Contribution of phosphate and FGF23 to CKD progression.

PURPOSE OF REVIEW: Progressive forms of chronic kidney disease (CKD) exhibit kidney inflammation and fibrosis that drive continued nephron loss; however, factors responsible for the development of these common pathologic features remain poorly defined. Recent investigations suggest pathways involved in maintaining urinary phosphate excretion in CKD may be contributing to kidney function decline. This review provides an update on recent evidence linking altered phosphate homeostasis to CKD progression. RECENT FINDINGS: High dietary phosphate intake and increased serum concentrations of fibroblast growth factor 23 (FGF23) both increase urinary phosphate excretion and are associated with increased risk of kidney function decline. Recent investigations have discovered high concentrations of tubular phosphate promote phosphate-based nanocrystal formation that drives tubular injury, cyst formation, and fibrosis. SUMMARY: Studies presented in this review highlight important scientific discoveries that have molded our current understanding of the contribution of altered phosphate homeostasis to CKD progression. The collective observations from these investigations implicate phosphaturia, and the resulting formation of phosphate-based crystals in tubular fluid, as unique risk factors for kidney function decline. Developing a better understanding of the relationship between tubular phosphate handling and kidney pathology could result in innovative strategies for improving kidney outcomes in patients with CKD.

Ouabain Regulates CFTR-Mediated Anion Secretion and Na,K-ATPase Transport in ADPKD Cells.

Cyst enlargement in autosomal dominant polycystic kidney disease (ADPKD) requires the transepithelial secretion of fluid into the cyst lumen. We previously showed that physiological amounts of ouabain enhance cAMP-dependent fluid secretion and cyst growth of human ADPKD cyst epithelial cells in culture and formation of cyst-like dilations in metanephric kidneys from Pkd1 mutant mice. Here, we investigated the mechanisms by which ouabain promotes cAMP-dependent fluid secretion and cystogenesis. Ouabain (3 nM) enhanced cAMP-induced cyst-like dilations in embryonic kidneys from Pkd1 (m1Bei) mice, but had no effect on metanephroi from Pkd1 (m1Bei) mice that lack expression of the cystic fibrosis transmembrane conductance regulator (CFTR). Similarly, ouabain stimulation of cAMP-induced fluid secretion and in vitro cyst growth of ADPKD cells were abrogated by CFTR inhibition, showing that CFTR is required for ouabain effects on ADPKD fluid secretion. Moreover, ouabain directly enhanced the cAMP-dependent Cl(-) efflux mediated by CFTR in ADPKD monolayers. Ouabain increased the trafficking of CFTR to the plasma membrane and up-regulated the expression of the CFTR activator PDZK1. Finally, ouabain decreased plasma membrane expression and activity of the Na,K-ATPase in ADPKD cells. Altogether, these results show that ouabain enhances net fluid secretion and cyst formation by activating apical anion secretion via CFTR and decreasing basolateral Na(+) transport via Na,K-ATPase. These results provide new information on the mechanisms by which ouabain affects ADPKD cells and further highlight the importance of ouabain as a non-genomic stimulator of cystogenesis in ADPKD.

The ability of massive osteochondral allografts from the medial tibial plateau to reproduce normal joint contact pressures after glenoid resurfacing: the effect of computed tomography matching.

BACKGROUND: Current techniques for resurfacing of the glenoid in the treatment of arthritis are unpredictable. Computed tomography (CT) studies have demonstrated that the medial tibial plateau has close similarity to the glenoid. The purpose of this study was to assess contact pressures of transplanted massive tibial osteochondral allografts to resurface the glenoid without and with CT matching. METHODS: Ten unmatched cadaveric tibiae were used to resurface 10 cadaveric glenoids with osteochondral allografts. Five cadaveric tibiae and glenoids were CT matched and studied. An internal control group of 4 matched pairs of glenoids, with the contralateral glenoid transplanted to the opposite glenoid, was also included as a best-case scenario to measure the effects of the surgical technique. All glenoids were tested before and after grafting at different abduction and rotation angles, with recording of peak contact pressures. RESULTS: Peak contact pressures were not different from the intact state in the autografted group but were increased in both allografted groups. CT-matched tibial grafts had lower peak pressures than unmatched grafts. Peak pressures were on average 24.8% (range [18.3%, 29.6%]) greater than in the native glenoids for unmatched allografts, 21.8% ([17.0%, 25.5%]) greater for the matched allografts, and 4.9% ([3.8%, 5.5%]) greater for matched autografts. CONCLUSION: Osteochondral grafting from the medial tibial plateau to the glenoid is feasible but results in increased peak contact pressures. The technique is reproducible as defined by the autografted group. Contact pressures between native and allografted glenoids were significantly different. The clinical significance remains unknown. Peak pressures experienced by the glenoid seem highly sensitive to deviations from the native glenoid shape.

Normal curvature of glenoid surface can be restored when performing an inlay osteochondral allograft: an anatomic computed tomographic comparison.

PURPOSE: The purpose of this study was to quantitatively measure the morphology of the glenoid and to assess feasibility of using the medial tibial plateau surface as a donor for osteoarticular allograft reconstruction of the glenoid. METHODS: Using computed tomography (CT), 10 tibias and 10 scapular models from our database (5 males and 5 females in each group) were randomly selected. Commercial software (Mimics, Materialize, Inc., Plymouth, MI) was used to extract the bone contours from the CT images and to reconstruct the 3-dimensional (3D) geometry of the scapula and tibia. By utilizing the software Creo Elements/Pro 5.0 (Parametric Technology Corp., Needham, MA), mean length and width of both the glenoid and medial tibial plateau were calculated. Radius of curvature was then measured in each 3D CT model at three intermediate segment points that were established within the length line at 25, 50, and 75 percent from superior to inferior in the glenoid and from posterior to anterior in the medial tibial plateau. Statistical analysis was performed and determined to be significant for P < 0.05. RESULTS: The mean (± SD) radius of curvature values at the established 25, 50, and 75 percent segments of the glenoid were 47.4 ± 17.5 mm, 51.2 ± 12.4 mm, and 45.9 ± 17.0 mm, respectively. For the medial tibial plateau, the radius of curvature at 25, 50, and 75 percent were 43.5 ± 9.7 mm, 37.4 ± 14.3 mm and 52.3 ± 21.5 mm, respectively. Values of the glenoid length were 34.0 ± 2.9 mm, and width values were 24.4 ± 2.3 mm. For the medial tibial plateau, the length was 42.6 ± 2.7 mm, and the width was 23.3 ± 4.3 mm. There was no statistical difference in the radius of curvature and dimensional surface area between the glenoid and medial tibial plateau surfaces. CONCLUSION: The 3D CT-based anatomic study found that there is a statistically similar relationship in the radius of curvature of the glenoid and the medial tibial plateau surface. This concept may allow the medial tibial plateau to be used as a donor for osteoarticular allograft reconstruction of the glenoid, especially in young patients where previous studies have demonstrated that the success rate in shoulder replacements is not as good as in older patients.

Functional bracing of ACL injuries: current state and future directions.

PURPOSE: Functional braces are commonly prescribed to treat anterior cruciate ligament (ACL) injury. The results of the existing literature on functional brace use are mixed. The purpose of this study was to evaluate the history and current state of functional ACL bracing and to identify design criteria that could improve upon current bracing technologies. METHODS: A literature search was performed through the PubMed MEDLINE database in April 2013 for the keywords "anterior cruciate ligament" and "brace". Articles published between January 1, 1980, and April 4, 2013, were retrieved and reviewed. Current functional braces used to treat ACL injury were identified. The function of the native ACL was carefully studied to identify design requirements that could improve upon current bracing technologies. RESULTS: Biomechanical evaluations of functional brace effects at time zero have been mixed. Functional brace use reportedly does not improve long-term patient outcomes following ACL reconstruction, but has been shown to reduce subsequent injury rates while skiing in both ACL-deficient and reconstructed skiers. In situ force in the ACL varies with flexion angle and activity. Currently, no brace has been designed and validated to replicate the force-flexion behavior of the native ACL. CONCLUSIONS: Biomechanical and clinical evidence suggests current functional bracing technologies do not sufficiently restore normal biomechanics to the ACL-deficient knee, protect the reconstructed ACL, and improve long-term patient outcomes. Further research into a functional brace designed to apply forces to the knee joint similar in magnitude to the native ACL should be pursued. LEVEL OF EVIDENCE: III.

Biomechanical evaluation of knotless anterior and posterior Bankart repairs.

PURPOSE: The value of modern tape-like suture materials and the influence of the number of anchors inserted for arthroscopic Bankart repairs compared to the intact state have yet to be investigated. It was hypothesised: (1) suture-tape repairs will show higher biomechanical strength than common suture repairs, (2) four anchors will be stronger than three, and (3) the strength of the native capsulolabral complex will be greater than repairs. METHODS: Six matched-paired cadaveric shoulders received Bankart lesions/reconstructions and three underwent intact state testing. Anteroinferior repairs compared suture and suture-tape repairs using three anchors, while posteroinferior repairs compared three and four suture anchors using common sutures. An established testing protocol was run for biomechanical testing. RESULTS: There was no significant difference in the maximum loads, loads at 2 mm displacement, stiffness or energy between repair groups or between repairs and the intact state (n.s.). However, failure modes were different: 16/24 (66.7%) of the repair groups showed glenoid labrum detachment compared to 2/12 (16.7%) within the intact state group (P = 0.012). CONCLUSIONS: While biomechanical parameters of repairs and intact states showed equivalence, failure-mode analysis reaffirms previous findings that capsulolabrum complex refixation is weaker than the native attachment. Therefore, in daily clinical practice, type of suture is secondary and insertion of a fourth anchor will be unlikely to add strength but may confer additional risk and cost.

The hip fluid seal--Part II: The effect of an acetabular labral tear, repair, resection, and reconstruction on hip stability to distraction.

PURPOSE: The acetabular labrum is theorized to be important to normal hip function by providing stability to distraction forces through the suction effect of the hip fluid seal. The purpose of this study was to determine the relative contributions of the hip capsule and labrum to the distractive stability of the hip, and to characterize hip stability to distraction forces in six labral conditions: intact labrum, labral tear, labral repair (looped vs. through sutures), partial resection, labral reconstruction with iliotibial band, and complete resection. METHODS: Eight cadaveric hips with a mean age of 47.8 years (SD 4.3, range 41-51 years) were included. For each condition, the hip seal was broken by distracting the hip at a rate of 0.33 mm/s while the required force, energy, and negative intra-articular pressure were measured. For comparisons between labral conditions, measurements were normalized to the intact labral state (percent of intact). RESULTS: The relative contribution of the labrum to distractive stability was greatest at 1 and 2 mm of displacement, where it was significantly greater than the role of the capsule and accounted for 77 % (SD 27 %, p = 0.006) and 70 % (SD 7 %, p = 0.009) of total distractive stability, respectively. The relative contribution of the capsule to distractive stability increased with progressive displacement, providing 41 % (SD 49 %) and 52 % (SD 53 %) of distractive stability at 3 and 5 mm of distraction, respectively. The maximal distraction force required to break the hip seal in the intact labral state (capsule removed) varied from 124 to 150 N. Labral tear, partial resection, and complete resection resulted in average maximal distraction forces of 76 % (SD 34 %), 29 % (SD 26 %), and 27 % (SD 22 %), respectively, compared to the intact state. Through type labral repairs resulted in significantly greater improvements (from the labral tear state) in maximal negative pressure generated, compared to looped type repairs (median increase; +32 vs. -9 %, p = 0.029). Labral reconstruction resulted in a mean maximal distraction force of 66 % (SD 35 %), with a significant improvement of 37 % compared to partial labral resection (p < 0.001). CONCLUSION: The acetabular labrum was the primary hip stabilizer to distraction forces at small displacements (1-2 mm). Partial labral resection significantly decreased the distractive strength of the hip fluid seal. Labral reconstruction significantly improved distractive stability, compared to partial labral resection. The results of this study may provide insight into the relative importance of the capsule and labrum to distractive stability of the hip and may help to explain hip microinstability in the setting of labral disease.

The hip fluid seal--Part I: the effect of an acetabular labral tear, repair, resection, and reconstruction on hip fluid pressurization.

PURPOSE: The acetabular labrum is theorized to be important to normal hip function by creating intra-articular fluid pressurization through the hip fluid seal. However, the effect of a labral tear or partial labral resection, and interventions including labral repair and labral reconstruction, on the hip fluid seal remains to be defined. The purpose of this study was to characterize intra-articular fluid pressurization in six labral conditions: intact, tear, repair (looped vs. through sutures), partial resection, reconstruction with iliotibial band, and complete resection. METHODS: Eight cadaveric hips with a mean age of 47.8 years (SD 4.3, range 41-51) were included in the study. For each labral condition, the hip was compressed with a force of 2.7 times body weight (2,118 N) while intra-articular pressure was continuously measured with 1.0 × 0.3 mm pressure transducers. Peak intra-articular pressure measurements for each condition were normalized relative to the intact state. Statistical analyses were performed utilizing linear mixed-effects models with repeated measures analysis. RESULTS: Intra-articular fluid pressurization of the intact state varied from 78 to 422 kPa (mean 188 kPa ± SD 120). Labral tear, partial resection, and complete resection resulted in average pressurization of 75 ± 33, 53 ± 37, and 24 ± 18 %, respectively compared with the intact state. Through type labral repair resulted in significantly greater increases in pressurization from the labral tear state, compared with the looped type repair (median increase; +46 vs. -12 %, p = 0.029). Labral reconstruction resulted in a mean pressurization of 110 ± 38 % relative to intact state, with a significant 56 ± 47 % improvement in pressurization compared with partial labral resection (p = 0.009). CONCLUSIONS: Partial labral resection caused significant decreases in intra-articular fluid pressurization. Through type labral suture repair restored the fluid pressurization better than looped type repairs. Labral reconstruction significantly improved pressurization to levels similar to the intact state. This study demonstrated the effect of labral tears and partial resections on intra-articular fluid pressurization via the hip fluid seal, and it also demonstrated improvements in pressurization seen with through type labral repairs and labral reconstructions.

Stability of mid-shaft clavicle fractures after plate fixation versus intramedullary repair and after hardware removal.

PURPOSE: Operative treatment for middle-third clavicle fractures has been increasing as recent data has demonstrated growing patient dissatisfaction and functional deficits after non-operative management. A controlled biomechanical comparison of the characteristics of locked intramedullary (IM) fixation versus superior pre-contoured plating for fracture repair and hardware removal is warranted. Therefore, the purpose of the present study was to investigate potential differences between these devices in a biomechanical model. METHODS: Thirty fourth-generation composite clavicles were randomized to one of five groups with 6 specimens each and tested in a random order. The groups tested were intact, repair with plate, repair with IM device, plate removal, and IM device removal. The lateral end of the clavicles was loaded to failure at a rate of 60 mm/min in a cantilever bending setup. Failure mechanism, energy (J), and torque (Nm) at the site of failure were recorded. RESULTS: Failure torque of the intact clavicle (mean ± standard deviation) was 36.5 ± 7.3 Nm. Failure torques of the IM repair (21.5 ± 9.0 Nm) and plate repair (18.2 ± 1.6 Nm) were not significantly different (n.s.) but were significantly less than the intact group (P < 0.05). Failure torque following IM device removal (30.2 ± 6.5 Nm) was significantly greater than plate removal (12.9 ± 2.0 Nm) (P < 0.05). No significant differences were observed between the intact and IM device removal groups (n.s.). CONCLUSION: The results of the current study demonstrate that IM and plate devices provide similar repair strength for middle-third clavicle fractures. However, testing of the hardware removal groups found the IM device removal group to be significantly stronger than the plate removal group.

Osteopontin deletion attenuates cyst growth but exacerbates fibrosis in mice with cystic kidney disease.

Osteopontin (OPN) is a multi-functional glycoprotein that coordinates the innate immune response, prevents nanocrystal formation in renal tubule fluid, and is a biomarker for kidney injury. OPN expression is markedly increased in cystic epithelial cells of polycystic kidney disease (PKD) kidneys; however, its role in PKD progression remains unclear. We investigated the in vitro effects of recombinant OPN on the proliferation of tubular epithelial cells from PKD and normal human kidneys and in vivo effects of OPN deletion on kidney cyst formation, fibrosis, and mineral metabolism in pcy/pcy mice, a non-orthologous model of autosomal-dominant PKD. In vitro studies revealed that OPN enhanced the proliferation of PKD cells but had no effect on normal kidney cells. Deletion of OPN in pcy/pcy mice significantly reduced kidney cyst burden; however, this was accompanied by increased fibrosis and no change in kidney function. The loss of OPN had no effect on kidney macrophage numbers, cyst epithelial cell proliferation, or apoptosis. Furthermore, there was no difference in kidney mineral deposition or mineral metabolism parameters between pcy/pcy mice with and without OPN expression. Global deletion of OPN reduced kidney cyst burden, while paradoxically exacerbating kidney fibrosis in mice with cystic kidney disease.

Overexpression of the polycystin-1 C-tail enhances sensitivity of M-1 cells to ouabain.

Cells derived from renal cysts of patients with autosomal dominant polycystic kidney disease (ADPKD) are abnormally sensitive to ouabain, responding to physiological ouabain concentrations with enhanced proliferation and increased forskolin-induced transepithelial fluid secretion. This requires activation of the epidermal growth factor receptor (EGFR), Src kinase and the extracellular signal-regulated kinases MEK and ERK. Here, we have determined if the ADPKD phenotype obtained in mouse cortical collecting duct cells by stable overexpression of the C-terminal domain of polycystin-1 (PC-1 C-tail) also elicits the ADPKD-like response to ouabain in the cells. M-1 C20 cells expressing the PC-1 C-tail and M-1 C17 cells lacking expression of this construct were treated with physiological concentrations of ouabain, and cell proliferation, activation of the EGFR-Src-MEK-ERK pathway, forskolin-induced transepithelial Cl(-) secretion and the sensitivity of Na,K-ATPase to ouabain were explored. M-1 C20 cells responded to ouabain with increased cell proliferation and ERK phosphorylation. Ouabain also augmented forskolin-induced and cystic fibrosis transmembrane conductance regulator-mediated apical secretion of Cl(-) in M-1 C20 cells. These effects required activation of EGFR, Src and MEK. In contrast, ouabain had no significant effects on M-1 C17 cells. Interestingly, approximately 20% of the Na,K-ATPase from M-1 C20 cells presented an abnormally increased sensitivity to ouabain. Overexpression of PC-1 C-tail in M-1 C20 cells is associated with an ouabain-sensitive phenotype and an increased ability of the cells to proliferate and secrete anions upon ouabain stimulation. This phenotype mimics the ouabain sensitivity of ADPKD cells and may help promote their cystogenic potential.

Cam lesion femoral osteoplasty: in vitro biomechanical evaluation of iatrogenic femoral cortical notching and risk of neck fracture.

PURPOSE: To investigate the effect of femoral cortical notching at different depths on the peak compressive load and energy required to cause a femoral neck fracture in composite femurs. METHODS: Thirty fourth-generation composite femurs were divided into 5 groups: (1) intact with an inherent alpha angle of 61°, (2) resection of inherent cam lesion by reducing the alpha angle from 61° to 45°, (3) cam resection and cortical notching of a 5.5-mm spherical diameter by 2.00-mm (grade I) depth, (4) cam resection with cortical notching of 4.00-mm (grade II) depth, and (5) cam resection with cortical notching of 6.00-mm (grade III) depth. The specimens were loaded in the position of midstance during gait and tested until failure using a dynamic tensile testing machine at a rate of 6 mm/min. RESULTS: Grade II and grade III cortical notching depths with cam resections resulted in a significant decrease in the ultimate load to failure and energy (P < .05) compared with the intact state. The grade II and grade III cortical notching groups with cam resection failed at a significantly lower ultimate load and with significantly lower energy when compared with the cam resection group alone. CONCLUSIONS: The findings of this study demonstrated significant decreases in ultimate load and energy to failure between the intact group and the grade II and grade III femoral cortical notching groups with cam resection. CLINICAL RELEVANCE: Iatrogenic cortical notching may lead to an increased risk of postsurgical complications, specifically femoral neck fracture. Thus, surgical intervention for a cam lesion femoral osteoplasty should strive for precision, especially around the femoral neck.

Biomechanical analysis of two-tendon posterosuperior rotator cuff tear repairs: extended linked repairs and augmented repairs.

PURPOSE: The purpose of this study was to compare single-row (SR), extended double-row (DR), and augmented, extended double-row (aDR) rotator cuff repairs in a two-tendon, posterosuperior rotator cuff tear (RCT) model with intact rotator cuff tendons. METHODS: RCTs were created and randomly assigned to SR, DR, or aDR repair (5 each) in 20 cadaveric shoulder specimens. A collagen scaffold was used for augmentation. In the remaining 5 specimens, the rotator cuffs were left intact. All specimens were cyclically loaded from 25 to 75 N for 50 cycles. Every 50 cycles, peak load was increased by 25 N until failure occurred. Cyclic stiffness and number of cycles were analyzed. RESULTS: The SR (72.9 ± 4.64 N/mm)- and aDR (72.6 ± 11.8 N/mm)-repaired specimens differed significantly in stiffness from the intact specimens (93.1 ± 14.8 N/mm) at ≥100 N (P < .05). The intact specimens and DR- and aDR-repaired specimens endured more cycles to failure (1,556 ± 677, 1,302 ± 248, and 1,211 ± 95, respectively) than the SR-repair specimens (388 ± 72 cycles, 260 ± 4 N) (P < .05 for all groups). CONCLUSIONS: Linked DR constructs were significantly stronger than SR repairs in this two-tendon RCT model and approached the strength of the intact rotator cuff. Augmentation with a collagen patch (aDR) did not influence biomechanical repair qualities in this model, but did result in less variability in failure load and more consistency in the mode of failure. CLINICAL RELEVANCE: The biomechanical properties of extended linked DR constructs are superior to those of SR constructs for repair of two-tendon RCTs, and are not compromised by graft augmentation.

A historical perspective of PCL bracing.

PURPOSE: Currently there are many functional knee braces but very few designed to treat the posterior cruciate ligament (PCL). No PCL braces have been biomechanically validated to demonstrate that they provide stability with proper force distribution to the PCL-deficient knee. The purpose of this review was to evaluate the history and current state of PCL bracing and to identify areas where further progress is required to improve patient outcomes and treatment options. METHODS: A PubMed search was conducted with the terms "posterior cruciate ligament", "rehabilitation", "history", "knee", and "brace", and the relevant articles from 1967 to 2011 were analysed. A review of the current available PCL knee bracing options was performed. RESULTS: Little evidence exists from the eight relevant articles to support the biomechanical efficacy of nonoperative and postoperative PCL bracing protocols. Clinical outcomes reported improvements in reducing PCL laxity with anterior directed forces to the tibia during healing following PCL tears. Biomechanics research demonstrates that during knee flexion, the PCL experiences variable tensile forces. One knee brace has been specifically designed and clinically validated to improve stability in PCL-deficient knees during rehabilitation. While available PCL braces demonstrate beneficial patient outcomes, they lack evidence validating their biomechanical effectiveness. CONCLUSIONS: There is limited information evaluating the specific effectiveness of PCL knee braces. A properly designed PCL brace should apply correct anatomic joint forces that vary with the knee flexion angle and also provide adjustability to satisfy the demands of various activities. No braces are currently available with biomechanical evidence that satisfies these requirements. LEVEL OF EVIDENCE: IV.

Endogenous concentrations of ouabain act as a cofactor to stimulate fluid secretion and cyst growth of in vitro ADPKD models via cAMP and EGFR-Src-MEK pathways.

In autosomal-dominant polycystic kidney disease (ADPKD), renal cysts develop by aberrant epithelial cell proliferation and transepithelial fluid secretion. We previously showed that ouabain increases proliferation of cultured human ADPKD cells via stimulation of the EGF receptor (EGFR)-Src-MEK/ERK signaling pathway. We examined whether ouabain affects fluid secretion and in vitro cyst growth of human ADPKD cell monolayers, ADPKD cell microcysts cultured in a three-dimensional collagen matrix, and metanephric organ cultures from Pkd1(m1Bei) mice. Physiological concentrations of ouabain alone did not affect net transepithelial basal-to-apical fluid transport in ADPKD monolayers or growth of cultured ADPKD microcysts. In contrast, in the presence of forskolin or 8-bromo-cAMP, ouabain significantly enhanced ADPKD fluid secretion and microcyst expansion. Ouabain exerted this effect by enhancing cAMP-dependent Cl(-) secretion via the CFTR. Similarly, ouabain accelerated cAMP-dependent cyst enlargement in Pkd1(m1Bei) mice metanephroi, with a more prominent response in homozygous than heterozygous mice. Ouabain had no effect on fluid secretion and cystogenesis of normal human kidney cells and caused only slight cystic dilations in wild-type mouse kidneys. The effects of ouabain in ADPKD cells and Pkd1(m1Bei) metanephroi were prevented by inhibitors of EGFR (AG1478), Src (PP2), and MEK (U0126). Together, our results show that ouabain, used in physiological concentrations, has synergistic effects on cAMP-mediated fluid secretion and cyst growth, via activation of the EGFR-Src-MEK pathway. These data provide important evidence for the role of ouabain as an endogenous hormone that exacerbates ADPKD cyst progression.

Prospective outcomes of young and middle-aged adults with medial compartment osteoarthritis treated with a proximal tibial opening wedge osteotomy.

PURPOSE: The purpose of this study was to conduct a prospective outcome analysis of proximal tibial opening wedge osteotomies performed in young and middle-aged patients (aged <55 years) for the treatment of symptomatic medial compartment osteoarthritis of the knee. METHODS: A consecutive series of young and middle-aged adults who underwent proximal tibial opening wedge osteotomies for symptomatic medial compartment osteoarthritis and genu varus alignment were prospectively followed up. Patients were evaluated with preoperative and postoperative modified Cincinnati Knee Scores and International Knee Documentation Committee objective knee subscores for knee effusions and the single-leg hop. Calculations were made of the preoperative and postoperative long-leg radiographic mechanical weight-bearing axis, patellar height (Insall-Salvati index), and tibial slope. A separate cohort of asymptomatic patients was used to quantify tibial plateau anatomy to provide an objective description of the lower extremity mechanical axis. RESULTS: There were 47 patients, with a mean age of 40.5 years, with a minimum of 2 years' follow-up, who formed this patient cohort. Modified Cincinnati Knee Scores improved significantly from 42.9 preoperatively to 65.1 at a mean of 3.6 years of follow-up. Radiographic analysis of a separate cohort showed the medial tibial eminence to be located at the 41% point along the tibial plateau from medial (0%) to lateral (100%). There was a significant improvement in malalignment: the mean mechanical axis passed through the tibial plateau at 23% of the distance along the proximal tibia preoperatively versus 54% postoperatively. The Insall-Salvati index decreased from 1.03 to 0.95 (P < .05), and posterior tibial slope increased from 9.4° to 11.7° (P < .05). Of the osteotomies, 3 (6%) were considered failures, defined by revision of the osteotomy or conversion to total knee arthroplasty. CONCLUSIONS: Performing proximal tibial opening wedge osteotomies to treat symptomatic medial compartment osteoarthritis in carefully selected patients leads to a significant improvement in subjective and objective clinical outcome scores with correction of malalignment at a mean of 3.6 years postoperatively. LEVEL OF EVIDENCE: Level IV, therapeutic case series.

Critical Role of Osteopontin in Maintaining Urinary Phosphate Solubility in CKD.

Background: Nephron loss dramatically increases tubular phosphate to concentrations that exceed supersaturation. Osteopontin (OPN) is a matricellular protein that enhances mineral solubility in solution; however, the role of OPN in maintaining urinary phosphate solubility in CKD remains undefined. Methods: Here, we examined (1) the expression patterns and timing of kidney/urine OPN changes in CKD mice, (2) if tubular injury is necessary for kidney OPN expression in CKD, (3) how OPN deletion alters kidney mineral deposition in CKD mice, (4) how neutralization of the mineral-binding (ASARM) motif of OPN alters kidney mineral deposition in phosphaturic mice, and (5) the in vitro effect of phosphate-based nanocrystals on tubular epithelial cell OPN expression. Results: Tubular OPN expression was dramatically increased in all studied CKD murine models. Kidney OPN gene expression and urinary OPN/Cr ratios increased before changes in traditional biochemical markers of kidney function. Moreover, a reduction of nephron numbers alone (by unilateral nephrectomy) was sufficient to induce OPN expression in residual nephrons and induction of CKD in OPN-null mice fed excess phosphate resulted in severe nephrocalcinosis. Neutralization of the ASARM motif of OPN in phosphaturic mice resulted in severe nephrocalcinosis that mimicked OPN-null CKD mice. Lastly, in vitro experiments revealed calcium-phosphate nanocrystals to induce OPN expression by tubular epithelial cells directly. Conclusions: Kidney OPN expression increases in early CKD and serves a critical role in maintaining tubular mineral solubility when tubular phosphate concentrations are exceedingly high, as in late-stage CKD. Calcium-phosphate nanocrystals may be a proximal stimulus for tubular OPN production.

Caspase-1 and the inflammasome promote polycystic kidney disease progression.

We and others have previously shown that the presence of renal innate immune cells can promote polycystic kidney disease (PKD) progression. In this study, we examined the influence of the inflammasome, a key part of the innate immune system, on PKD. The inflammasome is a system of molecular sensors, receptors, and scaffolds that responds to stimuli like cellular damage or microbes by activating Caspase-1, and generating critical mediators of the inflammatory milieu, including IL-1ß and IL-18. We provide evidence that the inflammasome is primed in PKD, as multiple inflammasome sensors were upregulated in cystic kidneys from human ADPKD patients, as well as in kidneys from both orthologous (PKD1 RC/RC or RC/RC) and non-orthologous (jck) mouse models of PKD. Further, we demonstrate that the inflammasome is activated in female RC/RC mice kidneys, and this activation occurs in renal leukocytes, primarily in CD11c+ cells. Knock-out of Casp1, the gene encoding Caspase-1, in the RC/RC mice significantly restrained cystic disease progression in female mice, implying sex-specific differences in the renal immune environment. RNAseq analysis implicated the promotion of MYC/YAP pathways as a mechanism underlying the pro-cystic effects of the Caspase-1/inflammasome in females. Finally, treatment of RC/RC mice with hydroxychloroquine, a widely used immunomodulatory drug that has been shown to inhibit the inflammasome, protected renal function specifically in females and restrained cyst enlargement in both male and female RC/RC mice. Collectively, these results provide evidence for the first time that the activated Caspase-1/inflammasome promotes cyst expansion and disease progression in PKD, particularly in females. Moreover, the data suggest that this innate immune pathway may be a relevant target for therapy in PKD.

Ouabain activates the Na-K-ATPase signalosome to induce autosomal dominant polycystic kidney disease cell proliferation.

The Na-K-ATPase is part of a cell signaling complex, the Na-K-ATPase signalosome, which upon activation by the hormone ouabain regulates the function of different cell types. We previously showed that ouabain induces proliferation of epithelial cells derived from renal cysts of patients with autosomal dominant polycystic kidney disease (ADPKD cells). Here, we investigated the signaling pathways responsible for mediating the effects of ouabain in these cells. Incubation of ADPKD cells with ouabain, in concentrations similar to those found in blood, stimulated phosphorylation of the epidermal growth factor receptor (EGFR) and promoted its association to the Na-K-ATPase. In addition, ouabain activated the kinase Src, but not the related kinase Fyn. Tyrphostin AG1478 and PP2, inhibitors of EGFR and Src, respectively, blocked ouabain-dependent ADPKD cell proliferation. Treatment of ADPKD cells with ouabain also caused phosphorylation of the caveolar protein caveolin-1, and disruption of cell caveolae with methyl-ß-cyclodextrin prevented Na-K-ATPase-EGFR interaction and ouabain-induced proliferation of the cells. Downstream effects of ouabain in ADPKD cells included activation of B-Raf and MEK and phosphorylation of the extracellular regulated kinase ERK, which translocated into the ADPKD cell nuclei. Finally, ouabain reduced expression of the cyclin-dependent kinase inhibitors p21 and p27, which are suppressors of cell proliferation. Different from ADPKD cells, ouabain showed no significant effect on B-Raf, p21, and p27 in normal human kidney epithelial cells. Altogether, these results identify intracellular pathways of ouabain-dependent Na-K-ATPase-mediated signaling in ADPKD cells, including EGFR-Src-B-Raf-MEK/ERK, and establish novel mechanisms involved in ADPKD cell proliferation.

Altered eicosanoid metabolism in the cystic fibrosis mouse small intestine.

OBJECTIVES: Imbalances in essential fatty acid levels have been reported in cystic fibrosis (CF), which may relate to elevated proinflammatory eicosanoid generation. The aim of this work was to better define eicosanoid metabolism in the CF intestine. MATERIALS AND METHODS: We used the small intestine of the cystic fibrosis transmembrane conductance regulator knockout mouse (CF mouse) to measure eicosanoid metabolic gene expression by quantitative reverse transcription polymerase chain reaction and Western blot, and eicosanoid levels by enzyme immunoassay, as compared with wild-type (WT) littermates. RESULTS: In the CF small intestine, expression of the secretory phospholipase A2 Pla2g5 mRNA was upregulated to 980% of WT levels. The following were downregulated: leukotriene C4 synthase Ltc4s (mRNA 55% of WT); omega-hydroxylase cytochrome P450s Cyp2c40 (mRNA 54% of WT), and Cyp4a10 (mRNA 4% of WT); and the major prostaglandin degradative enzymes prostaglandin dehydrogenase Hpgd (mRNA 27% of WT) and leukotriene B4 12-hydroxydehydrogenase/15-oxo-prostaglandin 13-reductase Ltb4dh (mRNA 64% and protein 30% of WT). The prostaglandins PGE2 and PGF2alpha were increased to 400% to 600% of WT levels in the CF mouse intestine, and the hydroxyeicosatetraenoic acids (HETEs) 12-, 15-, and 20-HETE were decreased to 3% to 20% of WT levels. CONCLUSIONS: There are changes in eicosanoid metabolic gene expression that are accompanied by significant changes in specific eicosanoid levels. These changes are expected to play important roles in the pathophysiology of CF in the intestine.