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A histological study of mineralised tissue formation around implants with 3D culture of HMS0014 cells in Cellmatrix Type I-A collagen gel scaffold in vitro. 细胞基质I-A型胶原凝胶支架中HMS0014细胞体外3D培养对植入物周围矿化组织形成的组织学研究
Pub Date : 2014-01-01 DOI: 10.2535/ofaj.91.57
Aiko Morishita, Shunji Kumabe, Michiko Nakatsuka, Yasutomo Iwai

We cultured HMS0014 Yub621b cells within a 3D collagen gel scaffold (Cellmatrix Type I-A) and aimed to study the fate and contribution of human bone-derived mesenchymal stem cells (MSCs) in the guided bone regeneration(GBR)-engineered tissue which has developed around the titanium (Ti) test dental implant (IP) in vitro. The light microscopy (LM) and transmission electron microscopy (TEM) results of the peri-IP tissue indicated that collagen fibrils of the Cellmatrix Type I-A gel were accumulated and fabricated to provide a 3D meshwork for proliferation and differentiation of the HMS0014 cells in the top (cell) layer; mineralisation of the GBR tissue had commenced since day 1 and became markedly deposited between days 7 and 14 of the experiment. TEM observation revealed sedimentation of cement line at the periphery of the interwoven Cellmatrix fibres and fibrils in the ECM scaffold of the GBR tissue; matrix vesicle-mediated and appositional collagen-mediated mineralisation were identified in the peri-IP ECM scaffold. The fine structure study of the plurimorphic osteoblast(Ob)-like osteogeneic cells demonstrated numerous membranous organelles related with vesicular trafficking, secretion and endocytosis in the cytoplasm; well-developed cytoskeleton networks and intercellular junctional complexes were also observed. The specimens on fluorescence immunohistochemistry (IHC) by confocal laser-scanning microscopy (LSM) showed the expression of LC3 and Cx43 associated with autophagic-lysosomal degeneration pathway and signal conduction mediated with gap junctions (GJS) in maintaining tissue homeostasis of the Ob-like cells which grew and degenerated in the 3D scaffold. Results from this in vitro study suggest that Ob-like HMS0014 cells actively regulate turnover of the peri-IP ECM to recapitulate the development and formation of osteoid tissue-engineered material which might contribute to augment osseointegration around the dental implant.

我们在3D胶原凝胶支架(Cellmatrix Type I-A)中培养HMS0014 Yub621b细胞,旨在研究人骨源性间充质干细胞(MSCs)在体外钛(Ti)试验牙种植体(IP)周围诱导骨再生(GBR)工程组织中的命运和贡献。通过光镜和透射电镜观察发现,细胞基质I-A型凝胶形成了胶原原纤维,为HMS0014细胞顶层的增殖和分化提供了三维网络;GBR组织矿化从第1天开始,并在实验的第7天至第14天之间明显沉积。透射电镜观察发现,GBR组织的ECM支架细胞基质纤维和原纤维交织的周围有水泥线沉积;在颈周ECM支架中发现了基质囊泡介导和相位胶原介导的矿化。多形性成骨细胞(Ob)样成骨细胞的精细结构研究表明,细胞质中有许多与囊泡运输、分泌和内吞作用有关的膜细胞器;还观察到发育良好的细胞骨架网络和细胞间连接复合物。共聚焦激光扫描显微镜(LSM)荧光免疫组化(IHC)显示,LC3和Cx43的表达与自噬-溶酶体退行性变途径和间隙连接介导的信号传导(GJS)有关,参与维持3D支架中ob样细胞生长和退行性变的组织稳态。这项体外研究的结果表明,ob样HMS0014细胞积极调节牙种植体周围ECM的周转,以重现类骨组织工程材料的发育和形成,这可能有助于增强牙种植体周围的骨整合。
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引用次数: 2
Morphology of the lingual papillae in the brush-tailed rat kangaroo. 刷尾鼠袋鼠舌乳头形态。
Pub Date : 2014-01-01 DOI: 10.2535/ofaj.90.85
Shoichi Emura, Toshihiko Okumura, Huayue Chen

We examined the dorsal lingual surface of an adult brush-tailed rat kangaroo (Bettongia penicillata) by scanning electron microscopy. The filiform and fungiform papillae on the lingual apex and body consisted of a main papilla and secondary papillae. The connective tissue core of the filiform papillae on the lingual apex was cylindrical in shape with a crushed top. The connective tissue core of the filiform papillae on the lingual body had one large and several small processes. The fungiform papillae were round in shape. The connective tissue core of the fungiform papillae had several depressions on its top. The surface of the vallate papillae was rough and the papillae were surrounded by a groove and a pad. Several long conical papillae derived from the posterolateral margin of the tongue where foliate papillae have been shown to be distributed in many other animal species. The long conical papillae were very similar to those of the koala and opossum.

我们用扫描电子显微镜检查了一只成年刷尾大鼠袋鼠(Bettongia penicillata)的舌背表面。舌尖和舌体上的丝状和真菌状乳头由主乳头和次乳头组成。舌尖丝状乳头的结缔组织核呈圆柱状,顶部压碎。舌体上丝状乳头的结缔组织核有一个大突和几个小突。菌状乳头呈圆形。真菌状乳头的结缔组织核心顶部有几个凹陷。凹状乳头表面粗糙,乳头周围有凹槽和衬垫。几个长锥形乳头起源于舌的后外侧边缘,其中叶状乳头已被证明分布在许多其他动物物种中。长而圆锥形的乳突与考拉和负鼠非常相似。
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引用次数: 5
GATA-dependent regulation of TPO-induced c-mpl gene expression during megakaryopoiesis. 巨核形成过程中tpo诱导的c-mpl基因表达的gata依赖性调控。
Pub Date : 2014-01-01 DOI: 10.2535/ofaj.90.101
Masataka Sunohara, Shigeru Morikawa, Akira Fuse, Iwao Sato

Thrombopoietin (TPO) and its receptor, c-Mpl, play the crucial role during megakaryocytopoiesis. Previously, we have shown that the promoter activity of c-mpl induced by TPO is modulated by transcription through a PKC-dependent pathway and that GATA(-77) is involved as a positive regulatory element in TPO-induced c-mpl gene expression in the megakaryoblastic CMK cells. In this research, to examine participating possibility of GATA promoter element in TPO- induced c-mpl gene expression through a PKC-independent pathway, the promoter activity of site-directed mutagenesis and the effect of potein kinase C modulator were measured by a transient transfection assay system. Together with our previous results on the TPO-induced c-mpl promoter, this study indicates destruction of -77GATA in c-mpl promoter decreased the activity by 47.3% under existence of GF109203. These results suggest that GATA promoter element plays significant role in TPO-induced c-mpl gene expression through a PKC-independent pathway.

血小板生成素(TPO)及其受体c-Mpl在巨核细胞生成过程中起着至关重要的作用。在此之前,我们已经证明TPO诱导的c-mpl启动子活性是通过pkc依赖途径通过转录调节的,并且GATA(-77)作为正调控元件参与了巨核母细胞CMK中TPO诱导的c-mpl基因表达。为了研究GATA启动子元件通过pkc非依赖性途径参与TPO诱导的C- mpl基因表达的可能性,本研究采用瞬时转染实验系统检测了位点定向诱变启动子活性和蛋白激酶C调节剂的作用。结合我们之前对tpo诱导的c-mpl启动子的研究结果,本研究表明,在GF109203存在的情况下,c-mpl启动子中-77GATA的破坏使其活性降低了47.3%。这些结果表明GATA启动子元件通过pkc非依赖性途径在tpo诱导的c-mpl基因表达中发挥重要作用。
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引用次数: 2
Human fetal topographical anatomy of the femoral triangle in relation with change in the hip joint position. 人胎股骨三角的地形解剖与髋关节位置变化的关系。
Pub Date : 2014-01-01 DOI: 10.2535/ofaj.91.5
Shin-Ichi Abe, Akimitsu Ishizawa, Hisayo Nasu, Tadashi Nakao, Takashi Umezawa, Hiroshi Abe, Jose Francisco Rodriguez-Vazquez, Gen Murakami

Fetal hip joint is characterized by its highly flexion and lateral rotation although adult anatomy of the femoral nerve and iliofemoral ligament suggested the medial rotation. To investigate topographical anatomy of the femoral nerve, artery and vein in the femoral triangle, we histologically examined 11 fetuses (15-37 weeks). The nerve-vessel topographical relation was basically similar to that in adults, but the fan-like nerve division was seen in the horizontal plane in the smaller specimens in contrast to that included in the sagittal plane in the larger specimens. The medial or internal rotation of the nerve division seemed to occur in late stage fetuses, at birth and at infancy. Blood supply to the head of the femur might be also accelerated by changes in the hip joint position.

胎儿髋关节的特点是高度屈曲和外侧旋转,而成人股骨神经和髂股韧带的解剖显示其内侧旋转。为了研究股神经、股三角动脉和静脉的地形解剖,我们对11例胎儿(15-37周)进行了组织学检查。神经血管的地形关系与成人基本相似,但较小标本在水平面上可见扇形神经分裂,而较大标本在矢状面可见扇形神经分裂。内侧或内部旋转的神经分裂似乎发生在晚期胎儿,在出生和婴儿期。股骨头部的血液供应也可能因髋关节位置的改变而加速。
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引用次数: 2
Crucial factor causing collapse and aggregation of cultured cells in epon resin. epon树脂中培养细胞塌缩聚集的关键因素。
Pub Date : 2014-01-01 DOI: 10.2535/ofaj.91.1
Prasarn Tangkawattana, Mamoru Yamaguchi, Wuthichai Klomkleaw, Hua Niu, Jun A Minaguchi, Kazushige Takehana

Ultrastructural artifacts regarding collapse and aggregation of cultured cells have been problematic, especially when investigated apoptotic cells. The infiltration process during sample preparation is considered to be the most crucial factor for this problem. This study was conducted using two culture systems: a suspension culture system of human T-lymphocyte Jurkat cells and rabbit mature dendritic cells and a monolayer culture system of human lung macrophages, human breast cancer cells (A-546 cells) and cat bone-invasive gingival cancer cells (sccf3 cells). Fixation was conducted prior to removing or detaching the cells from the culture dishes. Initial infiltration with a 1 : 3 volume ratio of epon resin : propylene oxide was found to be the most crucial step among these cultured cells. The improved epon-resin infiltration method could eliminate the artifacts. Thus, differentiation between artifactual images and true images is highly possible.

关于培养细胞的崩溃和聚集的超微结构伪影一直是有问题的,特别是在研究凋亡细胞时。样品制备过程中的渗透过程被认为是造成这一问题的最关键因素。本研究采用两种培养体系:人t淋巴细胞Jurkat细胞和兔成熟树突状细胞的悬浮培养体系和人肺巨噬细胞、人乳腺癌细胞(a -546细胞)和猫骨侵袭性牙龈癌细胞(sccf3细胞)的单层培养体系。将细胞从培养皿中取出或分离之前进行固定。epon树脂:环氧丙烷体积比为1:3的初始浸润是培养细胞中最关键的一步。改进的epon-resin浸渍法可以消除伪影。因此,区分人工图像和真实图像是非常可能的。
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引用次数: 0
Morphology of the lingual papillae of Egyptian buffalo (Bubalus bubalis). 埃及水牛舌乳头的形态。
Pub Date : 2014-01-01 DOI: 10.2535/ofaj.91.13
Shoichi Emura, Neveen E R El Bakary

The mammalian tongue has different functions for feeding. We investigated the morphological characteristics of the surface of the tongue of Egyptian buffalo (Bubalus bubalis) using scanning electron microscopy. The lingual surface exhibited different degrees of specialization. The tongue can be divided into the apex, body, and root. The lingual prominence was observed on the posterior area of the lingual body. The lenticular, conical, and vallate papillae were observed on the caudal part of the tongue. The filiform papillae were lingual papillae covering the entire dorsal and ventral surface of the tongue at the lingual apex; they consisted of a main process that functions to transport food materials towards the pharynx. The fungiform papillae were dispersed between the filiform papillae. Large lenticular papillae were found at the lingual prominence. The highly keratinized lenticular papillae aid the physical mastication of plant materials. Twelve vallate papillae were arranged in V shape on the lingual root. These anatomical characteristics of the lingual surface of Egyptian buffalo may enhance understanding of its feeding behavior adaptations.

哺乳动物的舌头有不同的进食功能。利用扫描电子显微镜对埃及水牛舌面形态特征进行了研究。舌面呈现不同程度的特化。舌可分为舌尖、舌体和舌根。舌突位于舌体后部。舌尾部可见透镜状、圆锥状和凹状乳头。丝状乳头为舌乳头,覆盖在舌尖的整个舌背面和腹面;它们由一个主要的过程组成,它的功能是将食物物质运送到咽部。真菌状乳头分散在丝状乳头之间。舌突处可见大透镜状乳头。高度角化的透镜状乳头有助于植物材料的物理咀嚼。12个凹状乳头呈V形排列在舌根上。埃及水牛舌表面的这些解剖特征可以增强对其进食行为适应的理解。
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引用次数: 12
Variation of the subscapularis tendon at the fetal glenohumeral joint. 胎儿肩关节肩胛下肌腱的变异。
Pub Date : 2014-01-01 DOI: 10.2535/ofaj.90.89
Shin-Ichi Abe, Mitsuhiro Aoki, Tadashi Nakao, Masaaki Kasahara, Jose Francisco Rodriguez-Vazquez, Gen Murakami, Baik Hwan Cho

We examined the topohistology of the subscapularis tendon at the glenohumeral joint in 10 mid-term (15-16 weeks of gestation) and 10 late-stage (27-32 weeks) human fetuses. At both stages, there were two patterns of terminal course of the subscapularis tendon: 1) the tendon was tightly attached to the medial part of the joint capsule and extended anterosuperiorly along the capsule to the lesser tubercle (7/10 mid-term fetuses; 5/10 late-stage fetuses); 2) the tendon passed superiorly through the joint cavity for a long distance in combination with the subcoracoid bursa opening widely to the joint cavity (3/10 mid-term fetuses; 5/10 late-stage fetuses). The lower glenoid labrum tended to be well developed in the former pattern because the subscapularis tendon did not interfere with the superior extension of the labrum. With only one exception (late stage), the capsule-attaching tendon was seen in fetuses in which the coracoid process was located on the superior side of the lesser tubercle, whereas the intra-articular tendon accompanied the coracoid process at the same supero-inferior level of the tubercle. Thus, the topographical relationship between the coracoid process and lesser tubercle in fetuses seemed to determine the courses of the subscapularis tendon at the glenohumeral joint. The present variation in the subscapularis tendon was likely connected with the adult morphologies of the middle and inferior glenohumeral ligaments or folds, whose variations are well known.

我们检查了10例中期(妊娠15-16周)和10例晚期(27-32周)人类胎儿肩胛下肌腱的拓扑组织学。在这两个阶段,肩胛下肌肌腱终末走向有两种模式:1)肌腱紧密附着于关节囊内侧,并沿关节囊向前向上延伸至小结节(7/10中期胎儿;5/10晚期胎儿);2)肌腱与喙下滑囊结合,较远地穿过关节腔(3/10中期胎儿;5/10晚期胎儿)。在前一种模式下,下盂唇往往发育良好,因为肩胛下肌肌腱不干扰上盂唇的伸展。只有一个例外(晚期),在胎儿中可以看到荚膜附着肌腱,其中喙突位于小结节的上侧,而关节内肌腱在结节的同一上下水平处伴随喙突。因此,胎儿喙突和小结节之间的地形关系似乎决定了肩胛下肌腱在肩关节处的运动轨迹。目前肩胛下肌腱的变化可能与成年后中、下盂肱韧带或褶皱的形态有关,其变化是众所周知的。
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引用次数: 3
Lectin-binding sites in epithelial cells of the mouse prostate gland. 小鼠前列腺上皮细胞中的凝集素结合位点。
Pub Date : 2014-01-01 DOI: 10.2535/ofaj.91.91
Kentaro Sakuda, Ayaka Yoshida, Ryoki Muragishi, Kazuya Yoshinaga

The prostate is an exocrine gland in the male reproductive tract that secretes seminal fluids. To gain insight into the cytochemical properties of prostatic epithelial cells, the characteristics of glycoconjugates in mouse prostate sections were examined by lectin histochemistry and immunohistochemistry. Characteristic staining patterns were observed, depending on the type of lectins present in the epithelia. Luminal cells reacted specifically with mannose-binding lectins (Galanthus nivalis lectin, Hippeastrum hybrid lectin, Narcissus pseudonarcissus lectin) and Maclura pomifera lectin in all lobes of the prostate. Luminal cells also expressed galactose, N-acetyl-D-galactosamine (GalNAc), N-acetyl-D-glucosamine (GlcNAc), and fucose residues in the lateral and ventral lobes. Basal cells expressed GlcNAc and fucose, and reacted with Datura stramonium lectin and Aleuria aurantia lectin in all lobes. These results indicate that in the mouse prostate, the selectivity of lectin-binding sites for distinct cell types and lobe-dependent staining may relate to cellular and regional differences in function. Furthermore, some lectins selectively bound to prostatic epithelial cells, indicating their potential use as markers for the histopathological evaluation of prostatic diseases, cancer diagnosis, or male infertility.

前列腺是男性生殖道内分泌精液的外分泌腺。为了进一步了解前列腺上皮细胞的细胞化学特性,我们采用凝集素组织化学和免疫组织化学方法检测了小鼠前列腺切片中糖缀合物的特征。根据上皮中存在的凝集素的类型,观察到特征性的染色模式。管腔细胞与甘露糖结合凝集素(Galanthus nivalis凝集素,Hippeastrum杂交凝集素,narissus pseudonarcissus凝集素)和Maclura pomifera凝集素在前列腺的所有叶中特异性反应。管腔细胞也表达半乳糖、n -乙酰- d -半乳糖胺(GalNAc)、n -乙酰- d -葡萄糖胺(GlcNAc),并在侧叶和腹叶中有聚焦残基。基底细胞表达GlcNAc和focal,并与曼陀罗凝集素和金百合凝集素发生反应。这些结果表明,在小鼠前列腺中,凝集素结合位点对不同细胞类型和叶依赖性染色的选择性可能与细胞和区域功能差异有关。此外,一些凝集素选择性地与前列腺上皮细胞结合,表明它们可能用作前列腺疾病、癌症诊断或男性不育的组织病理学评估标记物。
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引用次数: 1
Groped for a novel stimulation method for the prevention of lumbar vertebral compression pressure bone fracture and verification using a bone density drop model mouse. 探索一种预防腰椎压缩性骨折的新型刺激方法,并用骨密度下降模型小鼠进行验证。
Pub Date : 2014-01-01 DOI: 10.2535/ofaj.91.29
Kouji Yamada, Kazuhiro Nishii, Kazuyoshi Sakai, Toshio Teranishi, Mamoru Matsubara

Osteoporosis is leaving bones more fragile and susceptible to fracture. It has a massive impact, both physically and mentally, markedly diminishing quality of life. A new form of therapeutic exercise or physical therapy that mitigates the abrupt decrease in bone density in postmenopausal women must quickly be developed to avoid those problems. In this study, ovariectomy (OVX) mice were used as models to simulate the decrease in bone density observed in postmenopausal women. Physical therapy via a shaking stimulus, in the form of moving a platform that rotates in a roughly circular motion in the horizontal plane, was studied as a way to prevent the decrease in bone density of the lumbar vertebrae by analysis of bone histomorphometry, a feat that the stimulus from conventional therapeutic exercise and physical therapy have failed to achieve. Comparison of the stimulus/ovariectomized (+/+) group with the -/+ group indicated significant increases in ES (P < 0.01), N. Mu. Oc (P < 0.05), OV (P < 0.05), O. Th (P < 0.01), and L. Th (P < 0.01) in the +/+ group. If this finding is used clinically, we believe that it could lead to therapy that would prevent compression fractures of the lumbar vertebrae.

骨质疏松症使骨骼更加脆弱,容易骨折。它对身体和精神都有巨大的影响,显著降低了生活质量。必须尽快开发一种新的治疗性运动或物理疗法,以减轻绝经后妇女骨密度的突然下降,以避免这些问题。本研究以卵巢切除(OVX)小鼠为模型,模拟绝经后妇女骨密度的下降。通过移动在水平面上以大致圆周运动旋转的平台的形式进行震动刺激的物理治疗,研究了通过骨组织形态学分析来防止腰椎骨密度降低的方法,这是传统治疗运动和物理治疗的刺激未能实现的壮举。刺激/去卵巢(+/+)组与-/+组比较,ES显著升高(P < 0.01);+/+组Oc (P < 0.05)、OV (P < 0.05)、O. Th (P < 0.01)、L. Th (P < 0.01)。如果这一发现在临床上使用,我们相信它可能会导致治疗,以防止腰椎压缩性骨折。
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引用次数: 3
Secretion-related structures of hypothalamo-hypophysial terminals in the rat posterior pituitary. 大鼠垂体后叶下丘脑-垂体末梢的分泌相关结构。
Pub Date : 2013-01-01 DOI: 10.2535/ofaj.90.69
Yoshiki Takeuchi, Jun-Qian Liu, Yoshiki Matsumoto, Takanori Miki, Ken-ichi Ohta, Katsuhiko Warita, Shingo Suzuki, Motoki Tamai

Hypothalamic terminals were investigated in the rat posterior pituitary (PP). Injection of wheat germ agglutinin conjugated horseradish peroxidase (WGA-HRP) and co-injection of WGA-HRP with Rab3A-siRNA were made into the hypothalamus, respectively. Additional injection of WGA-HRP was made into the hypothalamus in the animals exposed to ethanol. These injections resulted in heavy labeling of fibers exclusively confined to the PP. Ultrastructural observations showed terminals, fibers, pituicytes, capillaries and vascular spaces in the PP. Although the majority of terminals were observed to contain large dense core vesicles (LDCVs) and HRP-reaction products (HRP-RPs), exocytosis of LDCVs in close proximity to cell membrane was not found. Interestingly, a few terminals showed alteration of cell membrane called "apocrine-like structure" containing LDCV and RP. The narrow neck portion of the structure gave the appearance that it may have been in some stage of separating from terminals. Other remarkable feature was that terminals occasionally reveal the structure of "leakage" of RP discharged into vascular spaces crossing cell membrane. Such hormone-releasing mechanism might be involved in one of "diacrine-like secretion". In the present study secretion-related structures of hypothalamic terminals in the PP are quite different from normal vesicular exocytosis.

对大鼠垂体后叶(PP)下丘脑末梢进行了研究。下丘脑分别注射小麦胚芽凝集素偶联的辣根过氧化物酶(WGA-HRP)和与Rab3A-siRNA共注射WGA-HRP。在暴露于乙醇的动物的下丘脑中额外注射WGA-HRP。超微结构观察显示了PP的末端、纤维、垂体细胞、毛细血管和血管间隙。尽管大多数末端被观察到含有大而致密的核心囊泡(ldcv)和酶反应产物(hrp - rp),但在靠近细胞膜的地方没有发现ldcv的胞外分泌。有趣的是,一些末端显示含有LDCV和RP的细胞膜被称为“apocine样结构”的改变。结构的窄颈部分使其看起来可能处于与末端分离的某个阶段。另一个显著特征是末端偶尔显示RP穿过细胞膜进入血管间隙的“渗漏”结构。这种激素释放机制可能与一种“尿样分泌”有关。在本研究中,下丘脑末梢的分泌相关结构与正常的水疱性胞吐有很大的不同。
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引用次数: 0
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Okajimas folia anatomica Japonica
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