Wake Forest Institute for Regenerative Medicine, Wake Forest University School of Medicine, Winston Salem, North Carolina 27157, USA.
Ann Surg. 2012 Aug;256(2):363-70. doi: 10.1097/SLA.0b013e31825a02ab.
It is important to identify new sources of transplantable organs because of the critical shortage of donor organs. Tissue engineering holds the potential to address this issue through the implementation of decellularization-recellularization technology.
To produce and examine acellular renal extracellular matrix (ECM) scaffolds as a platform for kidney bioengineering.
Porcine kidneys were decellularized with distilled water and sodium dodecyl sulfate-based solution. After rinsing with buffer solution to remove the sodium dodecyl sulfate, the so-obtained renal ECM scaffolds were processed for vascular imaging, histology, and cell seeding to investigate the vascular patency, degree of decellularization, and scaffold biocompatibility in vitro. Four whole renal scaffolds were implanted in pigs to assess whether these constructs would sustain normal blood pressure and to determine their biocompatibility in vivo. Pigs were sacrificed after 2 weeks and the explanted scaffolds were processed for histology.
Renal ECM scaffolds were successfully produced from porcine kidneys. Scaffolds retained their essential ECM architecture and an intact vascular tree and allowed cell growth. On implantation, unseeded scaffolds were easily reperfused, sustained blood pressure, and were tolerated throughout the study period. No blood extravasation occurred. Pathology of explanted scaffolds showed maintenance of renal ultrastructure. Presence of inflammatory cells in the pericapsular region and complete thrombosis of the vascular tree were evident.
Our investigations show that pig kidneys can be successfully decellularized to produce renal ECM scaffolds. These scaffolds maintain their basic components, are biocompatible, and show intact, though thrombosed, vasculature.
由于供体器官严重短缺,因此识别新的可移植器官来源非常重要。组织工程通过实施去细胞-再细胞化技术,具有解决这个问题的潜力。
制备和研究去细胞化肾脏细胞外基质(ECM)支架,作为肾脏生物工程的平台。
使用蒸馏水和基于十二烷基硫酸钠的溶液对猪肾进行去细胞处理。用缓冲液冲洗以除去十二烷基硫酸钠后,对所获得的肾 ECM 支架进行血管成像、组织学和细胞接种处理,以研究血管通畅性、去细胞化程度和体外支架生物相容性。将四个完整的肾支架植入猪体内,以评估这些构建体是否能维持正常血压,并确定其体内生物相容性。在 2 周后处死猪,对植入的支架进行组织学处理。
成功从猪肾中制备出肾 ECM 支架。支架保留了其基本的 ECM 结构和完整的血管树,并允许细胞生长。在植入时,未接种的支架很容易再灌注,维持血压,并在整个研究期间耐受。没有发生血液外渗。对植入支架的组织学检查显示保留了肾脏超微结构。可见包膜下区域存在炎症细胞和血管树完全血栓形成。
我们的研究表明,猪肾可以成功地去细胞化以制备肾 ECM 支架。这些支架保留了其基本成分,具有生物相容性,并且显示出完整的血管系统,尽管有血栓形成。