Effects of Fibrin Adhesive Material (Tissucol) Application on Furcation Defects in Dogs. Altan DOGAN1, Levent TANER1, Tiffin OYGUR2 and KOksal BALOS1

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1 J. Nihon Univ. Sch. Dent., Vol Effects of Firin Adhesive Mteril (Tissucol) Appliction on Furction Defects in Dogs Altn DOGAN1, Levent TANER1, Tiffin OYGUR2 nd KOksl BALOS1 (Received 19 June nd ccepted 1 Octoer 1991) Key words: firin seling, periodontl regenertion, new ttchment Astrct A histologicl study ws conducted to evlute the effects of firin dhesive mteril (FAM) ppliction on periodontl heling in seven experimentl dogs. Bilterl clss III furction defects were surgiclly creted t the second nd third premolrs, then orthodontic wires were plced round the teeth to induce periodontitis. Six weeks lter, the wires were removed nd the defects were treted y either surgery lone or surgery plus FAM ppliction. A totl of 21 specimens were otined for histologicl nd histometricl nlysis on dys 7, 21 nd 42. The Mnn-Whitney U test showed significntly more new ttchment nd one regenertion in the FAM-treted group compred to the control (p<0.05). Surgery plus FAM ppliction in the tretment of clss III furction defects seemed to e effective in promoting connective tissue ttchment nd one regenertion. Introduction Trditionl periodontl therpy results in only miniml regenertion of the periodontium in the most picl portion of the defect. The downgrowth of epithelil cells into the heling wound forms long junctionl epithelium preventing regenertion. The results of some experimentl studies suggest tht picl migrtion of the epithelium my not occur or my e limited under certin conditions[1-4]. The first requirement for successful regenertion my rest with clot dhesion to the root surfce, possily y virtue of firin linkge[5]. The rpid dhesion of lood clots to the root surfce my thus form sufficient rrier to epithelil picl migrtion[5,6]. Appliction of commercilly ville dhesive protein concentrte to plned root surfce my fcilitte erly dhesion of firin clot[7,8]. Moreover, regenertion of fier ttchment nd lveolr one in fenestrtion defects ppers to e enhnced y this mteril, which contins vrious plsm fctors nd highly 1 Deprtment of Periodontology, Fculty of Dentistry, Gzi University. 2 Deprtment of Pthology, Fculty of Medicine, Gzi University. To whom ll correspondence should e ddressed: Dr. Levent Tner, Gzi Universitesi, Dishekimligi Fkfiltesi, Emek 8. cd., Ankr, TURKEY

2 35 [9]. concentrted ttchment glycoproteins The purpose of this study ws to evlute the effect of firin dhesive mteril (FAM)(TISSUCOL KIT 0.5 ml, IMMUNO AG, VIENNA, AUSTRIA) on periodontl wound heling in furction defects. Mterils nd Methods Seven dult experimentl dogs in cliniclly helthy condition were used. During the opertive procedures, the dogs were sedted with intrvenously injected pentothl sodium. After clinicl nd rdiogrphicl exmintions, mucoperi- flps were rised on the uccl nd lingul surfces of the 2nd nd 3rd mndiulr premolr teeth on oth sides. Furction defects were surgiclly creted using slow-speed round urr nd continuous irrigtion with sline. Then wires were plced round the teeth to induce periodontl tissue rekdown round the furction res during period of six weeks. At the end of this period the orthodontic wires were removed nd rdiogrphs of the relted teeth were tken. After plning of the root surfces to remove remnnts of periodontl ligment nd cementum, reference notches were prepred t the level of the one mrgin. A totl of twenty-one defects were treted y either periodontl surgery lone or ppliction of commercilly ville firin dhesive mteril to the curetted root surfces nd mucoperiostel flps following surgery. The flps were then returned to their originl position nd sutured interproximlly. Plque control ws ccomplished y topicl ppliction of 0.2% chlorhexidine gluconte solution until the end of the experiment. The dogs were scrificed on dys 7, 21 nd 42 with n intrvenous overdose of pentothl sodium. Block iopsies were tken nd fixed in formlin for seven dys. Following declcifiction in formic cid-sodium citrte solution, the iopsy smples were dehydrted nd emedded in prffin. Step-seril mesio-distl sections were cut t 6 pm thickness, prllel to the long xis of ech root. The sections were stined with hemtoxylin nd eosin nd Crossmon's modified connective tissue stin. Histometricl nlysis ws performed on ech specimen lock using clirted grid mounted in light microscope (OLYMPUS) t x10 mgnifiction, nd the following liner distnces were mesured: ) from the picl order of the notch to the picl extent of the junctionl epithelium, ) from the picl order of the notch to the crest of the lveolr one. The Mnn-Whitney U test ws used for histometricl nlysis, nd differerces were considered significnt t p <0.05. Results Histologicl Oservtions Seven dys fter surgery. The grnultion tissue djcent to the experimentl root surfce ppered to consist of mononucler cells with occsionl polymorphonucler leucocytes, s well s numerous cpillries nd strnds of firin. Some loclized root resorption ctivity in the notch res nd newly deposited fine

3 36 collgen fiers nd osteoclstic ctivity in the periodontl spce were lso oserved. In ddition to these common findings, clot orgniztion nd mturtion of newly deposited collgen fiers were reltively pronounced in the FAM-treted group (group F) (Fig. l) thn tht in the control group (group C) (Fig. l). The young connective tissue filling the defect re ws in contct with the root surfce. Epitheliztion ws newly initited in some of the specimens. Twenty-one dys fter surgery. In the specimens of group F, more new ttchment formtion ws oserved in the suprcrestl re, where new lyer of cementum extending coronlly showed cellulr connective tissue with insertion of fiers to the root surfce (p < 0.05)(Tle 1)(Fig. 2,). Although sustntil mount of connective tissue ttchment hd formed in the notch, coronl to this region picl migrtion of the epithelium ws commonly seen in group C (Fig. 3,). In mny of the specimens in oth groups, infiltrtion of mononucler cells nd reduced edem were still present. A smll mount of one regenertion ws lso oserved in oth groups, remining round the notch region. Forty-two dys fter surgery. The sections from oth groups displyed further connective tissue mturtion. The ttchment of connective tissue to the root surfce nd formtion of new lveolr one were enhnced in group F (Fig. 4, ) compred with group C (Fig.5,)(p < 0.05)(Tle 1). Formtion of new lveolr one in oth groups hd incresed long the root surfces rther thn in the mid-portion of the furction defects. New cementum of vrying thickness ws clerly oserved with rtifctul splitting from the dentin surfce. Collgen fiers were functionlly oriented nd inserted into the cementum. Forty-two-dy specimens from group F demonstrted significntly more connective tissue nd one repir compred with 21-dy specimens(p<0.05). Tle 1 Results of histometricl mesurement nd levels of significnce Discussion The present study demonstrted significnt histometricl differences in the regenertion of connective tissue ttchment nd one in periodontl furction defects following periodontl flp surgery including topicl ppliction of firin dhesive mteril. For the purpose of the study, furction defects were surgiclly creted in the mndiulr premolrs of dogs nd exposed to the orl environment

4 37 for six weeks without plque control. Heling ws evluted t 7, 21 nd 42 dys fter surgery. A significntly incresed mount of connective tissue ttchment to the root surfce treted with FAM compred with roots treted y surgery lone my hve een due to the erly dhesion of protein concentrte to the root surfce. Thus, FAM might prevent the picl migrtion of epithelium t the root-soft tissue interfce nd mintin trctionl field for migrting firolsts[10,11]. The present results re in greement with the dt otined from previous studies[7,9,12,13]. However, it hs lso een reported tht the use of iologiclly ctive mterils nd ttchment glycoproteins did not enhnce connective tissue ttchment formtion nd might hve exerted n inhiitory rther thn n enhncing effect on firolst migrtion nd connective tissue ttchment[14]. In the present study, limited root resorption res were oserved in dy-21 specimens, lthough the root resorption res were replced with new cementum in dy-42 specimens. The tretment of one defects with FAM is usully followed y enhnced regrowth of new one[15-17]. This ws consistent with our study. Recently, it ws reported tht the use of FAM in flp model with reduced lveolr one nd fenestrtion defects effectively induced one regenertion[7,9]. The iologicl properties of firinogen, fironectin, thromin nd fctor XIII my hve contriuted to the regenertive process, fcilitting the new ttchment nd one formtion. It is postulted tht ny of the components present in FAM cting s lone or in comintion my ccount for the fvorle results[7,9,12,13]. Further experimenttion is needed to improve the dhesion nd strength of FAM on the root surfce nd to clrify the effects of individul components of FAM. References [1] CAFFESSE, R. G., HOLDEN, M. J., KON, S. nd NASJLETI, C. E.: The effect of citric cid nd fironectin ppliction on heling following surgicl tretment of nturlly occurring periodontl disese in Begle dogs, J. Clin. Periodontol., 12, , 1985 [2] CAFFESSE, R. G., KERRY, G. J., CHAVES, E. S., MCLEAN, T. N., MORRISON, E. C., LOPATIN, D. E., CAFFESSE, E. R. nd STULTS, D. L.: Clinicl evlution of the use of citric cid nd utologous fironectin in periodontl surgery, J. Periodontol., 59, , 1988 [3] CATON, J. G., POLSON, A. M., PINI PRATO, G., BARTOLUCCI, E. G. nd CLAUSER, C.: Heling fter ppliction of Tissue-Adhesive Mteril to denuded nd citric cid-treted root surfces, J. Periodontol., 57, , 1986 [4] PINI PRATO, G., CORTELLINI, P. nd CLAUSER, C.: Firin nd fironectin seling system in guided tissue regenertion procedure, J. Periodontol., 59, , 1988 [5] POLSON, A. M. nd PROYE, M.: Firin lekge: precursor for new ttchment, J. Periodontol., 54, 141, 1983 [6] WIKESJO, U. M. E., CLAFFEY, N. nd EGELBERG, J.: Periodontl repir in dogs. Effect of heprin tretment of the root surfces, J. Clin. Periodontol., 18, 60-64, 1991 [7] RIPAMONTI, V., PETIT, J. C., LEMMER, J. nd AUSTIN, J. C.: Regenertion of the connective tissue ttchment on surgiclly exposed roots using firin-fironectin dhesive system. An experimentl study on the oon (Pppio ursinus), J. Periodont. Res., 22, , 1987 [8] PINI PRATO, G., CORTELLINI, P., AGUDIO, G. nd CLAUSER, C.: Humn firin glue versus sutures in periodontl surgery, J. Periodontol., 58, , 1987 [9] BALOS, K., TANER, L., DOOAN, A. nd OYGUR, T.: The effect of firin dhesive mteril FAM (TISSUCOL) nd citric cid on periodontl wound heling. Presented t the 4th meeting of I. A. P., Istnul, 1990 (In press)

5 [10] SCHLAG, G., REDL, H., TURNHER, M. nd DINGER, H. P.: The importnce of firin in wound repir. In: Firin Selnt in Opertive Medicine, SCHLAG, G. nd REDL, H. (eds.), Vol. 4, 3, Plstic Surgery Mxillofcil nd Dentl Surgery, Springer Verlg, Heidelerg, Berlin, 1986 [11] REDL, H. nd SCHLAG, G.: Properties of different tissue selnts with specil emphsis on firinogen-sed preprtions. In: Firin Selnt in Opertive Medicine, SCHLAG, G. nd REDL, H. (eds.), Vol. 4, 27, Plstic Surgery Mxillofcil nd Dentl Surgery, Springer Verlg, Heidelerg, Berlin, 1986 [12] NASJLETI, C. E., CAFFESSE, R. G., CASTELLI, W. A., LOPATIN, D. E. nd KOWALSKI, C. J.: Effect of lyophilized utologous plsm on periodontl heling of replnted teeth, J. Periodontol., 57, , 1986 [13] NASJLETI, C. E., CAFFESSE, R. G., CASTDELLI, C. W., SIMITH, B. A., LOPATIN, D. E. nd KOWALSKI, C. J.: Effect of citric cid nd lyophilized utologous plsm on heling following periodontl flp surgery in monkeys, J. Periodontol., 58, , 1987 [14] RIPAMONTI, U. nd PETIT, J.-C.: Ptterns of heling on replnted oon incisors coted with n llogenic firin-fironectin protein concentrte, J. Periodont. Res., 24, , 1989 [15] PINI PRATO, G., CORTELLINI, P. nd CLAUSER, C.: Firin nd fironectin seling system in guided tissue regenertion procedure, A cse report, J. Periodontol., 59, , 1988 [16] ALBREKTSSON, T., BACH, A., EDSHAGE, S. nd JONSSON, A.: Firin Adhesive System (FAS), influence on one heling rte, Act Orthop. Scn., 53, 757, 1982 [17] ARBES, H., BOSCH, P., LINTNER, F. nd SALZER, M.: First clinicl experience with heterologous cncellous one grfting comined with the Firin Adhesive System (FAS), Arch. Orthop. Trumt. Surg., 98, 183, 1981

6 39 Fig. 1. Photomicrogrph of firin dhesive mteril (FAM)-treted specimen t 7 dys. Photomicrogrph of control specimen t 7 dys D, dentin; C, cementum; B, one; GT, grnultion tissue; A, rtifctul spce (connective tissue stin-cts x10) Fig. 2. Photomicrogrph showing 21-dy specimen treted with FAM (CTS x4). Higher-mgnifiction view of Fig. 2,. (CTS x20) D, dentin; C, new cementum; B, one; E, epithelium; CT, connective tissue; A, rtifctul spce

7 40 Fig. 3. Photomicrogrph of control specimen t 21 dys (CTS x4). Higher-mgnifiction view of Fig. 3, (CTS x10) D, dentin; C, cementum; B, one; CT, connective tissue; E, epithelium; A, rtifctul spce ; N, picl order of the notch Fig. 4. Photomicrogrph showing 42-dy specimen treted with FAM (CTS x4). Higher-mgnifiction view of the notch re (CTS x20) D, dentin; C, new cementum; B, one; E, epithelium; CT, connective tissue; A, rtifctul spce; N, picl order of the notch

8 41 Fig. 5. Photomicrogrph of control specimen t 42 dys (CTS x4). Higher-mgnifiction veiw of the notch re (CTS x10) D, dentin; C, new cementum; B, one; E, epithelium; CT, connective tissue; A, rtifctul spce ; N, picl order of the notch