Toni Roberto, Casa Claudia Della, Spaletta Giulia, Marchetti Giacomo, Mazzoni Perseo, Bodria Monica, Ravera Simone, Dallatana Davide, Castorina Sergio, Riccioli Vincenzo, Castorina Emilio Giovanni, Antoci Salvatore, Campanile Enrico, Raise Gabriella, Rossi Raffaella, Ugolotti Giorgio, Martorella Andrew, Roti Elio, Sgallari Fiorella, Pinchera Aldo
Department of Human Anatomy, University of Parma.
Acta Biomed. 2007;78 Suppl 1:129-55.
A new concept for ex situ endocrine organ bioengineering is presented, focused on the realization of a human bioartificial thyroid gland. It is based on the theoretical assumption and experimental evidence that symmetries in geometrical coordinates of the thyroid tissue remain invariant with respect to developmental, physiological or pathophysiological transformations occuring in the gland architecture. This topological arrangement is dependent upon physical connections established between cells, cell adhesion molecules and extracellular matrix, leading to the view that the thyroid parenchyma behaves like a deformable "putty", moulded onto an elastic stromal/vascular scaffold (SVS) dictating the final morphology of the gland. In particular, we have raised the idea that the geometry of the SVS per se provides pivotal epigenetic information to address the genetically-programmed, thyrocyte and endothelial/vascular proliferation and differentiation towards a functionally mature gland, making organ form a pre-requirementfor organ function. A number of experimental approaches are explored to obtain a reliable replica of a human thyroid SVS, and an informatic simulation is designed based on fractal growth of the thyroid intraparenchymal arterial tree. Various tissue-compatible and degradable synthetic or biomimetic polymers are discussed to act as a template of the thyroid SVS, onto which to co-seed autologous human thyrocyte (TPC) and endothelial/vascular (EVPC) progenitor cells. Harvest and expansion of both TPC and EVPC in primary culture are considered, with specific attention to the selection of normal thyrocytes growing at a satisfactory rate to colonize the synthetic matrix. In addition, both in vitro and in vivo techniques to authenticate TPC and EVPC lineage differentiation are reviewed, including immunocytochemistry, reverse trascriptase-polymerase chain reaction, flow cytomery and proteomics. Finally, analysis of viability of the thyroid construct following implantation in animal hosts is proposed, with the intent to obtain a bioartificial thyroid gland morphologically and functionally adequate for transplantation. We believe that the biotechnological scenario proposed herein may provide a template to construct other, more complex and clinically-relevant bioartificial endocrine organs ex situ, such as human pancreatic islets and the liver, and perhaps a new approach to brain bioengineering.
本文提出了一种异位内分泌器官生物工程的新概念,重点是实现人类生物人工甲状腺。它基于这样的理论假设和实验证据:甲状腺组织几何坐标中的对称性在腺体结构发生的发育、生理或病理生理转变方面保持不变。这种拓扑排列取决于细胞、细胞粘附分子和细胞外基质之间建立的物理连接,从而得出甲状腺实质表现得像一种可变形的“油灰”的观点,它被塑造在一个弹性的基质/血管支架(SVS)上,该支架决定了腺体的最终形态。特别是,我们提出了这样的观点,即SVS本身的几何形状提供了关键的表观遗传信息,以引导基因编程的甲状腺细胞和内皮/血管增殖以及向功能成熟腺体的分化,使器官形态成为器官功能的先决条件。探索了多种实验方法来获得人类甲状腺SVS的可靠复制品,并基于甲状腺实质内动脉树的分形生长设计了一个信息模拟。讨论了各种组织相容性和可降解的合成或仿生聚合物,以作为甲状腺SVS的模板,在其上共同接种自体人类甲状腺细胞(TPC)和内皮/血管(EVPC)祖细胞。考虑了原代培养中TPC和EVPC的收获和扩增,特别关注选择以令人满意的速率生长以在合成基质上定植的正常甲状腺细胞。此外,还综述了用于验证TPC和EVPC谱系分化的体外和体内技术,包括免疫细胞化学、逆转录聚合酶链反应、流式细胞术和蛋白质组学。最后,提出了在动物宿主中植入后对甲状腺构建体的活力分析,目的是获得形态和功能上适合移植的生物人工甲状腺。我们相信,本文提出的生物技术方案可能为异位构建其他更复杂且与临床相关的生物人工内分泌器官(如人类胰岛和肝脏)提供一个模板,也许还为脑生物工程提供一种新方法。