An Ovine Model of In Vivo Bioreactor-Based Bone Generation.
An Ovine Model of In Vivo Bioreactor-Based Bone Generation.
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基于体内生物反应器的骨生成的绵羊模型。
DOI：
10.1089/ten.tec.2020.0125
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Mikos,AntoniosG
中科院分区：
文献类型：
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作者：
Watson,Emma;Tatara,AlexanderM;vandenBeucken,JeroenJJP;Jansen,JohnA;Wong,MarkE;Mikos,AntoniosG
在不使用外源生长因子或外源细胞的情况下生成复杂几何形状的血管化矿化组织需要大型动物模型来重现临床中遇到的挑战。所提出的多功能绵羊模型可用于研究使用定制生物反应器生成矿化组织，在转移到另一个部位并整合之前匹配缺陷的大小和形状。该协议产生了生物反应器，可以收集这些生物反应器来研究不同生物材料对骨骼生成或生成适合修复骨缺损的组织的影响；该协议侧重于下颌骨的重建，但可以针对骨科应用进行修改。生物反应器的填充材料可以改变，从而可以研究各种市售或新型移植材料。外科手术需要约 1.5 小时才能在肋骨骨膜附近植入四个生物反应器。 9 周后，生物反应器组织的收获大约需要 1 小时。如果创建颅面缺损，则应额外花费 2 小时创建下颌缺损，并额外花费 2 至 3 小时进行重建。经过重建的绵羊通常会在 12 周后被安乐死，以便对转移的组织进行评估。在本协议中，我们讨论了确保评估骨再生的可重复性和分析技术的必要步骤，例如微计算机断层扫描、机械分析和组织学。影响陈述骨移植是骨科、耳鼻喉科和口腔颌面外科领域的常见手术。生成定制的、血管化的、机械坚固的骨组织，同时消除常见的并发症，例如自体移植物收获时的供体部位发病率或市售合成移植物缺乏合适的机械性能，将极大地改善患者的生活。需要大型动物模型来产生临床相关几何形状的组织。在本文中，提出了一种用于定制骨生成的体内生物反应器技术的可重复绵羊模型，该模型广泛应用于组织工程和再生医学。
The generation of vascularized mineralized tissues of complex geometry without the use of extrinsic growth factors or exogenous cells requires a large animal model to recapitulate the challenges seen in the clinic. The proposed versatile ovine model can be utilized to investigate the use of a customized bioreactor to generate mineralized tissue, matching the size and shape of a defect before transfer to and integration within another site. The protocol results in bioreactors that can be harvested for investigation of the effects of different biomaterials for the generation of bone or to generate tissues appropriate for repair of bony defects; this protocol focuses on reconstruction of the mandible but could be modified for orthopedic applications. The bioreactor packing material can be altered, allowing for the study of various commercially available or novel graft materials. The surgical procedure requires ∼1.5 h to implant four bioreactors adjacent to rib periosteum. After 9 weeks, the harvest of the bioreactor tissue takes approximately 1 h. If creating a craniofacial defect, an additional 2 h should be taken for mandibular defect creation and 2 to 3 h for the reconstruction. Sheep that have undergone reconstruction are typically euthanized after 12 weeks to allow for evaluation of transferred tissues. In this protocol, we discuss the necessary steps to ensure the reproducibility and analytical techniques to assess bone regeneration such as microcomputed tomography, mechanical analysis, and histology.Impact statementBone grafting is a frequent procedure in the fields of orthopedics, otolaryngology, and oral and maxillofacial surgery. Generating customized, vascularized, and mechanically robust bony tissues while eliminating common complications such as donor site morbidity with autograft harvest or lack of suitable mechanical properties with commercially available synthetic graft would greatly improve the lives of patients. A large animal model is necessary to generate tissues of clinically relevant geometries. In this article, a reproducible ovine model ofin vivobioreactor technology toward customized bone generation is presented with broad application to tissue engineering and regenerative medicine.