The effect of injectable platelet-rich fibrin in the initial therapy of chronic periodontitis

Parodontitis je hronično, inflamatorno i destruktivno oboljenje svih potpornih tkiva zuba: gingive, cementa korena zuba, periodontalnog ligamenta i alveolarne kosti. Inflamacija započinje u gingivi, a potom se, kako bolest napreduje, širi u dublja parodontalna tkiva. Kao posledica javlja se resorpcija alveolarne kosti koja može dovesti do gubitka zuba. Epidemiološki podaci iz čitavog sveta ukazuju na izuzetnu rasprostranjenost oboljenja potpornog aparata zuba – gingivitisa i parodontitisa. Pored eliminacije osnovnog etiološkog faktora, dentalnog plaka i inflamacije parodoncijuma, cilj inicijalne terapije (engl. scaling and root planing-SRP) je da se ostvari delimična ili potpuna regeneracija svih izgubljenih parodontalnih tkiva. Kako bi se unapredila SRP terapija mogu joj se pridotati pomoćni terapijski postupci. Regeneraciju kako mekih, tako i čvrstih tkiva parodoncijuma odlikuje kompleksna interakcija brojnih intracelularnih i ekstracelularnih signalnih mehanizama. Poznato je da trombociti kao rezervoari velikog broja faktora rasta i drugih bioaktivnih molekula, imaju značajnu ulogu u procesu zarastanja rana. Tečni oblik firbrina bogatog trombocititma, injektabilni fibrin bogat trombocitima (engl. injectable platelet-rich fibrin – i-PRF) predstavlja revolucionarni korak u terapijskom konceptu aktivne regeneracije tkiva. Ovaj autologi trombocitni koncentrat se dobija pažljivim centrifugiranjem venske krvi, bez dodavanja antikoagulantnih sredstava ili drugih biohemijski aktivnih suspstanci. Postepenim oslobađanjem bioaktivnih molekula porekla trombocita i leukocita zarobljenih u gustoj fibrinskoj mreži i-PRF -a, dolazi do pokretanja procesa regeneracije; sinteze DNK, diferencijacije pluripotentnih matičnih ćelija, proliferacije i migracije ćelija, hemotakse polimorfonuklearnih leukocita, kao i sinteze ekstracelularnog matriksa i angiogeneze. Cilj ove studije je bio da se ispita efekat aplikacije i-PRF-a u toku SRP tretmana u vidu uticaja na smanjenje inflamacije tkiva, koncentracije medijatora zapaljenja i mikroorganizama u sulkusnoj tečnosti (engl. gingival crevicular fluid – GCF), kao i stepena uspešnosti procesa regeneracije. Kod pacijenata obolelih od hroničnog parodontitisa uključenih u studiju, evidentirani su bili klinički parametri: dubina sondiranja (engl. periodontal pocket depth – PPD) , nivo ivice gingice (engl. level of gingival margin – LGM), nivo pripojnog epitela (engl. clinical attachment level – CAL), krvarenje na provokaciju (engl. bleeding on probing – BOP) i plak indeks (engl. plaque index – PI). Uzorci GCF uzimani papirnim poenima (Periopaper, Amityville, NY, USA) su čuvani na -70 °C. Koncentracije parodontopatogenih mikroorganizama: Aggregatibacter actinomycetemcomitans (A. a.), Porphyromonas gingivalis (P. g.), Prevotella intermedia (P. i.) i Tannerella forsythia (T. f.) su analizirane ili određivane primenom lančane reakcije polimeraze (engl. polymerase chain reaction, PCR) nakon dodavanja reverznih visoko specifičnih prajmera i univerzalnog „forward“ prajmera za 16s rRNK. Takođe, ukupna koncentracija mikroorganizama izračunavana je qPCR-om u realnom vremenu (engl. PCR-real time). Uzorkovanje GCF korišćeno je i za utvrđivanje nivoa matriksne-metaloproteinaze-8 (MMP-8), faktora nekroze tumora-α (TNF-α) i alkalne fosfataze (ALP). Analiza njihove koncentracije u ovoj tečnosti je sprovodena upotrebom odgovarajućih komercijalnih ELISA (engl. enzyme-linked immunosorbent assay) kompleta. Statistički paket za društvene nauke (softverski paket SPSS, verzija 20.0; SPSS Inc., Čikago, IL, SAD) korišćen je za analizu svih podataka...Periodontitis is a chronic, inflammatory and destructive disease of all tooth-supporting tissues: gingiva, root cementum, periodontal ligament, and alveolar bone. The inflammation begins in the gingiva. Afterwards, as the disease progresses, it spreads to deeper periodontal tissues. As the consequence, the resorption of the alveolar bone occurs which can lead to tooth loss. Epidemiological data from all over the world indicate exceptional prevalence of gingivitis and periodontitis. In addition to the elimination of the primary etiological factor - dental plaque and inflammation of the periodontium, the goal of the initial therapy (scaling and root planing-SRP) is to achieve partial or full regeneration of all lost periodontal tissues. In order to improve SRP therapy, adjunct therapeutic procedures can be included. The regeneration of both soft and hard periodontal tissues is characterized by a complex interaction of numerous intracellular and extracellular signalling mechanisms. Platelets are known to have a significant effect on wound healing, as reservoirs of a large number of growth factors and other bioactive molecules. The liquid form of platelet-rich fibrin, injectable platelet-rich fibrin – i-PRF, represents a revolutionary step in the therapeutic concept of active tissue regeneration. This autologous platelet concentrate is obtained by careful centrifugation of venous blood, without the addition of anticoagulants or other biochemically active substances. With the gradual release of bioactive molecules of platelet and leukocyte origin, trapped in the dense fibrin network of i-PRF, the regeneration process is initiated, such as DNA synthesis, pluripotent stem cell differentiation, cell proliferation and migration, polymorphonuclear leukocyte chemotaxis, along with the extracellular matrix synthesis and angiogenesis. The aim of the study was to explore the effect of the application of i-PRF during SRP treatment on reducing tissue inflammation, concentration of inflammatory mediators and microorganisms in gingival crevicular fluid - GCF, and the success of regeneration itself. In patients with chronic periodontitis included in the study, the following clinical parameters were recorded: periodontal pocket depth- PPD, level of gingival margin - LGM, clinical attachment level - CAL, bleeding on probing – BOP, and plaque index - PI. The samples of GCF collected by paper strips (Periopaper, Amityville, NY, USA) were stored at -70 °C. Having added reverse specific primers and a universal forward primer for 16s rRNA, the concentrations of periodontopathogenic microorganisms, i.e. Aggregatibacter actinomycetemcomitans (A. a.), Porphyromonas gingivalis (P. g.), Prevotella intermedia (P. i.) and Tannerella forsythia (T. f.), were analysed by using polymerase chain reaction (PCR). Moreover, the total concentration of microorganisms was calculated by PCR-real time. The samples of GCF were also used to determine the levels of matrix metalloproteinase-8 (MMP-8), tumor necrosis factor-α (TNF-α), and alkaline phosphatase (ALP). The analysis of their concentrations in GCF was performed by using appropriate commercial ELISA (enzyme-linked immunosorbent assay) kits..