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The Effects of Ischemia and Hyperoxygenation on Hair Growth and Cycle. 缺血和高氧对头发生长和周期的影响。
IF 2.3 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2020-07-02 Epub Date: 2020-07-30 DOI: 10.1080/15476278.2020.1794271
Harunosuke Kato, Kahori Kinoshita, Natsumi Saito, Koji Kanayama, Masanori Mori, Natsumi Asahi, Ataru Sunaga, Katsutoshi Yoshizato, Satoshi Itami, Kotaro Yoshimura

Alopecia has several causes, but its relationship with ischemia/hypoxia has not yet been investigated in detail. In this study, we studied the changes of hair follicles induced by ischemia and potential effects of normobaric hyperoxygenation (NBO) on the hair cycle and growth. We found that skin ischemia reduced hair growth rate, hair shaft size, and its pigmentation in the anagen phase of mice, which may reflect an aspect of pathophysiology of hair loss (alopecia) and depigmentation (gray/white hairs). Hyperoxygenation increased hair growth rate in organ culture of both human and murine hair follicles. Systemic NBO promoted hair growth in early anagen and mid-anagen, and delayed catagen onset in mice. However, telogen-to-anagen transition was not affected by NBO as far as non-ischemic skin is concerned. The results of this study indicated that the hair follicle is very sensitive to oxygen tension and oxygen tension affects the regulation of hair growth and cycle in vitro and in vivo. It was suggested that systemic NBO can be safely applied for a long period and can be a noninvasive therapeutic approach to alter hair growth and cycle by manipulating the microenvironment of hair follicles.

脱发有多种原因,但其与缺血/缺氧的关系尚未详细研究。在本研究中,我们研究了缺血诱导毛囊的变化以及正压高氧(NBO)对头发周期和生长的潜在影响。我们发现,皮肤缺血降低了毛发生长速率、毛干大小及其在毛发生长初期的色素沉着,这可能反映了脱发(脱发)和色素沉着(白发)的病理生理方面。在人体和小鼠毛囊器官培养中,高氧可提高毛发生长速率。全身NBO可促进小鼠毛发生长早期和中期的生长,并可延缓毛发生长的发生。然而,就非缺血性皮肤而言,NBO不影响休止期到生长期的过渡。本研究结果表明,毛囊对氧张力非常敏感,氧张力在体外和体内影响毛发生长和周期的调节。结果表明,系统NBO可以长期安全应用,并且可以通过控制毛囊微环境来改变头发生长和周期,是一种无创治疗方法。
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引用次数: 8
Surgical Models to Explore Acellular Liver Scaffold Transplantation: Step-by-Step. 探索脱细胞肝支架移植的外科模型:一步一步。
IF 2.3 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2020-07-02 Epub Date: 2020-08-15 DOI: 10.1080/15476278.2020.1801273
Marlon L Dias, Cíntia M P Batista, Victor J K Secomandi, Alexandre C Silva, Victoria R S Monteiro, Lanuza A Faccioli, Regina C S Goldenberg

Acellular liver scaffolds (ALS) have arisen as potential candidates for transplantation. Until now, all reports involving ALS transplantation failed in surgical method descriptions and do not offer support to scientists to reproduce the procedures used in experimental microsurgery to make the results comparable to literature. To overcome the lack of detail information, we described surgical steps details to perform heterotopic and partial orthotopic surgical models to promote ALS transplantation. After preservation and vessel cannulation steps, the liver grafts were decellularized. In addition, ex vivo blood perfusion tests were performed to obtain a successful anticoagulation treatment prior in vivo transplantation. Then, methods of partial liver resection, combination of hand-suture and cuff techniques to complete end-to-end anastomosis between the scaffold and the recipient animal were performed. These procedures which take 30-60 min and were efficient to allow acellular liver scaffold viability and recellularization of different types of cell post-surgery. In conclusion, our methods are practical and simple promising approach that provides the opportunity to investigate ways to achieve sufficient liver function post-transplantation in vivo.

无细胞肝支架(ALS)已成为移植的潜在候选者。到目前为止,所有涉及ALS移植的报道都没有对手术方法进行描述,也没有为科学家重现实验显微外科手术过程以使结果与文献相媲美提供支持。为了克服细节信息的缺乏,我们详细描述了实施异位和部分原位手术模型以促进ALS移植的手术步骤。在保存和血管插管步骤后,肝移植物被脱细胞。此外,还进行了体外血液灌注试验,以在体内移植前获得成功的抗凝治疗。然后,采用部分肝切除、手缝合结合袖带技术完成支架与受体动物的端到端吻合。这些过程需要30-60分钟,并且有效地允许无细胞肝支架的生存和术后不同类型细胞的再细胞化。总之,我们的方法是实用和简单的有前途的方法,为研究如何在体内移植后获得足够的肝功能提供了机会。
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引用次数: 3
Extracellular Matrix Remodeling During Palate Development. 腭发育过程中的细胞外基质重塑。
IF 2.3 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2020-04-02 Epub Date: 2020-03-31 DOI: 10.1080/15476278.2020.1735239
Xia Wang, Chunman Li, Zeyao Zhu, Li Yuan, Wood Yee Chan, Ou Sha

The morphogenesis of the mammalian secondary plate is a series of highly dynamic developmental process, including the palate shelves vertical outgrowth, elevation to the horizontal plane and complete fusion in the midline. Extracellular matrix (ECM) proteins not only form the basic infrastructure for palatal mesenchymal cells to adhere via integrins but also interact with cells to regulate their functions such as proliferation and differentiation. ECM remodeling is essential for palatal outgrowth, expansion, elevation, and fusion. Multiple signaling pathways important for palatogenesis such as FGF, TGF β, BMP, and SHH remodels ECM dynamics. Dysregulation of ECM such as HA synthesis or ECM breakdown enzymes MMPs or ADAMTS causes cleft palate in mouse models. A better understanding of ECM remodeling will contribute to revealing the pathogenesis of cleft palate.

哺乳动物次级板的形态发生是一系列高度动态的发育过程,包括腭架垂直生长、向水平面上升和中线完全融合。细胞外基质(Extracellular matrix, ECM)蛋白不仅是腭间充质细胞通过整合素粘附的基础,而且还与细胞相互作用,调节细胞的增殖和分化等功能。ECM重塑对腭生长、扩张、抬高和融合至关重要。FGF、TGF β、BMP和SHH等多种对腭发育重要的信号通路可重塑ECM动力学。在小鼠模型中,如HA合成或ECM分解酶MMPs或ADAMTS等ECM失调导致腭裂。更好地了解ECM重塑有助于揭示腭裂的发病机制。
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引用次数: 8
Development of Bioengineered Organ Using Biological Acellular Rat Liver Scaffold and Hepatocytes. 利用生物脱细胞大鼠肝支架和肝细胞制备生物工程器官。
IF 2.3 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2020-04-02 Epub Date: 2020-05-03 DOI: 10.1080/15476278.2020.1742534
Tanya Debnath, Chandra Shekar Mallarpu, Lakshmi Kiran Chelluri

The increasing demand for organs for transplantation necessitates the development of substitutes to meet the structural and physiological functions. Tissue decellularization and recellularization aids in retaining the three-dimensional integrity, biochemical composition, tissue ultra-structure, and mechanical behavior, which makes them functionally suitable for organ transplantation. Herein, we attempted to rebuild functional liver grafts in small animal model (Wistar rat) with a potential of translation. A soft approach was adopted using 0.1% SDS (Sodium Dodecyl Sulfate) for decellularization and primary hepatocytes were used as a potential cell source for recellularization. The decellularization process was evaluated and confirmed using histology, DNA content, ultra-structure analysis. The resultant scaffold was re-seeded with the rat hepatocytes and their biocompatibility was assessed by its metabolic functions and gene expression. The structural components of the Extracellular matrix (ECM) (Laminins, Collagen type I, Reticulins) were conserved and the liver cell-specific proteins like CK-18, alpha-fetoprotein, albumin were expressed in the recellularized scaffold. The functionality and metabolic activity of the repopulated scaffold were evident from the albumin and urea production. Expression of Cytokeratin-19 (CK-19), Glucose 6-Phosphatase (G6P), Albumin, Gamma Glutamyl Transferase (GGT) genes has distinctly confirmed the translational signals after the repopulation process. Our study clearly elucidates that the native extracellular matrix of rat liver can be utilized as a scaffold for effective recellularization for whole organ regeneration.

随着对移植器官需求的不断增长,需要开发能够满足器官结构和生理功能的替代品。组织脱细胞和再细胞化有助于保持三维完整性、生化组成、组织超结构和力学行为,使其在功能上适合器官移植。在此,我们尝试在具有翻译潜力的小动物模型(Wistar大鼠)中重建功能性肝移植物。采用0.1%十二烷基硫酸钠(SDS)软法进行脱细胞,原代肝细胞作为潜在的细胞源进行再细胞化。用组织学、DNA含量、超微结构分析对脱细胞过程进行评价和确认。用大鼠肝细胞重新植入支架,并通过其代谢功能和基因表达评估其生物相容性。细胞外基质(ECM)的结构成分(层粘连蛋白、I型胶原蛋白、网状蛋白)被保留,肝细胞特异性蛋白如CK-18、甲胎蛋白、白蛋白在再细胞化支架中表达。从白蛋白和尿素的产生可以看出重建支架的功能和代谢活性。细胞角蛋白-19 (CK-19)、葡萄糖6-磷酸酶(G6P)、白蛋白、γ谷氨酰转移酶(GGT)基因的表达清楚地证实了繁殖过程后的翻译信号。我们的研究清楚地表明,天然的大鼠肝脏细胞外基质可以作为一种支架,用于整个器官再生的有效再细胞化。
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引用次数: 14
Development of Small-diameter Polyester Vascular Grafts Coated with Silk Fibroin Sponge 丝素海绵包覆小直径聚酯血管移植物的研制
IF 2.3 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2020-01-02 DOI: 10.1080/15476278.2019.1686295
Takashi Tanaka, R. Tanaka, Yoko Ogawa, Yoshihide Takagi, T. Asakura
ABSTRACT In recent years, the demand for functional small-diameter (< 6 mm) artificial vascular grafts has greatly increased due to an increase in the number of patients with vascular heart disease. However, currently, there are no available commercial small-diameter grafts. The objective of this research was to develop a porous silk fibroin (SF)-coated poly(ethylene terephthalate) (PET) graft with a diameter < 6 mm. The graft was compared with a gelatin-coated PET graft because the latter PET graft with a diameter ~ 6 mm was widely used as a commercial vascular graft. Initially, porous SF was prepared using Glyc as the porogen [termed SF(Glyc)] and the PET grafts were prepared through the double-Raschel knitting method. Subsequently, the degradation of the SF coating was monitored using protease XIV in vitro and was compared with that observed in gelatin-coated PET grafts. Finally, these grafts were also implanted into rats for an in vivo comparison. In degradation experiments, after 7 days, the SF was clearly digested by protease XIV, but the gelatin on the graft was still remained at the outer surface. In implantation experiments in rats, the SF(Glyc)-coated PET graft was rapidly degraded in vivo and remodeling to self-tissues was promoted compared with the gelatin-coated PET graft. Thrombus formation and intimal hyperplasia were observed in the gelatin-coated PET graft; however, such side reactions were not observed in the SF(Glyc)-coated PET graft. Thus, the porous SF(Glyc)-coated PET graft with a small diameter < 6 mm may be useful as a commercial vascular graft.
摘要近年来,由于血管性心脏病患者数量的增加,对功能性小直径(<6mm)人工血管移植物的需求大幅增加。然而,目前还没有可用的商业小直径移植物。本研究的目的是开发直径<6mm的多孔丝素蛋白(SF)涂层聚对苯二甲酸乙二醇酯(PET)移植物。将该移植物与明胶涂层PET移植物进行比较,因为后者直径~6mm的PET移植物被广泛用作商业血管移植物。最初,使用甘氨酸作为成孔剂[称为SF(甘氨酸)]制备多孔SF,并通过双拉舍尔编织法制备PET移植物。随后,使用蛋白酶XIV在体外监测SF涂层的降解,并将其与明胶涂层的PET移植物中观察到的降解进行比较。最后,这些移植物也被植入大鼠体内进行体内比较。在降解实验中,7天后,SF被蛋白酶XIV清楚地消化,但移植物上的明胶仍然保留在外表面。在大鼠植入实验中,与明胶涂层的PET移植物相比,SF(Gly)涂层的PET移植植物在体内快速降解,并促进了自身组织的重塑。在明胶包被的PET移植物中观察到血栓形成和内膜增生;然而,在SF(甘氨酸)包被的PET移植物中没有观察到这样的副反应。因此,小直径<6mm的多孔SF(甘氨酸)涂层PET移植物可能可用作商业血管移植物。
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引用次数: 11
Evaluation of Radiosterilized Glyercerolated Amniotic Membranes as a Substrate for Cultured Human Epithelial Cells. 放射性灭菌甘油三酯羊膜作为培养人上皮细胞底物的评价。
IF 2.3 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2020-01-01 Epub Date: 2020-02-15 DOI: 10.1080/15476278.2020.1723366
André O Paggiaro, Monica B Mathor, Walcy R Teodoro, Cesár Isaac, Vera L Capelozzi, Rolf Gemperli

Human amniotic membrane (HAM) is a biomaterial with biological properties beneficial to tissue repair, serving as a substrate for cell cultivation. Irradiation is used for tissue sterilization, but can damage the HAM structure. The objective of this paper was to construct a skin substitute, composed of human keratinocytes cultured on glycerolated HAMs, and to evaluate the influence radiation on subsequent cell culture growth. Four batches of HAMs were glycerolated, and half of them were radio-sterilzed with 25 kGy. Non-irradiated glycerolated HAM (ni-HAM) and irradiated glycerolated HAM (i-HAM) samples were then de-epithelized and analyzed using optical microscopy (Picrossirius staining), immunofluorescence and electron microscopy. Subsequently, keratinocytes were cultured on ni- and i-HAMs, and either immersed or positioned at the air-liquid interface. The basement membranes of the ni-HAM group remained intact following de-epithelialization, whereas the i-HAM group displayed no evidence or remnant presence of these membranes. Concerning the keratinocyte cultures, the ni-HAM substrate promoted the growth of multi-layered and differentiated epithelia. Keratinocytes cultured on i-HAM formed epithelium composed of three layers of stratification and discrete cell differentiation. The glycerolated HAM was compatible with cultured epithelia, demonstrating its potential as a skin substitute. Irradiation at 25 kGy caused structural damage to the amnion.

人羊膜是一种具有有利于组织修复的生物学特性的生物材料,是细胞培养的底物。辐照用于组织灭菌,但会破坏火腿结构。本文的目的是构建一种由人角质形成细胞组成的皮肤替代物,并评估辐射对随后细胞培养生长的影响。将4批火腿甘油化,其中一半用25 kGy进行放射性灭菌。然后将未辐照的甘油化火腿(ni-HAM)和辐照的甘油化火腿(i-HAM)样品去上皮并使用光学显微镜(picrossius染色)、免疫荧光和电子显微镜进行分析。随后,在ni-和i- ham上培养角质形成细胞,并将其浸入或置于气液界面。ni-HAM组的基底膜在去上皮化后保持完整,而i-HAM组则没有这些膜的证据或残余存在。在角质细胞培养中,ni-HAM底物促进了多层分化上皮的生长。在i-HAM上培养的角化细胞形成由三层分层和离散细胞分化组成的上皮。甘油化的HAM与培养的上皮相容,显示其作为皮肤替代品的潜力。25kgy辐照对羊膜造成结构性损伤。
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引用次数: 4
Influence of Different Cell Types and Sources on Pre-Vascularisation in Fibrin and Agarose–Collagen Gels 不同细胞类型和来源对纤维蛋白和琼脂糖-胶原凝胶血管形成前的影响
IF 2.3 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2019-12-06 DOI: 10.1080/15476278.2019.1697597
Caroline Kniebs, F. Kreimendahl, M. Köpf, H. Fischer, S. Jockenhoevel, A. Thiebes
ABSTRACT Vascularisation is essential for the development of tailored, tissue-engineered organs and tissues due to diffusion limits of nutrients and the lack of the necessary connection to the cardiovascular system. To pre-vascularize, endothelial cells and supporting cells can be embedded in the scaffold to foster an adequate nutrient and oxygen supply after transplantation. This technique is applied for tissue engineering of various tissues, but there have been few studies on the use of different cell types or cells sources. We compare the effect of supporting cells from different sources on vascularisation. Fibrin gels and agarose-collagen hydrogels were used as scaffolds. The supporting cells were primary human dermal fibroblasts (HDFs), human nasal fibroblasts (HNFs), human mesenchymal stem cells from umbilical cord’s Wharton’s jelly (WJ MSCs), adipose-derived MSCs (AD MSCs) and femoral bone marrow-derived MSCs (BM MSCs). The tissue constructs were incubated for 14 days and analyzed by two-photon laser scanning microscopy. Vascularisation was supported by all cell types, forming branched networks of tubular vascular structures in both hydrogels. In general, fibrin gels present a higher angiogenic promoting environment compared to agarose-collagen hydrogels and fibroblasts show a high angiogenic potential in co-culture with endothelial cells. In agarose-collagen hydrogels, vascular structures supported by AD MSCs were comparable to our HDF control in terms of volume, area and length. BM MSCs formed a homogeneous network of smaller structures in both hydrogels. This study provides data toward understanding the pre-vascularisation properties of different supporting cell types and sources for tissue engineering of different organs and tissues.
摘要由于营养物质的扩散限制以及缺乏与心血管系统的必要连接,血管化对于定制、组织工程化器官和组织的发展至关重要。为了预血管化,可以将内皮细胞和支持细胞嵌入支架中,以在移植后培养足够的营养和氧气供应。该技术适用于各种组织的组织工程,但很少有关于使用不同细胞类型或细胞来源的研究。我们比较了来自不同来源的支持细胞对血管形成的影响。纤维蛋白凝胶和琼脂糖-胶原水凝胶用作支架。支持细胞是原代人真皮成纤维细胞(HDFs)、人鼻腔成纤维细胞、脐带沃顿氏果冻中的人间充质干细胞(WJ MSCs)、脂肪来源的MSCs(AD MSCs)和股骨来源的MSC(BM MSCs)。将组织构建体孵育14天,并通过双光子激光扫描显微镜进行分析。血管形成受到所有细胞类型的支持,在两种水凝胶中形成管状血管结构的分支网络。通常,与琼脂糖-胶原水凝胶相比,纤维蛋白凝胶具有更高的血管生成促进环境,成纤维细胞在与内皮细胞共培养中显示出高的血管形成潜力。在琼脂糖胶原水凝胶中,AD MSC支持的血管结构在体积、面积和长度方面与我们的HDF对照组相当。BM-MSC在两种水凝胶中都形成了较小结构的均匀网络。这项研究为理解不同支持细胞类型和来源的血管形成前特性提供了数据,用于不同器官和组织的组织工程。
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引用次数: 16
Characterization and in vivo study of decellularized aortic scaffolds using closed sonication system 应用封闭超声系统对脱细胞主动脉支架的表征和体内研究
IF 2.3 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2019-09-07 DOI: 10.1080/15476278.2019.1656997
A. Hazwani, M. Sha'ban, A. Azhim
ABSTRACT Extracellular matrix (ECM) based bioscaffolds prepared by decellularization has increasingly emerged in tissue engineering application because it has structural, biochemical, and biomechanical cues that have dramatic effects upon cell behaviors. Therefore, we developed a closed sonication decellularization system to prepare ideal bioscaffolds with minimal adverse effects on the ECM. The decellularization was achieved at 170 kHz of ultrasound frequency in 0.1% and 2% Sodium Dodecyl Sulphate (SDS) solution for 10 hours. The immersion treatment as control was performed to compare the decellularization efficiency with our system. Cell removal and ECM structure were determined by histological staining and biochemical assay. Biomechanical properties were investigated by the indentation testing to test the stiffness, a residual force and compression of bioscaffolds. Additionally, in vivo implantation was performed in rat to investigate host tissue response. Compared to native tissues, histological staining and biochemical assay confirm the absence of cellularity with preservation of ECM structure. Moreover, sonication treatment has not affected the stiffness [N/mm] and a residual force [N] of the aortic scaffolds except for compression [%] which 2% SDS significantly decreased compared to native tissues showing higher SDS has a detrimental effect on ECM structure. Finally, minimal inflammatory response was observed after 1 and 5 weeks of implantation. This study reported that the novelty of our developed closed sonication system to prepare ideal bioscaffolds for tissue engineering applications.
摘要通过脱细胞制备的基于细胞外基质(ECM)的生物支架越来越多地出现在组织工程应用中,因为它具有对细胞行为有显著影响的结构、生化和生物力学线索。因此,我们开发了一种封闭的超声脱细胞系统,以制备对ECM的不良影响最小的理想生物支架。在超声频率为170kHz的0.1%和2%十二烷基硫酸钠(SDS)溶液中进行10小时的脱细胞。以浸渍处理作为对照,将脱细胞效率与我们的系统进行比较。通过组织学染色和生化分析测定细胞去除和ECM结构。通过压痕试验研究了生物支架的生物力学性能,以测试生物支架的刚度、残余力和压缩力。此外,在大鼠体内进行植入,以研究宿主组织的反应。与天然组织相比,组织学染色和生化测定证实了ECM结构的保留,没有细胞性。此外,超声处理没有影响主动脉支架的硬度[N/mm]和残余力[N],除了压缩[%],与表现出较高SDS的天然组织相比,2%SDS显著降低,这对ECM结构具有不利影响。最后,在植入1周和5周后观察到最小的炎症反应。本研究报告了我们开发的封闭超声处理系统的新颖性,该系统用于制备组织工程应用的理想生物支架。
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引用次数: 9
Advances of Wnt signalling pathway in dental development and potential clinical application Wnt信号通路在口腔发育中的研究进展及临床应用前景
IF 2.3 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2019-09-04 DOI: 10.1080/15476278.2019.1656996
Xi Lu, Jun Yang, Shouliang Zhao, Shangfeng Liu
ABSTRACT Wnt signalling pathway is widely studied in many processes of biological development, like embryogenesis, tissue homeostasis and wound repair. It is universally known that Wnt signalling pathway plays an important role in tooth development. Here, we summarized the function of Wnt signalling pathway during tooth initiation, crown morphogenesis, root formation, and discussed the therapeutic potential of Wnt modulators.
Wnt信号通路在胚胎发生、组织稳态和伤口修复等生物发育过程中被广泛研究。众所周知,Wnt信号通路在牙齿发育过程中起着重要作用。本文总结了Wnt信号通路在牙齿起始、牙冠形态发生、牙根形成过程中的作用,并讨论了Wnt调节剂的治疗潜力。
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引用次数: 11
Specification of Sprouty2 functions in osteogenesis in in vivo context Sprouty2在体内成骨中的功能规范
IF 2.3 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2019-09-04 DOI: 10.1080/15476278.2019.1656995
B. Veselá, E. Svandova, M. Hovořáková, R. Peterkova, A. Kratochvílová, Martina Pasovská, A. Ramešová, H. Lesot, E. Matalova
ABSTRACT Sprouty proteins are modulators of the MAPK/ERK pathway. Amongst these, Sprouty2 (SPRY2) has been investigated as a possible factor that takes part in the initial phases of osteogenesis. However, the in vivo context has not yet been investigated and the underlying mechanisms taking place in vitro remain unknown. Therefore, in this study, the impact of Spry2 deficiency was examined in the developing tibias of Spry2 deficient (-/-) mouse. The investigation was performed when the osteogenic zone became clearly visible and when all three basic bone cells types were present. The main markers of osteoblasts, osteocytes and osteoclasts were evaluated by immunohistochemistry and RT-PCR. RT-PCR showed that the expression of Sost was 3.5 times higher in Spry2-/- than in the wild-type bone, which pointed to a still unknown mechanism of action of SPRY2 on the differentiation of osteocytes. The up-regulation of Sost was independent of Hif-1α expression and could not be related to its positive regulator, Runx2, since none of these factors showed an increased expression in the bone of Spry2-/- mice. Regarding the RANK/RANKL/OPG pathway, the Spry2-/- showed an increased expression of Rank, but no significant change in the expression of Rankl and Opg. Thanks to these results, the impact of Spry2 deletion is shown for the first time in the developing bone as a complex organ including, particularly, an effect on osteoblasts (Runx2) and osteocytes (Sost). This might explain the previously reported decrease in bone formation in postnatal Spry2-/- mice.
发芽蛋白是MAPK/ERK通路的调节剂。其中,Sprouty2 (SPRY2)已被研究为参与成骨初始阶段的可能因素。然而,体内环境尚未被研究,体外发生的潜在机制仍然未知。因此,在本研究中,我们研究了Spry2缺失对Spry2缺失(-/-)小鼠胫骨发育的影响。当成骨带清晰可见并且所有三种基本骨细胞类型都存在时进行调查。采用免疫组化和RT-PCR检测成骨细胞、骨细胞和破骨细胞的主要标志物。RT-PCR结果显示,Spry2-/-中Sost的表达量是野生型骨的3.5倍,提示Spry2对骨细胞分化的作用机制尚不清楚。Sost的上调与Hif-1α的表达无关,与其阳性调节因子Runx2无关,因为这些因子在Spry2-/-小鼠骨中的表达均未增加。在RANK/RANKL/OPG通路中,Spry2-/-表达RANK增加,而RANKL和OPG表达无明显变化。由于这些结果,Spry2缺失的影响首次显示在发育中的骨作为一个复杂的器官,特别是对成骨细胞(Runx2)和骨细胞(Sost)的影响。这可能解释了先前报道的Spry2-/-小鼠出生后骨形成减少的原因。
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引用次数: 2
期刊
Organogenesis
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