Glomerular Elasticity and Gene Expression Patterns Define Two Phases of Alport Nephropathy.

Rationale: We take a unique approach to understanding the causes of podocyte injury in collagen IV nephropathies, a crucial step in developing targeted therapies for conditions like Alport Syndrome. Objectives: We characterize the structural, functional, and biophysical properties of glomerular capillaries and podocytes in Col4α3 -/- mice and analyze kidney cortex transcriptional profiles at various disease stages. We investigate the effects of the ER stress mitigator TUDCA on these parameters. Furthermore, we used human FSGS associated podocyte enriched genes to identify molecular pathways rescued by TUDCA thereby offering potential therapeutic targets for Alport Syndrome. Findings: We find a clear disease progression timeline in Col4α3 -/- mice. Podocyte injury develops by 3 months, with glomeruli reaching maximum deformability at 4 months, associated with a 40% loss of podocytes. This is followed by progressive stiffening of glomerular capillaries, increasing proteinuria, reduced renal function, inflammatory infiltrates, and fibrosis from months 4 to 8. Bulk RNA sequencing at 2, 4, and 7 months reveals a progressive increase in expression of genes related to cytokine and chemokine signaling, matrix and cell injury, and activation of the TNF pathway, similar to observations in a NEPTUNE FSGS cohort. Podocyte-enriched genes from FSGS patients mapped to mice found that TUDCA, which mitigated glomerular and renal injury suppressed molecular pathways associated with extracellular matrix and basement membrane synthesis, podocyte stress and hypertrophy. Conclusions: We uncover two distinct phases of Col4α3 -/- nephropathy progression. The first is characterized by podocytopathy, increased glomerular capillary deformability and accelerated podocyte loss, and the second by increased capillary wall stiffening and renal inflammatory and profibrotic pathway activation. The response of podocytes to TUDCA treatment provides novel insights into downstream signaling pathways, offering potential therapeutic targets for treating Alport and related nephropathies.

Prediction of lifespan and assessing risk factors of large-sample implant prostheses: a multicenter study.

PURPOSE: This study aimed to analyze factors influencing the success and failure of implant prostheses and to estimate the lifespan of prostheses using standardized evaluation criteria. An online survey platform was utilized to efficiently gather large samples from multiple institutions. MATERIALS AND METHODS: During the one-year period, patients visiting 16 institutions were assessed using standardized evaluation criteria (KAP criteria). Data from these institutions were collected through an online platform, and various statistical analyses were conducted. Risk factors were assessed using both the Cox proportional hazard model and Cox regression analysis. Survival analysis was conducted using Kaplan-Meier analysis and nomogram, and lifespan prediction was performed using principal component analysis. RESULTS: The number of patients involved in this study was 485, with a total of 841 prostheses evaluated. The median survival was estimated to be 16 years with a 95% confidence interval. Factors found to be significantly associated with implant prosthesis failure, characterized by higher hazard ratios, included the 'type of clinic', 'type of antagonist', and 'plaque index'. The lifespan of implant prostheses that did not fail was estimated to exceed the projected lifespan by approximately 1.34 years. CONCLUSION: To ensure the success of implant prostheses, maintaining good oral hygiene is crucial. The estimated lifespan of implant prostheses is often underestimated by approximately 1.34 years. Furthermore, standardized form, online platform, and visualization tool, such as nomogram, can be effectively utilized in future follow-up studies.

Standardized multi-institutional data analysis of fixed and removable prosthesis: estimation of life expectancy with regards to variable risk factors.

PURPOSE: This study aims to assess and predict lifespan of dental prostheses using newly developed Korean Association of Prosthodontics (KAP) criteria through a large-scale, multi-institutional survey. MATERIALS AND METHODS: Survey was conducted including 16 institutions. Cox proportional hazards model and principal component analysis (PCA) were used to find out relevant factors and predict life expectancy. RESULTS: 1,703 fixed and 815 removable prostheses data were collected and evaluated. Statistically significant factors in fixed prosthesis failure were plaque index and material type, with a median survival of 10 to 18 years and 14 to 20 years each. In removable prosthesis, factors were national health insurance coverage, antagonist type, and prosthesis type (complete or partial denture), with median survival of 10 to 13 years, 11 to 14 years, and 10 to 15 years each. For still-usable prostheses, PCA analysis predicted an additional 3 years in fixed and 4.8 years in removable prosthesis. CONCLUSION: Life expectancy of a prosthesis differed significantly by factors mostly controllable either by dentist or a patient. Overall life expectancy was shown to be longer than previous research.

Tooth loss and dementia amongst older adults residing in long-term care facilities in Vancouver: A case-control study.

OBJECTIVE: The purpose of this case-control study was to determine the association between dementia and the number of missing teeth, functional occlusal units and denture use in older adults residing in Long-Term Care (LTC) facilities. BACKGROUND: Many studies have shown an association between dementia and tooth loss. However, few studies with a large sample size have been reported describing the relationship between dementia and the number of missing teeth, remaining teeth and functional occlusal units. METHODS: An oral health assessment database of 2160 older adults admitted to LTC facilities in Vancouver, Canada, between 2015-2019 was utilised. Participants with a diagnosis of dementia in their medical records (N = 1174) were compared to those without dementia (N = 986). Multiple logistic regression analysis was used to explore a potential association between the number of missing teeth, functional occlusal units and the use of dentures and dementia. RESULTS: The number of remaining teeth (OR = 1.0, 95% Confidence Interval = 1.0-1.0; P = .054) and number of functional occlusal units (OR = 1.0, 95% CI = 1.0-1.0; P = .059) were not associated with dementia after adjusting for age, sex, oral self-care and systemic conditions. Denture use (OR = 1.1, 95% CI = 0.5-2.4; P = .790) was not associated with dementia in edentulous patients. CONCLUSION: There was no association between dementia and the number of remaining teeth, functional occlusal units or wearing dentures.

Retrofitting of an implant-supported surveyed crown under an existing removable partial denture by using an acrylic resin template.

Retrofitting a crown to an existing removable partial denture (RPD) is a complex process and requires additional clinical and laboratory procedures. Various methods have been described for retrofitting a new tooth-supported crown. However, if an abutment tooth has to be extracted, descriptions of techniques for restoring a new edentulous site with an implant-supported crown retrofitted to an existing RPD are lacking. Therefore, this technical report describes a straightforward approach to fabricating an implant-supported surveyed crown fitted to an existing RPD by using an acrylic resin template.

Value of Additional Instrumented Fusion in the Treatment of Thoracic Ossification of the Ligamentum Flavum.

OBJECTIVE: The ossification of the ligamentum flavum (OLF) is one of the major causes of thoracic myelopathy. Surgical decompression with or without instrumented fusion is the mainstay of treatment. However, few studies have reported on the added effect of instrumented fusion. The objective of this study was to compare clinical and radiological outcomes between surgical decompression without instrumented fusion (D-group) and that with instrumented fusion (F-group). METHODS: A retrospective review was performed on 28 patients (D-group, n=17; F-group, n=11) with thoracic myelopathy due to OLF. The clinical parameters compared included scores of the Japanese Orthopedic Association (JOA), the Visual analogue scale of the back and leg (VAS-B and VAS-L), and the Korean version of the Oswestry disability index (K-ODI). Radiological parameters included the sagittal vertical axis (SVA), the pelvic tilt (PT), the sacral slope (SS), the thoracic kyphosis angle (TKA), the segmental kyphosis angle (SKA) at the operated level, and the lumbar lordosis angle (LLA; a negative value implying lordosis). These parameters were measured preoperatively, 1 year postoperatively, and 2 years postoperatively, and were compared with a linear mixed model. RESULTS: After surgery, all clinical parameters were significantly improved in both groups, while VAS-L was more improved in the Fgroup than in the D-group (-3.4±2.5 vs. -1.3±2.2, p=0.008). Radiological outcomes were significantly different in terms of changes in TKA, SKA, and LLA. Changes in TKA, SKA, and LLA were 2.3°±4.7°, -0.1°±1.4°, and -1.3°±5.6° in the F-group, which were significantly lower than 6.8°±6.1°, 3.0°±2.8°, and 2.2°±5.3° in the D-group, respectively (p=0.013, p<0.0001, and p=0.037). Symptomatic recurrence of OLF occurred in one patient of the D-group at postoperative 24 months. CONCLUSION: Clinical improvement was achieved after decompression surgery for OLF regardless of whether instrumented fusion was added. However, adding instrumented fusion resulted in better outcomes in terms of lessening the progression of local and regional kyphosis and improving leg pain. Decompression with instrumented fusion may be a better surgical option for thoracic OLF.

Recurrent Cervical Spontaneous Spinal Epidural Hematoma with Conservative Management: A Case Report.

Cervical spontaneous spinal epidural hematoma (CSSEH) is a rare condition that can be potentially fatal if not properly diagnosed and managed. While prompt surgical decompression and evacuation of the hematoma are generally considered as the first line of treatment, mild cases that were managed through observation and conservative treatment have been reported. Our patient was a 24-year-old man who experienced two CSSEH events 8 months apart, both of which were managed conservatively. This was a rare case of recurrent CSSEH in which recovery was achieved without surgical intervention. We believe conservative treatment with close observation may be effective in CSSEH patients presenting with mild neurologic symptoms who have a tendency towards spontaneous neurologic improvement.

Physiologic Regulation of Systemic Klotho Levels by Renal CaSR Signaling in Response to CaSR Ligands and pH o.

BACKGROUND: The kidney is the source of sKlotho and kidney-specific loss of Klotho leads to a phenotype resembling the premature multiorgan failure phenotype in Klotho-hypomorphic mice ( kl/kl mice). Klotho and the Ca-sensing receptor (CaSR) are highly expressed in the distal convoluted tubule (DCT). The physiologic mechanisms that regulate sKlotho levels are unknown. METHODS: We measured sKlotho in WT and tubule-specific CaSR -/- (TS-CaSR -/- ) mice treated with calcimimetics, alkali, or acid, and Klotho shed from minced mouse kidneys, and from HEK-293 cells expressing the CaSR and Klotho, in response to calcimimetics, calcilytics, alkalotic and acidic pH, and ADAM protease inhibitors. The CaSR, Klotho, and ADAM10 were imaged in mouse kidneys and cell expression systems using confocal microscopy. RESULTS: The CaSR, Klotho, and ADAM10 colocalize on the basolateral membrane of the DCT. Calcimimetics and HCO 3 increase serum sKlotho levels in WT but not in CaSR -/- mice, and acidic pH suppresses sKlotho levels in WT mice. In minced kidneys and cultured cells, CaSR activation with high Ca, calcimimetics, or alkali increase shed Klotho levels via ADAM10, as demonstrated using the ADAM10 inhibitor GI254023X and siRNA. In cultured cells, the CaSR, Klotho, and ADAM10 form cell surface aggregates that disperse after CaSR activation. CONCLUSIONS: We identify a novel physiologic mechanism for regulation of sKlotho levels by the renal CaSR-ADAM10-Klotho pathway. We show that CaSR activators, including alkali, increase renal CaSR-stimulated Klotho shedding and predict that this mechanism is relevant to the effects of acidosis and alkali therapy on CKD progression.

Posterior Approach in C2-3 Disc Herniation: C1 Laminectomy, C2-3 Laminoplasty and Posterior Fixation in C2-3 Disc Herniation.

C2-3 disc herniation is rare and a definitive treatment of choice has not been established. The purpose of this case report is to suggest posterior approach as one of the best options. A 49-year-old man visited our clinic with a 7-year history of neck pain and occipital headache and a 2-month history of right arm pain. C2-3 intervertebral disc herniation of the central type was diagnosed on magnetic resonance imaging (MRI), and surgery was performed, including C1 laminectomy, C2-3 laminoplasty, and C2-3 posterior fixation. The posterior approach was used because the patient's neck was difficult to operate anteriorly. After 3 months postoperatively, MRI showed widened cerebrospinal fluid space at the C2-3 level. The visual analogue scale score for pain improved in the occipital area and right arm. However, the untouched protruded central disc, subjective weakness in right hand grasping, and numbness persisted. In conclusion, this case highlights posterior decompression and fixation as a good treatment of choice for decompression at the C2-3 level disc herniation, from where it is difficult to remove compressive lesions directly via the anterior corridor.

Control of Podocyte and Glomerular Capillary Wall Structure and Elasticity by WNK1 Kinase.

Cytoskeletal structure and its regulation are essential for maintenance of the differentiated state of specific types of cells and their adaptation to physiologic and pathophysiologic conditions. Renal glomerular capillaries, composed of podocytes, endothelial cells, and the glomerular basement membrane, have distinct structural and biophysical properties and are the site of injury in many glomerular diseases. Calcineurin inhibitors, immunosuppressant drugs used for organ transplantation and auto-immune diseases, can protect podocytes and glomerular capillaries from injury by preserving podocyte cytoskeletal structure. These drugs cause complications including hypertension and hyperkalemia which are mediated by WNK (With No Lysine) kinases as well as vasculopathy with glomerulopathy. WNK kinases and their target kinases oxidative stress-responsive kinase 1 (OSR1) and SPS1-related proline/alanine-rich kinase (SPAK) have fundamental roles in angiogenesis and are activated by calcineurin inhibitors, but the actions of these agents on kidney vasculature, and glomerular capillaries are not fully understood. We investigated WNK1 expression in cultured podocytes and isolated mouse glomerular capillaries to determine if WNK1 contributes to calcineurin inhibitor-induced preservation of podocyte and glomerular structure. WNK1 and OSR1/SPAK are expressed in podocytes, and in a pattern similar to podocyte synaptopodin in glomerular capillaries. Calcineurin inhibitors increased active OSR1/SPAK in glomerular capillaries, the Young's modulus (E) of glomeruli, and the F/G actin ratio, effects all blocked by WNK inhibition. In glomeruli, WNK inhibition caused reduced and irregular synaptopodin-staining, abnormal capillary and foot process structures, and increased deformability. In cultured podocytes, FK506 activated OSR1/SPAK, increased lamellipodia, accelerated cell migration, and promoted traction force. These actions of FK506 were reduced by depletion of WNK1. Collectively, these results demonstrate the importance of WNK1 in regulation of the podocyte actin cytoskeleton, biophysical properties of glomerular capillaries, and slit diaphragm structure, all of which are essential to normal kidney function.

Application of the functionally generated path technique in designing an implant-supported fixed prosthesis with CAD-CAM technology: A dental technique.

A method involving the digital application of the functionally generated path (FGP) technique to fabricate an implant-supported fixed prosthesis is described. It uses an intraoral optical scanner, a dental design software program, and a specially designed removable FGP table. With this method, clinicians can design custom implant-supported fixed prostheses without occlusal interferences, reducing the chairside time required to deliver the prosthesis.

The socioeconomic impact of Korean dental health insurance policy on the elderly: a nationwide cohort study in South Korea.

PURPOSE: The purpose of this retrospective study was to investigate the relationships of types of dental insurance coverage in Korea with sociodemographic characteristics and the prevalence of systemic and oral diseases, as well as to evaluate the socioeconomic impact of Korean dental insurance policies. METHODS: Sample cohort data from 2006 to 2015 were obtained from the National Health Insurance Service. Patients were divided into 2 groups. The exposed group comprised patients who received insurance benefits for complete dentures, removable partial dentures, and implant care, while the control group comprised patients who did not receive these benefits. The type of insurance coverage and the prevalence of systemic and oral diseases were compared between the 2 groups. RESULTS: Patients who received benefits in the form of complete dentures, removable partial dentures, and implants had similar sociodemographic characteristics in terms of sex, age, income quintile, and type of insurance coverage to the control group. The prevalence of hypertension, anemia, renal disease, rheumatoid arthritis, osteoporosis, asthma, and cerebral infarction was higher in the exposed group than in the control group (P<0.05). The prevalence of periodontal diseases and dental caries was also higher in the exposed group. CONCLUSIONS: Korean dental health insurance policy has been beneficial for the medical expenses of low-income and elderly people suffering from a cost burden due to systemic diseases. However, since there is a tendency to avoid invasive interventions in older patients due to the high risk of systemic diseases, insurance coverage of dentures may be more helpful from a socioeconomic perspective than coverage of dental implant treatments.

The effect of missing teeth on dementia in older people: a nationwide population-based cohort study in South Korea.

BACKGROUND: To determine the effect of missing teeth on the risk of dementia onset among individuals who received tooth extractions and those who did not, based on the number of missing teeth. METHODS: We selected individuals who had not been diagnosed or treated for dementia between 2002 to 2011 from the National Health Insurance Service-Elderly Cohort Database (NHIS-ECD). We divided participants into two cohorts, a tooth extraction and non-extraction cohort, based on tooth loss from 2002 to 2011. After propensity score matching, there were 104,903 individuals in each cohort, and we included a total of 209,806 individuals in this study. Each cohort was grouped by sex, age, residential area, health insurance eligibility, income level, history of dental caries, history of periodontal treatment, and number of extracted teeth. We analyzed the relationship between dementia onset and these variables using logistic regression analysis. RESULTS: Individuals with tooth loss had a higher risk for dementia than those without tooth loss (odds ratio [OR] = 1.18; 95% confidence interval [CI]: 1.146-1.215). Regarding the incidence of dementia, the OR increased as the number of missing teeth and age increased, and the OR was higher for women (OR = 1.33; 95% CI: 1.286-1.367) than for men, and this difference was statistically significant (P < 0.01). The incidence of dementia decreased with periodontal treatment (OR = 0.96; 95% CI: 0.932-0.992) and increased with dental caries (OR = 1.07; 95% CI: 1.035-1.101). CONCLUSIONS: These results suggest that it is important to delay tooth loss and preserve the stable remaining teeth to help prevent dementia.

The effect of diabetes on tooth loss caused by periodontal disease: A nationwide population-based cohort study in South Korea.

BACKGROUND: Our purpose was to compare the risk of tooth loss caused by periodontal disease between diabetic and nondiabetic individuals and to estimate the relative risk of tooth loss according to the severity and control of diabetes. METHODS: We selected 10,215 individuals who were diagnosed as diabetics in the Korean National Health Insurance Service-National Sample Cohort (NHIS-NSC) database in 2003 and the same number of individuals who had never been diagnosed with diabetes during the period covered by the NHIS-NSC (2002-2013) with propensity score matching. Diabetic individuals were divided into two groups according to the treatment modality. We counted the number of teeth lost in each group to determine the difference in the risk of tooth loss among groups and used the Cox proportional hazards model to calculate hazard ratios (HRs) of tooth loss caused by periodontitis. RESULTS: Diabetic individuals had a higher risk of tooth loss than nondiabetic individuals (HR = 1.298, 95% confidence interval [CI]: 1.233 ≤ HR ≤ 1.366; P < 0.01). The severer the degree of diabetes, the higher the risk of tooth loss. As the number of dental visits increased, the risk of tooth loss declined (HR = 0.998, 95% CI: 0.996 ≤ HR ≤ 0.999; P < 0.01). CONCLUSIONS: Within the limits of this retrospective cohort study, the risk of tooth loss among diabetic individuals was higher than that among nondiabetic individuals. The risk of tooth loss declined with increasing numbers of dental visits and increased with the severity of diabetes.

Similar Biophysical Abnormalities in Glomeruli and Podocytes from Two Distinct Models.

Background FSGS is a pattern of podocyte injury that leads to loss of glomerular function. Podocytes support other podocytes and glomerular capillary structure, oppose hemodynamic forces, form the slit diaphragm, and have mechanical properties that permit these functions. However, the biophysical characteristics of glomeruli and podocytes in disease remain unclear.Methods Using microindentation, atomic force microscopy, immunofluorescence microscopy, quantitative RT-PCR, and a three-dimensional collagen gel contraction assay, we studied the biophysical and structural properties of glomeruli and podocytes in chronic (Tg26 mice [HIV protein expression]) and acute (protamine administration [cytoskeletal rearrangement]) models of podocyte injury.Results Compared with wild-type glomeruli, Tg26 glomeruli became progressively more deformable with disease progression, despite increased collagen content. Tg26 podocytes had disordered cytoskeletons, markedly abnormal focal adhesions, and weaker adhesion; they failed to respond to mechanical signals and exerted minimal traction force in three-dimensional collagen gels. Protamine treatment had similar but milder effects on glomeruli and podocytes.Conclusions Reduced structural integrity of Tg26 podocytes causes increased deformability of glomerular capillaries and limits the ability of capillaries to counter hemodynamic force, possibly leading to further podocyte injury. Loss of normal podocyte mechanical integrity could injure neighboring podocytes due to the absence of normal biophysical signals required for podocyte maintenance. The severe defects in podocyte mechanical behavior in the Tg26 model may explain why Tg26 glomeruli soften progressively, despite increased collagen deposition, and may be the basis for the rapid course of glomerular diseases associated with severe podocyte injury. In milder injury (protamine), similar processes occur but over a longer time.

Interdisciplinary treatment of a patient with multiple missing teeth and periodontitis.

A 49-year-old woman with several missing and periodontically compromised teeth was referred to the orthodontic department of National Health Insurance Service Ilsan Hospital by the periodontic department for interdisciplinary treatment. Multiple posterior teeth had been extracted 10 days earlier. Her chief complaint was crowding of the anterior teeth, and she wanted to improve both esthetics and function. Orthodontic, periodontic, and prosthodontic treatments were undertaken in the proper timing and sequence with an interdisciplinary approach. As a result, improved periodontal health and a stable occlusion and vertical dimension were achieved. Although there were limited teeth and alveolar bone for anchorage, good esthetic and functional treatment results were obtained through the application of temporary anchorage devices and proper biomechanics.

The Effect of Fibronectin-Immobilized Microgrooved Titanium Substrata on Cell Proliferation and Expression of Genes and Proteins in Human Gingival Fibroblasts.

BACKGROUND: We aimed to determine the effect of fibronectin (FN)-immobilized microgrooved titanium (Ti) on human gingival fibroblast proliferation, gene expression and protein expression. METHODS: Photolithography was used to fabricate the microgrooved Ti, and amine funtionalization (silanization) was used for FN immobilization on titanium surfaces. Cell proliferation, gene expression and protein expression were analyzed, followed by multiple regression analysis for determining the influential factors on cell proliferation. RESULTS: FN-immobilized microgrooved Ti significantly enhanced the fibroblast proliferation in various timelines of culture, among which a burst of fivefold increase is induced at 96 h of culture compared to that on the control smooth Ti. We suggest a presence of the synergistic promotion effect of microgrooves and FN immobilization on fibroblast proliferation. Through a series of analyses on the expression of various genes and proteins involved in cell adhesion and proliferation, cyclin-dependent kinase 6, cyclin D1, integrin α5, oncogene c-Src, osteonectin, paxillin and talin-2 were determined as influential factors on promoting fibroblast proliferation induced by FN-immobilized microgrooved Ti. CONCLUSION: FN-immobilized microgrooved Ti can act as an effective surface for enhancing fibroblast proliferation, and can be used for promoting soft tissue response on the connective tissue attachment zone of biomaterial surfaces.

Modeled structural basis for the recognition of α2-3-sialyllactose by soluble Klotho.

Soluble Klotho (sKlotho) is the shed ectodomain of antiaging membrane Klotho that contains 2 extracellular domains KL1 and KL2, each of which shares sequence homology to glycosyl hydrolases. sKlotho elicits pleiotropic cellular responses with a poorly understood mechanism of action. Notably, in injury settings, sKlotho confers cardiac and renal protection by down-regulating calcium-permeable transient receptor potential canonical type isoform 6 (TRPC6) channels in cardiomyocytes and glomerular podocytes. Inhibition of PI3K-dependent exocytosis of TRPC6 is thought to be the underlying mechanism, and recent studies showed that sKlotho interacts with α2-3-sialyllactose-containing gangliosides enriched in lipid rafts to inhibit raft-dependent PI3K signaling. However, the structural basis for binding and recognition of α2-3-sialyllactose by sKlotho is unknown. Using homology modeling followed by docking, we identified key protein residues in the KL1 domain that are likely involved in binding sialyllactose. Functional experiments based on the ability of Klotho to down-regulate TRPC6 channel activity confirm the importance of these residues. Furthermore, KL1 domain binds α2-3-sialyllactose, down-regulates TRPC6 channels, and exerts protection against stress-induced cardiac hypertrophy in mice. Our results support the notion that sialogangliosides and lipid rafts are membrane receptors for sKlotho and that the KL1 domain is sufficient for the tested biologic activities. These findings can help guide the design of a simpler Klotho mimetic.-Wright, J. D., An, S.-W., Xie, J., Yoon, J., Nischan, N., Kohler, J. J., Oliver, N., Lim, C., Huang, C.-L. Modeled structural basis for the recognition of α2-3-sialyllactose by soluble Klotho.

Soluble klotho binds monosialoganglioside to regulate membrane microdomains and growth factor signaling.

Soluble klotho, the shed ectodomain of the antiaging membrane protein α-klotho, is a pleiotropic endocrine/paracrine factor with no known receptors and poorly understood mechanism of action. Soluble klotho down-regulates growth factor-driven PI3K signaling, contributing to extension of lifespan, cardioprotection, and tumor inhibition. Here we show that soluble klotho binds membrane lipid rafts. Klotho binding to rafts alters lipid organization, decreases membrane's propensity to form large ordered domains for endocytosis, and down-regulates raft-dependent PI3K/Akt signaling. We identify α2-3-sialyllactose present in the glycan of monosialogangliosides as targets of soluble klotho. α2-3-Sialyllactose is a common motif of glycans. To explain why klotho preferentially targets lipid rafts we show that clustering of gangliosides in lipid rafts is important. In vivo, raft-dependent PI3K signaling is up-regulated in klotho-deficient mouse hearts vs. wild-type hearts. Our results identify ganglioside-enriched lipid rafts to be receptors that mediate soluble klotho regulation of PI3K signaling. Targeting sialic acids may be a general mechanism for pleiotropic actions of soluble klotho.

Promotion of osteoblastic differentiation and osteogenic transcription factor expression on a microgroove titanium surface with immobilized fibronectin or bone sialoprotein II.

We demonstrate that a composite surface of microgroove titanium (Ti) with immobilized fibronectin (FN) or bone sialoprotein II (BSP2) promotes osteoblastic differentiation and osteogenic transcription factor expression in human bone marrow-derived mesenchymal stem cells (MSCs). Comparisons made between smooth microgrooves, microgrooves with silanization and microgrooves with matrix protein (FN or BSP2)-immobilization Ti surfaces revealed a significant promotion of in vitro osteogenic activity and osteoblastic differentiation at various timelines of culture. An even more significant increase was verified on microgrooves with a matrix protein-immobilization Ti surface in 28 d time-dependent gene expression of the main osteogenic transcription factors, such as ARF4, FRA1, RUNX2, and OSX. As a result, a synergestic effect regarding the promotion of osteogenic transcription factor expression and osteoblastic differentiation in the matrix protein-microgroove Ti composite surface was confirmed. From a multiple regression analysis using various timelines of osteogenic culture as independent variables, day 13 was verified as the most prominent influential timeline for the promotion of osteoblastic differentiation induced by the matrix protein-microgroove Ti composite surface. The FN- or BSP2-microgroove Ti composite surface resulting from silanization can strongly induce the promotion of osteoblastic differentiation in human MSCs. The proposed surface is expected to be useful in the development of a variety of osteogenic biomaterial surfaces.