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Excessive hepatic glucose production (HGP) is a key factor promoting hyperglycemia in diabetes. Hepatic cryptochrome 1 (CRY1) plays an important role in maintaining glucose homeostasis by suppressing forkhead box protein O1 (FOXO1)-mediated HGP. Although downregulation of hepatic CRY1 appears to be associated with increased HGP, the mechanism(s) by which hepatic CRY1 dysregulation confers hyperglycemia in diabetic subjects is largely unknown. In this study, we demonstrate that a reduction in hepatic CRY1 protein is stimulated by elevated E3 ligase F-box and leucine-rich repeat protein 3 (FBXL3)-dependent proteasomal degradation in diabetic mice. In addition, we found that GSK3b-induced CRY1 phosphorylation potentiates FBXL3-dependent CRY1 degradation in the liver. Accordingly, in diabetic mice, GSK3b inhibitors effectively decreased HGP by facilitating the effect of CRY1-mediated FOXO1 degradation on glucose metabolism. Collectively, these data suggest that tight regulation of hepatic CRY1 protein stability is crucial for maintaining systemic glucose homeostasis.

Nonalcoholic fatty liver disease (NAFLD) is the most common cause of chronic liver disease and is strongly associated with obesity-related ectopic fat accumulation in the liver. Hepatic lipid accumulation encompasses a histological spectrum ranging from simple steatosis to nonalcoholic steatohepatitis (NASH), which can progress to cirrhosis and hepatocellular carcinoma. Given that dysregulated hepatic lipid metabolism may be an onset factor in NAFLD, understanding how hepatic lipid metabolism is modulated in healthy subjects and which steps are dysregulated in NAFLD subjects is crucial to identify effective therapeutic targets. Additionally, hepatic inflammation is involved in chronic hepatocyte damage during NAFLD progression. As a key immune signaling hub that mediates NF-κB activation, the IκB kinase (IKK) complex, including IKKα, IKKβ, and IKKγ (NEMO), has been studied as a crucial regulator of the hepatic inflammatory response and hepatocyte survival. Notably, TANK-binding kinase 1 (TBK1), an IKK-related kinase, has recently been revealed as a potential link between hepatic inflammation and energy metabolism. Here, we review (1) the biochemical steps of hepatic lipid metabolism; (2) dysregulated lipid metabolism in obesity and NAFLD; and (3) the roles of IKKs and TBK1 in obesity and NAFLD., Obesity: tracing the inflammation underlying liver disease A deeper understanding of the molecular processes underlying dysfunctional fatty-acid metabolism in nonalcoholic fatty liver disease (NAFLD) could help protect patients from severe liver damage. NAFLD is a common consequence of obesity, and severe disease can ultimately give rise to liver cancer or cirrhosis. Alan Saltiel of the University of California, San Diego, USA, and Jin Young Huh of Seoul National University, South Korea, have reviewed current knowledge of the mechanisms governing fatty acid metabolism in the liver, and how they are disturbed in obesity and NAFLD. The various IκB kinase (IKK) complexes and a protein known as TANK-binding kinase 1 (TBK1) play particularly prominent roles in the inflammatory response associated with NAFLD pathogenesis. Better insights into IKK- and TBK1-associated regulatory pathways could inform the development of new interventions for this currently untreatable condition.

In mammals, brown adipose tissue (BAT) is specialized to conduct non-shivering thermogenesis for survival under cold acclimation. Although emerging evidence suggests that lipid metabolites are essential for heat generation in cold-activated BAT, the underlying mechanisms of lipid uptake in BAT have not been thoroughly understood. Here, we show that very-low-density lipoprotein (VLDL) uptaken by VLDL receptor (VLDLR) plays important roles in thermogenic execution in BAT. Compared with wild-type mice, VLDLR knockout mice exhibit impaired thermogenic features. Mechanistically, VLDLR-mediated VLDL uptake provides energy sources for mitochondrial oxidation via lysosomal processing, subsequently enhancing thermogenic activity in brown adipocytes. Moreover, the VLDL-VLDLR axis potentiates peroxisome proliferator activated receptor (PPAR)β/δ activity with thermogenic gene expression in BAT. Accordingly, VLDL-induced thermogenic capacity is attenuated in brown-adipocyte-specific PPARβ/δ knockout mice. Collectively, these data suggest that the VLDL-VLDLR axis in brown adipocytes is a key factor for thermogenic execution during cold exposure.

Macrophage proinflammatory activation is an important etiologic component of the development of insulin resistance and metabolic dysfunction in obesity. However, the underlying mechanisms are not clearly understood. Here, we demonstrate that a mitochondrial inner membrane protein, adenine nucleotide translocase 2 (ANT2), mediates proinflammatory activation of adipose tissue macrophages (ATMs) in obesity. Ant2 expression was increased in ATMs of obese mice compared with lean mice. Myeloid-specific ANT2-knockout (ANT2-MKO) mice showed decreased adipose tissue inflammation and improved insulin sensitivity and glucose tolerance in HFD/obesity. At the molecular level, we found that ANT2 mediates free fatty acid-induced mitochondrial permeability transition, leading to increased mitochondrial reactive oxygen species production and damage. In turn, this increased HIF-1α expression and NF-κB activation, leading to proinflammatory macrophage activation. Our results provide a previously unknown mechanism for how obesity induces proinflammatory activation of macrophages with propagation of low-grade chronic inflammation (metaflammation).

Decreased adipose tissue oxygen tension and increased HIF-1α expression can trigger adipose tissue inflammation and dysfunction in obesity. Our current understanding of obesity-associated decreased adipose tissue oxygen tension is mainly focused on changes in oxygen supply and angiogenesis. Here, we demonstrate that increased adipocyte O2 demand, mediated by ANT2 activity, is the dominant cause of adipocyte hypoxia. Deletion of adipocyte Ant2 improves obesity-induced intracellular adipocyte hypoxia by decreasing obesity-induced adipocyte oxygen demand, without effects on mitochondrial number or mass, or oligomycin-sensitive respiration. This led to decreased adipose tissue HIF-1α expression and inflammation with improved glucose tolerance and insulin resistance in both a preventative or therapeutic setting. Our results suggest that ANT2 may be a target for the development of insulin sensitizing drugs and that ANT2 inhibition might have clinical utility.

White adipose tissue (WAT) is a key regulator of systemic energy metabolism, and impaired WAT plasticity characterized by enlargement of preexisting adipocytes associates with WAT dysfunction, obesity, and metabolic complications. However, the mechanisms that retain proper adipose tissue plasticity required for metabolic fitness are unclear. Here, we comprehensively showed that adipocyte-specific DNA methylation, manifested in enhancers and CTCF sites, directs distal enhancer-mediated transcriptomic features required to conserve metabolic functions of white adipocytes. Particularly, genetic ablation of adipocyte Dnmt1, the major methylation writer, led to increased adiposity characterized by increased adipocyte hypertrophy along with reduced expansion of adipocyte precursors (APs). These effects of Dnmt1 deficiency provoked systemic hyperlipidemia and impaired energy metabolism both in lean and obese mice. Mechanistically, Dnmt1 deficiency abrogated mitochondrial bioenergetics by inhibiting mitochondrial fission and promoted aberrant lipid metabolism in adipocytes, rendering adipocyte hypertrophy and WAT dysfunction. Dnmt1-dependent DNA methylation prevented aberrant CTCF binding and, in turn, sustained the proper chromosome architecture to permit interactions between enhancer and dynamin-1-like protein gene Dnm1l (Drp1) in adipocytes. Also, adipose DNMT1 expression inversely correlated with adiposity and markers of metabolic health but positively correlated with AP-specific markers in obese human subjects. Thus, these findings support strategies utilizing Dnmt1 action on mitochondrial bioenergetics in adipocytes to combat obesity and related metabolic pathology.

Accumulating evidence reveals that adipose tissue is an immunologically active organ that exerts multiple impacts on the regulation of systemic energy metabolism. Adipose tissue immunity is modulated by the interactions between adipocytes and various immune cells. Nevertheless, the underlying mechanisms that control inter-cellular interactions between adipocytes and immune cells in adipose tissue have not been thoroughly elucidated. Recently, it has been demonstrated that adipocytes utilize lipid metabolites as a key mediator to initiate and mediate diverse adipose tissue immune responses. Adipocytes present lipid antigens and secrete lipid metabolites to determine adipose immune tones. In addition, the interactions between adipocytes and adipose immune cells are engaged in the control of adipocyte fate and functions upon metabolic stimuli. In this review, we discuss an integrated view of how adipocytes communicate with adipose immune cells using lipid metabolites. Also, we briefly discuss the newly discovered roles of adipose stem cells in the regulation of adipose tissue immunity.

In mammals, white adipose tissues are largely divided into visceral epididymal adipose tissue (EAT) and subcutaneous inguinal adipose tissue (IAT) with distinct metabolic properties. Although emerging evidence suggests that subpopulations of adipose stem cells (ASCs) would be important to explain fat depot differences, ASCs of two fat depots have not been comparatively investigated. Here, we characterized heterogeneous ASCs and examined the effects of intrinsic and tissue micro-environmental factors on distinct ASC features. We demonstrated that ASC subpopulations in EAT and IAT exhibited different molecular features with three adipogenic stages. ASC transplantation experiments revealed that intrinsic ASC features primarily determined their adipogenic potential. Upon obesogenic stimuli, EAT-specific SDC1

In obesity, adipose tissue undergoes dynamic remodeling processes such as adipocyte hypertrophy, hypoxia, immune responses, and adipocyte death. However, whether and how invariant natural killer T (iNKT) cells contribute to adipose tissue remodeling are elusive. In this study, we demonstrate that iNKT cells remove unhealthy adipocytes and stimulate the differentiation of healthy adipocytes. In obese adipose tissue, iNKT cells were abundantly found nearby dead adipocytes. FasL-positive adipose iNKT cells exerted cytotoxic effects to eliminate hypertrophic and pro-inflammatory Fas-positive adipocytes. Furthermore, in vivo adipocyte-lineage tracing mice model showed that activation of iNKT cells by alpha-galactosylceramide promoted adipocyte turnover, eventually leading to potentiation of the insulin-dependent glucose uptake ability in adipose tissue. Collectively, our data propose a novel role of adipose iNKT cells in the regulation of adipocyte turnover in obesity.

Decreased adipose tissue oxygen tension and increased HIF-1α expression can trigger adipose tissue inflammation and dysfunction in obesity. Our current understanding of obesity-associated decreased adipose tissue oxygen tension is mainly focused on changes in oxygen supply and angiogenesis. Here, we demonstrate that increased adipocyte O2 demand, mediated by ANT2 activity, is the dominant cause of adipocyte hypoxia. Deletion of adipocyte Ant2 improves obesity-induced intracellular adipocyte hypoxia by decreasing obesity-induced adipocyte oxygen demand, without effects on mitochondrial number or mass, or oligomycin-sensitive respiration. This led to decreased adipose tissue HIF-1α expression and inflammation with improved glucose tolerance and insulin resistance in both a preventative or therapeutic setting. Our results suggest that ANT2 may be a target for the development of insulin sensitizing drugs and that ANT2 inhibition might have clinical utility.

Hepatic TANK (TRAF family member associated NFκB activator)-binding kinase 1 (TBK1) activity is increased during obesity, and administration of a TBK1 inhibitor reduces fatty liver. Surprisingly, liver-specific TBK1 knockout in mice produces fatty liver by reducing fatty acid oxidation. TBK1 functions as a scaffolding protein to localize acyl-CoA synthetase long-chain family member 1 (ACSL1) to mitochondria, which generates acyl-CoAs that are channeled for β-oxidation. TBK1 is induced during fasting and maintained in the unphosphorylated, inactive state, enabling its high affinity binding to ACSL1 in mitochondria. In TBK1-deficient liver, ACSL1 is shifted to the endoplasmic reticulum to promote fatty acid re-esterification in lieu of oxidation in response to fasting, which accelerates hepatic lipid accumulation. The impaired fatty acid oxidation in TBK1-deficient hepatocytes is rescued by the expression of kinase-dead TBK1. Thus, TBK1 operates as a rheostat to direct the fate of fatty acids in hepatocytes, supporting oxidation when inactive during fasting and promoting re-esterification when activated during obesity.

Recent findings, notably on adipokines and adipose tissue inflammation, have revised the concept of adipose tissues being a mere storage depot for body energy. Instead, adipose tissues are emerging as endocrine and immunologically active organs with multiple effects on the regulation of systemic energy homeostasis. Notably, compared with other metabolic organs such as liver and muscle, various inflammatory responses are dynamically regulated in adipose tissues and most of the immune cells in adipose tissues are involved in obesity-mediated metabolic complications, including insulin resistance. Here, we summarize recent findings on the key roles of innate (neutrophils, macrophages, mast cells, eosinophils) and adaptive (regulatory T cells, type 1 helper T cells, CD8 T cells, B cells) immune cells in adipose tissue inflammation and metabolic dysregulation in obesity. In particular, the roles of natural killer T cells, one type of innate lymphocyte, in adipose tissue inflammation will be discussed. Finally, a new role of adipocytes as antigen presenting cells to modulate T cell activity and subsequent adipose tissue inflammation will be proposed.

Systemic low-grade chronic inflammation has been intensively investigated in obese subjects. Recently, various immune cell types, such as macrophages, granulocytes, helper T cells, cytotoxic T cells, and B cells, have been implicated in the pathogenesis of adipose tissue inflammation. However, the roles of invariant natural killer T cells (iNKT cells) and the regulation of iNKT cell activity in adipose tissue are not thoroughly understood. Here, we demonstrated that iNKT cells were decreased in number in the adipose tissue of obese subjects. Interestingly, CD1d, a molecule involved in lipid antigen presentation to iNKT cells, was highly expressed in adipocytes, and CD1d-expressing adipocytes stimulated iNKT cell activity through physical interaction. iNKT cell population and CD1d expression were reduced in the adipose tissue of obese mice and humans compared to those of lean subjects. Moreover, iNKT cell-deficient Jα18 knockout mice became more obese and exhibited increased adipose tissue inflammation at the early stage of obesity. These data suggest that adipocytes regulate iNKT cell activity via CD1d and that the interaction between adipocytes and iNKT cells may modulate adipose tissue inflammation in obesity.

Adipose tissue inflammation is an important factor in obesity that promotes insulin resistance. Among various cell types in adipose tissue, immune cells actively regulate inflammatory responses and affect whole-body energy metabolism. In particular, invariant natural killer T (iNKT) cells contribute to mitigating dysregulation of systemic energy homeostasis by counteracting obesity-induced inflammation in adipose tissue. However, the molecular mechanisms by which adipose iNKT cells become activated and mediate anti-inflammatory roles in obese adipose tissue have not been thoroughly understood yet. In the current study, we demonstrate that adipocyte CD1d plays a key role in the stimulation of adipose iNKT cells, leading to anti-inflammatory responses in high-fat diet (HFD)–fed mice. Accordingly, adipocyte-specific CD1d-knockout (CD1dADKO) mice showed reduced numbers of iNKT cells in adipose tissues and decreased responses to α-galactosylceramide–induced iNKT cell activation. Additionally, HFD-fed CD1dADKO mice revealed reduced interleukin-4 expression in adipose iNKT cells and aggravated adipose tissue inflammation and insulin resistance. Collectively, these data suggest that adipocytes could selectively stimulate adipose iNKT cells to mediate anti-inflammatory responses and attenuate excess proinflammatory responses in obese adipose tissue.

Orosomucoid (ORM), also called alpha-1 acid glycoprotein, is an abundant plasma protein that is an immunomodulator induced by stressful conditions such as infections. In this study, we reveal that Orm is induced selectively in the adipose tissue of obese mice to suppress excess inflammation that otherwise disturbs energy homeostasis. Adipose Orm levels were elevated by metabolic signals, including insulin, high glucose, and free fatty acid, as well as by the proinflammatory cytokine tumor necrosis factor-alpha, which is found in increased levels in the adipose tissue of morbid obese subjects. In both adipocytes and macrophages, ORM suppressed proinflammatory gene expression and pathways such as NF-kappaB and mitogen-activated protein kinase signalings and reactive oxygen species generation. Concomitantly, ORM relieved hyperglycemia-induced insulin resistance as well as tumor necrosis factor-alpha-mediated lipolysis in adipocytes. Accordingly, ORM improved glucose and insulin tolerance in obese and diabetic db/db mice. Taken together, our results suggest that ORM integrates inflammatory and metabolic signals to modulate immune responses to protect adipose tissue from excessive inflammation and thereby from metabolic dysfunction.

Berberine (BBR) has been shown to improve several metabolic disorders, such as obesity, type 2 diabetes, and dyslipidemia, by stimulating AMP-activated protein kinase (AMPK). However, the effects of BBR on proinflammatory responses in macrophages are poorly understood. Here we show that BBR represses proinflammatory responses through AMPK activation in macrophages. In adipose tissue of obese db/db mice, BBR treatment significantly downregulated the expression of proinflammatory genes such as TNF-α, IL-1β, IL-6, monocyte chemoattractant protein-1 (MCP-1), inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2). Consistently, BBR inhibited LPS-induced expression of proinflammatory genes including IL-1β, IL-6, iNOS, MCP-1, COX-2, and matrix metalloprotease-9 in peritoneal macrophages and RAW 264.7 cells. Upon various proinflammatory signals including LPS, free fatty acids, and hydrogen peroxide, BBR suppressed the phosphorylation of MAPKs, such as p38, ERK, and JNK, and the level of reactive oxygen species in macrophages. Moreover, these inhibitory effects of BBR on proinflammatory responses were abolished by AMPK inhibition via either compound C, an AMPK inhibitor, or dominant-negative AMPK, implying that BBR would downregulate proinflammatory responses in macrophages via AMPK stimulation.

The aim of this study was to evaluate the use of autogenous bone in combination with platelet-enriched fibrin glue as a grafting material for maxillary sinus augmentation with simultaneous implant placement in dogs.The mucous membranes of 12 sinuses in 6 dogs were elevated bilaterally. In the right sinus, autogenous bone mixed with platelet-enriched fibrin glue was grafted into the space between the membrane and the sinus wall. In the left sinus, autogenous bone alone was grafted as a control. At the same time, 2 dental implants were inserted into the grafting material through the maxillary sinus floor. The animals were killed 6 months after surgery.The mean bone-implant contact was 40.5% on the fibrin glue side and 32.3% on the control side (P.05). The mean height of newly formed bone in the augmented area was 12.2 mm on the fibrin glue side and 10.7 mm on the control side (P.05).The results indicate that the use of autogenous bone mixed with platelet-enriched fibrin glue can achieve results superior to those for grafts of autogenous bone alone. The specific improvements of this technique include enhanced osseointegration of dental implants and increased height of new bone.

Objective The aim of this study was to investigate whether dental implant exposure to the maxillary sinus cavity increases the risk of maxillary sinus complications. Study design An implant was placed bilaterally in the maxillary sinus of 8 adult female mongrel dogs in a way that it penetrated the bone and mucous membrane of the maxillary sinus floor to the extent of 2 mm, 4 mm, or 8 mm. The implants were left in place for 6 months. Results Radiographic and histologic examinations did not show any signs of pathologic findings in the maxillary sinus of the 8 dogs. Conclusion This study indicates that implant protrusion into the maxillary sinus cavity is not related to the development of sinus complications in canines.

Summary Aim The aim of this study was to develop a novel artificial nerve conduit and to evaluate its efficiency based on the promotion of peripheral nerve regeneration in rabbits. Material and methods The nerve conduit was made of a poly (l-lactide- co -glycolic acid)-coated collagen tube filled with collagen gel. The conduits were implanted into a 15mm gap in the peroneal nerves of five rabbits. On the contralateral side, the defects were bridged with collagen-filled vein grafts. Results Twelve weeks postoperatively nerve regeneration was superior to the vein graft in the PLGA-coated collagen tube, both morphologically and electrophysiologically. Conclusion The results indicate the superiority of the PLGA-coated collagen tube over vein grafts. Furthermore, they show that entubulation repair with this type of tube can support nerve regeneration over a nerve gap distance of at least 15mm.

Objective The purpose of this study was to determine the minimum size defect in a canine mandible that would not spontaneously heal during the dog's natural life (the critical size defect). Study design Sixteen adult female mongrel dogs underwent continuity resection on both sides of the mandible to create bilateral defects. In 8 dogs, mandibular defects ranging from 5 to 20 mm were created with periosteal resection. In the other 8 dogs, mandibular defects ranging from 30 to 60 mm were created preserving the periosteum. The dogs were then killed at 6 months and the defects examined using radiographs and histologic analysis. Results When the periosteum was removed, mandibular defects greater than 15 mm failed to heal across the entire defect. However, when the periosteum was preserved, mandibular defects needed to be greater than 50 mm in order to fail to heal. Conclusion The critical size defect in a canine mandible model is 15 mm when the periosteum is removed and 50 mm when the periosteum is preserved.

We have developed a reliable method of microneural anastomosis using cyanoacrylate adhesives. This method involves overlapping the epineuriums of the two nerve ends and then applying two or three microdrops of cyanoacrylate adhesive on the surface of the epineurium just where the epineuriums overlap. The sciatic nerves of Sprague-Dawley rats were transected and repaired either with 10-0 nylon sutures or by using the described method. Histological evaluation showed no significant difference in the outcome of nerve regeneration between the two groups. It was concluded that the cyanoacrylate repair deserves to be considered as an alternative to the conventional suture technique in microneural anastomosis.

Introduction: Vertical augmentation of the alveolar ridge is necessary for extensive resorption of the alveolar ridge. Aim: To evaluate treatment outcome after alveolar ridge augmentation by a sandwich osteotomy combined with an interpositional allograft. Patients and Methods: The deficient alveolar ridges were augmented by a sandwich osteotomy combined with bovine collagen matrix as an interpositional allograft placed between the basal bone and the osteotomized fragment without fixation. Standardized lateral cephalographs were taken of nine patients, before surgery, immediately postoperatively and 3 months after augmentation to evaluate the level of augmentation, bone loss and stability of the osteotomized fragment. Results: The augmentation ranged between 8.4 and 11.0 mm (mean 9.8 mm). Bone resorption in the crestal bone height ranged from 1.5 to 3.0 mm (mean 2.1 mm) after 3 months. Bone resorption in thickness of the osteotomized fragment ranged from 0.3 to 2.0 mm (mean 1.0 mm). Conclusion: Although there was some resorption of the superior and anterior parts of the reconstructed alveolar process, it was concluded that this procedure of augmentation is safe.

SREBP1c is a key transcription factor that regulates de novo lipogenesis during anabolic periods. However, the molecular mechanisms involved in the suppression of SREBP1c under nutritional deprivation are largely unknown. In this study, we demonstrate that the small ubiquitin-related modifier (SUMO) E3 ligase, a protein inhibitor of activated STAT Y (PIASy), sumoylates SREBP1c at Lys98, leading to suppression of the hepatic lipogenic program upon fasting-induced signals. In primary hepatocytes, ablation of PIASy stimulated intracellular lipid accumulation through the induction of SREBP1c and its target genes. Given that protein kinase A (PKA) plays important roles in catabolic responses, activated PKA enhances the sumoylation of SREBP1c and potentiates the interaction between SREBP1c and PIASy. Notably, overexpression of PIASy in obese db/db mice ameliorated hepatic steatosis, while suppression of PIASy in lean (wild-type) mice stimulated hepatic lipogenesis with increased expression of SREBP1c target genes. Furthermore, PKA-mediated SREBP1c phosphorylation augmented SREBP1c sumoylation, subsequently leading to degradation of SREBP1c via ubiquitination. Together, these data suggest that PKA-induced SREBP1c sumoylation by PIASy is a key regulatory mechanism to turn off hepatic lipogenesis during nutritional deprivation.

OBJECTIVE Tissue inflammation is a key factor underlying insulin resistance in established obesity. Several models of immuno-compromised mice are protected from obesity-induced insulin resistance. However, it is unanswered whether inflammation triggers systemic insulin resistance or vice versa in obesity. The purpose of this study was to assess these questions. RESEARCH DESIGN AND METHODS We fed a high-fat diet (HFD) to wild-type mice and three different immuno-compromised mouse models (lymphocyte-deficient Rag1 knockout, macrophage-depleted, and hematopoietic cell-specific Jun NH2-terminal kinase–deficient mice) and measured the time course of changes in macrophage content, inflammatory markers, and lipid accumulation in adipose tissue, liver, and skeletal muscle along with systemic insulin sensitivity. RESULTS In wild-type mice, body weight and adipose tissue mass, as well as insulin resistance, were clearly increased by 3 days of HFD. Concurrently, in the short-term HFD period inflammation was selectively elevated in adipose tissue. Interestingly, however, all three immuno-compromised mouse models were not protected from insulin resistance induced by the short-term HFD. On the other hand, lipid content was markedly increased in liver and skeletal muscle at day 3 of HFD. CONCLUSIONS These data suggest that the initial stage of HFD-induced insulin resistance is independent of inflammation, whereas the more chronic state of insulin resistance in established obesity is largely mediated by macrophage-induced proinflammatory actions. The early-onset insulin resistance during HFD feeding is more likely related to acute tissue lipid overload.

The aim of this study was to compare the effects of platelet-enriched fibrin glue and platelet-rich plasma (PRP) on the repair of bone defects adjacent to titanium dental implants.In 6 mongrel dogs, 3 screw-shaped titanium dental implants per dog were placed into the osteotomy sites in the tibia. Before implantation, a standardized gap (2.0 mm) was created between the implant surface and the surrounding bone walls. Six gaps were left empty (control group), 6 gaps were filled with autogenous particulate bone mixed with PRP (PRP group), and 6 gaps were filled with autogenous particulate bone mixed with platelet-enriched fibrin glue (fibrin glue group).After 6 weeks, the bone-implant contact was 59.7% in the fibrin glue group, 29.2% in the PRP group, and 10.2% in the control defects; this difference was statistically significant (P.05).Greater bone-implant contact was achieved with platelet-enriched fibrin glue than with PRP. The results indicate that platelet-enriched fibrin glue can induce a stronger peri-implant bone reaction than PRP in the treatment of bone defects adjacent to titanium dental implants.

Objective The aim of this study was to evaluate the combined use of autogenous bone and platelet-enriched fibrin glue as grafting material for vertical alveolar ridge augmentation with simultaneous implant placement in a canine alveolar ridge defect model. Study design In 6 mongrel dogs, bilateral vertical alveolar ridge defects were created in the mandible. After 3 months of healing, 2 dental implants were placed in each defect of the mandible, creating 6-mm supra-alveolar peri-implant defects. The 2 implants per defect were subjected to surgical treatments involving either a combination of autogenous bone grafts and platelet-enriched fibrin glue, or a conventional flap procedure only (control). After a healing period of 6 months, the dogs were humanely killed for histological and histometric analyses. Results Implant placement alone produced limited vertical alveolar height (0.6 ± 0.4 mm). However, alveolar augmentation including a combination of autogenous bone grafts and platelet-enriched fibrin glue with simultaneous implant placement resulted in alveolar ridge augmentation amounting to 4.2 ± 1.0 mm, comprising 63% of the defect height. New bone-implant contact was 40.5% in the defects treated with combined autogenous bone grafts and platelet-enriched fibrin glue, and was 48.4% in the resident bone; this difference was not statistically significant. Conclusion The present study demonstrates that vertical alveolar ridge augmentation using autogenous bone grafts and platelet-enriched fibrin glue with simultaneous implant placement might effectively increase vertical alveolar ridge height and allow for an acceptable level of osseointegration.

Objective The purpose of this study was to evaluate the effects of autogenous bone grafts and platelet-enriched fibrin glue in the treatment of peri-implantitis. Study design Thirty-six screw-type commercially pure titanium implants with rough acid-etched surfaces were inserted into 6 mongrel dogs 3 months after extraction of mandibular premolars. After 3 months of healing, peri-implantitis was induced by placing gauze and wire around the implants. Once peri-implantitis was created, surgical treatments involving a combination of autogenous bone grafts and platelet-enriched fibrin glue, autogenous bone grafts alone, or a conventional flap procedure only (control) were carried out. Six months later, biopsies of the implant sites were taken and prepared for ground sectioning and analysis. Results The amount of reosseointegration was significantly higher in peri-implantitis defects treated with combined autogenous bone grafts and platelet-enriched fibrin glue as compared with the other 2 treatment procedures. A mean bone-to-implant contact of 50.1% was obtained in the peri-implantitis lesions treated with combined autogenous bone grafts and platelet-enriched fibrin glue. The corresponding values for the autogenous bone grafts and control groups were 19.3% and 6.5%, respectively. Conclusion The present study demonstrates that surgical treatment involving the combined use of autogenous bone grafts and platelet-enriched fibrin glue might effectively promote reosseointegration in lesions resulting from peri-implantitis.

The activity of several growth factors on peripheral nerve regeneration is reported. Autologous fibrin glue contains a large number of platelets, which release significant quantities of growth factors. In order to understand the role of autologous fibrin glue in peripheral nerve regeneration, a 15-mm rabbit peroneal nerve defect was repaired using a vein graft filled with autologous fibrin glue. Axonal regeneration was examined using histological and electrophysiological methods. The extent of axonal regeneration was superior when treated with autologous fibrin glue. Our data suggest that fibrin nets formed by fibrinogen, in combination with growth factors present in autologous fibrin glue, might effectively promote peripheral nerve regeneration in nerve defects.

Various miniplate fixation techniques were studied in dogs to determine whether the use of miniplate fixation provides sufficient stability in mandibular continuity defects. Continuity resections measuring 15 mm were made on the mandibles of 16 dogs and were bridged using 4 miniplate fixation techniques, each replicated 4 times. All dogs were placed on a normal diet throughout the postoperative period. Clinical and radiological examinations were carried out 6 weeks later. We found that the group with both double miniplates and bicortical screws was stable, whereas the groups utilizing either a single miniplate or monocortical screws were not. The results of this study indicate that using a combination of double miniplates and bicortical screws to bridge defects after mandibular resection produces stable and predictable results.

Objective The aim of this study was to examine the ability of platelet-enriched fibrin glue to enhance bone formation in critically sized defects in the dog mandible. Study design Seven adult female mongrel dogs underwent continuity resections on both sides of the mandible; 1 defect was reconstructed with the original particulate bone mixed with platelet-enriched fibrin glue, and as a control the contralateral defect was reconstructed with the original particulate bone alone. Results Biopsies after 6 weeks showed that the addition of platelet-enriched fibrin glue enhanced new bone formation in the autogenous bone grafts. Conclusion Our data suggest that fibrin nets formed by fibrinogen, in combination with growth factors present in platelet-enriched fibrin glue, might effectively promote bone healing at bone graft sites.

Sinus lift procedures depend greatly on fragile structures and anatomical variations. These procedures may cause sinus membrane perforations, which can lead to graft infection and early failure. The aim of this study was to assess the efficacy of autologous fibrin glue in the management of large perforations of the maxillary sinus membrane occurring during sinus lifts. After elevating the sinus membrane in the bilateral maxillary sinuses of 6 adult female mongrel dogs, a laceration (about 2.0 cm in length) was made in the membrane and either repaired with autologous fibrin glue or covered with a bioabsorbable collagen membrane as a control. Wounded areas were biopsied 2 weeks after the operation. Wounds repaired with autologous fibrin glue showed newly formed continuous epithelium across the previous perforation site. However, extensive fibrosis, inflammatory infiltration, and absent epithelium were observed in wounds treated with the collagen membrane control. Our results support the clinical use of autologous fibrin glue for repairing sinus membrane perforations.

For tissue-engineered bone formation, autogenous osteogenic cells are of paramount importance for successful bone formation. In order to investigate the donor cell-related differences in tissue-engineered bone, cultured bone marrow mesenchymal stem cells, cultured alveolar bone cells, and cultured periosteal cells were examined for their in vivo potential to form bone. These cells were isolated from dogs, expanded in vitro, mixed with autologous fibrin glue and BMP-2, and then injected into the subcutaneous space on the dorsum of nude mice. Bone formation was evaluated at 12 weeks. Histomorphometric analysis demonstrated that the subcutaneous nodules formed in nude mice contained 26.9% newly formed bone when using the bone marrow mesenchymal stem cells, 41.1% newly formed bone when using the alveolar bone cells, and 58.2% newly formed bone when using the periosteal cells. The results suggest that periosteal cells are the best choice for enhancing bone formation in tissue engineering of bone regeneration.

Summary Introduction Sinus lift procedures depend greatly on fragile structures and anatomical variations. The procedure may cause sinus membrane perforations, which can lead to graft infection and early failure. Aim To assess the efficacy of cyanoacrylate adhesive in the management of large perforations of the maxillary sinus membrane during sinus lifts. Material and methods Six rabbits were used in the study. Sinus membrane perforations (about 1.5 cm in length) were repaired with cyanoacrylate adhesive on one side of the maxillary sinus. On the contralateral side, an identical laceration was not repaired. Histological evaluation was performed 2 weeks after the operation. Results Wounds repaired with cyanoacrylate adhesive showed newly formed continuous epithelium across the previous perforation site and there was sinusitis on the contralateral side. Conclusion These results support the clinical use of cyanoacrylate adhesive for repairing sinus membrane perforations.

This paper describes a new technique for microvascular anastomosis, which involves the overlapping of the adventitias of the two ends of a severed blood vessel, and then painting a cyanoacrylate adhesive on the outer surface of the vessel around the anastomosed part. Sixteen anastomoses were performed in both radial arteries and cephalic veins in eight dogs. All vessels were patent without thrombus. The described method of anastomosis was faster and easier to perform than the conventional suture anastomosis. Histologic studies revealed that the adhesive did not flow into the lumen, and that normal healing of the endothelium and of the internal elastic lamina occurred across the anastomotic site, even though the adhesive remained on the adventitial side of the vessel at 4 weeks. This technique deserves to be considered as an alternative to conventional suture anastomosis.

The role of cultured bone marrow stromal cells (BMSCs) in peripheral nerve regeneration was examined using an established rabbit peroneal nerve regeneration model. A 15-mm peroneal nerve defect was bridged with a vein filled with BMSCs (1×10 6 ), which had been embedded in collagen gel. On the contralateral side, the defect was bridged with a vein filled with collagen gel alone. When the regenerated tissue was examined 4, 8 and 12 weeks after grafting, the number and diameter of the myelinated fibers in the side with the BMSCs were significantly higher than in the control side without the BMSCs. This demonstrates the potential of using cultured BMSCs in peripheral nerve regeneration.

Previous studies have shown that a combination of platelet-rich plasma (PRP) and autogenous bone graft can increase the rate of osteogenesis and enhance bone formation qualitatively. However, contradictory results were reported in a recent animal study. In order to clarify this inconsistency, this study examined the influence of the PRP concentrations on the viability and proliferation of alveolar bone cells in vitro. Bone cells obtained from the alveolar bone chips were exposed to various PRP concentrations. After a culture period of 7 days, cellular viability and proliferation were evaluated by counting the number of cells and a MTT assay. The results showed that the viability and proliferation of alveolar bone cells were suppressed by high PRP concentrations, but were stimulated by low PRP concentrations (1-5%). These in vitro results support the view that variations in the PRP concentrations might influence the bone formation within the PRP-treated bone grafts.

In this study, we evaluated the effect of platelet-rich plasma (PRP) on bone regeneration in an autogenous bone graft in a canine model. The mandibular premolar teeth had been bilaterally extracted previously, and the ridges had been allowed to heal for 3 months. After this period, continuity resection was performed on both sides of the mandible. One defect (the PRP group) was reconstructed with the original particulate bone mixed with PRP. As a control, the contralateral defect (non-PRP group) was reconstructed with the original particulate bone alone. Biopsies after 6 weeks showed lower levels of bone formation in the PRP group than in the non-PRP group, and fluorescence microscopy revealed a delay in the remodelling of grafts loaded with PRP. These findings suggest that the addition of PRP does not appear to enhance new bone formation in autogenous bone grafts.

Summary Aim Several injectable materials have been used as osteogenic bone substitutes. However, none has gained universal acceptance. This study was performed to investigate whether or not chitosan-alginate gel/mesenchymal stem cells/bone morphogenetic protein-2 composites are potentially injectable materials for new bone formation. Material and methods The composites were injected into the subcutaneous space on the dorsum of nude mice to investigate new bone tissue formation. The composites were examined histologically over a 12-week period. Results The composites injected into the mouse were able to stimulate new bone formation, which was trabecular in type. Conclusions This study showed that chitosan-alginate gel/MSCs/BMP-2 composites could become clinically useful injectable materials to generate new bone.

This study was undertaken to evaluate the biomechanical stability of various miniplate fixation techniques in varying degrees of atrophy and to determine optimal fixation techniques for fractures of the atrophic edentulous mandible. A total of 78 bovine ribs were randomly divided into 13 groups of six each; one non-atrophic group and twelve atrophic groups. Each one of the 13 sets of six ribs was formed based on the vertical dimension (40, 20, 15 or 10 mm) and osteotomized. The segments so produced were then reduced and fixed using various miniplate fixation techniques. The stability of various miniplate fixations in ribs showing varying degrees of atrophy (10, 15 and 20 mm) was compared with that of one miniplate fixation in non-atrophic ribs (40 mm), used as a standard. Atrophic groups utilizing single miniplate were significantly less stable than the non-atrophic group, whereas atrophic groups fitted with double miniplates, such as two 4-hole or two 6-hole miniplates, were significantly more stable than the non-atrophic group. The two miniplate fixation technique is recommended for the provision of adequate fracture site stability when open reduction is indicated in cases of atrophic edentulous mandibular fractures.