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Histologic Analysis (histologic + analysis)
Selected AbstractsHistologic Analysis of Clinical Biopsies Taken 6 Months and 3 Years after Maxillary Sinus Floor Augmentation with 80% Bovine Hydroxyapatite and 20% Autogenous Bone Mixed with Fibrin GlueCLINICAL IMPLANT DENTISTRY AND RELATED RESEARCH, Issue 2 2001Mats Hallman DDS Abstract: Background: Bovine hydroxyapatite (Bio-Oss®, Geistlich Pharmaceutical, Wollhausen, Switzerland) has been suggested to be used in maxillary sinus floor augmentation procedures prior to or in conjunction with implant placement. However, the long-term histologic fate of this material is not well understood. Purpose: The aim with this study was to histologically evaluate the tissue response in patients to a mixture of bovine hydroxyapatite (BH), autogenous bone, and fibrin glue 6 months and 3 years after a maxillary sinus floor augmentation procedure. Materials and Method: Biopsies were taken from a group of 20 consecutive patients 6 months (n = 16) and 3 years (n = 12) after maxillary sinus floor augmentation with a mixture of BH (80%), autogenous bone (20%), and fibrin glue and prepared for histologic analysis. Results: Light microscopy and morphometry from biopsies taken after 6 months showed various amounts of mineralized bone tissue. The specimen area was occupied by 54.1 ± 12.6% nonmineralized tissue, followed by 21.2 ± 24.5% lamellar bone, 14.5 ± 10.3% BH particles, and 10.2 ± 13.4% woven bone. The nonmineralized tissue seen in bone-forming areas consisted of a loose connective tissue, rich with vessels and cells. There were no signs of resorption of the BH particles. The lamellar bone appeared to have originated from the recipient site and was seldom in contact with the BH particles. After 3 years, the nonmineralized tissue area had decreased to 36.0 ± 19.0% (p > .05) and consisted mainly of bone marrow tissue. The surface area of lamellar bone had increased to 50.7 ± 22.8% (p > .05), and there was almost no immature bone. The mean specimen area occupied by BH particles, was 12.4 ± 8.7% and had not changed from 6 months (not significant). Moreover, the sizes of the particles were similar after 6 months and 3 years. The degree of BH particle,bone contact had increased from 28.8%± 19.9% after 6 months to 54.5 ± 28.8% after 3 years (p > .05). Conclusion: Histology of specimens from maxillary sinuses augmented with 80% BH particles, 20% autogenous bone, and fibrin glue showed a positive bone tissue response after 6 months and 3 years after augmentation of the maxillary sinus floor prior to implant placement in a group fo 20 patients. The bone surrounding and in contact with the BH particles after 6 months was mainly immature woven bone, which with time was replaced by mature lamellar bone filling the interparticle space as observed in the 3-year specimens. Moreover, bone-integrated BH particles seem to be resistant to resorption. The results indicate that the procedure may be considered when only small amounts of intraoral autogenous bone graft are available. [source] Induction of an antiinflammatory effect and prevention of cartilage damage in rat knee osteoarthritis by CF101 treatmentARTHRITIS & RHEUMATISM, Issue 10 2009S. Bar-Yehuda Objective Studies have suggested that rheumatoid arthritis (RA) and osteoarthritis (OA) share common characteristics. The highly selective A3 adenosine receptor agonist CF101 was recently defined as a potent antiinflammatory agent for the treatment of RA. The purpose of this study was to examine the effects of CF101 on the clinical and pathologic manifestations of OA in an experimental animal model. Methods OA was induced in rats by monosodium iodoacetate, and upon disease onset, oral treatment with CF101 (100 ,g/kg given twice daily) was initiated. The A3 adenosine receptor antagonist MRS1220 (100 ,g/kg given twice daily) was administered orally, 30 minutes before CF101 treatment. The OA clinical score was monitored by knee diameter measurements and by radiographic analyses. Histologic analyses were performed following staining with hematoxylin and eosin, Safranin O,fast green, or toluidine blue, and histologic changes were scored according to a modified Mankin system. Signaling proteins were assayed by Western blotting; apoptosis was detected via immunohistochemistry and TUNEL analyses. Results CF101 induced a marked decrease in knee diameter and improved the changes noted on radiographs. Administration of MRS1220 counteracted the effects of CF101. CF101 prevented cartilage damage, osteoclast/osteophyte formation, and bone destruction. In addition, CF101 markedly reduced pannus formation and lymphocyte infiltration. Mechanistically, CF101 induced deregulation of the NF-,B signaling pathway, resulting in down-regulation of tumor necrosis factor ,. Consequently, CF101 induced apoptosis of inflammatory cells that had infiltrated the knee joints; however, it prevented apoptosis of chondrocytes. Conclusion CF101 deregulated the NF-,B signaling pathway involved in the pathogenesis of OA. CF101 induced apoptosis of inflammatory cells and acted as a cartilage protective agent, which suggests that it would be a suitable candidate drug for the treatment of OA. [source] Presurgical Curettage Appropriately Reduces the Number of Mohs Stages by Better Delineating the Subclinical Extensions of Tumor MarginsDERMATOLOGIC SURGERY, Issue 9 2005Vinh Q. Chung MD Background. Whether presurgical curettage (PC), light curettage performed before Mohs surgery to delineate tumor margin, is appropriate or causes unnecessary removal of normal tissue has not been well established. Objective. We aim to determine histologically whether PC appropriately increases the size of the stage I specimen or causes unnecessary removal of healthy tissue. Methods. Before a surgical margin guided by PC was taken, a hypothetical margin determined by visual and tactile assessment alone (no curettage [NC]) was marked outside the clinically defined tumor. Histologic analysis at the NC and the PC margins revealed whether the increase in the stage I specimen as a result of PC was appropriate. Results. PC appropriately increased the stage I specimen in 21 cases and unnecessarily removed normal tissue in only 1 case. The estimation of tumor margins with PC was 15 times more accurate than with NC (p value = .0012). Conclusion. For basal cell and squamous cell carcinomas at least 4 mm in diameter, light curettage performed prior to Mohs surgery could better delineate subclinical extensions of the tumor margin and appropriately increase the size of the stage I specimen. [source] Integration and differentiation of human embryonic stem cells transplanted to the chick embryoDEVELOPMENTAL DYNAMICS, Issue 1 2002Ronald S. Goldstein Abstract Human embryonic stem (ES) cells are pluripotent cells that can differentiate into a large array of cell types and, thus, hold promise for advancing our understanding of human embryology and for contributing to transplantation medicine. In this study, differentiation of human ES cells was examined in vivo by in ovo transplantation to organogenesis-stage embryos. Colonies of human ES cells were grafted into or in place of epithelial-stage somites of chick embryos of 1.5 to 2 days of development. The grafted human ES cells survived in the chick host and were identified by vital staining with carboxyfluorescein diacetate or use of a green fluorescent protein,expressing cells. Histologic analysis showed that human ES cells are easily distinguished from host cells by their larger, more intensely staining nuclei. Some grafted cells differentiated en masse into epithelia, whereas others migrated and mingled with host tissues, including the dorsal root ganglion. Colonies grafted directly adjacent to the host neural tube produced primarily structures with the morphology and molecular characteristics of neural rosettes. These structures contain differentiated neurons as shown by ,-3-tubulin and neurofilament expression in axons and cell bodies. Axons derived from the grafted cells penetrate the host nervous system, and host axons enter the structures derived from the graft. Our results show that human ES cells transplanted in ovo survive, divide, differentiate, and integrate with host tissues and that the host embryonic environment may modulate their differentiation. The chick embryo, therefore, may serve as an accessible and unique experimental system for the study of in vivo development of human ES cells. © 2002 Wiley-Liss, Inc. [source] Benign parathyroid cyst causing vocal fold paralysis: A case report and review of the literatureHEAD & NECK: JOURNAL FOR THE SCIENCES & SPECIALTIES OF THE HEAD AND NECK, Issue 6 2006Daniel H. Coelho MD Abstract Background. Parathyroid cysts are uncommon, frequently asymptomatic lesions of the neck and superior mediastinum. Symptomatic parathyroid cysts are very rare, with roughly only 200 cases reported in the literature. Of these, only nine cases have been reported with recurrent laryngeal nerve (RLN) paralysis Methods. We report a case of a 49-year-old man initially seen with a 6-month history of worsening hoarseness. Physical examination revealed a palpable 3-cm, firm, smooth, nontender mass of the right thyroid lobe. Fiberoscopic laryngoscopy showed right vocal cord immobility consistent with RLN paralysis. After CT and fine-needle aspiration of the mass, the patient underwent a right thyroid lobectomy. During surgery, the recurrent laryngeal nerve was found to be stretched and adherent to a right inferior lobe mass. Results. Histologic analysis of the surgical specimen revealed a benign parathyroid adenomatous cyst. Postoperatively, the patient's voice improved markedly. This case represents an extremely rare return of function of the RLN after cyst removal. Conclusion. Parathyroid cysts should be included in the differential diagnosis for vocal fold paralysis. © 2006 Wiley Periodicals, Inc. Head Neck 28:564,566, 2006 [source] Age-Related Increase in Atrial Fibrillation Induced by Transvenous Catheter-Based Atrial Burst Pacing: An In Vivo Rat Model of Inducible Atrial FibrillationJOURNAL OF CARDIOVASCULAR ELECTROPHYSIOLOGY, Issue 1 2010DONGZHU XU M.D. AF Rat Model Induced by Transvenous Catheter Pacing.,Introduction: Large animal models of atrial fibrillation (AF) are well established, but limited experimental reports exist on small animal models. We sought to develop an in vivo rat model of AF using a transvenous catheter and to evaluate the model's underlying characteristics. Methods and Results: Echocardiogram, surface electrocardiogram (ECG), and atrial effective refractory period (AERP) were recorded at baseline in young (3 months) and middle-aged (9 months) Wistar rats. AF inducibility and duration were measured through transvenous electrode catheter in young (n = 11) and middle-aged rats (n = 11) and middle-aged rats treated with either pilsicainide (1 mg/kg iv, n = 7) or amiodarone (10 mg/kg iv, n = 9). Degrees of interstitial fibrosis and cellular hypertrophy in the atria were assessed histologically. The P-wave duration and AERP were significantly longer and echocardiographic left atrial dimension significantly larger in middle-aged versus young rats. AF was inducible in >90% of all procedures in both untreated rat groups, whereas AF inducibility was reduced by the antiarrhythmic drugs. The AF duration was significantly longer in middle-aged than in young rats and was significantly shortened by treatment with either pilsicainide or amiodarone. Histologic analysis revealed significant increases in atrial interstitial fibrosis and cellular diameter in middle-aged versus young rats. Conclusions: Transvenous catheter-based AF is significantly longer in middle-aged than in young rats and is markedly reduced by treatment with antiarrhythmic drugs. This rat model of AF is simple, reproducible, and reliable for examining pharmacologic effects on AF and studying the process of atrial remodeling.(J Cardiovasc Electrophysiol, Vol. 21, pp. 88,93, January 2010) [source] Transatrial Access to the Normal Pericardial Space For Local Cardiac Therapy: Preclinical Safety Testing with Aspirin and Pulmonary Artery HypertensionJOURNAL OF INTERVENTIONAL CARDIOLOGY, Issue 5 2001TODD C. PULERWITZ M.D. The reliability, rapidity, and safety of nonsurgical, transatrial pericardial access for local cardiac therapy have been demonstrated in healthy animals. Since many patients take aspirin or have increased right-sided pressures, we evaluated the procedure's safety under these conditions. Transatrial pericardial access was performed in anesthetized pigs following aspirin administration (162 mg po, n = 6) or during experimental pulmonary artery hypertension (n = 4 different animals) and required only 3 minutes following guide catheter positioning. Platelet aggregability testing with arachidonic acid confirmed aspirin effectiveness. Mean pericardial fluid hematocrit was 0.1 ± 0.1% after 2 days of aspirin therapy and 1.9 ± 1.1% at sacrifice 24 hours later (NS). Mean pericardial fluid hematocrit was 1.0 ± 0.5% after 45 minutes of pulmonary artery hypertension and 4.3 ± 0.8% at sacrifice 30 minutes later (NS). Histologic analysis in both groups revealed a small thrombus and localized inflammation at the site of puncture. Neither aspirin use nor pulmonary artery hypertension causes significant bleeding into the pericardial space following transatrial access and thus does not preclude this route for local cardiac drug delivery. [source] Joint degeneration following closed intraarticular fracture in the mouse knee: A model of posttraumatic arthritisJOURNAL OF ORTHOPAEDIC RESEARCH, Issue 5 2007Bridgette D. Furman Abstract Posttraumatic arthritis is one of the most frequent causes of disability following joint trauma. The objective of this study was to develop a model of a closed articular fracture in the mouse knee joint to quantify the temporal sequence of joint degeneration in a model of posttraumatic arthritis. Closed intraarticular fractures were created in the tibial plateau of adult mice (C57BL/6) using a computer-controlled materials testing system and a custom-built indenter tip. Tibial plateau fractures were classified and imaged over time using high-resolution digital radiography. Animals were sacrificed at 2, 4, 8, and 50 weeks following fracture, and the experimental and contralateral control limbs were harvested for histology and micro-computed tomography (microCT) analysis. By radiographic analysis, tibial plateau fractures closely resembled clinical fractures. More complex and comminuted fractures correlated to significantly higher fracture energies. Histologic analysis demonstrated progressive joint degeneration as measured by a modified Mankin scale, with fibrillation and loss of proteoglycan in the articular cartilage. Subchondral bone thickening was also observed in experimental joints. The induction of a closed intraarticular fracture of the mouse tibial plateau generated a reproducible and clinically relevant joint injury that progressed to osteoarthritis-like changes by histologic and microCT evaluations. The ability to induce joint degeneration without an osteotomy or open arthrotomy provides a valuable new model for studying the natural sequelae of posttraumatic arthritis. Notably, the use of a murine model will facilitate the use of genetically modified animals for the investigation of specific genes implicated in the pathology of posttraumatic arthritis. © 2007 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 25:578,592, 2007 [source] Effects of intense pulsed light on sun-damaged human skin, routine, and ultrastructural analysis,LASERS IN SURGERY AND MEDICINE, Issue 2 2002Victor G. Prieto MD Abstract Background and Objective New, non-ablative methods can be used in skin rejuvenation. Histologic analysis of non-ablative IPL effects on facial, sun-damaged skin. Study Design/Materials and Methods Five female subjects, wrinkle class I or II and Fitzpatrick skin types I, II, and III. IPL treatment: once monthly, 560-nm cut-off filters, spot size 8×35 mm, 28,36 J/cm. Routine histology or electron microscopy on 2-mm punches, before treatment and then 1 week, 3 months, and 12 months. Results Pre-treatment specimens contained solar elastosis and perifollicular lymphoid infiltrates. Collagen and elastic fibers appeared unaffected by treatment. At 1-week, Demodex organisms appeared coagulated. Conclusions Under these conditions, IPL induces minimal morphologic changes in mildly sun-damaged skin. Some esthetic improvement may be secondary to clearing of Demodex organisms and reduction of associated lymphocytic infiltrate. Lasers Surg. Med. 30:82,85, 2002. © 2002 Wiley-Liss, Inc. [source] Short-term administration of (-)-epigallocatechin gallate reduces hepatic steatosis and protects against warm hepatic ischemia/reperfusion injury in steatotic miceLIVER TRANSPLANTATION, Issue 3 2005Ryan N. Fiorini Hepatic steatosis increases the extent of cellular injury incurred during ischemia/reperfusion (I/R) injury. (-)-Epigallocatechin gallate (EGCG), the major flavonoid component of green tea (camellia sinensis) is a potent antioxidant that inhibits fatty acid synthase (FAS) in vitro. We investigated the effects of EGCG on hepatic steatosis and markers of cellular damage at baseline and after I/R injury in ob/ob mice. Animals were pretreated with 85 mg/kg EGCG via intraperitoneal (ip) injection for 2 days or oral consumption in the drinking water for 5 days before 15 minutes of warm ischemia and 24 hours of reperfusion. After EGCG administration, total baseline hepatic fat content decreased from baseline. Palmitic acid and linoleic acid levels also were reduced substantially in all ECGC-treated animals before I/R. Alanine aminotransferase (ALT) levels decreased in all EGCG-treated animals compared with control animals after I/R. Histologic analysis demonstrated an average decrease of 65% necrosis after EGCG administration. EGCG administration also increased resting hepatic energy stores as determined by an increase in cellular adenosine triphosphate (ATP) with a concomitant decrease in uncoupling protein 2 (UCP2) before I/R. Finally, there was an increased level of glutathione (GSH) in the EGCG-treated mice compared with the vehicle-treated mice both at baseline and after I/R. In conclusion, taken together, this study demonstrates that treatment with ECGC by either oral or ip administration, significantly protects the liver after I/R, possibly by reducing hepatic fat content, increasing hepatic energy status, and functioning as an antioxidant. (Liver Transpl 2005;11:298,308.) [source] Factitious Purpura in a 10-Year-Old GirlPEDIATRIC DERMATOLOGY, Issue 5 2009Kayo Yamada M.D. Both congenital and autoimmune hemorrhagic disorders were excluded based on her past medical history and physical and laboratory findings. Child abuse was also ruled out as purpura continued to develop after child,family separation. Histologic examination of the skin lesions revealed disruption of collagen fiber bundles. This finding indicated application of external force, leading to a definitive diagnosis of factitious purpura. Although it is very rare in school-age children, the diagnosis of factitious purpura should be included in the differential diagnosis of purpura in children. Histologic analysis of skin biopsies may aid in establishing the diagnosis. [source] Cutaneous Crohn Disease in a ChildPEDIATRIC DERMATOLOGY, Issue 1 2006Anna Luisa Pinna M.D. It is a very rare condition, especially in children, and is easily misdiagnosed when characteristic gastrointestinal symptoms are absent. We report a 10-year-old Caucasian girl with a 6-month history of erythematous, firm tumescence of the left labium majus pudendi and moist vegetations circumscribing the anal ostium. Histologic analysis of skin biopsy specimens from both types of lesions showed a characteristic granulomatous noncaseating infiltrate throughout the dermis. Endoscopic examination and a colon biopsy specimen showed chronic granulomatous inflammation consistent with Crohn disease. Treatment with prednisolone 20 mg daily, metronidazole 250 mg three times daily, topical corticosteroids, and mupirocin ointment produced marked improvement of the vulvar edema, whereas the perianal lesion had a relapsing course. Early recognition of extra-intestinal manifestations of Crohn disease, which are extremely rare in children, may be difficult. Coexistence of contiguous and noncontiguous lesions, representing the very first signs of the disease, are further peculiarities in our patient. [source] The Effect of Hyperbaric Oxygen Therapy on Erectile Function Recovery in a Rat Cavernous Nerve Injury ModelTHE JOURNAL OF SEXUAL MEDICINE, Issue 3 2008Alexander Müller MD ABSTRACT Introduction., Cavernosal oxygenation appears to be important for preservation of erectile tissue health. Hyperbaric oxygen therapy (HBOT) has been shown to improve tissue oxygenation and has neuromodulatory effects. Aim., This study was designed to define the effects of HBOT on erectile function (EF) and cavernosal tissue in the rat cavernous nerve (CN) injury model. Methods., Four groups of Sprague-Dawley rats were studied: rats with bilateral CN crush, HBOT treated (Crush+/HBOT+); bilateral CN-crush/no HBOT (C+/H,); no crush/no HBOT (C,/H,); and no crush/HBOT (C,/H+). HBOT was delivered daily for 90 minutes at three atmospheres for 10 days commencing the day of CN crush. Main Outcome Measures., Ten days after CN injury, the animals underwent CN stimulation measuring the maximal intracavernosal pressure/mean arterial pressure (ICP/MAP) ratios. Corporal tissue was harvested pre-sacrifice, and immunohistochemically stained for nerve growth factor (NGF), endothelial nitric oxide synthase (eNOS), and cluster of differentiation molecule (CD31). Histologic analysis was performed for Masson's trichrome to assess the smooth muscle,collagen ratio. Terminal deoxynucleotidyl transferase Biotin-dUTP Nick End Labeling assay was used to define apoptotic indices (AIs). Results., The C+/H, group had significantly lower ICP/MAP ratios compared with C,/H, rats, (31% vs. 70%, P < 0.001). C+/H+ rats had significantly higher ICP/MAP ratio recovery compared with the C+/H, group (55% vs. 31%, P = 0.005). NGF and eNOS staining densities were higher in C+/H+ rats compared with C+/H, rats (P < 0.05 and P < 0.001, respectively). No difference was seen in CD31 expression. Staining density for MT displayed a trend toward higher smooth muscle preservation after HBOT. AIs were significantly increased by HBOT (P < 0.05). Conclusion., HBOT following a CN injury improved EF preservation in this model, supporting the cavernosal oxygenation concept as protective mechanism for EF. The effects appear to be mediated via preservation of neurotrophic and endothelial factor expression. Müller A, Tal R, Donohue JF, Akin-Olugbade Y, Kobylarz K, Paduch D, Cutter SC, Mehrara BJ, Scardino PT, and Mulhall JP. The effect of hyperbaric oxygen therapy on erectile function recovery in a rat cavernous nerve injury model. J Sex Med 2008;5:562,570. [source] Viscoelastic and Histologic Properties in Scarred Rabbit Vocal Folds After Mesenchymal Stem Cell Injection,THE LARYNGOSCOPE, Issue 7 2006S Hertegård MD Abstract Objective/Hypothesis: The aim of this study was to analyze the short-term viscoelastic and histologic properties of scarred rabbit vocal folds after injection of human mesenchymal stem cells (MSC) as well as the degree of MSC survival. Because MSCs are antiinflammatory and regenerate mesenchymal tissues, can MSC injection reduce vocal fold scarring after injury? Study Design: Twelve vocal folds from 10 New Zealand rabbits were scarred by a localized resection and injected with human MSC or saline. Eight vocal folds were left as controls. Material and Methods: After 4 weeks, 10 larynges were stained for histology and evaluation of the lamina propria thickness. Collagen type I content was analyzed from six rabbits. MSC survival was analyzed by fluorescent in situ hybridization staining from three rabbits. Viscoelasticity for 10 vocal folds was analyzed in a parallel-plate rheometer. Results: The rheometry on fresh-frozen samples showed decreased dynamic viscosity and lower elastic modulus (P < .01) in the scarred samples injected with MSC as compared with the untreated scarred group. Normal controls had lower dynamic viscosity and elastic modulus as compared with the scarred untreated and treated vocal folds (P < .01). Histologic analysis showed a higher content of collagen type 1 in the scarred samples as compared with the normal vocal folds and with the scarred folds treated with MSC. MSCs remained in all samples analyzed. Conclusions: The treated scarred vocal folds showed persistent MSC. Injection of scarred rabbit vocal folds with MSC rendered improved viscoelastic parameters and less signs of scarring expressed as collagen content in comparison to the untreated scarred vocal folds. [source] Critical Role for IL-6 in Hypertrophy and Fibrosis in Chronic Cardiac Allograft RejectionAMERICAN JOURNAL OF TRANSPLANTATION, Issue 8 2009J. A. Diaz Chronic cardiac allograft rejection is the major barrier to long term graft survival. There is currently no effective treatment for chronic rejection except re-transplantation. Though neointimal development, fibrosis, and progressive deterioration of graft function are hallmarks of chronic rejection, the immunologic mechanisms driving this process are poorly understood. These experiments tested a functional role for IL-6 in chronic rejection by utilizing serial echocardiography to assess the progression of chronic rejection in vascularized mouse cardiac allografts. Cardiac allografts in mice transiently depleted of CD4+ cells that develop chronic rejection were compared with those receiving anti-CD40L therapy that do not develop chronic rejection. Echocardiography revealed the development of hypertrophy in grafts undergoing chronic rejection. Histologic analysis confirmed hypertrophy that coincided with graft fibrosis and elevated intragraft expression of IL-6. To elucidate the role of IL-6 in chronic rejection, cardiac allograft recipients depleted of CD4+ cells were treated with neutralizing anti-IL-6 mAb. IL-6 neutralization ameliorated cardiomyocyte hypertrophy, graft fibrosis, and prevented deterioration of graft contractility associated with chronic rejection. These observations reveal a new paradigm in which IL-6 drives development of pathologic hypertrophy and fibrosis in chronic cardiac allograft rejection and suggest that IL-6 could be a therapeutic target to prevent this disease. [source] Monoclonal anti-CD8 therapy induces disease amelioration in the K/BxN mouse model of spontaneous chronic polyarthritisARTHRITIS & RHEUMATISM, Issue 10 2010Bruno R. Raposo Objective CD8+ T cells are part of the T cell pool infiltrating the synovium in rheumatoid arthritis (RA). However, their role in the pathogenesis of RA has not been fully delineated. Using the K/BxN mouse model of spontaneous chronic arthritis, which shares many similarities with RA, we studied the potential of CD8+ T cell depletion with monoclonal antibodies (mAb) to stop and reverse the progression of experimental arthritis. Methods CD8+ T cells from the blood and articular infiltrate of K/BxN mice were characterized for cell surface phenotypic markers and for cytokine production. Additionally, mice were treated with specific anti-CD8 mAb (YTS105 and YTS169.4), with and without thymectomy. Results CD8+ T cells from the peripheral blood and joints of K/BxN mice were mainly CD69+ and CD62L,CD27+ T cells expressing proinflammatory cytokines (interferon-, [IFN,], tumor necrosis factor , [TNF,], interleukin-17a [IL-17A], and IL-4), and granzyme B. In mice receiving anti-CD8 mAb, the arthritis score improved 5 days after treatment. Recovery of the CD8+ T cells was associated with a new increase in the arthritis score after 20 days. In thymectomized and anti-CD8 mAb,treated mice, the arthritis score improved permanently. Histologic analysis showed an absence of inflammatory infiltrate in the anti-CD8 mAb,treated mice. In anti-CD8 mAb,treated mice, the serologic levels of TNF,, IFN,, IL-6, and IL-5 normalized. The levels of the disease-related anti,glucose-6-phosphate isomerase antibodies did not change. Conclusion These results indicate that synovial activated effector CD8+ T cells locally synthesize proinflammatory cytokines (IFN,, TNF,, IL-17, IL-6) and granzyme B in the arthritic joint, thus playing a pivotal role in maintaining chronic synovitis in the K/BxN mouse model of arthritis. [source] In vivo microfocal computed tomography and micro,magnetic resonance imaging evaluation of antiresorptive and antiinflammatory drugs as preventive treatments of osteoarthritis in the ratARTHRITIS & RHEUMATISM, Issue 9 2010Michael D. Jones Objective To determine whether treatment with an antiresorptive drug in combination with an antiinflammatory drug reduces periarticular bone and soft tissue adaptations associated with the progression of posttraumatic secondary osteoarthritis (OA). Methods We used in vivo microfocal computed tomography (micro-CT) to map bony adaptations and in vivo micro,magnetic resonance imaging (micro-MRI) to examine joint inflammation in a rat model of surgically induced OA secondary to knee triad injury. We examined the arthroprotective effects of the bisphosphonates alendronate and risedronate and the nonsteroidal antiinflammatory drug (NSAID) meloxicam. Results Micro-CT revealed reduced levels of periarticular trabecular bone loss in animals with knee triad injury treated with the bisphosphonate drugs alendronate or risedronate, or the NSAID meloxicam, compared with untreated animals. Alendronate treatment reduced bony osteophyte development. While risedronate as a monotherapy did not positively impact osteophytogenesis, combination therapy with risedronate and meloxicam reduced osteophyte severity somewhat. Micro-MRI revealed an increased, diffuse water signal in the epiphyses of untreated rats with knee triad injury 8 weeks after surgery, suggestive of a bone marrow lesion,like stimulus. In contrast, meloxicam-treated rats showed a significant reduction in fluid signal compared with both bisphosphonate-treated groups 8 weeks after surgery. Histologic analysis qualitatively confirmed the chondroprotective effect of both bisphosphonate treatments, showing fewer degradative changes compared with untreated rats with knee triad injury. Conclusion Our findings indicate that select combinations of bisphosphonate and NSAID drug therapy in the early stages of secondary OA preserve trabecular bone mass and reduce the impact of osteophytic bony adaptations and bone marrow lesion,like stimulus. Bisphosphonate and NSAID therapy may be an effective disease-modifying drug regimen if administered early after the initial injury. [source] Prevention of cartilage degeneration and restoration of chondroprotection by lubricin tribosupplementation in the rat following anterior cruciate ligament transectionARTHRITIS & RHEUMATISM, Issue 8 2010Gregory D. Jay Objective To investigate whether cartilage degeneration is prevented or minimized following intraarticular injections of lubricin derived from human synoviocytes in culture, recombinant human PRG4 (rhPRG4), or human synovial fluid (SF) in a rat model of anterior cruciate ligament (ACL) injury. Methods Unilateral ACL transection (ACLT) was performed in Lewis rats (n = 45). Nine animals were left untreated. The remaining rats were given intraarticular injections (50 ,l/injection) of either phosphate buffered saline (PBS) (n = 9), human synoviocyte lubricin (200 ,g/ml; n = 9), rhPRG4 (200 ,g/ml; n = 9), or human SF lubricin (200 ,g/ml; n = 9) twice weekly beginning on day 7 after injury. Joints were harvested on day 32 after injury. Histologic analysis was performed using Safranin O,fast green staining, and articular cartilage degeneration was graded using the Osteoarthritis Research Society International (OARSI),modified Mankin criteria. Histologic specimens were immunoprobed for lubricin and sulfated glycosaminoglycans. A 24-hour urine collection was performed on days 17 and 29 postinjury, and urinary C-terminal telopeptide of type II collagen (CTX-II) levels were measured. Results Treatment with human synoviocyte lubricin resulted in significantly lower OARSI scores for cartilage degeneration compared with no treatment or PBS treatment (P < 0.05). Increased immunostaining for lubricin in the superficial zone chondrocytes and on the surface of cartilage was observed in lubricin-treated, but not untreated or PBS-treated, joints. On day 17, urinary CTX-II levels in human synoviocyte lubricin, and human SF lubricin,treated animals were significantly lower than those in untreated animals (P = 0.005 and P = 0.002, respectively) and in PBS-treated animals (P = 0.002 and P < 0.001, respectively). Conclusion After treatment with any of the 3 types of lubricin evaluated in this study, a reduction in cartilage damage following ACLT was evident, combined with a reduction in type II collagen degradation. Our findings indicate that intraarticular lubricin injection following an ACL injury may be beneficial in retarding the degeneration of cartilage and the development of posttraumatic OA. [source] Loss of cartilage structure, stiffness, and frictional properties in mice lacking PRG4ARTHRITIS & RHEUMATISM, Issue 6 2010Jeffrey M. Coles Objective To assess the role of the glycoprotein PRG4 in joint lubrication and chondroprotection by measuring friction, stiffness, surface topography, and subsurface histology of the hip joints of Prg4,/, and wild-type (WT) mice. Methods Friction and elastic modulus were measured in cartilage from the femoral heads of Prg4,/, and WT mice ages 2, 4, 10, and 16 weeks using atomic force microscopy, and the surface microstructure was imaged. Histologic sections of each femoral head were stained and graded. Results Histologic analysis of the joints of Prg4,/, mice showed an enlarged, fragmented surface layer of variable thickness with Safranin O,positive formations sometimes present, a roughened underlying articular cartilage surface, and a progressive loss of pericellular proteoglycans. Friction was significantly higher on cartilage of Prg4,/, mice at age 16 weeks, but statistically significant differences in friction were not detected at younger ages. The elastic modulus of the cartilage was similar between cartilage surfaces of Prg4,/, and WT mice at young ages, but cartilage of WT mice showed increasing stiffness with age, with significantly higher moduli than cartilage of Prg4,/, mice at older ages. Conclusion Deletion of the gene Prg4 results in significant structural and biomechanical changes in the articular cartilage with age, some of which are consistent with osteoarthritic degeneration. These findings suggest that PRG4 plays a significant role in preserving normal joint structure and function. [source] The critical role of kinase activity of interleukin-1 receptor,associated kinase 4 in animal models of joint inflammationARTHRITIS & RHEUMATISM, Issue 6 2009Magdalena Koziczak-Holbro Objective We have previously reported that the kinase activity of interleukin-1 receptor,associated kinase 4 (IRAK-4) is important for Toll-like receptor and interleukin-1 receptor signaling in vitro. Using mice devoid of IRAK-4 kinase activity (IRAK-4 KD mice), we undertook this study to determine the importance of IRAK-4 kinase function in complex disease models of joint inflammation. Methods IRAK-4 KD mice were subjected to serum transfer,induced (K/BxN) arthritis, and migration of transferred spleen lymphocytes into joints and cartilage and bone degradation were assessed. T cell response in vivo was tested in antigen-induced arthritis (AIA) by measuring the T cell,dependent antigen-specific IgG production and frequency of antigen-specific T cells in the spleen and lymph nodes. T cell allogeneic response was tested in vitro by mixed lymphocyte reaction (MLR). Results Lipopolysaccharide-induced local neutrophil influx into subcutaneous air pouches was impaired in IRAK-4 KD mice. These mice were also protected from inflammation in the K/BxN and AIA models, as shown by reduced swelling of joints. Histologic analysis of joints of K/BxN serum,injected mice revealed that bone erosion, osteoclast formation, and cartilage matrix proteoglycan loss were reduced in IRAK-4 KD mice. Assessment of T cell response by MLR, by frequency of antigen-specific clones, and by production of antigen-specific IgG did not reveal substantial differences between IRAK-4 KD and wild-type mice. Conclusion These results demonstrate that IRAK-4 is a key component for the development of proarthritis inflammation, but that it is not crucial for T cell activation. Therefore, the kinase function of IRAK-4 appears to be an attractive therapeutic target in chronic inflammation. [source] Two-Year Outcome with Nobel Direct® Implants: A Retrospective Radiographic and Microbiologic Study in 10 PatientsCLINICAL IMPLANT DENTISTRY AND RELATED RESEARCH, Issue 3 2009Tommie Van de Velde LA ABSTRACT Introduction: The Nobel Direct® implant (Nobel Biocare AB, Göteborg, Sweden) was developed to minimize marginal bone resorption and to result in "soft tissue integration" for an optimized aesthetic outcome. However, conflicting results have been presented in the literature. The aim of this present study was to evaluate the clinical and microbiologic outcomes of Nobel Direct implants. Materials and Methods: Ten partially edentulous subjects without evidence of active periodontitis (mean age 55 years) received 12 Nobel Direct implants. Implants were loaded with single crowns after a healing period of 3 to 6 months. Treatment outcomes were assessed at month 24. Routine clinical assessments, intraoral radiographs, and microbiologic samplings were made. Histologic analysis of one failing implant and chemical spectroscopy around three unused implants was performed. Paired Wilcoxon signed-rank test was used for the evaluation of bone loss; otherwise, descriptive analysis was performed. Results: Implants were functionally loaded after 3 to 6 months. At 2 years, the mean bone loss of remaining implants was 2.0 mm (SD ± 1.1 mm; range: 0.0,3.4 mm). Three out of 12 implants with an early mean bone loss >3 mm were lost. The surviving implants showed increasing bone loss between 6 and 24 months (p = .028). Only 3 out of the 12 implants were considered successful and showed bone loss of <1.7 mm after 2 years. High rates of pathogens, including Aggregatibacter actinomycetemcomitans, Fusobacterium spp., Porphyromonas gingivalis, Pseudomonas aeruginosa, and Tanerella forsythia, were found. Chemical spectroscopy revealed, despite the normal signals from Ti, O, and C, also peaks of P, F, S, N, and Ca. A normal histologic image of osseointegration was observed in the apical part of the retrieved implant. Conclusion: Radiographic evidence and 25% implant failures are indications of a low success rate. High counts and prevalence of significant pathogens were found at surviving implants. Although extensive bone loss had occurred in the coronal part, the apical portion of the implant showed some bone to implant integration. [source] Partial generation of periodontal ligament on endosseous dental implants in dogsCLINICAL ORAL IMPLANTS RESEARCH, Issue 4 2005Leila Jahangiri Abstract Objectives: The objective of this pilot study was to investigate the feasibility of periodontal ligament (PDL) generation on an implant surface by approximating a tooth-to-implant contact using orthodontics. Methods: Maxillary second premolars of six beagle dogs were extracted bilaterally. After 2 weeks of healing, hydroxyapatite (HA) coated titanium implants, 5 mm in length and 3.3 mm in diameter, were placed in the extraction sites. One side of the arch was used as control. Orthodontic tooth movement was initiated following implant placement to tip the first premolar roots into contact with the implant. This was achieved in 4,6 weeks as confirmed radiographically. Tooth-to-implant contact was maintained for further 6 weeks after which time, teeth were separated from implant contact orthodontically. After further 2 weeks of stabilization, the animals were sacrificed. Samples were analyzed by Faxitron radiographs before histology. Histology samples were prepared with Stevenel's Blue and Van Gieson stain and were subjected to polarized light microscopy. Results: Histologic analysis revealed transfer and formation of PDL-like structure with formation of cellular cementum on the implant surfaces, in four out of six animals, where tooth-to-implant contact had been achieved. Direct bone-to-implant contact was noted in the areas coronal to the PDL-like tissue, an important sign to distinguish between PDL-like tissue and connective tissue that could originate from the coronal portion of a failing implant. Additionally, at the site of contact, the implant surface revealed some resorption of the HA coating. Conclusion: An animal model was established in which the proximity of tooth-to-implant contact lead to partial generation of PDL on a bioactive implant surface in four out of six animals. Résumé L'objectif de cette étude pilote a été d'analyser la possibilité d'obtenir une régénération du ligament parodontal (PDL) sur une surface implantaire en rapprochant le contact implant-dent par l'orthodontie. Les deuxièmes prémolaires maxillaires de six chiens beagle ont été avulsées bilatéralement. Après deux semaines de guérison, des implants en titane recouverts d'hydroxyapatite de 5 mm de long et de 3,3 mm de diamètre ont été insérés dans les sites d'avulsion. Un site de chaque arche était utilisé comme contrôle. Le mouvement orthodontique de la dent était initialisé après le placement de l'implant pour mettre en contact les raçines des premières prémolaires avec l'implant. Ceci a été réalisé en quatre à huit semaines comme l'a confirmé la radiographie. Le contact dent-implant a été maintenu pour une durée supplémentaire de six semaines après lesquelles les dents ont été séparées du contact des implants de manière orthodontique. Après deux semaines de stabilisation, les animaux ont été euthanasiés. Les échantillons ont été analysés par radiographie Faxitron avant l'histologie. Les échantillons histologiques ont été préparés par les colorations du Bleu de Stevenel et de Van Gieson et ont été analysés au microscope optique à polarisation. L'histologiea révélé un transfert et une formation d'une structure ressemblant à PDL avec la formation d'un cément cellulaire à la surface des implants chez quatre des six animaux chez lesquels le contact dent-implant avaient été réalisés. Le contact direct os-implant a été noté dans les zones coronaires au tissu qui ressemblait à PDL, un signe important pour distinguer le tissu ressemblant à PDL et le tissu conjonctif qui pouvait provenir de la portion coronaire de l'implant qui échoue. De plus, au site de contact, la surface implantaire révélait quelques résorptions du recouvrement d'hydroxyapatite. Un modèle animal a donc étéétabli dans lequel la proximité du contact dent-implant était suivie d'une génération partielle de PDL sur une surface implantaire bioactive chez quatre des six animaux. Zusammenfassung Ziele: Das Ziel dieser Pilotstudie war, zu untersuchen, ob es möglich sei, ein parodontales Ligament (PDL) auf einer Implantatoberfläche durch Herstellung eines Zahn-Implantat-Kontakts mittels Orthodontie zu generieren. Methoden: Bei sechs Beagle-Hunden wurden bilateral die zweiten Prämolaren extrahiert. Nach einer Abheilzeit von zwei Wochen wurden Hydroxyapatit beschichtete Titanimplantate mit einer Länge von 5 mm und einem Durchmesser von 3.3 mm in die Extraktionsstellen eingesetzt. Eine Seite des Zahnbogens diente als Kontrolle. Nach der Implantatplatzierung wurde mit der orthodontischen Zahnbewegung begonnen, um die Wurzeln der ersten Prämolaren in Kontakt mit den Implantaten zu bringen. Es wurde radiologisch bestätigt, dass dies nach 4,6 Wochen erreicht wurde. Der Zahn-Implantat-Kontakt wurde für weitere sechs Wochen belassen, danach wurden die Zähne wieder orthodontisch vom Kontakt mit den Implantaten separiert. Nach weiteren zwei Wochen zur Stabilisierung wurden die Tiere geopfert. Die Proben wurden vor der Histologie mittels Faxitron Röntgen analysiert. Die histologischen Proben wurden mit Stevenel`s Blau und van Gieson Färbungen bearbeitet und durch Lichtmikroskopie mit polarisiertem Licht untersucht. Resultate: Die histologische Analyse zeigte einen Transfer und eine Formation von Parodontalligament ähnlichen Strukturen mit der Bildung von zellulärem Zement auf der Implantatoberfläche bei 4 von 6 Tieren, bei welchen ein Zahn-Implantat-Kontakt erreicht worden war. Koronal des PDL-ähnlichen Gewebes bestand ein direkter Knochen-Implantat-Kontakt. Dies war ein wichtiges Merkmal, um zwischen PDL-ähnlichem Gewebe und Bindegewebe, welches vom koronalen Anteil eines fehlgeschlagenen Implantats stammen könnte, zu unterscheiden. Zusätzlich zeigten die Implantatoberflächen im Bereich der Berührungsfläche etwas Resorption der Hydroxyapatit Beschichtung. Schlussfolgerung: Es wurde ein Tiermodell etabliert, in welchem ein Zahn-Implantat-Kontakt bei 4 von 6 Tieren zu einer partiellen Generation eines PDL auf einer bioaktiven Implantatoberfläche führte. Resumen Objetivos: El objetivo de este estudio piloto fue investigar la viabilidad de generación de ligamento periodontal (PDL) en la superficie de un implante por aproximación de un contacto diente a implante usando ortodoncia. Métodos: Se extrajeron los segundos premolares de seis perros beagle bilateralmente. Tras dos semanas de cicatrización, se colocaron en los lugares de extracción implantes de titanio cubiertos de hidroxiapatita, de 5 mm de longitud y 3.3 mm de diámetro. Un lado del arco se usó como control. El movimiento dental ortodóntico se inició tras la colocación del implante para apuntar las raíces del primer premolar hacia el contacto con el implante. Esto se logró en 4-6 semanas tal como se confirmó radiograficamente. El contacto diente a implante se mantuvo durante seis semanas mas tras lo cual, los dientes se separaron del contacto con el implante ortodonticamente. Tras dos semanas más de estabilización, los animales se sacrificaron. Las muestras se analizaron por medio de radiografías Faxitron antes de la histología. Las muestras histológicas se prepararon con azul de Stevenel y tinción de Van Gieson y se sometieron a microscopio de luz polarizada. Resultados: El análisis histológico reveló transferencia y formación de estructuras tipo ligamento periodontal con formación de cemento celular en las superficies del implante, en 4 de 6 animales, en los que se logró el contacto diente a implante. Se notó contacto en las áreas coronales al tejido tipo PDL, un signo importante para distinguir tejido tipo PDL y tejido conectivo que se podría originar de la parte coronal de un implante en vías de fracaso. Además, en el lugar del contacto, la superficie del implante reveló algo de reabsorción de la cubierta de hidroxiapatita. Conclusión: Se estableció un modelo animal en el que la proximidad del contacto diente a implante llevó a la generación parcial de PDL en una superficie de implante bioactiva en 4 de 6 animales. [source] Orthodontic movement after periodontal regeneration of class II furcation: a pilot study in dogsJOURNAL OF CLINICAL PERIODONTOLOGY, Issue 6 2006Vanessa Camila Da Silva Abstract Purpose: The effect of orthodontic movement on the periodontal tissues of maxillary second pre-molars, after regenerative treatment for class II furcations, was evaluated in four mongrel dogs. Material and Methods: Class II furcation lesions were created. After 75 days they were treated with bovine bone mineral matrix and guided tissue regeneration with absorbable membrane. After 2 months of daily plaque control, each of the dog's furcation pre-molars was randomly assigned to a test or control group. Orthodontic appliances were placed on both sides of the maxilla using third pre-molars and canines as anchorages. In the test group, bodily orthodontic movement of the second pre-molars was performed in the mesial direction for 3 months while control pre-molars remained unmoved. The dogs were sacrificed for histometric and histologic analyses. Results: There were no statistically significant differences between the two groups in total bone and biomaterial areas or linear extension of periodontal regeneration on the radicular surfaces. In the test group, however, there was a tendency to a greater quantity of bone and a lesser quantity of biomaterial. Conclusion: The orthodontic movement was not pre-judicial to the results obtained with the regenerative periodontal treatment. [source] Quantitation of myelin oligodendrocyte glycoprotein and myelin basic protein in the thymus and central nervous system and its relationship to the clinicopathologic features of autoimmune encephalomyelitisJOURNAL OF NEUROSCIENCE RESEARCH, Issue 3 2006Hiroshi Sakuma Abstract There is controversy whether the amount of autoantigens expressed in the thymus regulates negative selection of autoreactive T cells and determine susceptibility or resistance to experimental autoimmune encephalomyelitis (EAE). In the present study, we have addressed this issue by quantifying neuroantigens in the thymus of two EAE-susceptible (LEW and LEW.1AV1) and one EAE-resistant (BN) rat strains. We further examined whether amounts of neuroantigens in various parts of the central nervous system (CNS) affect the clinical course and lesion distribution of acute and chronic EAE. Real-time PCR and histologic analyses showed that there was no significant difference in the amount and distribution of myelin oligodendrocyte glycoprotein and myelin basic protein in the thymus and CNS among the three strains and that both acute and chronic EAE lesions in the CNS were preferentially distributed in the area where neuroantigens were abundantly present. These findings suggest that susceptibility or resistance to EAE is not regulated by the amount of the neuroantigens expressed in the thymus. Furthermore, the lesion distribution, but not the clinical course, of EAE is related to the neuroantigen expression in the CNS. © 2006 Wiley-Liss, Inc. [source] Bone healing in critical-size defects treated with platelet-rich plasma: a histologic and histometric study in rat calvariaJOURNAL OF PERIODONTAL RESEARCH, Issue 2 2008M. R. Messora Background and Objective:, The purpose of this study was to analyze histologically the influence of autologous platelet-rich plasma on bone healing in surgically created critical-size defects in rat calvaria. Material and Methods:, Thirty-two rats were divided into two groups: the control group (group C) and the platelet-rich plasma group. An 8-mm-diameter critical-size defect was created in the calvarium of each animal. In group C the defect was filled by a blood clot only. In the platelet-rich plasma group, 0.35 mL of platelet-rich plasma was placed in the defect and covered by 0.35 mL of platelet-poor plasma. Both groups were divided into subgroups (n = 8) and killed at either 4 or 12 wk postoperatively. Histometric (using image-analysis software) and histologic analyses were performed. The amount of new bone formed was calculated as a percentage of the total area of the original defect. Percentage data were transformed into arccosine for statistical analysis (analysis of variance, Tukey, p < 0.05). Results:, No defect completely regenerated with bone. The platelet-rich plasma group had a statistically greater amount of bone formation than group C at both 4 wk (17.68% vs. 7.20%, respectively) and 12 wk (24.69% vs. 11.65%, respectively) postoperatively. Conclusion:, Within the limits of this study, it can be concluded that platelet-rich plasma placed in the defects and covered by platelet-poor plasma significantly enhanced bone healing in critical-size defects in rat calvaria. [source] Attenuation of pain and inflammation in adjuvant-induced arthritis by the proteasome inhibitor MG132ARTHRITIS & RHEUMATISM, Issue 7 2010Aisha S. Ahmed Objective In rheumatoid arthritis (RA), pain and joint destruction are initiated and propagated by the production of proinflammatory mediators. Synthesis of these mediators is regulated by the transcription factor NF-,B, which is controlled by the ubiquitin proteasome system (UPS). The present study explored the effects of the proteasome inhibitor MG132 on inflammation, pain, joint destruction, and expression of sensory neuropeptides as markers of neuronal response in a rat model of arthritis. Methods Arthritis was induced in rats by injection of heat-killed Mycobacterium butyricum. Arthritis severity was scored, and nociception was evaluated by mechanical pressure applied to the hind paw. Joint destruction was assessed by radiologic and histologic analyses. NF-,B DNA-binding activity was analyzed by electromobility shift assay, and changes in the expression of the p50 NF-,B subunit and the proinflammatory neuropeptides substance P (SP) and calcitonin gene-related peptide (CGRP) were detected by immunohistochemistry. Results Arthritic rats treated with MG132 demonstrated a marked reduction in inflammation, pain, and joint destruction. The elevated DNA-binding activity of the NF-,B/p50 homodimer and p50, as well as the neuronal expression of SP and CGRP, observed in the ankle joints of arthritic rats were normalized after treatment with MG132. Conclusion In arthritic rats, inhibition of proteasome reduced the severity of arthritis and reversed the pain behavior associated with joint inflammation. These effects may be mediated through the inhibition of NF-,B activation and may possibly involve the peripheral nervous system. New generations of nontoxic proteasome inhibitors may represent a novel pharmacotherapy for RA. [source] Akt1 in murine chondrocytes controls cartilage calcification during endochondral ossification under physiologic and pathologic conditionsARTHRITIS & RHEUMATISM, Issue 3 2010Atsushi Fukai Objective To examine the role of the phosphoinositide-dependent serine/threonine protein kinase Akt1 in chondrocytes during endochondral ossification. Methods Skeletal phenotypes of homozygous Akt1-deficient (Akt1,/,) mice and their wild-type littermates were compared in radiologic and histologic analyses. An experimental osteoarthritis (OA) model was created by surgically inducing instability in the knee joints of mice. For functional analyses, we used primary costal and articular chondrocytes from neonatal mice and mouse chondrogenic ATDC5 cells with retroviral overexpression of constitutively active Akt1 or small interfering RNA (siRNA) for Akt1. Results Among the Akt isoforms (Akt1, Akt2, and Akt3), Akt1 was the most highly expressed in chondrocytes, and the total level of Akt protein was decreased in Akt1,/, chondrocytes, indicating a dominant role of Akt1. Akt1,/, mice exhibited dwarfism with normal proliferative and hypertrophic zones but suppressed cartilage calcification in the growth plate compared with their wild-type littermates. In mice with surgically induced OA, calcified osteophyte formation, but not cartilage degradation, was prevented in the Akt1,/, joints. Calcification was significantly suppressed in cultures of Akt1,/, chondrocytes or ATDC5 cells overexpressing siRNA for Akt1 and was enhanced in ATDC5 cells overexpressing constitutively active Akt1. Neither proliferation nor hypertrophic differentiation was affected by the gain or loss of function of Akt1. The expression of ANK and nucleotide pyrophosphatase/phosphodiesterase 1, which accumulate pyrophosphate, a crucial calcification inhibitor, was enhanced by Akt1 deficiency or siRNA for Akt1 and was suppressed by constitutively active Akt1. Conclusion Our findings indicate that Akt1 in chondrocytes controls cartilage calcification by inhibiting pyrophosphate during endochondral ossification in skeletal growth and during osteophyte formation in OA. [source] Amelioration of alphavirus-induced arthritis and myositis in a mouse model by treatment with bindarit, an inhibitor of monocyte chemotactic proteinsARTHRITIS & RHEUMATISM, Issue 8 2009Nestor E. Rulli Objective Alphaviruses such as chikungunya virus, Sindbis virus, o'nyong-nyong virus, Mayaro virus, and Ross River virus (RRV), are commonly associated with arthralgias and overt arthritides worldwide. Understanding the processes by which arthritogenic viruses cause disease is a prerequisite in the quest for better treatments. In this regard, we have recently established that monocyte/macrophages are mediators of alphavirus-induced arthritis in mice. We hypothesized that chemokines associated with monocyte/macrophage recruitment may play an important role in disease. The aim of the present investigations was to determine whether bindarit, an inhibitor of monocyte chemotactic protein (MCP) synthesis, could ameliorate alphavirus-induced rheumatic disease in mice. Methods Using our recently developed mouse model of RRV-induced arthritis, which has many characteristics of RRV disease (RRVD) in humans, the effects of bindarit treatment on RRVD in mice were determined via histologic analyses, immunohistochemistry, flow cytometry, real-time polymerase chain reaction analysis, enzyme-linked immunosorbent assay, and electrophoretic mobility shift assay. Results Bindarit-treated RRV-infected mice developed mild disease and had substantially reduced tissue destruction and inflammatory cell recruitment as compared with untreated RRV-infected mice. The virus load in the tissues was not affected by bindarit treatment. Bindarit exhibited its activity by down-regulating MCPs, which in turn led to inhibition of cell infiltration and lower production of NF-,B and tumor necrosis factor ,, which are involved in mediating tissue damage. Conclusion Our data support the use of inhibitors of MCP production in the treatment of arthritogenic alphavirus syndromes and suggest that bindarit may be useful in treating RRVD and other alphavirus-induced arthritides in humans. [source] Blocking vascular endothelial growth factor with soluble Flt-1 improves the chondrogenic potential of mouse skeletal muscle,derived stem cellsARTHRITIS & RHEUMATISM, Issue 1 2009Seiji Kubo Objective To investigate the effect of vascular endothelial growth factor (VEGF) stimulation and the effect of blocking VEGF with its antagonist, soluble Flt-1 (sFlt-1), on chondrogenesis, using muscle-derived stem cells (MDSCs) isolated from mouse skeletal muscle. Methods The direct effect of VEGF on the in vitro chondrogenic ability of mouse MDSCs was tested using a pellet culture system, followed by real-time quantitative polymerase chain reaction (PCR) and histologic analyses. Next, the effect of VEGF on chondrogenesis within the synovial joint was tested, using genetically engineered MDSCs implanted into rat osteochondral defects. In this model, MDSCs transduced with a retroviral vector to express bone morphogenetic protein 4 (BMP-4) were coimplanted with MDSCs transduced to express either VEGF or sFlt-1 (a VEGF antagonist) to provide a gain- and loss-of-function experimental design. Histologic scoring was used to compare cartilage formation among the treatment groups. Results Hyaline-like cartilage matrix production was observed in both VEGF-treated and VEGF-blocked (sFlt-1,treated) pellet cultures, but quantitative PCR revealed that sFlt-1 treatment improved the expression of chondrogenic genes in MDSCs that were stimulated to undergo chondrogenic differentiation with BMP-4 and transforming growth factor ,3 (TGF,3). In vivo testing of articular cartilage repair showed that VEGF-transduced MDSCs caused an arthritic change in the knee joint, and sFlt-1 improved the MDSC-mediated repair of articular cartilage, compared with BMP-4 alone. Conclusion Soluble Flt-1 gene therapy improved the BMP-4, and TGF,3-induced chondrogenic gene expression of MDSCs in vitro and improved the persistence of articular cartilage repair by preventing vascularization and bone invasion into the repaired articular cartilage. [source] Myopericytoma of the oral cavityHEAD & NECK: JOURNAL FOR THE SCIENCES & SPECIALTIES OF THE HEAD AND NECK, Issue 6 2007Vivekanand Datta MD Abstract Background. Myopericytoma is a rare mesenchymal neoplasm of pericytic cells demonstrating myoid differentiation. The lesion typically arises within the subcutaneous tissue of the extremities. We report a case that, to the best of our knowledge, is the first case of myopericytoma involving the soft tissue of the oral cavity. Methods. A 36-year-old woman had a 5-mm sessile, whitish-pink, firm tongue nodule. The patient underwent excisional biopsy, and histopathologic examination as well as immunohistochemical analysis were performed. Results. The differential diagnosis by histologic analysis included solitary fibrous tumor, myofibroma, glomus tumor, and myopericytoma. The results of immunohistochemical analysis, when combined with the histologic features, led to a diagnosis of myopericytoma. Conclusions. Applying strict morphologic criteria and appropriately selective immunohistochemical markers will help to distinguish myopericytoma in the oral cavity. © 2007 Wiley Periodicals, Inc. Head Neck, 2007 [source] | ||||||||||||||||||||||||||||