The effect of L.L.L.T. in the viability, proliferation and differentiation of premature osteoblasts, fibroblasts and in their co-culture

Postgraduate Thesis uoadl:2896678 217 Read counter

Unit:
Κατεύθυνση Περιοδοντολογία (Κλινικές Ειδικεύσεις)
Βιβλιοθήκη Οδοντιατρικής
Deposit date:
2020-02-05
Year:
2019
Author:
Bourouni Ioanna
Supervisors info:
Ιωάννης-Παναγιώτης Βρότσος, Καθηγητής, Τμήμα Οδοντιατρικής, Σχολή Επιστημών Υγείας, ΕΚΠΑ
Αλεξάνδρα Πανδή-Τσάμη, Καθηγήτρια, Τμήμα Οδοντιατρικής, Σχολή Επιστημών Υγείας, ΕΚΠΑ
Ιωάννης Καρούσης, Αναπληρωτής Καθηγητής, Τμήμα Οδοντιατρικής, Σχολή Επιστημών Υγείας, ΕΚΠΑ
Original Title:
Η επίδραση της L.L.L.T. στη βιωσιμότητα, πολλαπλασιασμό και τη διαφοροποίηση πρώιμων οστεοβλαστών, ουλικών ινοβλαστών και σε συνκαλλιέργειά τους
Languages:
Greek
Translated title:
The effect of L.L.L.T. in the viability, proliferation and differentiation of premature osteoblasts, fibroblasts and in their co-culture
Summary:
Introduction: Laser devices, after their introduction, find a wide range of applications in Health Sciences. Specifically in Dentistry, they can be used with excellent results in many dental specialties, such as, Restorative Dentistry, Maxillofacial Surgery, Stomatology, Endodontics and Periodontology. There are two main ways of operating these devices. First, they are applied for surgical purposes, where the laser devices are set to high power in order to cause thermomechanical ablation used for incisions and hard or soft tissue removal. Second, they are used for their bio-stimulatory capacity, in low power mode. In particular, Low Level Laser Therapy (LLLT), has been used for many years in a variety of medical and dental applications. Regarding laser activity, it is presented by photobiological phenomena, associated with the reaction of organisms to monochromatic laser radiation. Radiation photons, cause intracellular changes in cells (mainly in mitochondria), and this affects their mimetics, but also the secretion of several growth factors. However, the precise biological mechanisms that interpret the biological effect of LLLT have not yet been established. The radiation applied with LLLT using Diode Laser has wavelength near the red and the infrared area of the spectrum. This means that radiation with such a wavelength can activate the mitochondrial respiratory chain and initiate a series of reactions that ultimately will favor cell proliferation and the production of growth factors. Studies related to the effect of LLLT on osteoblasts and human gingival fibroblasts are limited. However, it appears that the application of LLLT to these cells has a positive effect in terms of proliferation and differentiation. To date, there is no study conducted to examine the effect of LLLT with a Diode laser (810nm) on immature osteoblasts and human gingival fibroblasts.
Aim: The aim of this study was to evaluate the effect of LLLT with Diode laser (810nm) on osteoblasts, human gingival fibroblasts and their co-culture, in terms of viability, proliferation and differentiation.
Materials and Methods: Primary cultures of human gingival fibroblasts (hGF), cultures of immature osteoblasts (MG63) and their co-cultures, were irradiated by a Diode laser (810nm) with a density of 12J/cm2. Controls were not irradiated. Cultures were examined for viability, cell proliferation and differentiation after 24, 48 and 72 hours. Statistical analysis was performed using graph pad prism. The cell viability after irradiation was calculated by the MTT test and the cell apoptosis was calculated by FDI/PI. The quantification of gene expression of ALP and COL1a was performed using Real Time PCR.
Results: In all laser-irradiaton groups was observed increased cell proliferation with respect to the control group. Statistically significant differences (p<0.001) were observed for the specific energy densities in all experimental groups in the first 24 hours. Also, at 48 hours, a statistically significant difference (p<0.01) was found for both irradiated and not irradiated hGF, as well as for co-cultures. A statistically significant difference (p<0.05) is observed for the experimental group of early osteoblasts. Furthermore, a statistically significant difference (p<0.0001) was observed for all experimental groups 72 hours after radiation. Finally, during gene expression of ALP for the radiating experimental groups, 7 days after irradiation, a statistically significant superiority was observed for early osteoblasts (p<0.005). Also, a statistically significant superiority was observed for the co-cultures (p<0.005) on the gene expression of COL1a for the experimental groups.
Conclusion: The LLLT application with Diode laser (810nm) seems to have a positive effect on the proliferation of osteoblasts, human gingival fibroblasts and their co-cultures, which was evident at 24,48 and 72 hours after irradiation. Finally, in the present study there was a significant increase in the production of ALP and COL1a from the cells mentioned above.
Main subject category:
Health Sciences
Keywords:
Differentiation, Gene expression, Low-level laser therapy, Gingival fibroblasts, Osteoblasts, Diode, Co-culture
Index:
No
Number of index pages:
0
Contains images:
Yes
Number of references:
211
Number of pages:
162
File:
File access is restricted only to the intranet of UoA.

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