Development of vascularized nerve scaffold using perfusion-decellularization and recellularization

Wüthrich, Tsering; Leşe, Ioana; Haberthür, David; Zubler, Cédric; Hlushchuk, Ruslan; Hewer, Ekkehard; Maistriaux, Louis; Gianello, Pierre; Lengelé, Benoît; Rieben, Robert; Vögelin, Esther; Olariu, Radu; Duisit, Jérôme; Taddeo, Adriano

doi:10.1016/j.msec.2020.111311

Cited by 22 publications

(22 citation statements)

References 47 publications

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“…The ideal graft must preserve the extracellular matrix and basal membrane since they provide attachment and proliferation of cells and maintain its morphological and biochemical properties. 12–14…”

Section: Discussionmentioning

confidence: 99%

In vivo performance of decellularized tracheal grafts in the reconstruction of long length tracheal defects: Experimental study

et al. 2021

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Background: The repair of long-segment tracheal lesions remains an important challenge. Nowdays no predictable and dependable substitute has been found. Decellularized tracheal scaffolds have shown to be a promising graft for tracheal transplantation, since it is non-immunogenic. Objective: Evaluate in vivo decellularized tracheal allografts performance to replace long tracheal segment. Methods: Forty-five swines underwent surgery as follows: Fifteen trachea donors and 30 receptors of decellularized trachea allografts. The receptors were randomly divided in five groups ( n = 6). In groups I and II, donor trachea segment was decellularized by 15 cycles with sodium deoxycholate and deoxyribonuclease, in group II, the allograft was reinforced with external surgical steel wire. Groups, III, IV, and V decellularization was reduced to seven cycles, supplemented with cryopreservation in group IV and with glutaraldehyde in group V. A 10 rings segment was excised from the receptor swine and the decellularized trachea graft was implanted to re-establish trachea continuity. Results: Both decellularization cycles caused decreased stiffness. All trachea receptors underwent euthanasia before the third post-implant week due to severe dyspnea and trachea graft stenosis, necrosis, edema, inflammation, hemorrhage, and granulation tissue formation in anastomotic sites. Histologically all showed total loss of epithelium, separation of collagen fibers, and alterations in staining. Conclusions: Both decellularization techniques severely damaged the structure of the trachea and the extracellular matrix of the cartilage, resulting in a no functional graft, in spite of the use of surgical wire, cryopreservation or glutaraldehyde treatment. An important drawback was the formation of fibrotic stenosis in both anastomosis.

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Section: Discussionmentioning

confidence: 99%

In vivo performance of decellularized tracheal grafts in the reconstruction of long length tracheal defects: Experimental study

et al. 2021

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“…Wüthrich et al in particular assessed the preservation several growth factors post-decellularization whereby detectable levels could be observed. While they did not focus on characterizing this profile of growth factors, their preliminary work elucidated that some of the key growth factors remain post-decellularization and could be beneficial in promoting nerve cell growth ( 67 ). Additionally, the interplay of the ECM with nerve function cannot be missed where components such as GAGs have known interactions with neurotropic factors such as promoting neural adhesion and migration ( 67 , 84 ).…”

Section: Challenges In Composite Tissues Recellularizationmentioning

confidence: 99%

Recellularization of Bioengineered Scaffolds for Vascular Composite Allotransplantation

Adil

Haykal

2022

Front. Surg.

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Traumatic injuries or cancer resection resulting in large volumetric soft tissue loss requires surgical reconstruction. Vascular composite allotransplantation (VCA) is an emerging reconstructive option that transfers multiple, complex tissues as a whole subunit from donor to recipient. Although promising, VCA is limited due to side effects of immunosuppression. Tissue-engineered scaffolds obtained by decellularization and recellularization hold great promise. Decellularization is a process that removes cellular materials while preserving the extracellular matrix architecture. Subsequent recellularization of these acellular scaffolds with recipient-specific cells can help circumvent adverse immune-mediated host responses and allow transplantation of allografts by reducing and possibly eliminating the need for immunosuppression. Recellularization of acellular tissue scaffolds is a technique that was first investigated and reported in whole organs. More recently, work has been performed to apply this technique to VCA. Additional work is needed to address barriers associated with tissue recellularization such as: cell type selection, cell distribution, and functionalization of the vasculature and musculature. These factors ultimately contribute to achieving tissue integration and viability following allotransplantation. The present work will review the current state-of-the-art in soft tissue scaffolds with specific emphasis on recellularization techniques. We will discuss biological and engineering process considerations, technical and scientific challenges, and the potential clinical impact of this technology to advance the field of VCA and reconstructive surgery.

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“…The available decellularization protocols have important limitations. Most of them require prolonged periods, taking from 6–7 h ( Rosmark et al, 2018 ) to several days ( Wishart et al, 2020 ; Wüthrich et al, 2020 ). Additionally, most of the available methods fail in the preservation of some ECM components.…”

Section: Introductionmentioning

confidence: 99%

“…Additionally, most of the available methods fail in the preservation of some ECM components. For instance, significant decreases in elastin ( Maghsoudlou et al, 2013 ; Alshaikh et al, 2020 ), collagen ( Hill et al, 2015 ; Alshaikh et al, 2020 ), glycosaminoglycans ( Mendoza-Novelo et al, 2011 ; Lü et al, 2014 ), laminins ( Hill et al, 2015 ; Wüthrich et al, 2020 ) and proteoglycans ( Hill et al, 2015 ; Calle et al, 2016 ) have been described. In addition to these limitations, the current protocols are still not particularly flexible or accessible.…”

Section: Introductionmentioning

confidence: 99%

Novel Decellularization Method for Tissue Slices

Narciso

Ulldemolins

Júnior

et al. 2022

Front. Bioeng. Biotechnol.

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Decellularization procedures have been developed and optimized for the entire organ or tissue blocks, by either perfusion of decellularizing agents through the tissue’s vasculature or submerging large sections in decellularizing solutions. However, some research aims require the analysis of native as well as decellularized tissue slices side by side, but an optimal protocol has not yet been established to address this need. Thus, the main goal of this work was to develop a fast and efficient decellularization method for tissue slices—with an emphasis on lung—while attached to a glass slide. To this end, different decellularizing agents were compared for their effectiveness in cellular removal while preserving the extracellular matrix. The intensity of DNA staining was taken as an indicator of remaining cells and compared to untreated sections. The presence of collagen, elastin and laminin were quantified using immunostaining and signal quantification. Scaffolds resulting from the optimized protocol were mechanically characterized using atomic force microscopy. Lung scaffolds were recellularized with mesenchymal stromal cells to assess their biocompatibility. Some decellularization agents (CHAPS, triton, and ammonia hydroxide) did not achieve sufficient cell removal. Sodium dodecyl sulfate (SDS) was effective in cell removal (1% remaining DNA signal), but its sharp reduction of elastin signal (only 6% remained) plus lower attachment ratio (32%) singled out sodium deoxycholate (SD) as the optimal treatment for this application (6.5% remaining DNA signal), due to its higher elastin retention (34%) and higher attachment ratio (60%). Laminin and collagen were fully preserved in all treatments. The SD decellularization protocol was also successful for porcine and murine (mice and rat) lungs as well as for other tissues such as the heart, kidney, and bladder. No significant mechanical differences were found before and after sample decellularization. The resulting acellular lung scaffolds were shown to be biocompatible (98% cell survival after 72 h of culture). This novel method to decellularize tissue slices opens up new methodological possibilities to better understand the role of the extracellular matrix in the context of several diseases as well as tissue engineering research and can be easily adapted for scarce samples like clinical biopsies.

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Development of vascularized nerve scaffold using perfusion-decellularization and recellularization

Cited by 22 publications

References 47 publications

In vivo performance of decellularized tracheal grafts in the reconstruction of long length tracheal defects: Experimental study

In vivo performance of decellularized tracheal grafts in the reconstruction of long length tracheal defects: Experimental study

Recellularization of Bioengineered Scaffolds for Vascular Composite Allotransplantation

Novel Decellularization Method for Tissue Slices

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