Integrated Automatic Bioreactor of Human-Sized Lungs: Achievement of One Decade’s Efforts to Engineering Whole Lung Organs

سال انتشار: 1397
نوع سند: مقاله کنفرانسی
زبان: انگلیسی
مشاهده: 525

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شناسه ملی سند علمی:

NSCMRMED03_109

تاریخ نمایه سازی: 30 دی 1397

چکیده مقاله:

Background and Aim: Engineered lungs are an excellent solutionfor dealing with the increasing demand for transplantation. There is a wide range of hurdles which must be overcome to advance the fieldand facilitate moving toward the clinic. To do so, we tried these steps:i) engineering rodent lungs using a custom-designed bioreactor as anecessary element to guide the development of tissues, ii) establishing anon-invasive online method to monitor cell viability within lung scaffoldsas an essential step toward a functional tissue, iii) building an integratedbioreactor to support large-scale tissue engineering.Methods: In the step (i), lungs were harvested from 2-month-old male rats.The lungs were decellularized with liquid detergent. Hematoxylin andeosin staining were used to verify efficient removal of cell components ofacelluar tissues. Furthermore, scanning electron microscopy proved themaintenance of the extracellular matrix architecture. Human umbilicalcord vein endothelial cells (HUVECs) were cultured on acellular tissueusing a perfusion-based bioreactor. In the step (ii), an online monitoringsystem based on respiration activity was developed to monitor cellviability within acellular lung scaffolds. Acellular lung scaffolds wererecellularized with HUVECs, and then, cell viability was monitoredduring a 5-day period. In the step (iii), sheep lungs were decellularizedusing a 20-L chamber with an SDS-based protocol. The chamber witha custom-designed cap was modified to contain a human-sized lung,providing necessary connections to trachea and pulmonary artery viaembedded ports. The decellularization was performed up to 24 hours atthe perfusion rate of 1.1-1.4 L/min for different stages.Results: In this study, (i) H&E staining and SEM analysis indicated thatthe entire pulmonary airway architecture and alveolar structure in theacellular scaffolds were intact. DNA assay indicated that the removalof the DNA content of the decellularized scaffold was approximately94.9% (compared to the native lung), and the ultimate tensile strengthvalues of decellularized and native samples were very similar; therewas no decrease in UTS for the acellular lung scaffold. (ii) The realtimemonitoring system produced a cell growth profile representinginvaluable information on cell viability during the culture period. Thecell growth profile obtained by the monitoring system was consistentwith MTT analysis and glucose consumption measurement. (iii) Thearchitecture of native and decellularized lungs was analyzed with H&Estaining and SEM. Removal of nuclear materials was proved by H&Estaining; also, SEM imaging demonstrated a total removal of cells andpreserved structural architecture. Furthermore, the MTT assay showedthe biocompatibility of the decellularized matrix.Conclusion: To move faster toward the clinic, the integration ofengineering and biological parameters in bioreactors is needed. Thiscan lead to more consistent products, reduce the process time and laborintensity, maintain sterility and necessary information on tissue growthand maturity, which is required to have a functional tissue needed forcommercialization.

نویسندگان

Ghassem Amoabediny

School of Chemical Engineering, University of Tehran, Tehran, Iran

Seyed Hossein Mahfouzi

Department of Life Science Engineering, University of Tehran, Tehran, Iran

Seyed Hamid Safiabadi Tali

Department of Life Science Engineering, University of Tehran,Tehran, Iran

Mostafa Ghanei

Department of Pulmonary Medicine, Baqiyatallah University of Medical Sciences, Tehran, Iran