The growth of long bones in human embryological and fetal upper limbs and its relationship to other developmental patterns

Measurements were made of the long bones of the upper limbs (humerus, ulna, radius) of 58 aborted embryos and fetuses, developmental age from 8 to 14 weeks, crown-rump length (CRL) between 38 and 116 mm. The specimens were cleared and double-stained, using alcian blue and alizarin red S for a differential detection of cartilage and bone. The values of both the total length (TL) and the ossified part (OL) of each long bone were related to the fetal developmental age previously estimated by freshly measured CRL. The relationship to another developmental pattern, i.e. the number of ossified centres in the vertebral column, suggested that the OL values could be much more significant than TL for the assessment of fetal growth.     

Osteology and Radiological Anatomy of the Thoracic Limbs of Temminck's Ground Pangolin (Smutsia temminckii)

Temminck's ground pangolin is the only pangolin present in South Africa. It is a myrmecophagous mammal with a bipedal gait. The thoracic limbs are used to break open ant nests, dig for food, and expand previously occupied burrows. This study describes the osteology and radiological anatomy of the thoracic limbs of this threatened species. Thoracic limbs from four Temminck's ground pangolins, which succumbed from electrocution or natural causes, were digitally radiographed in situ. The individual bones were then cleaned, described and digitally radiographed. The skeleton of the thoracic limbs revealed a similar number and arrangement of bones compared to that of domestic carnivores. The bones were robust and displayed numerous open epiphyseal lines. The latter provide an estimate of sexual maturity and should not be confused with fractures in injured ground pangolins. The scapula was broad and triangular‐shaped. The humerus displayed a massive medial epicondyle. The radius and ulna were similarly sized, and displayed a broad radial trochlea and large olecranon tuber, respectively. The manus was composed of seven carpal bones, five short metacarpal bones and five digits of which the three central digits were the best developed. The unguicular process of the distal phalanx was bifid and elongated. The osteological characteristics indicate that the thoracic limbs of Temminck's ground pangolin are specifically adapted for protraction and retraction, strong elbow extension, flexion of the carpus and digits as well as pronation and supination of the antebrachium, as opposed to weight‐bearing. These functions are likewise documented for other scratch‐digging species. Anat Rec, 301:624–635, 2018. © 2017 Wiley Periodicals, Inc.     

Morphometry by computerized three-dimensional reconstruction of the human carpal bones during embryogenesis

Carpal skeleton shows drastic developmental changes during embryogenesis. At this stage, the cartilaginous matrices appear and later form models of the limb bones. The purpose of this study was to investigate the morphometry of carpal bones in humans during embryological development. We obtained digitalized histological serial sections of 18 human embryos and early fetuses from the Institute of Anatomy in Paris. Surfdriver and MSC.Patran software were used for three-dimensional reconstruction and morphometry. There was a strong correlation between the volume of the carpal cartilaginous structure and the size of the embryos (P<0.001) and an exponential correlation between the carpal volume and the percentage of volume presented by the proximal carpal row (P=0.005). According to inertia parameters, the geometry of carpal cartilaginous structure, initially plane, becomes curved during embryogenesis. Carpal bones growth follows non-homothetic transformation. The innovations in embryo reconstruction serve as new tool for scientific investigation. A hypothesis of carpal development is proposed.     

Ossification Pattern in Forelimbs of the Siamese Crocodile (Crocodylus siamensis): Similarity in Ontogeny of Carpus Among Crocodylian Species

Crocodylians have highly derived elongated carpus, which is related to their use of forelimbs in many types of gaits as well as in burrowing. The objective of present study was to describe the ossification of the forelimb in five stages of Siamese crocodile (Crocodylus siamensis). The ossification begins approximately at stage 20 in arm and forearm bones moving sequentially to the metacarpal elements. The first carpal elements with ossification centers are radiale + intermedium and ulnare (stage 22–23), and their ossification mode is typical of long bones. Between stages 22 and 24 distal carpals 3, 4, and 5 fuse together to a single formation. In the stage 25, the ossification proceeds to the pisiform, which starts ossifying late during the embryogenesis. The phalangeal formula of the digits is 2,3,4,5,3. Although there are some interspecific differences, it appears that all crocodylians have similarly uniform skeletal pattern, the process of ossification, number of carpal elements and phalangeal formulas probably due to their similar lifestyles. Anat Rec, 301:1159–1168, 2018. © 2018 Wiley Periodicals, Inc.     

Role of fasciae around the median nerve in pathogenesis of carpal tunnel syndrome: microscopic and ultrasound study

This study investigated the connections between the median nerve paraneural sheath and myofascial structures near it, from both macroscopic and microscopic points of view. Four samples of median nerve and surrounding tissues were excised from nine non-embalmed upper limbs for microscopic analysis. Ultrasound images were analysed in 21 healthy subjects and 16 carpal tunnel syndrome patients to evaluate median nerve transversal displacement during finger motion at carpal tunnel and forearm levels. An anatomical continuity between epimysium and paraneural sheath and a reduction of paraneural fat tissue from proximal to distal was found in all samples. Median nerve displacements at both levels were significantly reduced in carpal tunnel syndrome subjects (P < 0.001). It was observed that the median nerve is not an isolated structure but is entirely connected to myofascial structures. Therefore, unbalanced tension of epimysial fasciae can affect the paraneural sheath, limiting nerve displacement, and consequently this must be included in carpal tunnel syndrome pathogenesis.     

Histamine dilutions modulate basophil activation

Background:In order to demonstrate that high dilutions of histamine are able to inhibit basophil activation in a reproducible fashion, several techniques were used in different research laboratories.Objective:The aim of the study was to investigate the action of histamine dilutions on basophil activation.Methods:Basophil activation was assessed by alcian blue staining, measurement of histamine release and CD63 expression. Study 1 used a blinded multi-centre approach in 4 centres. Study 2, related to the confirmation of the multi-centre study by flow cytometry, was performed independently in 3 laboratories. Study 3 examined the histamine release (one laboratory) and the activity of H₂ receptor antagonists and structural analogues (two laboratories).Results:High dilutions of histamine (10^–30–10^–38 M) influence the activation of human basophils measured by alcian blue staining. The degree of inhibition depends on the initial level of anti-IgE induced stimulation, with the greatest inhibitory effects seen at lower levels of stimulation. This multicentre study was confirmed in the three laboratories by using flow cytometry and in one laboratory by histamine release. Inhibition of CD63 expression by histamine high dilutions was reversed by cimetidine (effect observed in two laboratories) and not by ranitidine (one laboratory). Histidine tested in parallel with histamine showed no activity on this model.Conclusions:In 3 different types of experiment, it has been shown that high dilutions of histamine may indeed exert an effect on basophil activity. This activity observed by staining basophils with alcian blue was confirmed by flow cytometry. Inhibition by histamine was reversed by anti-H2 and was not observed with histidine these results being in favour of the specificity of this effect We are however unable to explain our findings and are reporting them to encourage others to investigate this phenomenon.Received 11 December 2002; returned for revision 29 January 2003; accepted by A. Falus 12 November 2003     

Skeletal advance and arrest in giant non-metamorphosing African clawed frog tadpoles (Xenopus laevis: Daudin)

This study examines the skeletons of giant non‐metamorphosing (GNM) Xenopus laevis tadpoles, which arrest their development indefinitely before metamorphosis, and grow to excessively large sizes in the absence of detectable thyroid glands. Cartilage growth is isometric; however, chondrocyte size is smaller in GNM tadpoles than in controls. Most cartilages stain weakly with alcian blue, and several cartilages are calcified (unlike controls). However, cartilages subjacent to periosteum‐derived bone retain strong affinities for alcian blue, indicating a role for periosteum‐derived bone in the retention of glycosaminoglycans during protracted larval growth. Bone formation in the head, limb, and axial skeletons is advanced in comparison with stage‐matched controls, but arrests at various mid‐metamorphic states. Both dermal and periosteum‐derived bones grow to disproportionately large sizes in comparison to controls. Additionally, mature monocuspid teeth form in several GNM tadpoles. Advances in skeletal development are attributable to the old ages and large sizes of these tadpoles, and reveal unexpected developmental potentials of the pre‐metamorphic skeleton.     

Morphological observations and morphometric analysis in three human fetuses with bilateral cervical cystic hygroma

Three human fetuses (crown-rump length, CRL, ranging from 71 to 77 mm), presenting bilateral cervical cystic hygroma were examined. The specimens were cleared and double-stained with alcian blue and alizarin red S for detecting the ossification growth patterns in the vertebral column, ribs, ischium, limbs, and face. Longitudinal measurements of some long bones in the upper (humerus, ulna, radius) and lower (femur, tibia, fibula) limb were taken. The values of both the total length (TL) and the ossified part (OL) of each long bone, as well as the OL/TL per cent ratio were considered. Reference points were located on the mandible, i.e. condylar process (Pcl), coronoid process (Pco), gnathion (GN), gonion (GO), inferior interdental point (IDI) for measuring linear dimensions. All values obtained were related with those relative to a group of fetuses, without any detectable malformation and chromosomal abnormalities, with CRL mean value 75 mm, in order to assess the presence of further anomalies, besides the cystic hygroma, in the three fetuses considered.     

孕鼠子宫肥大细胞分布、数量和组化性质的动态变化

目的：探讨小鼠妊娠期子宫内肥大细胞的分布、数量和组化性质的动态变化。方法：采用甲苯胺益等染色法研究了孕１ｄ、３ｄ、５ｄ、７ｄ、９ｄ、１１ｄ、１３ｄ、１５ｄ、１７ｄ、１９ｄ时子宫内的肥大细胞。结果：（１）肥大细胞主要分布于子宫肌层,子宫内膜较少,且胚泡植入部位的肥大细胞明显少于非植入部位;（２）妊娠早期（孕１～５ｄ）和妊娠后期（孕１５～１９ｄ）肥大细胞数显著多于妊娠中期（Ｐ〈０．０５）,;（３）Ａｌ     

Animal model: skeletal anomalies in mice with cleidocranial dysplasia

Cleidocranial dysplasia in mice, a radiation-induced skeletal mutation, showed striking homology with cleidocranial dysplasia in humans. Genetic studies indicated that the condition in mice is inherited as an autosomal dominant trait with variable expressivity and almost complete penetrance. The homozygous condition was lethal in utero. Radiographic and alcian blue/alizarin red S-stained whole-skeletal preparation studies were used to determine the extent, pattern, incidence, and distribution of skeletal abnormalities in heterozygous mice. Cleidocranial dysplasia in mice was characterized by variable clavicular hypoplasia, delayed closure of cranial fontanelles and sutures, and variable hypoplasia of pelvic bones, in particular ischiopubic rami. The gene symbol Ccd is proposed for the cleidocranial dysplasia mutation in mice and humans.     

Chondrodysplasia in the Norwegian Elkhound.

Dwarfism in the Norwegian Elkhound occurred as a result of a generalized disturbance in endochondral ossification. Radiographic changes included flaring and increased width of the distal metaphyses of the radius and ulna, delayed ossification of the cuboid bones of the carpus, and reduction in length of the vertebral bodies. The zone of chondrocyte proliferation was decreased in width and contained areas of abnormal cell column formation alternated with wide areas of matrix. Chondrocytes in all zones contained one or more inclusions bounded by a smooth discontinuous membrane. The material within the inclusions appeared homogeneous and stained blue-green with Movat's pentachrome and deep blue with alcian blue-periodic acid-Schiff at pH 1.0 and 2.6. The distribution of ruthenium red granules in the matrix frequently revealed poor differentiation into territorial and interterritorial zones. Twenty-four-hour urine samples were negative for glucose, and the glycosaminoglycan excretion pattern was normal.     

人体腕骨显微骨硬度分布特征的研究

目的 探讨人体腕骨显微骨硬度的分布特征及其临床意义。方法 纳入3具新鲜冰冻成人尸体标本(62岁男性、58岁男性、45岁女性),将右侧腕骨软组织剥离后,用慢速锯分别在舟骨、月骨、头状骨、钩骨、大多角骨和小多角骨切取厚3 mm的骨组织标本。舟骨选取舟骨结节、腰部内外侧和舟骨体部,月骨选取腕关节面、掌侧面、背侧面和远端,头状骨、钩骨、大多角骨和小多角骨选取外层皮质不同区域,应用德国KB-5型显微维氏硬度仪测试标本不同区域的硬度值,采用50 gf力加载50 s、维持12 s的标准操作方法进行硬度值测定,每个区域选取 5 个有效值,全体有效值的平均值作为该部位的骨硬度值。观察不同骨骼间及骨骼内部不同区域的骨硬度值差异。结果 3具标本中共取得舟骨、月骨、头状骨、钩骨、大多角骨和小多角骨标本切片18片,测量位点255个。腕骨不同骨骼骨硬度从高到低依次为钩状骨(39.04±5.79)HV、头状骨(38.98±6.17)HV、舟骨(37.72±5.85)HV、大多角骨(35.89±4.75)HV、月骨(33.65±5.42)HV及小多角骨(31.82±5.54)HV,不同骨骼间总体骨硬度差异有统计学意义(F=10.783,P＜0.01)。舟骨、月骨内部不同区域骨硬度分布近似,舟骨结节、腰部外侧、腰部内侧和舟骨体部骨硬度分别为(37.07±5.77)、(37.51±6.39)、(40.00±5.64)、(36.31±5.47)HV,其中腰部内侧骨硬度最大,舟骨体部骨硬度最小,4部位间骨硬度比较差异无统计学意义(F=1.129,P＞0.05)。月骨腕关节面、掌侧面、背侧面和远端骨硬度分别为(33.57±3.61)、(34.58±6.04)、(35.47±5.24)、(30.97±5.88)HV,其中背侧骨硬度最大,远端骨硬度最小, 4个部位间骨硬度比较差异无统计学意义(F=2.040,P＞0.05)。结论 健康人腕骨不同骨骼间硬度各有不同,舟骨和月骨内部各部位骨硬度分布均匀一致。测量腕骨显微骨硬度值,了解其分布特征,有助于了解腕骨微观生物力学性能,亦可指导腕骨骨折内固定方法的选择,设计制作更加符合人体生理状态下的腕部骨骼假体及建立腕部肌骨组织的有限元模型。临床试验注册 中国临床试验注册中心,注册号为ChiCTR-BPR-17010818。     

Effects of dividing the transverse carpal ligament on the mechanical behavior of the carpal bones under axial compressive load: a finite element study

Transecting the transverse carpal ligament (TCL) is a routine procedure to surgically treat carpal tunnel syndrome; yet, its mechanical consequences on carpal bones are unclear. In this study, our intent was to perform a computational analysis of carpal biomechanics resulting from TCL release. A three-dimensional finite element model of the wrist was constructed, which included all the carpal bones, the distal ulna and radius, the proximal metacarpals and the interosseous ligaments. Cartilage layers of each bone were modeled manually according to anatomic visualization software. The TCL was also modeled in three dimensions and added to the bone model. A 100-Newton axial load was applied to the upper section of the second and third metacarpals. The effects of dividing the TCL on the displacements of the carpal bones and the contact stress distribution in the midcarpal joints were studied using a finite element analysis method. When the TCL was divided, the axial compressive load resulted in the carpal bones deviating more radially. More specifically, the carpal bones on the radial side of the capitate and lunate (i.e. the trapezium, trapezoid, and scaphoid) moved further toward the radius, and the carpal bones on the ulnar side of the capitate and lunate (i.e. hamate, triquetrum, and pisiform) moved further toward the metacarpals. The contact stresses and contact locations in the midcarpal joints changed as a result of dividing the TCL. The changes in displacements of carpal bones and the contact stress distributions in the midcarpal joints due to TCL release may be implicated for some of the postoperative complications associated with carpal tunnel release.     

Piepkorn type of osteochondrodysplasia: Defining the severe end of FLNB‐related skeletal disorders in three fetuses and a 106‐year‐old exhibit

The Piepkorn type of lethal osteochondrodysplasia (POCD) is a rare and lethal dwarfing condition. Four cases have been reported to date. The characteristic features are distinctly shortened “flipper‐like” limbs, polysyndactyly, excessive underossification, especially of the limb bones and vertebrae, and large (giant) chondrocytes in the cartilaginous bone primordia. These characteristics allowed the diagnosis of Piepkorn type of osteochondrodysplasia in four new cases, three fetuses of 15 to 22 weeks and one 106‐year‐old museum exhibit. Piepkorn type of osteochondrodysplasia has been assigned to the giant cell chondrodysplasias such as atelosteogenesis type 1 (AO1) and boomerang dysplasia (BD). Analysis of the Filamin B gene in 3p14.3, which is associated with these disorders, allowed the identification of the first FLNB mutations in Piepkorn type of osteochondrodysplasia. The heterozygous missense mutations, found in the three fetuses, were located in exons 28 and 29, encoding the immunoglobulin‐like repeat region R15, one of three mutational hot spots in dominant FLNB‐related skeletal disorders. Direct preparations and alcian blue staining revealed single upper and lower arm and leg bone primordia, preaxial oligodactyly, and polysyndactyly with complete fusion and doubling of the middle and end phalanges II–V to produce eight distal finger rays. Considering the unique clinical features and the extent of underossification, Piepkorn type of osteochondrodysplasia can be regarded as a distinct entity within the AO1‐BD‐POCD continuum.     

Novel Melt-Processable Chitosan–Polybutylene Succinate Fibre Scaffolds for Cartilage Tissue Engineering

Novel chitosan/polybutylene succinate fibre-based scaffolds (C-PBS) were seeded with bovine articular chondrocytes in order to assess their suitability for cartilage tissue engineering. Chondrocytes were seeded onto C-PBS scaffolds using spinner flasks under dynamic conditions, and cultured under orbital rotation for a total of 6 weeks. Non-woven polyglycolic acid (PGA) felts were used as reference materials. Tissue-engineered constructs were characterized by scanning electron microscopy (SEM), hematoxylin–eosin (H&E), toluidine blue and alcian blue staining, immunolocalization of collagen types I and II, and dimethylmethylene blue (DMB) assay for glycosaminoglycans (GAG) quantification at different time points. SEM showed the chondrocytes' typical morphology, with colonization at the surface and within the pores of the C-PBS scaffolds. These observations were supported by routine histology. Toluidine blue and alcian blue stains, as well as immunohistochemistry for collagen types I and II, provided qualitative information on the composition of the engineered extracellular matrix. More pronounced staining was observed for collagen type II than collagen type I. Similar results were observed with constructs engineered on PGA scaffolds. These also exhibited higher amounts of matrix glycosaminoglycans and presented a central region which contained fewer cells and little matrix, a feature that was not detected with C-PBS constructs.     

Quantitative in vivo analysis of the kinematics of carpal bones from three-dimensional CT images using a deformable surface model and a three-dimensional matching technique

The purpose of this study was to obtain quantitative information of the relative displacements and rotations of the carpal bones during movement of the wrist. Axial helical CT scans were made of the wrists of 11 volunteers. The wrists were imaged in the neutral position with a conventional CT technique, and in 15-20 other postures (flexion-extension, radial-ulnar deviation) with a low-dose technique. A segmentation of the carpal bones was obtained by applying a deformable surface model to the regular-dose scan. Next, each carpal bone, the radius, and ulna in this scan was registered with the corresponding bone in each low-dose scan using a three-dimensional matching technique. A detailed definition of the surfaces of the carpal bones was obtained from the regular-dose scans. The low-dose scans provided sufficient information to obtain an accurate match of each carpal bone with its counterpart in the regular-dose scan. Accurate estimates of the relative positions and orientations of the carpal bones during flexion and deviation were obtained. This quantification will be especially useful when monitoring changes in kinematics before and after operative interventions, like mini-arthrodeses. This technique can also be applied in the quantification of the movement of other bones in the body (e.g., ankle and cortical spine).     

新疆哈萨克族儿童青少年手和腕部骨骼发育情况

目的：了解哈萨克族儿童青少年手和腕部骨骼发育情况,探讨其发育规律。并为制定哈萨克族儿童青少年骨龄标准提供科学依据。同时也为临床医学和人类学提供资料。材料和方法：选择哈萨克族儿童青少年１５１５人（男７４２,女７７３）其年龄为新生儿、３个月、６个月、１～２０岁共２３个年龄组,利用摄片法拍摄双手、腕部Ｘ片,进行观察和统计分析。结果：得出哈萨克族正常手、腕部各骨骨化中心出现及干骺愈合顺序。结论：手、腕部双     

New form of idiopathic osteolysis: nodulosis, arthropathy and osteolysis (NAO) syndrome

We describe 10 patients (6 females and 4 males) from 6 unrelated families with an autosomal recessive disease characterized by simultaneous presentation of nodulosis, arthropathy and osteolysis. They were followed up regularly at King Faisal Specialist Hospital and Research Center in Saudi Arabia for clinical evaluation, serial blood work-up, and evaluating radiological changes. Nodulosis and arthropathy were the clinical criteria for inclusion in this study, and the ten patients fulfilled these criteria. All patients had nodulosis and distal arthropathy. Eight patients (80%) presented with deformed hands and four (40%) with painful hands. All patients had parents who were first cousins and three families had more than one affected child, the finding suggesting autosomal recessive inheritance. Osteopenia and undertubulation of bones distally more than proximally, and upper limbs affected more often than lower limbs, were found in all patients. Osteolysis was seen in carpal and tarsal bones. Other common findings were sclerotic cranial sutures, brachycephaly, and broad medial clavicles. This novel phenotype should be considered in the differential diagnosis of chronic arthritis. Familial arthropathies are more often seen in communities where interfamilial marriage is common. Such a collection of patients is ideal for homozygosity mapping of the disease locus.     

掌向拉伸力作用下腕管的三维有限元分析

腕管综合征是腕部正中神经受到卡压而引起的一种神经病症.目前对腕管综合征的生物力学研究大都集中在对尸体标本的实验测试与测量方面,而少数数值模型则单独集中在对腕骨的载荷传递及应力分析方面,缺乏针对整个腕管结构的三维有限元建模.本研究从生物力学角度,建立了基于CT扫描图像数据的腕管三维有限元模型,首次立体构架了包括腕骨、腕横韧带和腕骨间韧带在内的三维实体腕管管道模型,计算并分析了掌向拉伸力作用下腕横韧带和腕管典型截面的变形情况.通过与前人文献数据相对照,证明本模型能够对腕管的生物力学研究提供较为准确的理论数据,从而对腕管综合征的临床诊断、临床手术以及术后康复等具有一定的指导意义.