Calcium Hydroxide Promotes Cementogenesis and Induces Cementoblastic Differentiation of Mesenchymal Periodontal Ligament Cells in a CEMP1-and ERK-Dependent Manner

Periodontal tissue engineering is a complex process requiring the regeneration of bone, cementum, and periodontal ligament (PDL). Since cementum regeneration is poorly understood, we used a dog model of dental pulpal necrosis and in vitro cellular wounding and mineralization assays to determine the mechanism of action of calcium hydroxide, Ca(OH)(2), in cementogenesis. Laser capture microdissection (LCM) followed by qRT-PCR were used to assay responses of periapical tissues to Ca(OH)(2) treatment. Additionally, viability, proliferation, migration, and mineralization responses of human mesenchymal PDL cells to Ca(OH)(2) were assayed. Finally, biochemical inhibitors and siRNA were used to investigate Ca(OH)(2)-mediated signaling in PDL cell differentiation. In vivo, Ca(OH)(2)-treated teeth formed a neocementum in a STRO-1- and cementum protein-1 (CEMP1)-positive cellular environment. LCM-harvested tissues adjacent to the neocementum exhibited higher mRNA levels for CEMP1, integrin-binding sialoprotein, and Runx2 than central PDL cells. In vitro, Ca(OH)(2) and CEMP1 promoted STRO-1-positive cell proliferation, migration, and wound closure. Ca(OH)(2) stimulated expression of the cementum-specific proteins CEMP1 and PTPLA/CAP in an ERK-dependent manner. Lastly, Ca(OH)(2) stimulated mineralization by CEMP1-positive cells. Blocking CEMP1 and ERK function abolished Ca(OH)(2)-induced mineralization, confirming a role for CEMP1 and ERK in the process. Ca(OH)(2) promotes cementogenesis and recruits STRO-1-positive mesenchymal PDL cells to undergo cementoblastic differentiation and mineralization via a CEMP1- and ERK-dependent pathway.

Vimentin Expression and Myofibroblast Infiltration Are Early Markers of Renal Dysfunction in Kidney Transplantation: An Early Stage of Chronic Allograft Dysfunction?

Introduction. The objective of this study was to show the morphologic characteristics of allograft renal biopsies in renal transplant patients with stable renal function, which can potentially be early markers of allograft dysfunction, after 5 years of follow-up. Methods. Forty-nine renal transplant patients with stable renal function were submitted to renal biopsies and simultaneous measurement of serum creatinine (Cr). Histology was evaluated using Banff scores, determination of interstitial fibrosis by Sirius red staining and immunohistochemical study of proximal tubule and interstitial compartment (using cytokeratin, vimentin, and myofibroblasts as markers). Biopsies were evaluated according to the presence or absence of the epitheliomesenchymal transition (EMT). The interstitial presence of myofibroblasts and tubular presence of vimentin was also analyzed simultaneously. Renal function was measured over the follow-up period to estimate the reduction of graft function. Results. Median posttransplant time at enrollment was 105 days. Patients were followed for 64.3 +/- 8.5 months. The mean Cr at biopsy time was 1.44 +/- 0.33 mg/dL, and after the follow-up it was 1.29 +/- 0.27 mg/dL. Nine patients (19%) had a reduction of their graft function. Eleven biopsies (22%) had tubulointerstitial alterations according to Banff score. Seventeen biopsies (34%) presented EMT. Fifteen biopsies (32%) had high interstitial expression of myofibroblasts and tubular vimentin. Using Cox multivariate analysis, HLA and high expression of interstitial myofibroblasts and tubular vimentin were associated with reduction of graft function, yielding a risk of 3.3 (P = .033) and 9.8 (P = .015), respectively. Conclusion. Fibrogenesis mechanisms occur very early after transplantation and are risk factors for long-term renal function deterioration.

Functional and morphologic evaluation of kidney proximal tubuli and correlation with renal allograft prognosis

P>Renal transplant patients with stable graft function and proximal tubular dysfunction (PTD) have an increased risk for chronic allograft nephropathy (CAN). In this study, we investigated the histologic pattern associated with PTD and its correlation with graft outcome. Forty-nine transplant patients with stable graft function were submitted to a biopsy. Simultaneously, urinary retinol-binding protein (uRBP) was measured and creatinine clearance was also determined. Banff`s score and semi-quantitative histologic analyses were performed to assess tubulointerstitial alterations. Patients were followed for 24.0 +/- 7.8 months. At biopsy time, mean serum creatinine was 1.43 +/- 0.33 mg/dl. Twelve patients (24.5%) had uRBP >= 1 mg/l, indicating PTD and 67% of biopsies had some degree of tubulointerstitial injury. At the end of the study period, 18 (36.7%) patients had lost renal function. uRBP levels were not associated with morphologic findings of interstitial fibrosis and tubular atrophy (IF/TA), interstitial fibrosis measured by Sirius red or tubulointerstitial damage. However, in multivariate analysis, the only variable associated with the loss of renal function was uRBP level >= 1 mg/l, determining a risk of 5.290 of loss of renal function (P = 0.003). Renal transplant patients who present PTD have functional alteration, which is not associated with morphologic alteration. This functional alteration is associated to progressive decrease in renal function.

Detection of the Tim-3 ligand, galectin-9, inside the allograft during a rejection episode

Introduction: Tim-3 is a Th1 lymphocytes membrane protein with inhibitory function. Its ligand, galectin-9, was recently identified and it is expressed in some lymphocyte subpopulation. In addition, endothelial cells and fibroblasts can also express galectin-9 according to the local cytokine milieu. Both molecules can act as important regulatory tools in the immune system. Aim: Evaluate the expression of these immunoregulatory molecules inside kidney allografts during acute rejection episodes. Methods: By using a quantitative polymerase chain reaction assay, we measured the levels of messenger RNA (mRNA) for galectin-9 and Tim-3 in 21 samples obtained at allograft nephrectomy. Five samples received the histological diagnosis of acute non-vascular rejection (ANVR), twelve of acute vascular rejection (AVR), and five of loss of non-immune cause (LNIC; as control). As cytolytic response markers we measured mRNA levels of granzyme B, interferon-gamma and perforin. The statistic analysis was performed using one way analysis of variance (ANOVA) and Pearson correlation. Results: The mean levels of Tim-3 mRNA expression were 13.99 +/- 6.99 for LNIC, 48.13 +/- 54.47 for RACNV and 238.63 +/- 333.14 for RAV (p = 0.004). For galectin-9, the mean values were 0.57 +/- 0.49 for LNIC, 0.66 +/- 0.36 for RACNV and 2.34 +/- 1.62 for RAV (p = 0.006). Furthermore, there was a positive correlation between both molecules (r = 0.526, p = 0.016). Also. granzyme B, perforin and interferon-gamma mRNA expression were different among the three groups. Conclusion: Messenger RNA level expressions of all the studied molecules were higher inside allografts with more severe rejection. Moreover, there was a positive correlation between galectin-9 and Tim-3 mRNA levels. The simultaneous expression of galectin-9 and Tim-3 may indicate an immunoregulatory function, during the ongoing cytotoxic response. (C) 2008 Elsevier B.V. All rights reserved.

Toll-like receptors-related genes in kidney transplant patients with chronic allograft nephropathy and acute rejection

Introduction: Toll-like receptors (TLR) comprehend an emerging family of receptors that recognize pathogen-associated molecular patterns and promote the activation of leukocytes. Surgical trauma and ischemia-reperfusion injury are likely to provide exposure to endogenous ligands for TLR in virtually all kidney transplant recipients. Methods: Macroarray (GEArray OHS-018.2 Series-Superarray) analyses of 128 genes involved in TLR signaling pathway were performed in nephrectomy samples of patients with chronic allograft nephropathy (CAN) and acute rejection (AR, vascular and non vascular). The analysis of each membrane was performed by GEArray Expression Analysis Suite 2.0. Results: Macroarray profile identified a gene expression signature that could discriminate CAN and AR. Three genes were significantly expressed between CAN and vascular AR: Pellino 2; IL 8 and UBE2V1. In relation to vascular and non-vascular AR, there were only two genes with statistical significance: IL-6 and IRAK-3. Conclusion: Vascular and non-vascular AR and CAN showed different expression of a few genes in TLR pathway. The analysis of nephrectomy showed that activation of TLR pathway is present in AR and CAN. (C) 2008 Elsevier B.V. All rights reserved.

Rupture of the cranial cruciate ligament associated with deformity of the proximal tibia in five dogs

Excessive mechanical stress due to caudal sloping of the tibial plateau may result In early breakdown of the cranial cruciate ligament (CrCL). Five dogs with CrCL rupture associated with caudal sloping of the proximal tibial plateau are described. All were small dogs, of between three and six years of age, with a mean bodyweight of 9.3 kg, which had acute hindlimb lameness. Radiographic examination revealed cranial displacement of the tibia, with a tibial angle varying from 58 to 60 degrees. All cases were treated with a lateral fabellotibial suture and cranial cuneiform osteotomy of the proximal tibia. All dogs were using the operated limb three days after surgery, with normal gait re-established after a mean period of 10 days. Excessive tibial plateau sloping is not a frequent cause of hindlimb lameness In small animals, although it Is Important to consider it as a predisposing factor for rupture of the CrCL.

Clinical and radiographic evaluation of a polyester prosthesis in dogs with cranial cruciate ligament rupture

Os achados clínicos e radiográficos após reparo intra-articular do ligamento cruzado cranial com prótese de poliéster, utilizando a técnica over-the-top modificada, foram avaliada em seis cães. Sete cirurgias foram realizadas devido ao acometimento bilateral em um dos animais. A avaliação clínica foi realizada ao 3º, 10º e 40º dias de pós-operatório, e a avaliação radiográfica realizada ao 5º e 24º meses após a cirurgia em cinco cães, por meio da qual se confirmou a progressão da doença articular degenerativa. A resolução dos sinais clínicos foi observada entre o 25º e 68º dias após a cirurgia, segundo avaliação realizada pelos proprietários. A função do membro operado foi considerada boa. Dois cães apresentaram desgaste e ruptura da prótese após a cirurgia. Concluiu-se que a prótese de poliéster, da forma como foi implantada neste estudo, não pode ser considerada como substituto satisfatório, uma vez que resultados superiores podem ser obtidos com ténicas menos invasivas e mais simples.

Role of the oblique ligament in the integrity of the medial collateral ligament of the canine elbow joint

Foram estudados o arranjo e o diâmetro médio das fibrilas colágenas do ligamento colateral medial da articulação do cotovelo do cão, isolado ou associado ao ligamento oblíquo e tracionado até a ruptura. Dezoito articulações foram divididas em três grupos. O primeiro grupo teve o ligamento colateral medial coletado, mas não tracionado; o segundo grupo teve o ligamento colateral medial tracionado isoladamente; o terceiro grupo teve os ligamentos colateral medial e oblíquo tracionados associadamente. O ligamento colateral medial não submetido ao ensaio de tração apresentou um padrão ondulado das fibras colágenas, o qual não foi totalmente destruído quando foi tracionado, associado ao ligamento oblíquo, e perdeu totalmente o padrão reticular das fibras colágenas quando testado isoladamente. Quando o ligamento colateral medial foi submetido à tensão isoladamente, o diâmetro médio das fibrilas colágenas aumentou em relação ao grupo não submetido à tensão. Associado ao ligamento oblíquo, o diâmetro médio das fibrilas colágenas foi o maior na região de inserção e o menor na região média, em relação aos outros grupos. Concluiu-se que o ligamento oblíquo pode favorecer a integridade da região de inserção do ligamento colateral medial, aumentando a eficácia de sua reconstrução após a lesão.

Gastric Torsion from an Intestinal Hernia Through a Rent in the Gastrosplenic Ligament in a Horse

The present report describes an 8-year-old gelding presenting with signs of severe abdominal pain. After performing a thorough physical examination, including rectal palpation and additional diagnostic tests, an exploratory laparotomy was recommended. The jejunum was found herniated through the gastrosplenic ligament, and the stomach was severely distended with gas. Given a poor prognosis, the horse was euthanized on the table. At necropsy, the stomach appeared dilated, with an 180 horizontal gastric torsion, from left (lateral) to right (medial), dividing the organ into dorsal and ventral compartments. We believe that the chronic traction exerted by an incarcerated and distended loop of jejunum, in the dorsal aspect of the gastrosplenic ligament, associated with trauma during episodes of intense rolling, enlarged the rent until it ruptured. Because of this rupture, the lateral dorsal aspect of the stomach became unattached, predisposing it to the torsion. (C) 2012 Elsevier B.V. All rights reserved.

Role of Peritubular Capillaries and Vascular Endothelial Growth Factor in Chronic Allograft Nephropathy

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Modified stabilization method for the tibial tuberosity advancement technique: a biomechanical study

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Mechanical removal of necrotic periodontal ligament by either Robinson bristle brush with pumice or scalpel blade. Histomorphometric analysis and scanning electron microscopy

One of the important factors accounting for successful delayed replantation of avulsed teeth is seemingly the type of root surface treatment. Removal of necrotic cemental periodontal ligament remnants may prevent the occurrence of external root resorption, which is the major cause of loss of teeth replanted in such conditions. The purpose of this study was to compare the efficacy of two mechanical techniques for removal of root-adhered periodontal ligament. Preservation or removal of the cementum layer concomitantly with these procedures was also assessed. Forty-five roots of healthy premolars extracted for orthodontic purposes were selected. After extraction, the teeth were kept dry at room temperature for 1 h and then immersed in saline for rehydration for an additional 10 min. Thereafter, the roots were assigned to three groups, as follows: group 1 (control) - the cemental periodontal ligament was preserved; group 2 - removal of the periodontal ligament by scraping root surface with a scalpel blade (SBS); group 3 - periodontal ligament remnants were removed using a Robinson bristle brush at low-speed with pumice/water slurry (RBP). The specimens were analysed histomorphometrically and examined by scanning electron microscopy. The quantitative and qualitative analyses of the results showed that the RBP technique was significantly more effective than the SBS technique for removal of the periodontal ligament remnants adhered to root surface. Both techniques preserved the cementum layer.

Dentists' level of knowledge of the treatment plans for periodontal ligament injuries after dentoalveolar trauma

This study investigated the level of knowledge held by dentists about the possible treatment plan procedures for periodontal ligament injuries after dentoalveolar trauma. A 5-item self-applied questionnaire was prepared with questions referring to the professional profile of the interviewees and to the treatment plan they would propose for periodontal ligament injuries secondary to dentoalveolar trauma. The questionnaires were filled out by 693 dentists attending the 23rd Annual Meeting of the Brazilian Society for Dental Research, and the data obtained were subjected to descriptive analysis. Either the chi-square test or Fisher's exact test was applied to assess associations among variables, at a 5% level of significance. The results revealed that dentists experienced difficulty in establishing a treatment plan for subluxation, and for extrusive, lateral and intrusive luxations. In general, holding a dental specialty degree had no influence on the knowledge about treatment plan procedures for the most severe injuries. It could be concluded that the dentists participating in this study, whether specialists or not, did not have sufficient knowledge to treat most of the periodontal ligament injuries resulting from dentoalveolar trauma adequately.

Mechanics of the maxillary central incisor. Influence of the periodontal ligament represented by beam elements

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)