Hipogalaktozilacija imunoglobulina G (IgG) gingivalne tečnosti i pljuvačke u obolelih od parodontopatije

Changed glycosylation of immunoglobulin G (IgG), above all, the expression of thermal galactose, influence to numerous functions of those immunoglobulin and correlate with the inflammatory level in a number of diseases. Aim: This work analyses the distribution of IgG subclasses and the content of thermal galactose in them, in saliva and gingival fluid of the patients with periodontal disease and different gum inflammatory level. Materials and methods: It was used saliva and gingival fluid of 30 adults with clinical picture of periodontal disease and 20 persons with healthy periodontium. The qualification of IgG was done by “dot-blot” procedure and the, and thermal galactose was determined by lectin immunoblot procedure. Results: The division of IgG subclasses in both fluids was different in the patients with periodontal disease and in control samples. In saliva and gingival fluid of the diseased quantitatively dominated IgG2 subclasses, independently from periodontal status. In IgG of both fluids, thermal galactose was exprimated at the healthy periodontium persons (control) and with the person with initial periodontal disease, while at the person with increased periodontal disease the expression of this saccharide wasn’t registered in neither of fluids. Conclusion: The results showed that there is a shift towards hypogalactosyled IgG glikoforms during the process of gum inflammation at the periodontal disease patients.Promenjena glikozilacija imunoglobulina G (IgG), pre svega ekspresija terminalne galaktoze, utiče na brojne funkcije ovih imunoglobulina i korelira sa stepenom zapaljenja u mnogim bolestima. Cilj rada: U ovom radu analizirana je distribucija IgG podklasa i sadržaj terminalne galaktoze u njima, u pljuvačci i gingivalnoj tečnosti bolesnika sa parodontopatijom različitog stepena inflamacije gingive. Materijal i metod: Kao materijal u ispitivanjima korišćena je gingivalna tečnost i pljuvačka 30 odraslih osoba sa kliničkom dijagnozom parodontopatija i 20 osoba sa zdravim parodoncijumom. Kvantifikacija IgG urađena je “dot-blot” postupkom, a određivanje terminalne galaktoze lektinskim imunoblot postupkom. Rezultati: Raspodela IgG podklasa u obe tečnosti se razlikovala kod parodontopatija i u kontrolnim uzorcima. I u pljuvačci i u gingivalnoj tečnosti obolelih, kvantitativno je dominirala IgG2 podklasa, nezavisno od parodontalnog statusa. U IgG obe oralne tečnosti terminalna galaktoza je bila eksprimirana kod osoba sa zdravim parodoncijumom (kontrola) i kod osoba sa početnom (inicijalnom) parodontopatijom, dok kod osoba sa uznapredovalom parodontopatijom ekspresija ovog šećera nije registrovana ni u jednoj od ove dve tečnosti. Zaključak: Rezultati ovih istraživanja ukazuju da postoji pomeranje prema hipogalaktozilovanim IgG glikoformama tokom procesa inflamacije gingive u obolelih od parodontopatije

Mogućnosti upotrebe pljuvačke kao dijagnostičke tečnosti u stomatologiji

Saliva is a secretion of the salivary and mucous glands and is of major importance in the maintainance of oral health. Over the last few decades, saliva has been evaluated as a diagnostic fluid in medicine for determining systemic disease markers as well as for monitoring numerous drugs, narcotics, and hormones. The biochemical analysis of saliva is particularly important in dentistry. The estimation of the risk of appearance and diagnosis of disease, monitoring of disease progression, evaluation of therapy efficacy for caries, periodontitis, premalignant and malignant oral lesions, as well as infectious diseases of the oral cavity, can be assessed by analyzing different constituent: of saliva, individuals at risk of caries can be identified using test: that determine saliva flow rate, saliva buffer capacity, and colonization of the oral cavity by cariogenic bacteria. Today, these rapid and simple diagnostic tests are used routinely in caries risk determination. The study and use of saliva-based diagnostics have increased over the last few decades. Clinical testing of saliva shows much promise. However, there is a need for much additional research in this area, before the true clinical value of saliva as a diagnostic fluid in dentistry can be determined.Pljuvačka je sekret pljuvačnih i mukoznih žlezda koji je značajan za održavanje oralnog zdravlja. Poslednjih decenija sve više se razmatra mogućnost njene primene kao zamena krvi, odnosno krvnom serumu i plazme, radi dokazivanja pokazatelja sistemskih oboljenja, kao i za nadgledanje primene lekova, droga i hormona. Analiziranje biohemijskog sastava pljuvačke posebno je značajno u stomatologiji za procenu rizika nastanka, praćenja toka bolesti i kontrole rezultata primenjenog lečenja zubnog karijesa, parodontopatije, oralnih karcinoma i infektivnih oboljenja usne duplje. U proceni rizika nastanka karijesa na zubima rutinski se izvode ispitivanja kojima se određuju količina izlučene pljuvačke, njen puferski kapacitet i stepen kolonizacije usne duplje kariogenim vrstama bakterija. Jednostavnost izvođenja ovih ispitivanja omogućava da ih u stomatološkoj ordinaciji primenjuje lekar praktičar, što predstavlja veliku pomoć u proceni rizika za nastanak zubnog karijesa. Zbog bliskog kontakta s oralnim tkivima, pljuvačka sadrži brojne biohemijske pokazatelje patoloških procesa lokalizovanih u ovim tkivima (enzimi, imunoglobulini, ostali proteini, fenotipski pokazatelji). Ispitivanje pomenutih pokazatelja je značajno kao dopuna kliničkom dijagnostikovanju ukoliko ono ne pruža dovoljno informacija

Identifikacija parodontopatogenih mikroorganizama PCR tehnikom

INTRODUCTION Periodontitis is an inflammatory disease of the supporting tissues of teeth and is a major cause of tooth loss in adults. The onset and progression of periodontal disease is attributed to the presence of elevated levels of a consortium of pathogenic bacteria. Gram negative bacteria, mainly strict anaerobes, play the major role. OBJECTIVE The present study aimed to assess the presence of the main types of microorganisms involved in the aetiopathogenesis of periodontal disease: Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Eikenella corrodens, Treponema denticola, Tanerella forsythia and Prevotella intermedia in different samples collected from the oral cavity of 90 patients diagnosed with periodontitis. METHOD Bacterial DNA detection was performed in diverse biological materials, namely in dental plaque, gingival tissue and saliva, by means of multiplex PCR, a technique that allows simultaneous identification of two different bacterial genomes. RESULTS In the dental plaque of the periodontitis patients, Treponema denticola dominated. In the gingival tissue, Tannerella forsythia and Treponema denticola were the microbiota most frequently detected, whilst in saliva Treponema denticola and Eikenella corrodens were found with the highest percentage. CONCLUSION The identification of microorganisms by multiplex PCR is specific and sensitive. Rapid and precise assessment of different types of periodontopathogens is extremely important for early detection of the infection and consequently for the prevention and treatment of periodontal disease. In everyday clinical practice, for routine bacterial evaluation in patients with periodontal disease, the dental plaque is the most suitable biological material, because it is the richest in periodontal bacteria.Uvod Epidemiološki podaci iz čitavog sveta ukazuju na veliku rasprostranjenost gingivitisa i parodontopatije, oboljenja potpornog aparata zuba. U etiopatogenezi oboljenja parodoncijuma ključnu ulogu igraju različiti rodovi Gram-negativnih bakterija, ponajviše striktnih anaeroba. Cilj rada Cilj rada je bio da se ispita postojanje genoma glavnih parodontopatogenih mikroorganizama Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Eikenella corrodens, Treponema denticola, Tanerella forsythia i Prevotella intermedia u različitim uzorcima poreklom iz usne duplje pacijenata s klinički dijagnostikovanom parodontopatijom. Metod rada Kao biološki materijal u kojem je dokazivano postojanje DNK mikroorganizama korišćeni su zubni plak, tkivo zapaljene gingive i pljuvačka. Za otkrivanje bakterijskog genoma primenjena je multipleks tehnika reakcije lančanog umnožavanja (engl. polymerase chain reaction PCR), odnosno simultana amplifikacija gena dve različite bakterije. Rezultati S manjom ili većom učestalošću, u svim ispitanim uzorcima utvrđeno je postojanje parodontopatogenih mikroorganizama. U zubnom plaku osoba obolelih od parodontopatije najčešći je bio genom vrste Treponema denticola. U tkivu parodoncijuma otkriveno je u najvećem procentu postojanje genoma vrsta Tannerella forsythia i Treponema denticola, što je odlika hroničnog oblika parodontopatije, a u pljuvački ispitanika dominirale su Treponema denticola i Eikinella corrodens. Najmanje ukupno postojanje bakterija je zapaženo u pljuvački. Zaključak Primenjeni metod PCR ima veliku osetljivost i specifičnost. Brzo i precizno otkrivanje mikroorganizama je veoma važno za pravovremeno dijagnostikovanje infekcije, a samim tim i za prevenciju i lečenje parodontopatija. U svakodnevnoj kliničkoj praksi optimalan biološki materijal za dokazivanje parodontopatogena kod osoba obolelih od parodontopatije je zubni plak, koji se smatra pouzdanim pokazateljem zastupljenosti pojedinih bakterija u obolelom parodoncijumu

Interaction between fibronectin fragments and immunoglobulin G in gingival crevicular fluid of patients with periodontal disease

Introduction Fibronectin (FN) can interact with immunoglobulin G (IgG) molecules affecting the process of physiological elimination and causing abnormal deposition of immune complexes. The aim of the study was to analyze interaction between FN fragments and IgG molecules with different glycosylation profiles in gingival crevicular fluid (GCF) of patients with periodontal disease and healthy controls. Material and Methods The study included 30 patients with moderate and advanced periodontitis and 22 healthy subjects. IgG and FN content in GCF were measured as well as the presence of FN and galactose expression on IgG molecules. Results IgG content in GCF was five times higher in patients with moderate (p<0.01) and eight time higher in patients with advanced periodontitis (p<0.001) compared to healthy subjects. Also, hypogalactosylated forms of IgG were found in higher concentration in GCF of patients with advanced periodontitis compared to moderate periodontitis and healthy subjects (p<0.05). FN fragments of molecular mass 48 - 53 kDa were the most commonly found fragments in all three groups. Furthermore, in patients with advanced periodontitis, fibronectin fragments were attached to IgG molecules. Conclusion IgG and FN fragments form complexes in GCF in patients with periodontal disease and healthy subjects

Kompleksi fibronektinskih fragmenata i imunoglobulina G u gingivalnoj tečnosti osoba obolelih od parodontopatije

Introduction Fibronectin (FN) can interact with immunoglobulin G (IgG) molecules affecting the process of physiological elimination and causing abnormal deposition of immune complexes. The aim of the study was to analyze interaction between FN fragments and IgG molecules with different glycosylation profiles in gingival crevicular fluid (GCF) of patients with periodontal disease and healthy controls. Material and Methods The study included 30 patients with moderate and advanced periodontitis and 22 healthy subjects. IgG and FN content in GCF were measured as well as the presence of FN and galactose expression on IgG molecules. Results IgG content in GCF was five times higher in patients with moderate (p lt 0.01) and eight time higher in patients with advanced periodontitis (p lt 0.001) compared to healthy subjects. Also, hypogalactosylated forms of IgG were found in higher concentration in GCF of patients with advanced periodontitis compared to moderate periodontitis and healthy subjects (p lt 0.05). FN fragments of molecular mass 48 - 53 kDa were the most commonly found fragments in all three groups. Furthermore, in patients with advanced periodontitis, fibronectin fragments were attached to IgG molecules. Conclusion IgG and FN fragments form complexes in GCF in patients with periodontal disease and healthy subjects.Uvod Fibronektin može da interreaguje s molekulima imunoglobulina G (IgG) i utiče na normalan klirens ili poremećeno deponovanje imunskih kompleksa. Cilj ovog rada je bio da se ispita veza između fibronektina i IgG različitih glikoformi u gingivalnoj tečnosti osoba obolelih od parodontopatije i parodontalno zdravih ispitanika. Materijal i metode rada U studiju je uključeno 30 pacijenata s umerenom i uznapredovalom parodontopatijom i 22 parodontalno zdrave osobe. U gingivalnoj tečnosti određivan je sadržaj IgG i fibronektina dot blot i imunoblot tehnikama. IgG iz gingivalnih tečnosti su afinitetno izolovani i analizirani na prisustvo fibronektina i ekspresiju galaktoze. Rezultati Sadržaj IgG u gingivalnoj tečnosti osoba s umerenom parodontopatijom bio je oko pet puta veći u odnosu na sadržaj IgG kod zdravih osoba (p lt 0,01), dok je kod uznapredovalih oblika bio oko osam puta veći (p lt 0,001). Takođe, hipogalaktozilovane forme IgG su većoj meri postojale kod osoba sa uznapredovalom parodontopatijom u odnosu na zdrave i osobe s umerenom parodontopatijom (p lt 0,05). U sve tri analizirane grupe dominirali su fibronektinski fragmenti od 48 do 53 kDa. Uočeno je da su IgG izolovani iz gingivalne tečnosti vezani za fragmente fibronektina, pri čemu su IgG osoba sa uznapredovalom parodontopatijom, imali najveću količinu ovih vezanih fragmenata. Zaključak Dobijeni rezultati pokazuju da IgG gingivalne tečnosti zdravih i osoba s parodontopatijom formiraju komplekse sa fibronektinom

The effects of sorbitol on autotomy in rats after peripheral nerve lesions

The autotomy phenomenon is an animal model that is used for studying chronic pain. It is based on the observation that transection of peripheral nerve in animals triggers self-mutilation of denervated limb. Autotomy phenomenon today presents the most reliable and useful tool for experimental investigation of pain-suppressing effect of some substances on experimentally induced pain. Organic alcohol glycerol has been used for peripheral blocks in treating paroxysmal trigeminal neuralgia. There is evidence that sorbitol, also an organic alcohol, disturbs nerve function in humans. Therefore, the present study was aimed to investigate the effects of 2 organic alcohols on autotomy phenomenon after peripheral nerve lesions. After inducing autotomy, 12 rats were divided into 3 groups, each comprising 4 animals, according to the substance applied in order to suppress autotomy (sorbitol solution in the 1st, absolute glycerol in the 2nd, and saline in the 3rd, the control group). Degree of self-mutilation was measured for 8 weeks. Results indicated the efficacy of glycerol and sorbitol in relieving experimentally induced pain

Hypogalactosylation of salivary and gingival fluid immunoglobulin G in patients with advanced periodontitis

Background: Altered glycosylation of immunoglobulin G (IgG) has been found to affect certain immunological activities of IgG and to correlate with increased inflammation in various disease states. This work deals with the changes in distribution and galactosylation of IgG subclasses present in saliva and gingival crevicular fluid (GCF) of patients with initial and advanced periodontitis and of normal controls. Methods: IgG subclasses were quantified by dot-blot assay, and the degrees of expression of galactose in the total IgG and its individual subclasses were estimated by lectin immunoblot assay after sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE) separation of IgG and by capture enzyme-linked immunosorbent assay (ELISA), using biotinylated Ricinus communis (RCA-1) and Bandeiraea simplicifolia (BS-II) lectins. Results: The distribution of IgG subclasses in both fluids was found to differ in periodontal patients compared to normal controls. In the periodontitis saliva and GCF, the IgG2 subclass dominated quantitatively, regardless of periodontal status. However, galactose was found to be expressed in IgG heavy chains in normal controls and patients with initial periodontitis but not, or at barely detectable levels, in advanced periodontitis. Conclusion: The results suggest that the shift toward hypo-galactosylated glycoforms may occur during the process of inflammation of the gingiva

Identification of periodontopathogen microorganisms by PCR technique

INTRODUCTION Periodontitis is an inflammatory disease of the supporting tissues of teeth and is a major cause of tooth loss in adults. The onset and progression of periodontal disease is attributed to the presence of elevated levels of a consortium of pathogenic bacteria. Gram negative bacteria, mainly strict anaerobes, play the major role. OBJECTIVE The present study aimed to assess the presence of the main types of microorganisms involved in the aetiopathogenesis of periodontal disease: Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Eikenella corrodens, Treponema denticola, Tanerella forsythia and Prevotella intermedia in different samples collected from the oral cavity of 90 patients diagnosed with periodontitis. METHOD Bacterial DNA detection was performed in diverse biological materials, namely in dental plaque, gingival tissue and saliva, by means of multiplex PCR, a technique that allows simultaneous identification of two different bacterial genomes. RESULTS In the dental plaque of the periodontitis patients, Treponema denticola dominated. In the gingival tissue, Tannerella forsythia and Treponema denticola were the microbiota most frequently detected, whilst in saliva Treponema denticola and Eikenella corrodens were found with the highest percentage. CONCLUSION The identification of microorganisms by multiplex PCR is specific and sensitive. Rapid and precise assessment of different types of periodontopathogens is extremely important for early detection of the infection and consequently for the prevention and treatment of periodontal disease. In everyday clinical practice, for routine bacterial evaluation in patients with periodontal disease, the dental plaque is the most suitable biological material, because it is the richest in periodontal bacteria

Association of Fibronectin With Hypogalactosylated Immunoglobulin G in Gingival Crevicular Fluid in Periodontitis

Background: Fibronectin (FN) can bind to immunoglobulins (Ig), influencing both the normal clearance and abnormal deposition of circulating immune complexes. This study focuses on the possible interaction between FN and IgG present in gingival crevicular fluid (GCF) of periodontitis patients and periodontally healthy controls, with the aim to determine whether such interaction may be connected with the glycosylation profile of IgG and, thus, consequentional in accumulation of proinflammatory IgG in periodontal pockets. Methods: The study included 30 patients with initial or advanced periodontitis, and 13 periodontally healthy subjects. GCF IgG was purified and analyzed for the presence of FN and its fragments and for galactose expression. Results: IgG isolated from GCF contained FN, which was bound to the IgG heavy chains. IgG from GCF of advanced periodontitis patients contained high levels of hypogalactosylated IgG, and bound more FN than IgG from GCF of the other two groups, which contained low levels of this glycoform. FN was in a degraded form in GCF from all studied patients, and a fragment of 48- to 53-kDa molecular mass seemed to be the sole one linked to IgG. Conclusions: IgG and the FN fragment of 48 to 53 kDa in GCF of periodontitis patients and periodontally healthy subjects are physically connected. This fragment was linked to the heavy chains of IgG and the reaction seemed to be particularly intensive with IgG from advanced periodontitis, which contained significantly less galactose in its sugar chains. J Periodontol 2010;81:1472-1480