EXPRESSION OF ENZYME HEPARANASE 1 IN PATIENTS WITH ADVANCED GENERALIZED PERIODONTITIS

Cilj istraživanja: Cilj istraživanja bio je ispitati moguću ulogu HPSE1, enzima za modifikaciju HS GAG lanaca, u regulaciji upalnog odgovora tijekom uznapredovalog generaliziranog parodontitisa. Materijali i metode: U svrhu istraživanja uzeti su uzorci gingive zdravih ispitanika (kontrolna skupina; n = 20) i ispitanika s dijagnozom uznapredovalog generaliziranog parodontitisa (ispitna skupina; n = 20) koji su obrađeni za imunohistokemijsko bojenje. Preparati uzoraka bojeni su orijentacijski (H/E bojenje) te primarnim protutijelima na HPSE1 i opći upalni biljeg CD45 (imunofluorescencija) i potom fotografirani tehnikom panoramskog snimanja. Kvantifikacijska različitih atributa bojenja (histomorfometrijski parametri gingive, površine izražajnih domena i prostorni gradijenti intenziteta HPSE1 i CD45) napravljena je računalno korištenjem programa AdobePhotoshop® CC i ImageJ za obradu slika, odnosno Microsoft Office Excel® 2016 (Microsoft Corp, Redmond, WA, USA) i GraphPad v8 (GraphPad Software, La Jolla, CA, USA) za statističku analizu. Rezultati: Pokazana je statistički značajna razlika u izražaju HPSE1 u zdravoj gingivi i gingivi oboljelih od parodontitisa. Izražaj HPSE1 je negativno korelirao sa prisutnošću upalnog infiltrata u subepitelnoj stromi zdrave gingive i gingive oboljelih od parodontitisa. S druge strane, izražaj HPSE1 u gingivi oboljelih od uznapredovalog generaliziranog parodontitisa je pozitivno korelirao sa nekim strukturnim promjenama u tkivu (ukupna i stromalna celularnost). Zaključak: HPSE1 sudjeluje u regulaciji upalnog odgovora tijekom uznapredovalog generaliziranog parodontitisa. Moduliranje enzimatske aktivnosti HPSE1 se može iskoristiti u budućnosti kao jedan od terapijskih pristupa u liječenju uznapredovalog generaliziranog parodontitisa, ali potrebna su daljnja istraživanja kako bi se bolje razjasnila priroda uloge HPSE1 u patogenezi ove bolestiObjective: The aim of the study was to examine if HPSE-1, the enzyme for modification of HS GAG chains, could be considered as a regulator of inflammatory response in advanced generalized periodontitis. Materials and methods: For the purpose of the study, the samples of healthy gingiva (controls; n = 20) and diseased gingiva from patients diagnosed with advanced generalized periodontitis (test group; n = 20) were taken and processed for immunhistochemistry (H/E staining and immunofluorescence). For immunofluorescence, the primary antibodies against HPSE1 and general inflammatory cell marker CD45 were applied. Quantification of various staining properties (histomorphometric parameters of gingival tissue, expression domains and intensity gradients of immunofluorescence staining) was performed on high-resolution panoramic images by using AdobePhotoshop® CC and ImageJ (image editing), as well as Microsof Office Excel® 2016 (Microsoft Corp, Redmond, WA, USA) and GraphPad v8 (GraphPad Software, La Jolla, CA, USA) for statistical analysis. Results: The expression of HPSE1 in healthy and diseased gingiva displayed statistically significant difference. The expression of HPSE1 was negatively correlated with the presence of inflammatory infiltrate in subepithelial stroma of both healthy and diseased gingiva. On the other hand, the expression of HPSE1 was correlated positively with specific structural changes in diseased gingiva (total and stromal celularity). Conclusion: HPSE1 could be considered as a regulator of inflammatory response in advanced generalized periodontitis. Modulation of enzymatic activity of HPSE1 could be exploited in the future as a possible approach for treatment of advanced generalized periodontitis. However, further studies are still needed in order to fully elucidate the scope of regulatory roles of HPSE1 in pathogenesis of advanced generalized periodontiti