This thesis aims to deepen our knowledge of the strategies used for whole tooth regeneration, which include dental tissue engineering, stimulation of third dentition formation, cell-tissue recombination, and gene-manipulated tooth regeneration. In doing so, a more specific focus is placed on adult human dental pulp stem cells whose regenerative potential declines with age. Our study investigated if it were possible to rejuvenate aged human DPSCs (hDPSCs) using the microenvironment of young hDPSCs. To accomplish this, hDPSCs from young donors (YDPSCs) and old donors (ODPSCs) were first isolated, cultured and characterized. Following the collection of their respective conditioned medium, YDPSCs were treated with conditioned medium (CM) derived from ODPSCs (YDPSCs-OCM), ODPSCs treated with CM of YDPSCs (ODPSCs-YCM), YDPSCs replenished with basal medium (YDPSCs-DMEM/F12) and ODPSCs replenished with basal medium (ODPSCs-DMEM/F12). Each group was evaluated based on morphologic changes, proliferation, apoptosis, senescence and oxidative stress. The trophic factors secreted by the young CM exerted their rejuvenating effect via its antioxidative potential. Young CM significantly reduced ROS and superoxide generation in aged hDPSCs. Young CM also decreased nuclear DNA fragmentation in aged hDPSCs via a caspase-independent pathway. The antiproliferative and antisenescence effect of the conditioned medium derived from young hDPSCs on aged hDPSCs was not evidenced. Our results demonstrate that the CM of young hDPSCs may not rejuvenate aged hDPSCs fully and will require further investigation.
Questa tesi mira ad approfondire la nostra conoscenza delle strategie utilizzate per la rigenerazione del dente intero, che includono la "dental tissue engineering", la stimolazione della formazione della terza dentizione, la ricombinazione cellulo-tissutale e la rigenerazione del dente tramite manipolazione genica. A tal fine, un'attenzione più specifica viene posta sulle cellule staminali della polpa dentale adulta il cui potenziale rigenerativo diminuisce con l'età. Il nostro studio ha ulteriormente indagato se fosse possibile ringiovanire le cellule staminali della polpa dentale umani (hDPSC) invecchiate utilizzando il microambiente delle hDPSC giovani. A questo scopo, le hDPSC di donatori giovani (YDPSC) e anziani (ODPSC) sono state dapprima isolate, coltivate e caratterizzate. Dopo la raccolta dei rispettivi terreni condizionati (CM), le YDPSC sono state trattate con CM derivato da ODPSC (YDPSCs-OCM), ODPSC trattate con CM di YDPSC (ODPSCs-YCM), YDPSC coltivate in terreno basale (YDPSCs-DMEM/F12) e ODPSC coltivate in terreno basale (ODPSCs-DMEM/F12). Ogni gruppo è stato valutato sulla base di cambiamenti morfologici, proliferazione, apoptosi, senescenza e stress ossidativo. I fattori trofici secreti nel giovane CM hanno esercitato il loro effetto ringiovanente attraverso il suo potenziale antiossidante. Il giovane CM ha ridotto significativamente la generazione di ROS e superossido negli ODPSC. Il CM da YDPSC ha anche ridotto la frammentazione del DNA nucleare negli ODPSC invecchiati attraverso un percorso caspasi-indipendente. L'effetto antiproliferativo e anti-senescenza del CM da YDPSC su ODPSC non sono stati evidenziati. I nostri risultati hanno dimostrato che il CM degli YDPSC potrebbe non ringiovanire completamente gli ODPSC e richiederà ulteriori indagini.
Strategies for whole tooth regeneration and possible rejuvenation of adult dental pulp stem cells / Hosein, Andrell. - (2023 Jun 05).
Strategies for whole tooth regeneration and possible rejuvenation of adult dental pulp stem cells.
HOSEIN, ANDRELL
2023-06-05
Abstract
This thesis aims to deepen our knowledge of the strategies used for whole tooth regeneration, which include dental tissue engineering, stimulation of third dentition formation, cell-tissue recombination, and gene-manipulated tooth regeneration. In doing so, a more specific focus is placed on adult human dental pulp stem cells whose regenerative potential declines with age. Our study investigated if it were possible to rejuvenate aged human DPSCs (hDPSCs) using the microenvironment of young hDPSCs. To accomplish this, hDPSCs from young donors (YDPSCs) and old donors (ODPSCs) were first isolated, cultured and characterized. Following the collection of their respective conditioned medium, YDPSCs were treated with conditioned medium (CM) derived from ODPSCs (YDPSCs-OCM), ODPSCs treated with CM of YDPSCs (ODPSCs-YCM), YDPSCs replenished with basal medium (YDPSCs-DMEM/F12) and ODPSCs replenished with basal medium (ODPSCs-DMEM/F12). Each group was evaluated based on morphologic changes, proliferation, apoptosis, senescence and oxidative stress. The trophic factors secreted by the young CM exerted their rejuvenating effect via its antioxidative potential. Young CM significantly reduced ROS and superoxide generation in aged hDPSCs. Young CM also decreased nuclear DNA fragmentation in aged hDPSCs via a caspase-independent pathway. The antiproliferative and antisenescence effect of the conditioned medium derived from young hDPSCs on aged hDPSCs was not evidenced. Our results demonstrate that the CM of young hDPSCs may not rejuvenate aged hDPSCs fully and will require further investigation.File | Dimensione | Formato | |
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