lesions were observed among treated rats.nnnCONCLUSIONnOur results showed that progression of CRF in rats could be slowed/stabilized by intrarenal parenchymal injection of MoSCs. A trend toward reduction in the progression rate of CRF was also observed with injection of MSCs.

Nonsurgical Periodontal Therapy Combined with Laser and Photodynamic Therapies for Periodontal Disease in Immunosuppressed Rats

BACKGROUNDnPeriodontal disease is often associated with systemic diseases and is characterized by destruction of the tissues supporting the teeth. Patients using immunosuppressive drugs such as tacrolimus are among those who suffer from tissue destruction.nnnOBJECTIVEnWe sought to evaluate the effects of laser and photodynamic therapies (PDT; nonsurgical) as an adjunct to scaling and rootplaning (SRP) in the treatment of corona-induced periodontitis in rats immunosuppressed with tacrolimus (Prograf).nnnMATERIALS AND METHODSnThe animals were divided into 5 groups. Each groups had 6 rats. Group I, the control group, received only saline solution throughout the study period of 42 days and did not receive periodontal treatment; group II received saline solution and SRP; group III received tacrolimus (1 mg/kg per day) and was treated with SRP; group IV animals were treated identically to group III and then administered laser treatment; and in group V, the animals were treated identically to group III and then administered PDT.nnnRESULTSnStatistical analysis indicated decreased bone loss with the progression of time (P = .035). There was no difference between the bone loss associated with the types of treatment administered to groups I, II, and III (P > .9) or groups IV and V (P > .6). The analysis also indicated that immunosuppression was not a bone loss-determining factor.nnnCONCLUSIONnLaser and PDT therapies were effective as an adjunctive treatment to SRP in reducing bone loss caused by experimental periodontitis induced in animals being treated systemically with tacrolimus.

Clinical and histopathologic comparative analysis between kidney transplant recipients from expanded-criteria donors and standard-criteria donors.

Owing to the disparity between the supply of kidney donors and demand, the use of organs from older deceased donors was initiated in recent years. The potentially poor outcome of these grafts is a major concern. This retrospective study compares graft and patient 1-year survivals between recipients from expanded-criteria donors (ECD; n = 30) and standard-criteria donors (SCD; n = 104). Rates of delayed graft function (DGF), acute rejection (AR), and chronic injury in the pre-implantation biopsy were also assessed. Increasing donor age was associated with increased rates of DGF, and DGF correlated with AR. Cold ischemia time >30 hours was associated with worse graft outcomes. Induction with Simulect correlated with better patient survival compared with Timoglobulina. Chronic injury pre-implantation biopsy correlated with worse renal function, but graft survival was similar. Death-censored graft survival at 1 year was 90% and patient survival 82%, and these were similar in ECD and SCD recipients. Selection of transplant candidates for ECD kidneys must be performed with caution. One-year graft survival was similar to that of SCD kidneys, but kidney function was worse during the same period. This may result in poorer graft survival over longer follow-up.

Fetal microchimerism in kidney biopsies of lupus nephritis patients may be associated with a beneficial effect

IntroductionMicrochimeric male fetal cells (MFCs) have been associated with systemic lupus erythematosus, and published studies have further correlated MFC with lupus nephritis (LN). In the present study, we evaluated the frequency of MFC in the renal tissue of patients with LN.MethodsTwenty-seven renal biopsies were evaluated: Fourteen were from women with clinical and laboratory findings of LN, and thirteen were from controls. Genomic DNA was extracted from kidney biopsies, and the male fetal DNA was quantified using real-time quantitative polymerase chain reactions for the detection of specific Y chromosome sequences.ResultsMFCs were detected in 9 (64%) of 14 of patients with LN, whereas no MFCs were found in the control group (P = 0.0006). No differences in pregnancy history were found between patients with LN and the control group. Significantly higher amounts of MFCs were found in patients with LN with serum creatinine ≤1.5 mg/dl. Furthermore, women with MFCs had significantly better renal function at the time of biopsy (P = 0.03). In contrast, patients with LN without MFCs presented with more severe forms of glomerulonephritis (World Health Organization class IV = 60% and class V = 40%).ConclusionsOur data indicate a high prevalence of MFCs in renal biopsy specimens from women with LN, suggesting a role for MFCs in the etiology of LN. The present report also provides some evidence that MFCs could have a beneficial effect in this disease.

Altered Expression of Inflammation Genes in Preimplantation Kidney Biopsies of Extended Criteria Donors

Background Changes to an organ can occur at the time of brain death and a series of inflammatory proteins are generated within the organ. It is not known whether this inflammatory similarly affects the kidneys from extended (ECD) and standard (SCD) criteria donors and because we have previously reported that preimplantation kidney biopsies from ECD donors have a heavier inflammatory profile when compared with SCD donors. In the present study we extended our results using great number of genes and seeking to identify some immunologic pathways involved in the mechanism of sterile inflammation. Methods Pretransplant kidney biopsies (Bx) were obtained from ECD (n=40) and SCD (n=40). Gene expression profile measured by Real Time qPCR Array representing expression levels of genes indicative of inflammation (IL-10, IL-1b, TNF-&agr;, MCP-1, NFK-b, TLR-4, HMGB1, IFN-gamma, TGF-b, Myd-88), cytoprotection (HO-1, HIF-1a), apoptose (CASP-1) and intercellular adhesion (ICAM-1) and correlated with donor variables. Results ECD donors were older had more cerebrovascular accident, arterial hypertension and diabetes (p<0.01) and recipients of ECD kidneys had renal function and 24h proteinuria worst 1 year after transplantation (p<0.006). Genes IL-10, IL-1 &bgr;, TLR-4, HMGB-1, HIF-1 and CASP-1 were significantly more expressed in biopsies from ECD than SCD. Presence of DGF, acute rejection, was not associated with any individual transcript. Conclusions The present results confirm and expand our previous findings that ECD kidney is highly inflamed when compared with SCD and that a Myd-88 independent pathway of innate immunity may be activated. We appreciate the FAPESP (#2014/25831-5) for the financial support.

BIOMATERIAL SEEDED WITH BONE MARROW DERIVED CELLS DID RETARD PROGRESSION AND IMPACT DIFFERENTLY DISTINCT STAGES OF CHRONIC KIDNEY DISEASE: 1649

Introduction: Different routes for the administration of bone marrow derived cells (BMDC) have been proposed to treat the progression of chronic kidney disease (CKD). In this study, we investigated whether: 1) bovine pericardium-(BP) scaffold for cell therapy would retard progression of CKD; 2) cell therapy impacts differently distinct degrees of mass reduction (5/6 or 2/3 CKD models). Methods: Treatment consisted of BP seeded with either mesenchymal stem cells (BPMSC) or mononuclear cells (BPMO). Renal function and 24-hours proteinuria (PT24h) were measured at 0, 45, and 90 days after surgery. Histological evaluation was performed in all groups. Results: After 90 days animals the group CKD2/3 treated with BPMSC had sCr in levels similar to Sham (S) (BPMSC= 7.6% vs S= 5.2%; P=NS) and lower than the untreated groups (BPMSC= 7.6% vs CKD2/3= 48.2% vs BP= 39%; P=0.043). Following treatment with BPMSC CLcr was higher than in untreated animals (S=1.05 ± 0.20 vs CKD2/3= 0.40 ± 0.12 vs BP= 0.34 ± 0.09 vs BPMSC= 0.65 ± 0.22 vs BPMO= 0.50 ± 0.07ml/min; P< 0.0001) while PT24h was similar to S group (S= 5.4 ± 1.14 vs BPMSC= 4.92 ± 2.4mg/24h [P=NS] vs CKD2/3= 27 ± 16.7 vs BP= 20.2 ± 4.7 vs BPMO= 10 ± 2.8mg/24h; P=0.0005). Progression of CKD2/3 measured by 1/Cr slopes was reduced by BPMSC (S= 0.884 ± 0.91 vs BPMSC= 0.78 ± 0.81 [P=NS] vs CKD2/3= -0.24 ± 0.03 vs BP= -0.20 ± 0.09; P=0.0343). BPMO did not significantly affect sCr, Clcr, or PT24h of CKD2/3 animals. In the CKD5/6 model both treatments were effective to prevent increases in sCr (CKD5/6= 128% vs BP= 168% vs BPMSC= 78.6% vs BPMO= 72.4%; P<0.0001). PT24h decreased significantly in treated groups after 45 days (S= 2.8 ± 1.64 vs CKD5/6= 123.8 ± 112 vs BP= 51.6 ± 32 vs BPMSC= 20.8 ± 26.4 vs BPMO= 9.6 ± 6.6 mg/24h; P=0.0117). This effect seemed to persist at 90 day in spite of not reach statistical significance; (S= 5.4 ± 1.14 vs CKD5/6= 151 ± 109 vs BP= 116 ± 48,7 vs BPMSC= 46,5 ± 37.7 vs BPMO= 66.1 ± 61.38mg/24h; P=0.012) and decreases in Clcr was observed in untreated groups (S= 1.1 ± 0.2 vs CKD5/6= 0.25 ± 0.17 vs BP= 0.22 ± 0.09 vs BPMSC= 0.46 ± 0.10 vs BPMO= 0.53 ± 0.15ml/min; P<0.0001). Progression of disease in the CKD5/6 model was partially retarded also by both treatments (CKD5/6= -0.493 ± 0.04 vs BP= -0.53 ± 0.10 vs BPMSC= -0.35 ± 0.12 vs BPMO= -0.32 ± 0.13; P=0.016), although in a less intense way than that observed in the CKD2/3 model. Histological evaluation showed significantly less renal damage in the remnant kidney of treated rats in both models using both types of BMDC. Conclusions: Our results demonstrate that 1) BP combined to BMDC did retard the progression of experimental CKD; 2) cellular therapy seems to be more effective when given in less severe stages of CKD; 3) BP seeded with BMDC can be an alternative route to cellular therapy.