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Generation and evaluation of bioartificial substitutes for their use as advanced therapy in the surgical repair of critical lesions of the peripheral nerve
Spain (IFMSA Spain) - University of Granada, Granada
Department of Histology
Professor Antonio Campos Muñoz
Professor Victor Carriel
Type of Research Project
- Basic science
What is the background of the project?
Damaged nerves have a limited capacity for regeneration and, with substantial loss of nerve segment, a graft of nervous tissue is usually required. The gold standard of this treatment is an autologous transplant, although allogenic transplants have also been used with less success due to immune rejection and pathogen transmission. The transplants provide the cellular, extracellular and trophic factors necessary for the regeneration of the tissue, however, even the autotransplant brings with it a series of limitations: lack of donated material, morbidity, loss of sensitivity of the donor area. Given this situation, tissue engineering techniques have been used to develop different nerve substitutes combing different cell types and biomateriales. En recent years, our research group has designed, developed and evaluated different 3D artificial tissues, based on the use of fibrin-agarose (FA) hydrogels. These have shown to be of a high biocompatibility, in vitro differentiation and tissular regeneration in vivo. The research group has a recent, but productive experience basing our strategy on the incorporation of FA hydrogels, mesenquimal stem cells and neuropeptides.
What is the aim of the project?
1. Generate and evaluate new biomaterials that allow the reproduction of the neuroconductive perineural component and the endoneural component of the peripheral nerve. 2. Determine the therapeutic effectiveness in vivo of the new compound bioartifical substitutes generated in this project through the use of structural, ultrastructural, histologic, molecular, eletrophysiological, functional and clinical indicators.
What techniques and methods are used?
To create the different components, different types of hydrogels will be manafactured. The external perineural component materials will also be subjeceted to nanostructurization and Genipin crosslinking. Following surgical removal of 1cm of nerve in an animal model, effectiveness of reinnervation by these implants will be assessed by clinical evaluation for presence and size of neurotrophic ulcers, pinch test, determination of the nerve functional index, toe spread test, electromiography, analysis of the loss of muscle mass and volume, ultrastructural and histological (staining and immunohistochemistry) analysis.
What is the role of the student?
- The student will mainly observe
- The student will observe the practical experiments but will be highly involved in the analysis of the results
What are the tasks expected to be accomplished by the student?
The student will learn different techniques involved in tissular engineering, such as: • Management of cell cultures of human nerve cells and human mesenquimal cells. • Trypsinization, cell counting, analysis of cellular visibility, subcultures, freezing and criopreservation of cells. • Production of artifical tissues using the biomaterials fibrin and agarose and the process of nanostructurization. • Histological analysis of artifical nerves, Picrosirius staining for collagen, Alsian Blue staining for proteoglycans, Period acid-Schiff (PAS) staining for glycosaminoglycans.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
The student will be able to participate in the seminars that the group holds weekly. They will also have access to the department's bibliography.
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- No specific outcome is expected
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Knowledge of basic histology is necessary.
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students - Graduated students (less than 6 months) - Pre-Medical students from the American-British system - Students in biomedical fields - Dental medicine students (IADS members)
- Carriel V; Garzón I; Alaminos M; Campos A. Evaluation of myelin sheath and collagen reorganization pattern in a model of peripheral nerve regeneration using an integrated histochemical approach. Histochem Cell Biol. 2011 Dec;136(6):709-17. Epub 2011 Oct 29.
- Carriel V; Garrido-Gómez J; Hernández-Cortés P; Garzón I; García-García S; Sáez-Moreno JA; Del Carmen Sánchez-Quevedo M; Campos A; Alaminos M. Combination of fibrin-agarose hydrogels and adipose-derived mesenchymal stem cells for peripheral nerve regeneration. J Neural Eng. 2013 Apr;10(2):026022. doi: 10.1088/1741-2560/10/2/026022. Epub 2013 Mar 26
- Carriel V; Garzón I; Campos A; Cornelissen M; Alaminos M. Differential expression of GAP-43 and neurofilament during peripheral nerve regeneration through bio-artificial conduits. J Tissue Eng Regen Med. 2017 Feb;11(2):553-563. doi: 10.1002/term.1949. Epub 2014 Jul 31.
- Carriel V; Garzón I; Alaminos M; Cornelissen M. Histological assessment in peripheral nerve tissue engineering. Neural Regen Res. 2014 Sep 15;9(18):1657-60. doi: 10.4103/1673-5374.141798. Review.
- Carriel V; Scionti G; Campos F; Roda O; Castro B; Cornelissen M; Garzón I; Alaminos M. In vitro characterization of a nanostructured fibrin agarose bio-artificial nerve substitute. J Tissue Eng Regen Med. 2017 May;11(5):1412-1426. doi: 10.1002/term.2039. Epub 2015 Jul 14
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