Comparison of Decellularization Protocols to Generate Peripheral Nerve Grafts: A Study on Rat Sciatic Nerves

Soury, Marwa El; García-García, Óscar Darío; Moretti, Matteo; Perroteau, Isabelle; Raimondo, Stefania; Lovati, Arianna B.; Carriel, Víctor

doi:10.3390/ijms22052389

Cited by 22 publications

(28 citation statements)

References 42 publications

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“…After functionalization, SIRC and hADSCs were trypsinized and carefully seeded on the surface of the DL (170,000 cells per cm 2 of sample). To promote attachment, cells were resuspended in a minimal amount of medium (50 µL) and DLs were immobilized using agarose casts, as previously reported [38]. In order to induce epithelial differentiation of both cell types seeded on the RL, these tissues were cultured for 21 days in EM epithelial differentiation medium containing epithelial growth and differentiation factors, as previously described [39].…”

Section: Generation Of Recellularized Limbal Substitutes By Tissue Engineeringmentioning

confidence: 99%

Generation of a Biomimetic Substitute of the Corneal Limbus Using Decellularized Scaffolds

et al. 2021

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Patients with severe limbal damage and limbal stem cell deficiency are a therapeutic challenge. We evaluated four decellularization protocols applied to the full-thickness and half-thickness porcine limbus, and we used two cell types to recellularize the decellularized limbi. The results demonstrated that all protocols achieved efficient decellularization. However, the method that best preserved the transparency and composition of the limbus extracellular matrix was the use of 0.1% SDS applied to the half-thickness limbus. Recellularization with the limbal epithelial cell line SIRC and human adipose-derived mesenchymal stem cells (hADSCs) was able to generate a stratified epithelium able to express the limbal markers p63, pancytokeratin, and crystallin Z from day 7 in the case of SIRC and after 14–21 days of induction when hADSCs were used. Laminin and collagen IV expression was detected at the basal lamina of both cell types at days 14 and 21 of follow-up. Compared with control native limbi, tissues recellularized with SIRC showed adequate picrosirius red and alcian blue staining intensity, whereas limbi containing hADSCs showed normal collagen staining intensity. These preliminary results suggested that the limbal substitutes generated in this work share important similarities with the native limbus and could be potentially useful in the future.

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Section: Generation Of Recellularized Limbal Substitutes By Tissue Engineeringmentioning

confidence: 99%

Generation of a Biomimetic Substitute of the Corneal Limbus Using Decellularized Scaffolds

et al. 2021

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“…This glycoprotein supports Schwann cell migration and proliferation, being an essential substrate for the Büngner band formation process (Chernousov et al, 2008;Jessen and Mirsky, 2016). Here, an abundant and properly distributed preservation of laminin was successfully achieved with the three new decellularization techniques, being these results considerably better than SD group and comparable to use of similar procedures (El Soury et al, 2021;Garcia-Garcia et al, 2021). Finally, the wellpreservation of the ECM obtained with P1, P2 and SD, as well as the alterations in P3, were well-confirmed by ultrastructural analyses (SEM and TEM).…”

Section: Discussionmentioning

confidence: 74%

“…and atropine (Pfizer, New York, NY, United States, 0.05 μg/g body weight) and then euthanized by an intraperitoneal injection of Eutanex ® solution (0.5 mL/animal). Subsequently, ~3 cm of both sciatic nerves (n = 70 nerves) were removed and cryopreserved (10% DMSO in fetal bovine serum at −80 °C) until use as previously described (Chato-Astrain et al, 2020b;El Soury et al, 2021;Garcia-Garcia et al, 2021;Contreras et al, 2022).…”

Section: Nerve Isolation and Chemical Decellularizationmentioning

confidence: 99%

Comprehensive ex vivo and in vivo preclinical evaluation of novel chemo enzymatic decellularized peripheral nerve allografts

García-García

Soury²,

Campos³

et al. 2023

Front. Bioeng. Biotechnol.

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As a reliable alternative to autografts, decellularized peripheral nerve allografts (DPNAs) should mimic the complex microstructure of native nerves and be immunogenically compatible. Nevertheless, there is a current lack of decellularization methods able to remove peripheral nerve cells without significantly altering the nerve extracellular matrix (ECM). The aims of this study are firstly to characterize ex vivo, in a histological, biochemical, biomechanical and ultrastructural way, three novel chemical-enzymatic decellularization protocols (P1, P2 and P3) in rat sciatic nerves and compared with the Sondell classic decellularization method and then, to select the most promising DPNAs to be tested in vivo. All the DPNAs generated present an efficient removal of the cellular material and myelin, while preserving the laminin and collagen network of the ECM (except P3) and were free from any significant alterations in the biomechanical parameters and biocompatibility properties. Then, P1 and P2 were selected to evaluate their regenerative effectivity and were compared with Sondell and autograft techniques in an in vivo model of sciatic defect with a 10-mm gap, after 15 weeks of follow-up. All study groups showed a partial motor and sensory recovery that were in correlation with the histological, histomorphometrical and ultrastructural analyses of nerve regeneration, being P2 the protocol showing the most similar results to the autograft control group.

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“…Although SDS and SDC are the most common ionic detergents used for decellularization [ 31 ], we also tested the efficacy of the organic solvent tri-n-butyl phosphate (TnBP), which is known to dissolve lipids while minimally affecting proteins [ 32 ]. Nerve tissue is rich in lipids, mainly in a form of myelin produced by Schwann cells, and a recent study shows that TnBP is more effective than SDS for decellularization of the rat sciatic nerve [ 33 ]. Our data suggested that TnBP did not efficiently remove growth-inhibitory MAG [ 34 ] from the nerve tissue, and the resulting P3 hydrogels induced apoptosis of the Schwann cells in 3D cultures.…”

Section: Discussionmentioning

confidence: 99%

Efficacy of Nerve-Derived Hydrogels to Promote Axon Regeneration Is Influenced by the Method of Tissue Decellularization

Kuna

Lundgren

Anerillas

et al. 2022

IJMS

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Injuries to large peripheral nerves are often associated with tissue defects and require reconstruction using autologous nerve grafts, which have limited availability and result in donor site morbidity. Peripheral nerve-derived hydrogels could potentially supplement or even replace these grafts. In this study, three decellularization protocols based on the ionic detergents sodium dodecyl sulfate (P1) and sodium deoxycholate (P2), or the organic solvent tri-n-butyl phosphate (P3), were used to prepare hydrogels. All protocols resulted in significantly decreased amounts of genomic DNA, but the P2 hydrogel showed the best preservation of extracellular matrix proteins, cytokines, and chemokines, and reduced levels of sulfated glycosaminoglycans. In vitro P1 and P2 hydrogels supported Schwann cell viability, secretion of VEGF, and neurite outgrowth. Surgical repair of a 10 mm-long rat sciatic nerve gap was performed by implantation of tubular polycaprolactone conduits filled with hydrogels followed by analyses using diffusion tensor imaging and immunostaining for neuronal and glial markers. The results demonstrated that the P2 hydrogel considerably increased the number of axons and the distance of regeneration into the distal nerve stump. In summary, the method used to decellularize nerve tissue affects the efficacy of the resulting hydrogels to support regeneration after nerve injury.

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Comparison of Decellularization Protocols to Generate Peripheral Nerve Grafts: A Study on Rat Sciatic Nerves

Cited by 22 publications

References 42 publications

Generation of a Biomimetic Substitute of the Corneal Limbus Using Decellularized Scaffolds

Generation of a Biomimetic Substitute of the Corneal Limbus Using Decellularized Scaffolds

Comprehensive ex vivo and in vivo preclinical evaluation of novel chemo enzymatic decellularized peripheral nerve allografts

Efficacy of Nerve-Derived Hydrogels to Promote Axon Regeneration Is Influenced by the Method of Tissue Decellularization

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