Use of hybrid chitosan membranes and N1E-115 cells for promoting nerve regeneration in an axonotmesis rat model

Amado, Sandra; Simões, Maria João; Armada-da-Silva, Paulo; Luís, Ana Lúcia; Shirosaki, Yuki; Lopes, M. A.; Santos, J. D.; Fregnan, Federica; Gambarotta, Giovanna; Raimondo, Stefania; Fornaro, Michele; Veloso, António; Varejão, Artur S.P.; Maurício, Ana Colette; Geuna, Stefano

doi:10.1016/j.biomaterials.2008.07.043

Cited by 109 publications

(118 citation statements)

References 53 publications

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“…Intracellular free Ca2 þ concentration ([Ca2 þ ]i) was measured in Fura-2-loaded cells by using dual wavelength spectrofluorometry as previously described ( Amado et al, 2008). The measurements were performed on undifferentiated HMSCs after confluence was obtained and on neuroglial-differentiated HMSCs, cultured on Vivosorbs discs in order to correlate the HMSCs ability to differ-entiate and survival capacity in the presence of the Vivosorbs membrane.…”

Section: Cell Culture and In Vitro Differentiation Of Hmsc From Whartmentioning

confidence: 99%

“…Previous results obtained by our research group using N1E-115 cells in vitro differentiated into neuroglial-like cells to promote regeneration of axonotmesis and neurotm-esis lesions in the rat model showed that there was no significant effect in promoting axon regeneration and, when N1E-115 cells were cultured inside a PLGA scaffold used to bridge a nerve defect, they can even exert negative effects on nerve fiber regeneration ( Luis et al, 2008a;Maurı´cio et al, 2011). The presence of transplanted N1E-115 cells in nerve scaffolds com-peting for the local blood supply of nutrients and oxygen and by space-occupying effect could have hindered the positive effect of local neurotrophic factor release leading a negative outcome on nerve regeneration ( Amado et al, 2008Luis et al, 2008a;Maurı´cio et al, 2011). Thus, N1E-115 cells did not prove to be a suitable candidate cellular system for treatment of nerve injury after axonotmesis and neurotmesis ( Amado et al, 2008Luis et al, 2008a;Maurı´cio et al, 2011) and their application is limited only to research purposes as a basic scientific step for the development of other cell delivery systems, due to its neoplastic origin.…”

Section: Groupmentioning

confidence: 99%

“…The presence of transplanted N1E-115 cells in nerve scaffolds com-peting for the local blood supply of nutrients and oxygen and by space-occupying effect could have hindered the positive effect of local neurotrophic factor release leading a negative outcome on nerve regeneration ( Amado et al, 2008Luis et al, 2008a;Maurı´cio et al, 2011). Thus, N1E-115 cells did not prove to be a suitable candidate cellular system for treatment of nerve injury after axonotmesis and neurotmesis ( Amado et al, 2008Luis et al, 2008a;Maurı´cio et al, 2011) and their application is limited only to research purposes as a basic scientific step for the development of other cell delivery systems, due to its neoplastic origin. In this study, we used a PLC membrane to deliver undifferentiated and in vitro differentiated HMSCs from the UC Wharton jelly and we compared this delivery approach with direct infiltration of these undifferentiated HMSCs in suspension, in the rat sciatic nerve axonotmesis model.…”

Section: Groupmentioning

confidence: 99%

“…Because of its peripheral nerve size, the rat sciatic nerve has been the most commonly experimental model used in studies concerning the peripheral nerve regeneration and possible thera-peutic approaches . Although sciatic nerve injuries are rare in humans, this experimental model provides a very realistic testing bench for lesions involving plurifascicular mixed nerves with axons of different size and type competing to reach and reinnervate distal targets ( Amado et al, 2008;Mackinnon et al, 1985). Focal crush causes axonal interruption but preserves the connective sheaths (axonotmesis).…”

Section: Introductionmentioning

confidence: 99%

“…Tissue engineering of peripheral nerves associates biomaterials, like chitosan, PLC, and other biomaterials, some of them, previously studied by our group ( Amado et al, 2008;Luis et al, 2008a;Maurı´cio et al, 2011) to cellular systems, able to differentiate into neuroglial like cells or even by modulating the inflammatory process, which might improve nerve regeneration, in terms of motor and sensory recovery, and also, by shortening the healing period avoiding regional muscular atrophy. Cell transplantation has been proposed as a method of improving peripheral nerve regeneration ( Chen et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

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Use of poly(DL-lactide-ε-caprolactone) membranes and mesenchymal stem cells from the Wharton's jelly of the umbilical cord for promoting nerve regeneration in axonotmesis: In vitro and in vivo analysis

et al. 2012

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Cellular systems implanted into an injured nerve may produce growth factors or extracellular matrix molecules, modulate the inflammatory process and eventually improve nerve regeneration. In the present study, we evaluated the therapeutic value of human umbilical cord matrix MSCs (HMSCs) on rat sciatic nerve after axonotmesis injury associated to Vivosorbs membrane. During HMSCs expansion and differentiation in neuroglial-like cells, the culture medium was collected at 48, 72 and 96 h for nuclear magnetic resonance (NMR) analysis in order to evaluate the metabolic profile. To correlate the HMSCs ability to differentiate and survival capacity in the presence of the Vivosorbs membrane, the [Ca 2þ ]i of undifferentiated HMSCs or neuroglial-differentiated HMSCs was determined by the epifluorescence technique using the Fura-2AM probe. The Vivosorbs membrane proved to be adequate and used as scaffold associated with undiffer-entiated HMSCs or neuroglial-differentiated HMSCs. In vivo testing was carried out in adult rats where a sciatic nerve axonotmesis injury was treated with undifferentiated HMSCs or neuroglial differentiated HMSCs with or without the Vivosorbs membrane. Motor and sensory functional recovery was evaluated throughout a healing period of 12 weeks using sciatic functional index (SFI), extensor postural thrust (EPT), and withdrawal reflex latency (WRL).Stereological analysis was carried out on regenerated nerve fibers. In vitro investigation showed the formation of typical neuroglial cells after differentiation, which were positively stained for the typical specific neuroglial markers such as the GFAP, the GAP-43 and NeuN.NMR showed clear evidence that HMSCs expansion is glycolysis-dependent but their differentiation requires the switch of the metabolic profile to oxidative metabolism. In vivo studies showed enhanced recovery of motor and sensory function in animals treated with transplanted undifferentiated and differentiated HMSCs that was accompanied by an increase in myelin sheath. Taken together, HMSC from the umbilical cord Wharton jelly might be useful for improving the clinical outcome after peripheral nerve lesion.

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Section: Cell Culture and In Vitro Differentiation Of Hmsc From Whartmentioning

confidence: 99%

Section: Groupmentioning

confidence: 99%

Section: Groupmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Use of poly(DL-lactide-ε-caprolactone) membranes and mesenchymal stem cells from the Wharton's jelly of the umbilical cord for promoting nerve regeneration in axonotmesis: In vitro and in vivo analysis

et al. 2012

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Biocompatibility and hemocompatibility of polyvinyl alcohol hydrogel used for vascular grafting-In vitroandin vivostudies

Alexandre

Ribeiro

Gärtner

et al. 2014

J. Biomed. Mater. Res.

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Polyvinyl alcohol hydrogel (PVA) is a synthetic polymer with an increasing application in the biomedical field that can potentially be used for vascular grafting. However, the tissue and blood-material interactions of such gels and membranes are unknown in detail. The objectives of this study were to: (a) assess the biocompatibility and (b) hemocompatibility of PVA-based membranes in order to get some insight into its potential use as a vascular graft. PVA was evaluated isolated or in copolymerization with dextran (DX), a biopolymer with known effects in blood coagulation homeostasis. The effects of the mesenchymal stem cells (MSCs) isolated from the umbilical cord Wharton's jelly in the improvement of PVA biocompatibility and in the vascular regeneration were also assessed. The biocompatibility of PVA was evaluated by the implantation of membranes in subcutaneous tissue using an animal model (sheep). Histological samples were assessed and the biological response parameters such as polymorphonuclear neutrophilic leucocytes and macrophage scoring evaluated in the implant/tissue interface by International Standards Office (ISO) Standard 10993-6 (annex E). According to the scoring system based on those parameters, a total value was obtained for each animal and for each experimental group. The in vitro hemocompatibility studies included the classic hemolysis assay and both human and sheep bloods were used. Relatively to biocompatibility results, PVA was slightly irritant to the surrounding tissues; PVA-DX or PVA plus MSCs groups presented the lowest score according to ISO Standard 10993-6. Also, PVA was considered a nonhemolytic biomaterial, presenting the lowest values for hemolysis when associated to DX.

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Evaluation of PVA biodegradable electric conductive membranes for nerve regeneration in axonotmesis injuries: the rat sciatic nerve animal model

Ribeiro

Caseiro

Pereira

et al. 2017

J Biomedical Materials Res

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The therapeutic effect of three polyvinyl alcohol (PVA) membranes loaded with electrically conductive materials - carbon nanotubes (PVA-CNTs) and polypyrrole (PVA-PPy) - were tested in vivo for neuro-muscular regeneration after an axonotmesis injury in the rat sciatic nerve. The membranes electrical conductivity measured was 1.5 ± 0.5 × 10 S/m, 579 ± 0.6 × 10 S/m, and 1837.5 ± 0.7 × 10 S/m, respectively. At week-12, a residual motor and nociceptive deficit were present in all treated groups, but at week-12, a better recovery to normal gait pattern of the PVA-CNTs and PVA-PPy treated groups was observed. Morphometrical analysis demonstrated that PVA-CNTs group presented higher myelin thickness and lower g-ratio. The tibialis anterior muscle, in the PVA-PPy and PVA-CNTs groups showed a 9% and 19% increase of average fiber size area and a 5% and 10% increase of the "minimal Feret's diameter," respectively. No inflammation, degeneration, fibrosis or necrosis were detected in lung, liver, kidneys, spleen, and regional lymph nodes and absence of carbon deposits was confirmed with Von Kossa and Masson-Fontana stains. In conclusion, the membranes of PVA-CNTs and PVA-PPy are biocompatible and have electrical conductivity. The higher electrical conductivity measured in PVA-CNTs membrane might be responsible for the positive results on maturation of myelinated fibers. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1267-1280, 2017.

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Use of hybrid chitosan membranes and N1E-115 cells for promoting nerve regeneration in an axonotmesis rat model

Cited by 109 publications

References 53 publications

Use of poly(DL-lactide-ε-caprolactone) membranes and mesenchymal stem cells from the Wharton's jelly of the umbilical cord for promoting nerve regeneration in axonotmesis: In vitro and in vivo analysis

Use of poly(DL-lactide-ε-caprolactone) membranes and mesenchymal stem cells from the Wharton's jelly of the umbilical cord for promoting nerve regeneration in axonotmesis: In vitro and in vivo analysis

Biocompatibility and hemocompatibility of polyvinyl alcohol hydrogel used for vascular grafting-In vitroandin vivostudies

Evaluation of PVA biodegradable electric conductive membranes for nerve regeneration in axonotmesis injuries: the rat sciatic nerve animal model

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