FROM IN VITRO STUDIES TO A LARGE ANIMAL MODEL: A MULTISTEP DISSECTION ON THE FUTURE ROLE OF ADIPOSE-DERIVED STEM CELLS FOR MUSCULOSKELETAL TISSUE ENGINEERING.
Tesi di Dottorato
Data di Pubblicazione:
2014
Citazione:
FROM IN VITRO STUDIES TO A LARGE ANIMAL MODEL: A MULTISTEP DISSECTION ON THE FUTURE ROLE OF ADIPOSE-DERIVED STEM CELLS FOR MUSCULOSKELETAL TISSUE ENGINEERING / S. Niada ; tutor: A.T. Brini ; coordinatore: A. E. Panerai. DIPARTIMENTO DI SCIENZE BIOMEDICHE, CHIRURGICHE ED ODONTOIATRICHE, 2014 Jan 16. 26. ciclo, Anno Accademico 2013. [10.13130/niada-stefania_phd2014-01-16].
Abstract:
Tissue engineering is an emerging interdisciplinary field, born with the purpose to provide an alternative solution for the regeneration of lesioned or lost tissues, combining cells, biocompatible scaffolds and bioactive factors. The cells for this approach should be non-immunoreactive and non-tumorigenic. Moreover, they should be available in large amount and possess, or be able to acquire, a specific protein expression pattern similar to that of the damaged tissue and/or act as a pool of trophic factors for resident cells. All these reasons, make mesenchymal stem cells (MSCs) good candidates for applications in regenerative medicine. Although bone marrow is still the most common source of MSCs, these cells could be harvested from all vascularised tissues, and, interestingly, from tissues that are normally discarded, such as fat, placenta or umbilical cord. One of the most convenient source of MSCs, is unequivocally, the adipose tissue due to the easily accessible anatomical location and the abundance of subcutaneous adipose tissue. Adipose-derived stem cells (ASCs) are similar to MSCs isolated from bone marrow in morphology, immunophenotype, and differentiation ability, and own interesting features such as immunoregolatory and anti-inflammatory properties.
In the recent years, many strategies for the cure of musculoskeletal tissues critical lesions, mainly in orthopaedic, oral and maxillo-facial surgery, have been under investigations. In this contest, the regeneration of structures including different tissues, such as the periodontium and the osteochondral unit, are particularly challenging.
Periodontal regeneration is especially demanding, as it requires regeneration of three quite diverse and unique tissues such as the alveolar bone, the periodontal ligament and the cementum, that have to interface with each other to restore their complex structure. Since the promising results obtained with ASCs in preclinical studies of periodontal diseases arouse the curiosity of maxillofacial and dental surgeons, we decided to identify a novel source of ASCs, i.e, the buccal fat pad, convenient for these specialists. For this purpose, we studied human adipose derived-stem cells from buccal fat pad (BFP-ASCs), comparing them with cells from the subcutaneous adipose tissue (SC-ASCs) of the same donor (n=2). In parallel, considering the need for preclinical studies in which the effect of allogenic cells should be tested, and swine as an accepted animal model in tissue engineering applications, we also characterized porcine cells (n=6). With preclinical and clinical application prospective, we also investigated ASC interactions with oral tissues, natural and synthetic scaffolds and Amelogenin, an oral bioactive molecule. First of all, we showed that it is feasible to isolate ASCs even starting from very limited amounts of tissue (0,5 ml) and that the cellular yield is influenced by species, but not by the site of harvesting (1.1x105±1.4x104 human BFP-ASCs/ml and 1.15x105±7.1x103 human SC-ASCs/ml; 3.0x104±9.3x103 porcine BFP-ASCs/ml and 5.5x104±3.3x104 porcine SC-ASCs/ml). Despite the lower yield, the pASCs great proliferation rate allows to obtain high number of cells (potentially, 108 - 109) after few (3, 4) passages in culture. After the isolation, a great amount of cells deriving from all the tissues, adhered to cell culture plates showing the MSC fibroblast like morphology, with only mild shape differences constituted by the higher elongation and dimension of human SC-ASCs. Moreover, all the cells are easily expandable and showed good clonogenic ability at early passages. Cells of the same species, from both the harvesting site, displayed the same surface markers profile, that, in particular for human ASCs, was the typical one of hM
Tipologia IRIS:
Tesi di dottorato
Keywords:
mesenchymal stem cells ; buccal fat pad ; periodontal disease ; osteochondral defect ; tissue engineering ; large animal model
Elenco autori:
S. Niada
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