Effect of Osteon II Collagen with Hyaluronic Acid and Collagen Membrane on Bone Healing Process in Rabbits: A Radiograghical Study

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1 ISSN: , Vol. 5 Issue 8, ugust-2016 Effect of Osteon II Collgen with Hyluronic cid nd Collgen Memrne on Bone Heling Process in Rits: Rdiogrghicl Study Yusr H. L Mukhtr 1, Wf K.id 2 1 BDS, Deprtment of L Khns Hospitl - Mosul, Irq 2 sst.prof.deprtmentof orl nd mxillofcil surgery, college of dentistry, university of Mosul, Irq BSTRCT Ojective: To evlute the effectiveness of locl ppliction new injectle one grft sustitute osteon II collgen seprtely nd with ddition of hyluronic cid nd collgen memrne in surgiclly creted one defect in tii of rits. Mterils nd Methods: Twenty dult mle rits were used in this study. Two one defects were mde in the tii of ech rit. The defects were filled with Osteon II /hyluronic cid, Osteon II collgen with collgen memrne, Osteon II collgen lone nd the other left unfilled s control. Specimens were collected in one, two nd four weeks fter surgery. The rdiodensity nd the mount of one formtion were evluted rdiogrphiclly. Results: It ws found tht Osteon II collgen with Hyluronic cid comintion led to significnt increse in one density nd more mount of new one formtion (P 0.05) in 2 nd 4 weeks postopertivly thn other groups. On the other hnd Osteon II collgen with Collgen memrne showed etter results thn Osteon II collgen, nd control groups. Conclusions: It ws concluded tht topicl ppliction of Hyluronic cid with one sustitute mteril will improve heling process of the one represented with more new one formtion rdiogrphiclly nd densitometry compred with collgen memrne tht dded to one grft mteril or with one grft lone. Key wrds: Bone regenertion, osteon II, Collgen memrne, Hyluronic cid, Digitl rdiogrph, CT scn INTRODUCTION Bone grfting therpy hs ecome n integrl prt of dentistry; ptients re ecoming more wre of grfting s tretment modlity nd expect etter predictility, fit, function nd esthetics. Tody, with the introduction of dvnced one grfting techniques nd the use of sophisticted one replcement grft mterils, it is possile to increse the volume, width, nd height of one in deficient res to regenerte the tissues supporting (1). Osteon is one of lloplstic mterils composed of hydroxyptite70% nd et- triclcium phosphte 30% which re most close to mjor minerl components of humn one. Osteon is osteoconductive mteril tht cts s one growth scffold. It hs interconnected porosity structure which is similr to tht of humn cncellous one (2). Hyluronic cid (H) hs osteoconductive potentil; it ccelertes the one regenertion y mens of chemotxis, prolifertion nd successive differentition of mesenchyme cells. H my ct s iomteril scffold for other molecules, such s Bone Morphogenic Protein-2 (BMP-2) nd Trnsformtion Growth Fctor- β (TGF-β) used in guided one regenertion techniques nd tissue engineering reserch (3). Resorle collgen memrnes (RCMs) re mnufctured from llogeneic or xenogeneic sources to mnge orl wounds such s extrction sockets, for sinus-lift procedures nd repirs, nd for periodontl or endodontic surgeries (4). Brrier memrnes re mong the most widely studied scffolds for tissue regenertion, including one, nd the choice of type of memrne depends lrgely on the required durtion of memrne function (5) The purpose of the present study ws designed to investigte nd evlute the effectiveness of new injectle one grft sustitute osteon II collgen in one heling process when used seprtely nd to compre the effect of osteon II collgen when mixed with hyluronic cid once or comined with collgen memrnes in one regenertion process. Rits were used s niml model in this study ecuse of their short developmentl period nd their fster one turnover the rit hs fster skeletl chnge nd one turnover (significnt intrcorticl, Hversin remodeling nd it chieve skeletl mturity Pge 36

2 ISSN: , Vol. 5 Issue 8, ugust-2016 (closure of epiphysel pltes) shortly fter reching complete sexul development t pproximtely 6 months(6). lso there is miniml literture etween humn nd rit one composition nd one density, some similrities re reported in the one minerl density (7). There re mny study conducted to investigte the sole effect of locl ppliction of hyluronic cid, collgen memrne tht dded to the one grfting mteril on one regenertion process. ccording to our knowledge this is the first study tht compre etween these two mterils (hyluronic cid nd collgen memrne) when dding to osteon II collgen to fill the one defect. The one density nd the mount of new one formtion were determined y densitometric nd rdiogrphic nlysis. niml model nd Surgicl Procedure MTERIL ND METHODS The protocol of this study ws pproved y scientific Committee Deprtment of orl nd mxillofcil surgery college Dentistry of Mosul. The surgicl prt done in surgicl deprtment/veterinry Collge of Duhok University. This study ws crried out on locl twenty mle rits of (8 10 months) weighing g ws used s experimentl niml. During the entire period of the experiment the rit ws fed three times dily with greenery diet nd tp wter nd generl heth continuously monitored, the niml were housed in niml house of Veterinry Collge of Duhok University. nimls were qurntined for 7 dys prior to the surgicl procedure to check their generl sttement nd ensure the sence of generl infectious disese efore the opertion time, the niml legs were wshed with sop nd wter efore the surgicl procedure nd then the opertion side ws shved, nd disinfected with 10% povidin iodine. mixture contining (10mg/kg) ketmine hydrochloride generl nesthetic gent (Grcure phrceuticls Ltd.,Bhwdi,Indi) nd (2mg/kg) xylzine sedtive (Xyl - Interchemie, Hollnd) nd tropine 2mg/kg solution (phrmceuticls Ltd.,Bhwdi,Indi) were given intrmusculrly to chieved generl nesthesi. Complete nesthesi ws otined within 10 minutes, this dose kept the rit nesthetized for out hlf hour. Locl nesthesi, 2% lidocine HCL with epinephrine 1: locl nesthetic gent ws dministered y infiltrtion t the surgicl site prior the incision for hemostsis. The niml ws plced in lterl position during the procedure. 5cm skin incision ws mde long the longitudinl xis of lterl spect of ech tii, fsci ws dissected nd full- thickness flp ws reflected to expose the under lining one of ech tii under septic condition (Fig.1). Figure (1) The study mterils: ppliction of the mterils, two one defect D1 control group,d2 hyluronic cid mixed with osteon II collgen, on the right tii, ;defect osteon II collgen with collgen memrned3, osteon II collgen lone D4, on the left tii. fter exposure nd reflection of soft tissue, two monocorticl holes (cylindricl in shpe) on ech tii ws performed with trephine ur of 3mm in width (Dentium Compny- Mde in Kore), nd 4mm depth until rech the mrrow spce Pge 37

3 ISSN: , Vol. 5 Issue 8, ugust-2016 under constnt norml sline irrigtion on slow speed surgicl hnd piece t 1500 rpm to prevent therml one necrosis (SESHIN X-Cue Implnt surgery motor,south Kore). The distnce which were left etween ech hole out 1cm. Then the one cvities were crefully wshed with sterile norml sline to eliminte one deris efore eing filled with the mterils. The right tii hd 2 defect ;the first one serve s control which left empty, the second one filled with (0.2 ml of hyluronic cid gel(mnufctured y Bio Polymer, Germny) nd one grft mteril osteon II collgen, mnufctured y Dentium Compny (GENOSS Seoul, Kore.), the mixture ws left untouched for 5 minutes to chieve homogeneity. The mixture ws loded in ech defect nd gently pressed. The second opertion in the left tii hd 2 one defects lso; one of them filled with osteon II collgen lone, nother one hd osteon II collgen covered with collgen memrne (Resorle Memrne mde in Kore). t completion of mteril plcement, the flp ws gently pproximted nd primry wound closure ws performed, using 3-0 non resorle lck silk suture (Huin. M. instrument.,chin). which ws to e removed 10 dys post opertively. Finlly, topicl ntiiotic ws pplied t the wounds site nd dressed. The nimls were given oxytetrcycline hydrochloride injectle solution 50mg/Kg (Chongqing Fngtong niml phrmceuticl, Chin). The nimls were scrificed t 1, 2, nd 4 weeks postopertively (8). Rdiologicl nlysis The operted tiis were dissected su periostel to llow direct oservtion of newly formed one; nd fixed in uffered 10% formlin solution.. Computer Tomogrphy Scn ssessment: To mesure the one density ll specimens were sujected to CT scn rdiogrphicl exmintion in Wn- hospitl (CT scn Siemens G2002computer tomogrphy, Siemens TR. 1, D91301, Germny Dicum print4 system). ll imges were scn with 32 slices multidetector CT dentl protocol xil slices to ssess the mount of one density. Computed tomogrphy (CT) scns, increment thickness of 0.6 mm, nd multi-plnr coronl, sgittl reconstructions. Threedimensionl chnges in the one were evluted y mesuring the lrgest dimeter (mm) in the nterior posterior direction, medil lterl direction nd depth on ech scn. The nterior posterior size ws mesured t the level of the su chorl one plte on the sgittl CT reconstruction. The imge from CT scn ws loded to work sttion (Siemens G 2002, T R.1, D130, Germny) with work plce softwre (Synco CT). The cross section imges of tii were trnsferred nd reformed in three plnes (xil, coronl nd sgittl) through multi plnner reconstruction, drwing three points inside the defect nd tke the men for it. The nterior posterior size ws mesured t the level of the su chorl one plte on the sgittl CT reconstruction. The rdiogrphic computer tomogrphy ws mesured (3D) for ech defect fter mke three point, two in the lterl nd one medium position inside the defect then tke the men for ech defect. The results re expressed s the men ± stndrd devition (SD) for ech defect ws performed. B. Digitl dentl x-ry: The rdiogrphic exmintion ws crried out in digitl rdiogrphy which confirmed y correct positioning of the specimen in lterl rdiogrph. digitl rdiogrph ws tken to ll specimen using portle dentl x-ry system nd digitl sensor (Rextr X, mde in Jpn), ttched to it clirtion stinless steel wire of 10 mm in length. The tii of the rit ws plced in contct with the sensor nd the lterl order of the tii is prllel to the sensor nd the distnce etween the end of the long cone nd the sensor ws focused to 30 cm) nd the cone ws kept perpendiculr to the sensor ll the time. Using the following prmeters 8 m, 63Kv, nd exposure time 0.12 sec. nd the red out strts utomticlly, the imge ws displyed grdully on the computer screen, when the red out ws completed; the newly red imge ws stored. Mesurement the mount of one formtion: The rdiogrphic imge ws trnsferred to the Imge J 1.47v softwre (Ntionl Institute of Helth, US), which enle us to quntify the mount of one formtion. Then y selecting rectngulr re of pre- defined size (12mm2 ) to define originl size of one defect to ssess the newly formed one (Fig.2-). The region of defect ws delineted through drwing lines of 4 mm in length which represent the upper nd lower order of defect nd these lines re joined with two verticl lines of 3mm in height which represent the side wll oundries of the defect. Then strt to estimte the opcifiction re through using polygonl shped symol in selector tool of the min windows of imge J progrm to demonstrte the periphery of rdio dense re within the defect cvity tht representing ossifiction zone of newly formed one. The imge were clirted first to get reding in millimeters y mesuring the length of reference wire, so the reference distnce ws defined efore using the function of re mesure from nlyze tool in the progrm(fig.2-b). The mesurements were performed three times in ech defect in different dys to minimize interoserver mesurement error nd the men vlues of ll reding for ech defect were otined. Pge 38

4 ISSN: , Vol. 5 Issue 8, ugust-2016 Figure (2-).: Demonstrte the re of one formtion using ImgeJ softwre used for digitl ssessment of heling re which is displyed in the " result " window in mm. Figure (2-B): Demonstrte the re of one formtion using ImgeJ softwre used for digitl ssessment of heling re which is displyed in the "Result" window in mm. The verge vlue of one formtion ws mesured y two exminers who didn t know to which group the nimls elonged to the estimtion of new one formtion ws pplied y using n eqution s follow: re of new one formtion=totl defect re non formed one re Percentge of one formtion = re of new one formtion x 100 Totl defect re ll the incresed re of opcifiction from the externl nd internl surfce of originl defect size ws mesured. The men of vlue were sumitted to, one-wy nlysis of vrince (NOV, followed y Duncn test to identify significnt differences mong groups nd P vlue 0.05 were tken to e significnt. Pge 39

5 ISSN: , Vol. 5 Issue 8, ugust-2016 RESULTS Rdiologicl ssessment of one regenertion y Computer Tomogrphy (Mesurements of one Density): The men vlue of one density of one defects, stndrd devition, nd p vlues in ll experimentl groups t one, two nd four weeks intervls re summrized in (Tle 1).The nlysis of one density for ll test sites utilized y CT scn demonstrted t ech period intervls showed significnt increse in oth experimentl groups compred with control group long the periods of the study (chrt 1). Tle (1) Men nd P vlue rdiogrphic finding of one density evlution y CT scn of regenerted re in ll groups' rdio grphicl using NOV nd Duncn test. HELING PERIOD 1WEEK 2WEEKS 4WEEKS P vlue GROUPS D1 Control group 74.8± D2 Osteon with hyluronic cid D3 Osteon with Collgen group D4 Osteon group 104.2± ± ±60.28 BC ±46.35 C B 184.2±89.62 B B c c P vlue Men significnt t p Cpitl letters refer to comprison mong tretments (Verticlly).Smll letters men comprison etween periods of heling. (Horizontlly) Chrt (1): Men of CT Scn Rdiogrphicl finding using Duncn Test ; D1- Control group. D2- Osteon with hyluronic cid group.d3-osteon with collgen memrne group.d4-osteon group. Pge 40

6 ISSN: , Vol. 5 Issue 8, ugust-2016 Evlution of one density t one week postopertively: Fig. (3-) t the end of one week, the men of CT scn density of new one ws no significntly deference etween groups only slight superiority in the tretment groups (p vlue=.327). 1week D1, CONTROL D2, WITH HYLUR- ONIC CID D3, WITH CO- LLGEN D4, 2weeks D1, CONTROL D2, WITH HYLUR- ONIC CID D3, WITH CO- LLGEN D4, B 4weeks D1, CONTROL D2, WITH HYLUR- ONIC CID D3, WITH CO- LLGEN D4, C Figure ( 3 ) - CT Scn Rdiogrph: - one week D1,D2,D3,D4.ll groups showed no rdiogrphicl one heling; B-Two weeks D1,D2,D3,D4showed more one heling in D2,D3 more thn D1,D2 ; C- four weeks D1,D2,D3,D4,superiority in one density in D2,D3 more thn D1,D2; R(rdiogrphic tken to demonstrte the re of one density) Pge 41

7 ISSN: , Vol. 5 Issue 8, ugust-2016 Evlution of one density t two weeks postopertively: Fig. (3-B) t the end of two weeks, the density of one defects in CT scn, one heling ws superior in ll groups, nd exhiited one density in D3 Osteon II collgen with Collgen groups, ws (292.0±48.54) nd D2 Osteon II collgen with Hyluronic cid groups(215.6±46.35) nd D4 groups Osteon II group (184.20±89.63), significnt difference (p vlue= 0.000) Evlution of one density t four weeks postopertively: Fig. (3-C) The CT scn exmintion of heled one defects site demonstrted significnt progression one density in D2 Osteon II with Hyluronic cid groups ws ( ±258.08) nd D3 Osteon II with Collgen groups ws (603.0± ) nd Osteon II collgen group (375.4±159.51). Bone heling ws exhiited more dvnced phse of one formtion in groups Osteon II with hyluronic cid group nd Osteon II collgen with collgen memrne group, treted defects. (pvlue=.020). Computer Tomogrphy (CT) imging of one regenertion: The restortion of one t the defect ws oserved using X-ry- CT scn detiling the progressive one regenertion illustrted in Figure (3) the control group shows no signs of ridging ossifiction t one week thus heling of the defect ws representtive of incomplete heling. The defects were clerly detectle tile four weeks lthough one regenertion seems to e proceeding. The effectiveness of Osteon II collgen with Hyluronic cid nd osteon II collgen with collgen memrne showed t two weeks thin lyer of one developing t the defects were rely detectle from surrounding norml structure t four weeks lthough the defects seemed complete heling nd significnt difference in ll groups ws oservtion. Rdiogrphic evlution of one formtion (DIGITL): Rdiogrphic evlution for groups through different period: Tle (2), Chrt (2). Tle (2) Men & p-vlue of Imge J rdiogrphicl finding for one Formtion Using NOV nd Duncn Test. Heling period Groups 1week 2weeks 4weeks P vlue D1:control D2: Osteon II with H 22.69± ± ±7.561 C 54.96± ±13.18 c B 95.46±12.50 c D3:Osteon II With collgen 34.44±15.60 BC 45.34±7.44 B 86.12±10.92 c.000 D4: Osteon II 25.69±20.33 B 36.56± ±10.63 c.000 P vlue Men significnt t p 0.05 Cpitl letters refer to comprison mong tretment (verticlly). Smll letter refer to comprison mong period (Horizontlly). Pge 42

8 ISSN: , Vol. 5 Issue 8, ugust-2016 Chrt (2) Men of Digitl Rdiogrph using Duncn Test D1: Control group; D2: Osteon II with hyluronic group; D3: Osteon II with collgen memrne group; D4: OsteonII group ll groups showed Rdiogrphic evlution through different period postopertively there ws new on formtion nd significnt sttisticl difference found etween them (p vlue=.000). Rdiogrphic evlution t one week: Fig. (4) 1 Week 2 Week Pge 43

9 ISSN: , Vol. 5 Issue 8, ugust Week Figure (4) Digitl rdiogrph: show new one formtion t difference period: 1, 2, nd 4weeks D1: Control group; D2: Osteon II with hyluronic group;d3 :Osteon II with collgen memrne group;d4:osteonii group When rdiologicl comprison ws mde etween groups through 1 week period postopertively there were no significnt sttisticl difference found etween them t one week. (p vlue=.106). Rdiogrphic evlution of one heling ws superior in (D3) Osteon with collgen groups (34.44±15.60) nd D2 Osteon II with Hyluronic groups (30.44±15.60). Rdiogrphic evlution t two weeks: Fig. (4) Rdiogrphic evlution of one heling ws superior in (D2) Osteon II with Hyluronic cid (54.96±9.53) nd one formtion in D3 Osteon II with collgen (45.34±7.44) while Osteon II lone group 36.56± 7.32; p vlue=.001 Rdiogrphic evlution t four weeks: Fig. (4) Rdiogrphic evlution of one heling ws superior in (D2) Osteon II with Hyluronic cid (95.46 ±12.30) then (D3) Osteon II with collgen ws (86.12±10.92), Osteon II lone group ws (76.86±10.630). There were sttisticlly differences in the results of one formtion through the detection periods of evlution. P vlue =.022 DISCUSSION Tretment of lrge one defects represents gret chllenge, s one regenertion is required in lrge quntity nd my e eyond the potentil for self-heling. Lrge one defects include segmentl or lrge corticl defects creted y trum, infection, tumor resection, septic loosening round implnts nd skeletl normlities (9). In this study used new one grft mteril Osteon II collgen which is newly developed lloplstic mteril contining 70% H nd 30%- TCP which re quite close to mjor minerl components of the humn one (10).The ddition of collgen with one grft mteril to increse the osteoconductivity of Osteon II (11). No evidence of post-opertive infection ws noticed in this study. This my e due to the ppliction of sterile surgicl procedure nd the use of ntiiotic during postopertive cre in the form of intrmusculr injection nd topicl ntiiotic spry nd the high concentrtion of high moleculr weight of H hs the gretest cteriosttic effect prticulrly on ggregticter ctinomy cetemcomitns Prevotell nd Stphylococcus ureus strins commonly found in orl gingivl lesions nd periodontl wounds. Clinicl ppliction of H gels s surgicl therpy my reduce the cteril contmintion of surgicl wound site, therey, lessening the risk of post-surgicl infection nd promoting more predictle one genertion (12). The use of Collgen memrnes which ws designed in this study is to prevented the picl migrtion of epithelium nd supported new connective tissue ttchment nd tissue regenertion (13).The dditionl use of one-grfting mterils within the memrne to fill the defect should lso e evluted, iming to 'mimic' or even ccelerte the norml process of one formtion (14).The rit hs fster skeletl chnge nd one turnover (6) so it choses s niml model for this study. ccording to the scope of the present study it is the first time to evlute the effectiveness of loclly ppliction hyluronic cid nd collgen memrne with Osteon II collgen nd compre etween them on heling processes of the one. Pge 44

10 ISSN: , Vol. 5 Issue 8, ugust-2016 In the present experimentl investigtions were focused on mteril tht used for ccelerting one regenertion nd mturtion of one to shorten the tretment period nd improve the qulity of one mss. Groups using denstometoric nlysis y CT scn indictes more mturtion of the regenerted one in tested groups, s confirmed y incresed one density over ll time points. Oservtion nd ssessment of cllus density is fcilitted ecuse the holes of one defect re quickly distinguishle on the imges otined y CT scn clerer thn using DEX. lthough the rdiogrphic evlution showed tht the Osteon II collgen with hyluronic cid groups hve greter osteogenic potency thn Osteon II collgen with collgen memrne group; these two groups hve greter osteogenic potency thn Osteon II collgen lone nd control groups. These result ws gree with (sln et l., 2006) which demonstrted tht the Osteon II collgen with hyluronic cid groups hs superiority one heling histologiclly. (15). t the first week postopertively no significnt difference etween the four experimentl groups, lthough Osteon II collgen with hyluronic cid groups, Osteon II collgen with collgen memrne groups enhnce one formtion tht correlted to increse one heling. t the end of the second week postopertively, there were significnt incresed one formtion mong experimentl groups.t the end of four weeks the newly formtion one in Osteon II collgen with hyluronic cid showed more rdiodense nd greter mount of one formtion thn other experimentl groups.the result of this study ws greed with Kim (8) tht demonstrted the importnt of osteoconductive nd osteoinduction processes which re ctively ongoing. The explntion of Hylonect nd grfting tht significntly enhnce the heling process when used lone or together relted to increse morphogenesis nd tissue heling during one regenertion (8). The study done y sln et l (15) were compred the effects of utologous one grfting with or without H in rit tii defect model they reported tht H requires n osteoconductive scffold to e effective. The present study showed tht Hylonect or grfting significntly speeds the heling process with etter erly rdiologicl one heling in comintion group thn one grfting lone, especilly in the short term (16). t the end of four weeks the result showed more one formtion in Osteon II collgen with collgen memrne due to the ction of collgen memrne preventing the non osteogenic cells from prolifertion in the site where one formtion is wnted to grow (17). Resorle collgen memrnes re frequently used s wound dressings ecuse they ct s scffold, promote pltelet ggregtion, stilize clots, nd ttrct firolsts, fcilitting wound heling; therefore often used for GBR (17).Collgen memrne ct s scffolds for one deposition in guided one regenertion (GBR) fcilitting wound heling (18). CONCLUSION The use of osteon II collgen with hyluronic cid seemed to produce ppositive effect on rdiogrphicl ssessment of mount of one formtion t second nd fourth weeks of the study. good correltion etween osteon II collgen with hyluronic cid group compre with osteon II collgen with collgen memrne group rdiogrphiclly found superiority of osteon II collgen with hyluronic cid. The one density of regenerted one is greter in Hyluronic cid with Osteon II collgen nd in collgen memrne with Osteon II collgen compred to control groups nd Osteon II collgen groups long period of study. REFERENCES [1]. Hoexter DL. (2002). Bone regenertion grft mterils. J Orl Implntol. 28(6): [2]. Kim YK.,Yun PY., Lim SC., Kim SG.& Lee HJ,Ong JL.(2008). Clinicl evlution of Osteon s new lloplstic mteril in sinus ome grfting nd its effect on one heling. J Biomed. Mt. Res. B ppl Biometer 86: [3]. Bnsl J., Kedige SD.&nnd S.(2010). Hyluronic cid: promising meditor for periodontl regenertion. Indin J Dent Res. 21(4): [4]. Tschieri S., Corell S., Tsesis I., Bortolin M. & Del Fro M. (2007). Effect of guided tissue regenertion on the outcome of surgicl endodontic tretment of through-nd-through lesions: retrospective study t 4-yer follow-up. Orl Mxillofc Surg. 15: [5]. Mcllister BS.& Hghight K. (2007). Bone ugmenttion techniques.j Periodontol. 78(3): [6]. Gilsnz V., Roe TF., Giens DT., Schulz EE.; Crlson ME.; Gonzlez.;.&.;Boecht;MI.(1988).Effect of sex steroid pek one density of growing rits. m J Physiol. 255(4 Pt 1):E [7]. Wng X., Mrey JD.& grwl CM. (1998). n interspecies comprison of one frcture properties. Biomed Mter Eng. 8(1):1-9. [8]. Kim JM., Kim MH., Kng SS., Kim G.&, Choi SH.(2013). Comprle one heling cpcity of different one grft mtrices in rit segmentl defect model. J Vet Sci. 15(2): [9]. Dimitriou R., Jones E., McGongle D.& Ginnoudis PV. (2011). Bone regenertion: current concept nd future directions. BMC Med. 31;9:66.86/ [10]. Kim YK.,Yun PY., Lim SC., Kim SG.& Lee HJ,Ong JL.(2008). Clinicl evlution of Osteon s new lloplstic mteril in sinus ome grfting nd its effect on one heling. J Biomed. Mt. Res. B ppl Biometer 86: [11]. Keting J.F & McQueen M.M. (2001). Sustitutes for utologous one grft in orthopedic trum. J Bone Joint Surg Br. 83(1):3-8. [12]. Pirnzr P., Wolinsky L., Nchnni S., Hke S., Pilloni.& Bernrd GW.(1999). Bcteriosttic effects of hyluronic cid. J periodontl. 70(4): [13]. Gzdg R., Lne JM, Glser D.& Forster R.(1995). lterntives to utogenous Bone Grft: Efficcy nd Indictions. J m cd Orthop Surg. 3(1):1-8. Pge 45

11 ISSN: , Vol. 5 Issue 8, ugust-2016 [14]. Rozli Dimitriou., George I Mtliotkis., Giorgio Mri Clori eter V Ginnoudis Emil uthor Contri eqully. (2012). The role of rrier memrnes for guided one regenertion nd restortion of lrge one defects: current experimentl nd cliniclevidence. BMC Medicine 10:81. [15]. sln M.,Simsek G. & Dyi E.(2006). The effect of hyluronic cid-supplemented one grft in one heling: experimentl study in rits. J Biomter ppl. 20(3): [16]. Rhodes NP., Hunt J., Longinotti C.&, Pvesio. (2011). In vivo chrcteriztion of Hylonect, novel iodegrdle surgicl mesh J Surg Res. 1;168(1):e [17]. Bunyrtvej P& Wng HL. (2001). Collgen memrnes: review. J Periodontol. 72(2): [18]. Luitud C., Lflmme C., Semlli., Sidi S., Grenier G., Zkrzewski.&Rouhi. M.(2007). Development of n engineering utologous pltl mucos-like tissue for potentil clinicl pplictions. J Biomed Mter Res B ppl Biomter. 83(2): Pge 46