Three cases of multicentric carpotarsal osteolysis syndrome: a case series.

BACKGROUND: Multicentric carpotarsal osteolysis syndrome (MCTO) is characterized by progressive destruction and disappearance of the carpal and tarsal bones associated with nephropathy. MCTO is caused by loss-of-function mutations in the MAF bZIP transcription factor B (MAFB) gene. CASE PRESENTATION: This report describes three unrelated patients with MAFB mutations, including two male and one female patient. Osteolytic lesions in the carpal and tarsal bones were detected at 2 years, 12 years, and 14 months of age, respectively. Associated proteinuria was noted at 4 years, 12 years, and 3 months of age, respectively. Kidney biopsy was performed in two of them and revealed focal segmental glomerulosclerosis (FSGS). One patient showed progression to end-stage renal disease, that is by 1 year after the detection of proteinuria. The second patient had persistent proteinuria but maintained normal renal function. In the third patient, who did not undergo a kidney biopsy, the proteinuria disappeared spontaneously. The bony lesions worsened progressively in all three patients. Mutational study of MAFB revealed three different mutations, two novel mutations [c.183C > A (p.Ser61Arg) and c.211C > G (p.Pro71Ala)] and one known mutation [c.212C > T (p.Pro71Leu)]. CONCLUSION: We report three cases with MCTO and two novel MAFB mutations. The renal phenotypes were different among the three patients, whereas progressive worsening of the bony lesions was common in all patients. We also confirmed FSGS to be an early renal pathologic finding in two cases. A diagnosis of MCTO should be considered in patients with progressive bone loss concentrated primarily in the carpal and tarsal bones and kidney involvement, such as proteinuria.

Determination of reference intervals for fluid analysis and cytologic evaluation variables in synovial fluid samples obtained from carpal and tarsal joints in commercial nonlame growing swine.

OBJECTIVE To determine reference intervals for total nucleated cell count, total protein concentration, pH, RBC count, and percentages of neutrophils, lymphocytes, and large mononuclear cells in synovial fluid samples (SFSs) obtained from the carpal and tarsal joints of healthy swine. ANIMALS 54 healthy commercial finisher pigs that had no evidence of lameness or gross joint swelling. PROCEDURES Each pig was anesthetized, and SFSs were collected from 1 carpal and 1 tarsal joint for fluid analysis, cytologic evaluation, bacterial culture, and PCR analyses for common swine joint pathogens. Each pig was euthanized after SFS collection, and synovial tissue samples were collected for histologic assessment. If necessary, postmortem SFSs were collected. RESULTS Overall, 37 of 50 tarsal and 46 of 53 carpal SFSs met inclusion criteria of sufficient volume, no gross blood contamination, and negative results of bacterial culture and PCR analyses, and were from joints with histologically normal synovial tissues. For the carpal and tarsal joints, upper reference limits were as follows: total nucleated cell count, 3,281 cells/µL and 2,368 cells/µL, respectively; total protein concentration, 3.6 g/dL and 3.6 g/dL, respectively; pH, 7.2 and 7.0, respectively; RBC count, 0.8 × 106 cells/µL and 0.1 × 106 cells/µL, respectively; and percentage of neutrophils, 46.5% and 33.7%, respectively; percentage of lymphocytes, 40.6% and 56.3%, respectively; and percentage of large mononuclear cells, 92.0% and 95.3%, respectively. CONCLUSIONS AND CLINICAL RELEVANCE Results have provided reference intervals for selected variables in SFSs obtained from the carpal and the tarsal joints of healthy swine, which should be useful in diagnostic investigations of swine lameness and arthritis.

Characterization of Normal Murine Carpal Bone Development Prompts Re-Evaluation of Pathologic Osteolysis as the Cause of Human Carpal-Tarsal Osteolysis Disorders.

Multicentric carpal-tarsal osteolysis; multicentric osteolysis, nodulosis, and arthropathy; and Winchester syndromes, skeletal dysplasias characterized by carpal/tarsal and epiphyseal abnormalities, are caused by mutations in v-maf musculoaponeurotic fibrosarcoma oncogene ortholog B (MAFB), matrix metalloproteinase (MMP) 2, and MMP14, respectively; however, the underlying pathophysiology is unclear. Osteoclast-mediated osteolysis has been regarded as the main mechanism, but does not explain the skeletal distribution. We hypothesized that MAFB, MMP-2, and MMP-14 have integral roles in carpal/tarsal and epiphyseal bone development. Normal neonatal mouse forepaws were imaged by micro-computed tomography and examined histologically. Murine forepaw ossification occurred sequentially. Subarticular regions of endochondral ossification showed morphologic and calcification patterns that were distinct from archetypical physeal endochondral ossification. This suggests that two different forms of endochondral ossification occur. The skeletal sites showing the greatest abnormality in the carpal-tarsal osteolysis syndromes are regions of subarticular ossification. Thus, abnormal bone formation in areas of subarticular ossification may explain the site-specific distribution of the carpal-tarsal osteolysis phenotype. MafB, Mmp-2, and Mmp-14 were expressed widely, and tartrate-resistant acid phosphatase staining notably was absent in the subarticular regions of the cartilage anlagen and entheses at a time point most relevant to the human osteolysis syndromes. Thus, abnormal peri-articular skeletal development and modeling, rather than excessive bone resorption, may be the underlying pathophysiology of these skeletal syndromes.

Surgical Treatment of Symptomatic Congenital Type I Lunotriquetral Coalition: Technique and a Report of 4 Cases.

Lunotriquetral (LT) synchondrosis is an uncommon variant of congenital LT coalition. Unlike complete LT fusions, this incomplete pseudoarthrosis-like coalition can become symptomatic. Surgical LT arthrodesis is a described treatment for this variant of LT coalition (Minnaar type I). We report 4 patients who underwent LT fusion with a second-generation headless compression screw and distal radius cancellous autograft. Fusion was achieved in all cases at an average of 2.5 months postoperatively. All the patients were satisfied with complete pain relief in 2 patients and minimal pain in 2 patients, and all improved their range of motion. We recommend this procedure, and report the techniques for this limited wrist arthrodesis as treatment for the symptomatic LT synchondrosis.

A homozygous HOXD13 missense mutation causes a severe form of synpolydactyly with metacarpal to carpal transformation.

Synpolydactyly (SPD) is a rare congenital limb disorder characterized by syndactyly between the third and fourth fingers and an additional digit in the syndactylous web. In most cases SPD is caused by heterozygous mutations in HOXD13 resulting in the expansion of a N-terminal polyalanine tract. If homozygous, the mutation results in severe shortening of all metacarpals and phalanges with a morphological transformation of metacarpals to carpals. Here, we describe a novel homozygous missense mutation in a family with unaffected consanguineous parents and severe brachydactyly and metacarpal-to-carpal transformation in the affected child. We performed whole exome sequencing on the index patient, followed by Sanger sequencing of parents and patient to investigate cosegregation. The DNA-binding ability of the mutant protein was tested with electrophoretic mobility shift assays. We demonstrate that the c.938C>G (p.313T>R) mutation in the DNA-binding domain of HOXD13 prevents binding to DNA in vitro. Our results show to our knowledge for the first time that a missense mutation in HOXD13 underlies severe brachydactyly with metacarpal-to-carpal transformation. The mutation is non-penetrant in heterozygous carriers. In conjunction with the literature we propose the possibility that the metacarpal-to-carpal transformation results from a homozygous loss of functional HOXD13 protein in humans in combination with an accumulation of non-functional HOXD13 that might be able to interact with other transcription factors in the developing limb.

A New Subtype of Multiple Synostoses Syndrome Is Caused by a Mutation in GDF6 That Decreases Its Sensitivity to Noggin and Enhances Its Potency as a BMP Signal.

Growth and differentiation factors (GDFs) are secreted signaling molecules within the BMP family that have critical roles in joint morphogenesis during skeletal development in mice and humans. Using genetic data obtained from a six-generation Chinese family, we identified a missense variant in GDF6 (NP_001001557.1; p.Y444N) that fully segregates with a novel autosomal dominant synostoses (SYNS) phenotype, which we designate as SYNS4. Affected individuals display bilateral wrist and ankle deformities at birth and progressive conductive deafness after age 40 years. We find that the Y444N variant affects a highly conserved residue of GDF6 in a region critical for binding of GDF6 to its receptor(s) and to the BMP antagonist NOG, and show that this mutant GDF6 is a more potent stimulator of the canonical BMP signaling pathway compared with wild-type GDF6. Further, we determine that the enhanced BMP activity exhibited by mutant GDF6 is attributable to resistance to NOG-mediated antagonism. Collectively, our findings indicate that increased BMP signaling owing to a GDF6 gain-of-function mutation is responsible for loss of joint formation and profound functional impairment in patients with SYNS4. More broadly, our study highlights the delicate balance of BMP signaling required for proper joint morphogenesis and reinforces the critical role of BMP signaling in skeletal development.

Osteoclasts are recruited to the subchondral bone in naturally occurring post-traumatic equine carpal osteoarthritis and may contribute to cartilage degradation.

UNLABELLED: The role of osteoclasts in osteochondral degeneration in osteoarthritis (OA) has rarely been investigated in spontaneous disease or animal models of OA. OBJECTIVE: The objectives of the current study were to investigate osteoclast density and location in post-traumatic OA (PTOA) and control specimens from racehorses. METHOD: Cores were harvested from a site in the equine third carpal bone, that undergoes repetitive, high intensity loading. Histological and immunohistochemical (Cathepsin K and Receptor-activator of Nuclear Factor kappa-ß ligand (RANKL)) stained sections were scored (global and subregional) and the osteoclast density calculated. The cartilage histological scores were compared with osteoclast density and RANKL scores. RESULTS: There was a greater density of osteoclasts in PTOA samples and they were preferentially located in the subchondral bone plate. RANKL scores positively correlated to the scores of cartilage degeneration and the osteoclast density. The relationship between hyaline articular cartilage RANKL score and osteoclast density was stronger than that of the subchondral bone RANKL score suggesting that cartilage RANKL may have a role in recruiting osteoclasts. The RANKL score in the articular calcified cartilage correlated with the number of microcracks also suggesting that osteoclasts recruited by RANKL may contribute to calcified cartilage degeneration in PTOA. CONCLUSION: Our results support the hypothesis that osteoclasts are recruited during the progression of spontaneous equine carpal PTOA by cartilage RANKL, contributing to calcified cartilage microcracks and focal subchondral bone loss.

Functional analysis of limb transcriptional enhancers in the mouse.

Transcriptional enhancers are genomic sequences bound by transcription factors that act together with basal transcriptional machinery to regulate gene transcription. Several high-throughput methods have generated large datasets of tissue-specific enhancer sequences with putative roles in developmental processes. However, few enhancers have been deleted from the genome to determine their roles in development. To understand the roles of two enhancers active in the mouse embryonic limb bud we deleted them from the genome. Although the genes regulated by these enhancers are unknown, they were selected because they were identified in a screen for putative limb bud-specific enhancers associated with p300, an acetyltransferase that participates in protein complexes that promote active transcription, and because the orthologous human enhancers (H1442 and H280) drive distinct lacZ expression patterns in limb buds of embryonic day (E) 11.5 transgenic mice. We show that the orthologous mouse sequences, M1442 and M280, regulate dynamic expression in the developing limb. Although significant transcriptional differences in enhancer-proximal genes in embryonic limb buds accompany the deletion of M1442 and M280 no gross limb malformations during embryonic development were observed, demonstrating that M1442 and M280 are not required for mouse limb development. However, M280 is required for the development and/or maintenance of body size; M280 mice are significantly smaller than controls. M280 also harbors an "ultraconserved" sequence that is identical between human, rat, and mouse. This is the first report of a phenotype resulting from the deletion of an ultraconserved element. These studies highlight the importance of determining enhancer regulatory function by experiments that manipulate them in situ and suggest that some of an enhancer's regulatory capacities may be developmentally tolerated rather than developmentally required.

A comprehensive review of reported heritable noggin-associated syndromes and proposed clinical utility of one broadly inclusive diagnostic term: NOG-related-symphalangism spectrum disorder (NOG-SSD).

The NOG gene encodes noggin, a secreted polypeptide that is important for regulating multiple signaling pathways during human development, particularly in cartilage and bone. The hallmark of NOG-related syndromes is proximal symphalangism, defined by abnormal fusion of the proximal interphalangeal joints of the hands and feet. Many additional features secondary to NOG mutations are commonly but inconsistently observed, including a characteristic facies with a hemicylindrical nose, congenital conductive hearing loss due to stapes fixation, and hyperopia. The variable clinical presentations led to the designation of five different autosomal dominant syndromes, all subsequently found to have resulted from NOG mutations. These include (1) proximal symphalangism; (2) multiple synostoses syndrome 1; (3) stapes ankylosis with broad thumbs and toes; (4) tarsal-carpal coalition syndrome; and (5) brachydactyly type B2. Herein, we review the phenotypic features associated with mutations in the NOG gene, demonstrating the overlapping characteristics of these syndromes. Due to the variable phenotypic spectrum within families and among families with the same mutation, we propose a unifying term, NOG-related symphalangism spectrum disorder (NOG-SSD), to aid in the clinical recognition and evaluation of all affected individuals with these phenotypes. These NOG gene variants are available in a new locus-specific database (https://NOG.lovd.nl).

Circadian rhythm of osteocalcin in the maxillomandibular complex.

The human body displays central circadian rhythms of activity. Recent findings suggest that peripheral tissues, such as bone, possess their own circadian clocks. Studies have shown that osteocalcin protein levels oscillate over a 24-hour period, yet the specific skeletal sites involved and its transcriptional profile remain unknown. The current study aimed to test the hypothesis that peripheral circadian mechanisms regulate transcription driven by the osteocalcin promoter. Transgenic mice harboring the human osteocalcin promoter linked to a luciferase reporter gene were used. Mice of both genders and various ages were analyzed non-invasively at sequential times throughout 24-hour periods. Statistical analyses of luminescent signal intensity of osteogenic activity from multiple skeletal sites indicated a periodicity of ~ 24 hrs. The maxillomandibular complex displayed the most robust oscillatory pattern. These findings have implications for dental treatments in orthodontics and maxillofacial surgery, as well as for the mechanisms underlying bone remodeling in the maxillomandibular complex.

Automatic bone age assessment for young children from newborn to 7-year-old using carpal bones.

A computer-aided-diagnosis (CAD) method has been previously developed based on features extracted from phalangeal regions of interest (ROI) in a digital hand atlas, which can assess bone age of children from ages 7 to 18 accurately. Therefore, in order to assess the bone age of children in younger ages, the inclusion of carpal bones is necessary. However, due to various factors including the uncertain number of bones appearing, non-uniformity of soft tissue, low contrast between the bony structure and soft tissue, automatic segmentation and identification of carpal bone boundaries is an extremely challenging task. Past research works on carpal bone segmentation were performed utilizing dynamic thresholding. However, due to the limitation of the segmentation algorithm, carpal bones have not been taken into consideration in the bone age assessment procedure. In this paper, we developed and implemented a knowledge-based method for fully automatic carpal bone segmentation and morphological feature analysis. Fuzzy classification was then used to assess the bone age based on the selected features. This method has been successfully applied on all cases in which carpal bones have not overlapped. CAD results of total about 205 cases from the digital hand atlas were evaluated against subject chronological age as well as readings of two radiologists. It was found that the carpal ROI provides reliable information in determining the bone age for young children from newborn to 7-year-old.

Connexin 40, a target of transcription factor Tbx5, patterns wrist, digits, and sternum.

Haploinsufficiency of T-box transcription factor 5 (TBX5) causes human Holt-Oram syndrome (HOS), a developmental disorder characterized by skeletal and heart malformations. Mice carrying a Tbx5 null allele (Tbx5(+/Delta)) have malformations in digits, wrists, and sternum joints, regions where Tbx5 is expressed. We demonstrate that mice deficient in connexin 40 (Cx40), a Tbx5-regulated gap junction component, shared axial and appendicular skeletal malformations with Tbx5(+/Delta) mice. Although no role in skeleton patterning has been described for gap junctions, we demonstrate here that Cx40 is involved in formation of specific joints, as well as bone shape. Even a 50% reduction in either Tbx5 or Cx40 produces bone abnormalities, demonstrating their crucial control over skeletal development. Further, we demonstrate that Tbx5 exerts in part its key regulatory role in bone growth and maturation by controlling via Cx40 the expression of Sox9 (a transcription factor essential for chondrogenesis and skeleton growth). Our study strongly suggests that Cx40 deficiency accounts for many skeletal malformations in HOS and that Tbx5 regulation of Cx40 plays a critical role in the exquisite developmental patterning of the forelimbs and sternum.

The effects of reduced oxygen tension on cell proliferation and matrix synthesis in synovium and tendon explants from the rabbit carpal tunnel: an experimental study in vitro.

Local ischemia may play an important role in the development of tendon degeneration as well as entrapment neuropathies. In order to investigate the cellular effects of hypoxia on tendon and synovial tissue from the carpal canal, dose response effects of oxygen on cell proliferation and synthesis of matrix components were examined in segments of synovial and flexor digitorum profundus tendon from the carpal tunnel of rabbits during short term culture. Explants were incubated in airtight containers flushed with either 0%, 1%, 3%, 20% O2 plus 2% CO2 and N2 to balance and labeled with either 3H-thymidine or 3H-proline and 35S-sulfate. Cell proliferation was significantly inhibited by hypoxia in synovium but not in tendon (P = 0.03). In parallel, the synthesis of non-collagenous proteins was significantly reduced in synovium but not in tendon (P = 0.006). In both tissues hypoxia significantly inhibited collagen synthesis. On the other hand, hypoxia had no significant effect on the synthesis of new proteoglycans as determined by 35S-sulfate incorporation. Hypoxia can inhibit cell proliferation and alter synthesis of matrix components in synovial tissue, but may only have minor effects on non-collagen protein synthesis in tendon explants from the carpal canal of rabbit forepaws.

Experimental osteonecrosis of the lunate. Revascularization may cause collapse.

Is lunate collapse in Kienböck's disease a consequence of spontaneous revascularization, leading to focal osteolysis? A literature review of osteonecrosis in other locations such as the femoral head and bone allografts showed clearly that the loss of mechanical integrity is due to cellular processes which follow the spontaneous restoration of blood supply. We found no evidence in the literature that the lunate has been shown to be avascular at the time of collapse. On the contrary, increased osteoclastic activity has been reported. We excised and reimplanted the lunate in two monkeys, and found spontaneous revascularization, leading to increased osteoblastic activity. Other parts of the bone were destroyed by osteoclasts, leading to collapse. This histological example suggests that it may be possible to make an analogy with osteonecrosis in other locations. Thus, changes on plain radiography may indicate that the bone is revascularized spontaneously. Before performing operative revascularization of the lunate, one should consider that revascularization is a probable cause for collapse.

[Status and frequency of local juxta-articular demineralization and erosion in the wrist area in the early phase of chronic polyarthritis. Results based on 3-dimensional soft x-ray pictures]. / Lage und Häufigkeit lokaler gelenknaher Demineralisationen und Erosionen im Handgelenksbereich in der Frühphase der chronischen Polyarthritis. Ergebnisse nach Weichstrahlaufnahmen in 3 Ebenen.

In early rheumatoid arthritis the location and incidence of localized juxta-articular demineralizations and erosions were investigated at 53 points of the wrist. On the level of the metacarpal bases, the distal and proximal row of the carpal bones more changes are seen in the oblique vd. and the lateral view than in the dv. view. At the distal bones of the forearm more changes are seen at the radius than at the ulnar styloid. The most often changes at all occur at the volar middle third of the triquetrum in the oblique view and at the distal volar articular facet of the scaphoid in the lateral view. Close relations between localized juxta-articular demineralizations and erosions do not exist. If the early bone changes at the wrist in rheumatoid arthritis are to be detected additional oblique and lateral view are prerequisite.

Advantage of hand bone calcium content measurement by local neutron activation analysis for following up hemodialysis patients.

Hand bone calcium content [Ca] has been measured by local neutron analysis in hemodialyzed patients free of signs of osteomalacia before and 6-12 months after treatment with vitamin D metabolites, and it has been compared to some iliac bone histomorphometric parameters. With 1,25-(OH)2D3 alone, [Ca] increases significantly at the 6th month of treatment (p less than 0.001) but not from the 6th to the 12th month. With 1 alpha-(OH)D3 + 25-(OH)D3 the same phenomenon is observed at the 6th and 12th months of treatment. Before treatment, [Ca] was correlated negatively with the osteoid surfaces (OS) (r = -0.62, p = 0.01) and the number of osteoclasts per square millimeter (Ocl/mm2) (r = -0.79, p less than 0.001). At the 6th month of treatment, [Ca] was still correlated negatively with OS (r = -0.42, p = 0.05) and with Ocl/mm2 (r = -0.60, p less than 0.005). At the 12th month, the two negative correlations decreased but remained significant (r = -0.42, p = 0.05; r = -0.45, p = 0.05). These data suggest that (1) hand bone calcium analysis by neutron activation permits to follow up the peripheral bone mineral content in hemodialyzed patients and (2) the [Ca] increase only observed during the first months of treatment with vitamin D metabolites probably result from an attenuation of secondary hyperparathyroidism.