Assembly of discrete collagen-chitosan microenvironments into multiphase tissue constructs.
Assembly of discrete collagen-chitosan microenvironments into multiphase tissue constructs.
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DOI：
10.1002/adhm.201200346
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发表时间：
2013-05
影响因子：
10
通讯作者：
Stegemann, Jan P.
中科院分区：
文献类型：
--
作者：
Caldwell, David J.;Rao, Rameshwar R.;Stegemann, Jan P.
我们报告了两种新的方法，从离散的细胞外基质微环境中使用离心和真空成型创建复杂的多相组织结构。通常，组织工程的广泛领域包括用于产生用于替换或修复损伤或患病组织的生物组织的各种方法。模块化组织工程试图通过从重复的亚基或“模块”产生更大的结构来创建组织。[1]这些重复的亚基可以被设计成具有独特的微结构特征，并允许自下而上创建宏观组织。模块化组织工程方法的一个优点是能够创建具有精确设计的形态和空间控制的图案的复杂组织。然而，该技术目前的局限性是为细胞提供适当的功能性细胞外基质，这对于细胞发育和分化成所需的组织类型至关重要。本文介绍了一种简单的技术，通过该技术，由直径为200-300 μm的胶原-壳聚糖微珠组成的模块化微环境可以组装成具有限定空间排列的大规模结构。介绍了两种方法：真空成型法和离心法。以这种方式控制细胞周围环境的能力允许对细胞微环境进行图案化以研究细胞相互作用，并且这种方法可以扩展到复杂组织的产生。将细胞微环境组装成宏观的、图案化的、工程化的组织是一个不断发展的研究领域。以前探索的方法包括使用基于喷射的3D打印机的细胞打印，[2]自组装，[3]生物打印，[4]机器人3D打印，[5]微组织工程，[6]和细胞片工程。[7]含有多种细胞类型的成形的多层组织的产生在开发各种各样的复杂组织和器官中具有应用，其中第一努力被应用于产生血管化骨组织和骨软骨界面。[1，8]随着新材料、组装技术和细胞培养方法的开发，这些努力正在取得进展。然而，仍然需要更稳健的方法来构建具有限定结构的三维宏观组织构建体，并且在从生理学相关的细胞外基质组分产生这些结构中存在特别的挑战。我们的实验室已经创建了水凝胶“微珠”，由嵌入在确定的细胞外基质蛋白和多糖（包括胶原蛋白、纤维蛋白、琼脂糖和壳聚糖）的球形模块中的细胞组成。[10-12]壳聚糖和胶原蛋白是特别令人感兴趣的，因为这些天然来源的材料通常都是生物可降解和生物相容的，[13-15]并且已经在血管，[16]皮肤，[17]韧带，[18]和骨[19]组织工程中的许多应用中进行了研究。通常，可以通过限定细胞外基质组成、将生长因子掺入基质中以及通过控制基质的细胞密度和机械性质来引导包埋在微珠内的细胞的功能。此外，可以调节微珠本身的性质（密度、刚度、内聚性）以控制它们组装成更大规模的结构。在本研究中，使用图1中所示的过程从I型胶原蛋白和壳聚糖的复合物产生微珠。制备微珠、胶原蛋白和壳聚糖。
We report on two novel methods of creating complex multiphase tissue constructs from discrete extracellular matrix microenvironments using centrifugation and vacuum molding. In general, the broad field of tissue engineering encompasses a variety of methodologies for creating biological tissues for the replacement or repair of injured or diseased tissues. Modular tissue engineering attempts to create tissues by generating larger structures from repeating subunits, or “modules”.[1] These repeating subunits can be designed to have unique microarchitectural features, and allow for the bottom-up creation of macroscale tissues. One advantage presented by the modular tissue engineering method is the ability to create complex tissues with precisely designed morphologies and spatially controlled patterning. However, a current limitation of this technology is providing cells with appropriate functional extracellular matrices, which are critical for the development and differentiation of cells into desired tissue types. This paper presents facile techniques by which modular microenvironments comprised of collagen-chitosan microbeads 200–300 μm in diameter can be assembled into larger-scale constructs with defined spatial arrangement. Two methods are presented: vacuum molding and centrifugation. The ability to control the environment surrounding cells in this manner allows for patterning of cellular microenvironments for the study of cell interactions, and this method may be extended to generation of complex tissues. Assembly of cellular microenvironments into macroscopic, patterned, engineered tissues is a growing area of research. Previously explored methods include cell printing using jetbased 3D printers,[2] self-assembly,[3] bioprinting,[4] robotic 3D printing,[5] microtissue engineering,[6] and cell sheet engineering.[7] The generation of shaped, multilayered tissues containing multiple cell types has applications in developing a wide a variety of complex tissues and organs, with the first efforts being applied to generating vascularized bone tissue and osteochondral interfaces.[1, 8] These efforts are progressing as new materials, assembly techniques and cell culture methods are developed. However, there is still a need for more robust approaches to the construction of three dimensional macroscale tissue constructs with defined architecture, and there is a particular challenge in creating these structures from physiologically relevant extracellular matrix components.Microscale hydrogels [9] and similar cellular microenvironments have previously been used in the modular tissue engineering approach. Our lab has created hydrogel “microbeads” comprised of cells embedded in spherical modules of defined extracellular matrix proteins and polysaccharides, including collagen, fibrin, agarose, and chitosan.[10–12] Chitosan and collagen are of particular interest, as these naturally derived materials are both generally biodegradable and biocompatible,[13–15] and have been investigated for numerous applications in vascular,[16] skin,[17] ligament,[18] and bone [19] tissue engineering. In general, the function of cells embedded within microbeads can be guided by defining the extracellular matrix composition, incorporating growth factors into the matrix, and by controlling the cell density and mechanical properties of the matrix. In addition, the properties of the microbeads themselves (density, stiffness, cohesiveness) can be modulated to control their assembly into larger scale structures. In this study, microbeads were created from a composite of collagen Type I and chitosan using the process shown in Figure 1. To prepare microbeads, collagen and chitosan …
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影响因子：
3.4
作者：
Nichol JW;Khademhosseini A
通讯作者：
Khademhosseini A
影响因子：
9
作者：
Jakab K;Norotte C;Marga F;Murphy K;Vunjak-Novakovic G;Forgacs G
通讯作者：
Forgacs G
影响因子：
9.8
作者：
Rao, Rameshwar R.;Peterson, Alexis W.;Ceccarelli, Jacob;Putnam, Andrew J.;Stegemann, Jan P.
通讯作者：
Stegemann, Jan P.
影响因子：
9
作者：
Tejavibulya N;Youssef J;Bao B;Ferruccio TM;Morgan JR
通讯作者：
Morgan JR
影响因子：
11.1
作者：
Elloumi-Hannachi, I.;Yamato, M.;Okano, T.
通讯作者：
Okano, T.