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Adenovirus-mediated tissue inhibitor of metalloproteinase-3 gene transfection inhibits rabbit intervertebral

Xudong YU MM, Zengwu SHAO MD, Liming XIONG MD, Weiwei XU MM, Hezhong WANG MM, Huifa XU MM,

《医学前沿（英文）》 2009年第3卷第4期页码 415-420 doi: 10.1007/s11684-009-0072-z

摘要： The aim of this study was to investigate the inhibitory effects of recombinant adenovirus vector carrying tissue inhibitor of metalloproteinase-3 (RAdTIMP-3) against degeneration of rabbit intervertebral disc. Thirty Japanese white rabbits of 4 months old were randomly divided into 5 groups. Mild or moderate rabbit lumbar disc degeneration model was constructed with the controllable axial loading device by imposing 98N pressure at the discs for 2 weeks. Various doses of virus were injected into the degenerated discs as follows: 20μL of normal saline in group 1; 20μL of RAd66 (an empty adenovirus vector, 1.0×10OPU/mL) in group 2; and 20, 10, and 5μL of RAdTIMP-3 (1.0×10OPU/mL) in groups 3, 4, and 5, respectively. Two weeks after the injection, the discs were collected for investigations, including assessment of degeneration degrees according to the Thompson’s grading system, reverse-transcription polymerase chain reaction (RT-PCR) assay for TIMP-3 gene, Safranin O-Fast green staining, and immunohistochemical staining for TIMP-3 and type II collagen. According to Thompson’s criteria, the degeneration of groups 3, 4, and 5, especially group 3, was alleviated as compared with groups 1 and 2. RT-PCR revealed that the expression of TIMP-3 in groups 3, 4, and 5, especially in group 3, was significantly enhanced as compared with group 1 (<0.01). Both Safranin O-Fast green staining and type II collagen staining demonstrated better reserved integrity of disc matrix in groups 3, 4, and 5 than in groups 1 and 2. TIMP-3 staining exhibited an obvious increase of positive-staining rate in groups 3, 4, and 5 as compared with group 1. The positive-staining rate in group 3 (79.42%±1.35%) was about 3times that of group 1 (25.47%±5.46%, <0.01). RAdTIMP-3 can effectively protect the matrix of rabbit intervertebral disc against overloading-induced degeneration in a dose-dependent manner, resulting in the alleviation of disc degeneration.

关键词： tissue inhibitor of metalloproteinase-3 intervertebral disc rabbit gene therapy

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Expression of integrin in hepatic fibrosis and intervention of resveratrol

Jianye WU, Chuanyong GUO, Jun LIU, Xuanfu XUAN

《医学前沿（英文）》 2009年第3卷第1期页码 100-107 doi: 10.1007/s11684-009-0013-x

摘要： The aim of this study was to explore the expression of integrin-β1 in different stages of hepatic fibrosis and intervention of resveratrol as well as the way by which integrin-β1 promoted hepatic fibrosis. Hepatic fibrosis models of male Sprague Dawley (SD) rats were created and intragastric administration of resveratrol was given in low (40 mg/kg), middle (120 mg/kg) and high (200 mg/kg) dose groups. The expression of integrin-β1, tumor growth factor-β (TGF-β) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in different stages of hepatic fibrosis was detected by using RT-PCR. The expression of hexadecenoic acid (HA) and precollagen III (pc III) was assayed by radioimmunoassay. The expression of integrin-β1, TGF-β and TIMP-1 was determined in each group. Liver function and pathological sections of each group in different stages of hepatic fibrosis was tested to judge the therapeutic efficacy of resveratrol at different doses. The expression of integrin-β1 in normal control group was low and steady and was not increased as the development of hepatic fibrosis, but it is increased in other groups. The expression levels of integrin-β1 in the model control group (0.878±0.03, <0.01) and low dose group (0.855±0.04, <0.01) were higher than other groups, but there was no difference between model control group and low dose group ( >0.05). The expression levels of integrin-β1 and TGF-β in middle dose group and high dose group were higher than other groups ( <0.01). The expression levels of integrin-β1 and TGF-β in model control group and low dose group were lower than the normal control group ( <0.01). The expression levels of TIMP-1 in the model control and low dose groups were higher than the other groups ( <0.01). The expression levels of TIMP-1 in the middle dose group and the high dose group were lower than the normal control group ( <0.01). The expression of integrin-β1 existed in all stages of hepatic fibrosis of SD rats, and it was increased as the development of hepatic fibrosis. The expression of TGF-β and TIMP-1 was consistent with that of integrin-β1 in different stages of hepatic fibrosis. Resveratrol could improve the degree of hepatic fibrosis of SD rats and decrease the expression of integrin-β1 markedly at a dose of 120 mg/kg.

关键词： liver fibrosis integrin-β1 resveratrol tumor growth factor-β tissue inhibitor of metalloproteinase-1

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Endogenous tissue factor pathway inhibitor in vascular smooth muscle cells inhibits arterial thrombosis

null

《医学前沿（英文）》 2017年第11卷第3期页码 403-409 doi: 10.1007/s11684-017-0522-y

摘要：

Tissue factor pathway inhibitor (TFPI) is the main inhibitor of tissue factor-mediated coagulation. TFPI is expressed by endothelial and smooth muscle cells in the vasculature. Endothelium-derived TFPI has been reported to play a regulatory role in arterial thrombosis. However, the role of endogenous TFPI in vascular smooth muscle cells (VSMCs) in thrombosis and vascular disease development has yet to be elucidated. In this TFPI^Flox mice crossbred with Sma–Cre mice were utilized to establish TFPI conditional knockout mice and to examine the effects of VSMC-directed TFPI deletion on development, hemostasis, and thrombosis. The mice with deleted TFPI in VSMCs (TFPI^Sma) reproduced viable offspring. Plasma TFPI concentration was reduced 7.2% in the TFPI^Sma mice compared with TFPI^Flox littermate controls. Plasma TFPI concentration was also detected in the TFPI^Tie2 (mice deleted TFPI in endothelial cells and cells of hematopoietic origin) mice. Plasma TFPI concentration of the TFPI^Tie2 mice was 80.4% lower (P<0.001) than that of the TFPI^Flox mice. No difference in hemostatic measures (PT, APTT, and tail bleeding) was observed between TFPI^Sma and TFPI^Flox mice. However, TFPI^Sma mice had increased ferric chloride–induced arterial thrombosis compared with TFPI^Flox littermate controls. Taken together, these data indicated that endogenous TFPI from VSMCs inhibited ferric chloride–induced arterial thrombosis without causing hemostatic effects.

关键词： arterial thrombosis conditional knockout mice tissue factor pathway inhibitor vascular smooth muscle cells

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Decellularized extracellular matrix mediates tissue construction and regeneration

《医学前沿（英文）》 2022年第16卷第1期页码 56-82 doi: 10.1007/s11684-021-0900-3

摘要： Contributing to organ formation and tissue regeneration, extracellular matrix (ECM) constituents provide tissue with three-dimensional (3D) structural integrity and cellular-function regulation. Containing the crucial traits of the cellular microenvironment, ECM substitutes mediate cell–matrix interactions to prompt stem-cell proliferation and differentiation for 3D organoid construction in vitro or tissue regeneration in vivo. However, these ECMs are often applied generically and have yet to be extensively developed for specific cell types in 3D cultures. Cultured cells also produce rich ECM, particularly stromal cells. Cellular ECM improves 3D culture development in vitro and tissue remodeling during wound healing after implantation into the host as well. Gaining better insight into ECM derived from either tissue or cells that regulate 3D tissue reconstruction or organ regeneration helps us to select, produce, and implant the most suitable ECM and thus promote 3D organoid culture and tissue remodeling for in vivo regeneration. Overall, the decellularization methodologies and tissue/cell-derived ECM as scaffolds or cellular-growth supplements used in cell propagation and differentiation for 3D tissue culture in vitro are discussed. Moreover, current preclinical applications by which ECM components modulate the wound-healing process are reviewed.

关键词： decellularized extracellular matrix 3D culture organoids tissue repair

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Effects of phosphatidylinositol 3-kinase inhibitor on human cervical carcinoma cells

Yuan ZHANG MD , Xiaoyan ZHANG MM , Yanhui LI MM , Xuan DU MM , Zehua WANG MD, PhD , Hongbo WANG MD ,

《医学前沿（英文）》 2009年第3卷第3期页码 341-346 doi: 10.1007/s11684-009-0067-9

摘要： Phosphatidylinositol 3-kinase (PI3K) is a crucial cell survival pathway implicated in tumorigenesis because of its role in stimulating cell proliferation and suppressing apoptosis. This study was to investigate the regulation of proliferation and apoptosis by LY294002, an inhibitor of PI3K in cervical cancer cells and the expression of FLICE-like inhibitory protein (c-FLIP) . Human cervical cancer HeLa cells were used in this experiment and cultured. The cultured cells were treated with LY294002 at different concentrations (10, 25, 50 and 100µmol/L) for 6, 12, 24, and 48h before harvesting for evaluation. Cell viability was measured by 3-(4,5)-dimethylthiazol(-2-y1)-3,5-di-phenyltetrazoliumbromide (MTT) assay. Apoptosis was analyzed by flow cytometry. The expression of c-FLIP was detected by Western blot. Cell viability was inhibited by LY294002 significantly (<0.05). Flow cytometry analysis revealed that cell apoptosis was significantly increased in the presence of LY294002 as compared with the control group. Although the expression of c-FLIP was increased in a short time, the expression of c-FLIP was markedly suppressed after the treatment of LY294002 for 48h. These results suggested that the PI3K/Akt signal pathway might be involved in the regulation of cell apoptosis in cervical cancer cells. Moreover, the regulation of c-FLIP expression through PI3K/Akt signal pathway in cervical cancer cells was observed .

关键词： human cervical cancer cells apoptosis phosphatidylinositol 3-kinase (PI3K)/Akt FLICE-like inhibitory protein

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Cryo-copolymerization preparation of dextran-hyaluronate based supermacroporous cryogel scaffolds for tissue

Dongjiao ZHOU, Shaochuan SHEN, Junxian YUN, Kejian YAO, Dong-Qiang LIN

《化学科学与工程前沿（英文）》 2012年第6卷第3期页码 339-347 doi: 10.1007/s11705-012-1209-1

摘要： Dextran-hyaluronate (Dex-HA) based supermacroporous cryogel scaffolds for soft tissue engineering were prepared by free radical cryo-copolymerization of aqueous solutions containing the dextran methacrylate (Dex-MA) and hyaluronate methacrylate (HA-MA) at various macromonomer concentrations under the freezing condition. It was observed that the suitable total concentration of macromonomers for the preparation of Dex-HA cryogel scaffold with satisfied properties was 5% (w/w) at the HA-MA concentration of 1% (w/v), which was then used to produce the test scaffold. The obtained cryogel scaffold with 5% (w/w) macromonomer solution had high water permeability (5.1 × 10 m ) and high porosity (92.4%). The pore diameter examined by scanning electron microscopy (SEM) was in a broad range of 50–135 μm with the mean pore diameter of 91 μm. Furthermore, the cryogel scaffold also had good elastic nature with the elastic modulus of 17.47±1.44 kPa. The culture of 3T3-L1 preadipocyte within the scaffold was investigated and observed by SEM. Cells clustered on the pore walls and grew inside the scaffold indicating the Dex-HA cryogel scaffold could be a promising porous biomaterial for applications in tissue engineering.

关键词： cryogel scaffold tissue engineering dextran hyaluronate 3T3-L1 preadipocyte

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3D打印技术在医疗器械与组织工程中的应用——工艺、材料及监管要求 Article

Wei Long Ng, Jia An, Chee Kai Chua

《工程（英文）》 2024年第36卷第5期页码 146-166 doi: 10.1016/j.eng.2024.01.028

摘要：

三维（3D）打印是一种高度自动化的平台，能够以逐层方式沉积材料，按需制造预先确定的3D复杂结构。每种类型的3D打印技术都有其独特的优势和局限性，选择合适的3D打印技术在很大程度上取决于其预期用途。在这篇综述论文中，我们展示并强调了3D打印个性化医疗器械的一些关键工艺（打印参数、构建方向、构建位置和支撑结构）、材料（批次之间的一致性、回收利用、蛋白质吸附、生物相容性和降解性能）和监管考量（无菌性和力学性能本综述论文的目标是让读者对3D打印的各种关键考量（工艺、材料和监管）有很好的理解，这对于生产制造已经改善的、专用于患者的3D打印医疗器械和组织构造物至关重要。

关键词： 3D打印生物打印生物制造医疗器械组织结构

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关于3D打印技术在医学模具以及再生组织和器官方面的应用综述

Kan Wang, Chia-Che Ho, Chuck Zhang, Ben Wang

《工程（英文）》 2017年第3卷第5期页码 653-662 doi: 10.1016/J.ENG.2017.05.013

摘要：随着三维（3D）打印和3D 生物打印技术的快速发展，许多研究人员已经开始使用增材制造技术来生产具有多种功能的医学模具。本文综述了3D 打印和3D 生物打印技术在制作功能性医学模具和生物结构方面的应用。特别讨论了3D 打印功能性医学模具（即组织模拟医学模具、放射性医学模具和生理医学模具）及被用于再生组织和器官的3D 生物打印模具的制备（即混合模式支架材料、可转换支架和集成传感器）工艺、发展现状以及未来发展趋势

关键词： 3D打印 3D生物打印医学模具再生组织/器官支架

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Fabrication of scaffolds in tissue engineering: A review

Peng ZHAO, Haibing GU, Haoyang MI, Chengchen RAO, Jianzhong FU, Lih-sheng TURNG

《机械工程前沿（英文）》 2018年第13卷第1期页码 107-119 doi: 10.1007/s11465-018-0496-8

摘要：

Tissue engineering (TE) is an integrated discipline that involves engineering and natural science in the development of biological materials to replace, repair, and improve the function of diseased or missing tissues. Traditional medical and surgical treatments have been reported to have side effects on patients caused by organ necrosis and tissue loss. However, engineered tissues and organs provide a new way to cure specific diseases. Scaffold fabrication is an important step in the TE process. This paper summarizes and reviews the widely used scaffold fabrication methods, including conventional methods, electrospinning, three-dimensional printing, and a combination of molding techniques. Furthermore, the differences among the properties of tissues, such as pore size and distribution, porosity, structure, and mechanical properties, are elucidated and critically reviewed. Some studies that combine two or more methods are also reviewed. Finally, this paper provides some guidance and suggestions for the future of scaffold fabrication.

关键词： tissue engineering scaffolds electrospinning 3D printing molding techniques conventional methods

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Role of salivary matrix metalloproteinase-8 (MMP-8) in chronic periodontitis diagnosis

null

《医学前沿（英文）》 2015年第9卷第1期页码 72-76 doi: 10.1007/s11684-014-0347-x

摘要：

Periodontitis is an inflammatory disease of the periodontium. Any imbalance between the matrix metalloproteinases (MMPs) secreted by neutrophils and tissue inhibitors initiates the destruction of collagen in gum tissue, leading to chronic periodontitis. This study aimed to correlate salivary levels of MMP-8 and periodontal parameters of chronic periodontitis to establish MMP-8 as a noninvasive marker for the early diagnosis of chronic periodontitis. The study involved 40 subjects visiting the periodontic OPD of Dr. Ziauddin Ahmad Dental College and Hospital, located in Aligarh, U.P., India, from 2011 to 2012. The subjects were divided into two groups: group I consisted of 20 periodontally healthy subjects (controls) while group II consisted of 20 patients with chronic periodontitis. Chronic periodontitis was assessed on the basis of several periodontal parameters, including pocket probing depth (PPD), clinical attachment level (CAL), gingival index (GI), and plaque index (PI). Around 3 ml of unstimulated and whole expectorated saliva was collected for MMP-8 estimation by ELISA using Quantikine human total MMP-8 immunoassay kits. Data were analyzed using STATISTICA (Windows version 6) software. Salivary MMP-8 levels of groups I and II were 190.91±143.89 ng/ml and 348.26±202.1 ng/ml, respectively. The MMP-8 levels and periodontal status (PPD, CAL, GI, and PI) of groups I and II showed positive and significant correlations (for PPD, r = 0.63, P<0.001; for CAL, r = 0.54, P<0.001; for GI, r = 0.49, P<0.001; and for PI, r = 0.63, P<0.001). The results of this study demonstrate elevated concentrations of MMP-8 in individuals with chronic periodontitis.

关键词： matrix metalloproteinase-8 chronic periodontitis pocket probing depth clinical attachment level gingival index plaque index

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组织工程和给药技术中的三维光制造 Review

Rúben F. Pereira, Paulo J. Bártolo

《工程（英文）》 2015年第1卷第1期页码 90-112 doi: 10.15302/J-ENG-2015015

摘要：细胞和治疗分子可以包含在初始水凝胶前体的溶液中，并加工成三维（3D）结构。近来，光制造也已被开发用来动态调节水凝胶的实时特性，加强控制细胞寿命和生物活性物质的传递。本文聚焦于利用3D光制造技术为组织再生和给药技术生产先进结构的相关研究，同时介绍了目前最先进的光制造技术，重点放在控制细胞生物活性因子分布形式的工作原理和生物制造方法上。因光制造技术具有工艺快速、时空控制、高分辨率和高精度等特性，故其在复杂的3D结构设计中扮演着重要角色。这种技术同样能够为组织再生构建适当的环境，并可调节治疗方法的实施状况。

关键词：三维光制造生物材料组织工程给药

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可见光引发的3D 生物打印技术及其生物墨水材料在组织工程领域的研究进展 Review

郑子卓, David Eglin, Mauro Alini, Geoff R. Richards, 秦岭, 赖毓霄

《工程（英文）》 2021年第7卷第7期页码 966-978 doi: 10.1016/j.eng.2020.05.021

摘要：

3D生物打印技术是一种基于传统3D打印技术发展起来的新兴技术。在众多3D生物打印技术中，光固化3D生物打印技术在组织工程研究领域发挥着重要作用，而光固化3D生物打印技术的成功应用又与生物材料的光固化特性不可分割。然而，紫外线用于3D生物打印可能会造成细胞损伤，影响细胞的活力。而可见光在生物安全性方面具有独特的优势，所以，将可见光交联技术应用在3D生物打印领域极具应用前景。本文回顾了目前可应用的由可见光引发的3D生物打印技术，并对可见光交联生物墨水的交联机理、可见光引发剂类型和生物医学应用进行了介绍。本文最后对可见光引发的3D生物打印设备及水凝胶目前在生物医学领域应用中存在的挑战和前景进行了讨论。

关键词：医用增材制造生物墨水组织工程 3D生物打印

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Three-dimensional reconstruction of light microscopy image sections: present and future

null

《医学前沿（英文）》 2015年第9卷第1期页码 30-45 doi: 10.1007/s11684-014-0337-z

摘要：

Three-dimensional (3D) image reconstruction technologies can reveal previously hidden microstructures in human tissue. However, the lack of ideal, non-destructive cross-sectional imaging techniques is still a problem. Despite some drawbacks, histological sectioning remains one of the most powerful methods for accurate high-resolution representation of tissue structures. Computer technologies can produce 3D representations of interesting human tissue and organs that have been serial-sectioned, dyed or stained, imaged, and segmented for 3D visualization. 3D reconstruction also has great potential in the fields of tissue engineering and 3D printing. This article outlines the most common methods for 3D tissue section reconstruction. We describe the most important academic concepts in this field, and provide critical explanations and comparisons. We also note key steps in the reconstruction procedures, and highlight recent progress in the development of new reconstruction methods.

关键词： microtomy 3D imaging computer-assisted image processing 3D printing tissue scaffold

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支架与组织的设计及3D打印 Review

安佳, Joanne Ee Mei Teoh, Ratima Suntornnond, Chee Kai Chua

《工程（英文）》 2015年第1卷第2期页码 261-268 doi: 10.15302/J-ENG-2015061

摘要：

目前，组织工程学中应用3D打印工艺的情况越来越多。本文介绍3D打印技术在组织工程学应用领域的最前沿研究，尤其侧重于计算机辅助支架设计系统的开发、功能梯度支架的直接3D打印、选择性激光烧结 (SLS) 和熔融沉积成型 (FDM) 工艺建模、利用微观和宏观特征进行支架间接增材制造、生物反应器的开发，以及3D/4D生物打印。本文还讨论3D打印的技术限制，进而突出新3D打印方法在组织工程学领域得到进一步改善的可能性。

关键词：快速原型制造 3D打印增材制造组织工程生物打印

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Progress and perspectives of neural tissue engineering

null

《医学前沿（英文）》 2015年第9卷第4期页码 401-411 doi: 10.1007/s11684-015-0415-x

摘要：

Traumatic injuries to the nervous system lead to a common clinical problem with a quite high incidence and affect the patient’s quality of life. Based on a major challenge not yet addressed by current therapeutic interventions for these diseases, a novel promising field of neural tissue engineering has emerged, grown, and attracted increasing interest. This review provides a brief summary of the recent progress in the field, especially in combination with the research experience of the author’s group. Several important aspects related to tissue engineered nerves, including the theory on their construction, translation into the clinic, improvements in fabrication technologies, and the formation of a regenerative environment, are delineated and discussed. Furthermore, potential research directions for the future development of neural tissue engineering are suggested.

关键词： nerve injury tissue engineering nerve grafts