Effetti della biostimolazione laser sul tessuto epiteliale per il differenziamento dello strato cheratinizzato: studio in vitro

Background: The overall view of gingival augmentation techniques proposed in the international literature does not exclude a surgical component. The planning of one or more surgical phases with the consequent post-surgical discomfort and results not always predictable are required when setting up periodontal flaps combined eventually with free gingival grafts. The introduction in the last years of laser biostimulation has led to a less invasive approach, particularly in the treatment of periodontally compromised patients, limiting the surgical phase to seriously compromised cases, with regeneration techniques for the restoration of a correct periodontal tissue’s anatomy. Aim of this in vitro study is to establish the validity of laser biostimulation in order to develop the epithelial tissue’s keratinized layer, by stimulating fibroblasts-keratinocytes organotypic cultures obtained and fibroblasts and keratinocytes mono-cultures. Materials and Methods: We created two groups (test and control) each one of them composed by 3 fibroblast cultures, 3 keratinocyte cultures and 3 organotypic cultures. We performed laser irradiation of test group at 50 J/cm2 of fluency with one application every 48 hours for a total of five applications. 48 hours from last laser application we investigated the presence and amount of keratins 5 and 8 by performing citofluorymetric and western blotting analyses. Results: the analyses results showed an increase in keratin synthesis of the test groups’ cultures, particularly showing a remarkable increase in production of keratin 8 by test co-cultures. Conclusions: laser biostimulation proved to considerably enhance keratin synthesis when applied with high energy doses and repeated applications to keratinocytes-fibroblasts co-cultures.