Hatice ÖZER, Aysan LEKTEMUR ALPAN, Hülya TOKER

Ozonun deneysel diabetes mellitustta otojen greft iyileşmesini artırması: Morfometrik ve immünhistokimyasal çalışma

Amaç: Diabetes mellitus (DM), periodontitis patogenezinde önemli rol oynayan bir metabolik bozukluktur. Ozonun antimikrobiyal, immünostimülasyon, antihipoksik etkileri gibi çeşitli etkileri vardır ve antioksidan enzimleri ve anjiyogenezi aktive eder. Bu çalışmanın amacı otojen kemik grefti ile tedavi edilen diyabetik sıçan kalvaryal defektlerinde uygulanan ozonunun kemik iyileşmesi üzerine etkisini morfometrik ve immünhistokimyasal olarak incelemektir. Gereç ve Yöntemler: Sıçanlarda diyabet indüklendi ve kalvaryal defektler oluşturuldu. Çalışma grupları: 1-Boş defekt (Kontrol, n = 14) grubu, 2-Otogreft (AG, n = 14) grubu, 3-Boş defekt + ozon tedavisi (Kontrol + Ozon, n = 14) grubu, 4-Otogreft + Ozon uygulaması (AG + Ozon, n = 14) grubu. Operasyon gününde ve sonraki 2 haftada gaz ozon (140ppm2L / d, 2.24 mg) kalvaryal bölgeye uygulandı. Toplam kemik alanı ölçüldü. Osteokalsin ve Kemik morfojenik protein-2 protein ekspresyonları değerlendirildi. Bulgular: Kontrol ve Kontrol + Ozon gruplarında çalışma süresince osteoklast ve rezidüel lakün gözlenmedi. AG + Ozon grubunda osteoblastlar, 4. haftada AG grubundan daha fazla gözlendi (p> 0.05). AG + Ozon grubunun 4. haftada AG grubundan daha fazla toplam kemik alanı vardı. AG + Ozon grubu diğer gruplara göre daha fazla BMP-2 immün pozitifliği saptadı. AG gruplarında osteokalsin immün pozitifliği, kontrol gruplarından daha yüksekti. Sonuç: Bu çalışmanın kısıtlılıkları dahilinde, gaz ozon uygulaması, özellikle diyabetik sıçanlarda kemik rejenerasyonunun erken aşamalarında osteoklast sayısını azaltarak ve osteoblast sayısını artırarak kemik rejenerasyonunu artırmıştır.

Ozone improves autogenous graft healing in experimental diabetes mellitus: A morphometric and immunohistochemical study

Background: Diabetes mellitus (DM) is a metabolic disorder whichplays crucial role in the pathogenesis of periodontitis. Ozone haveseveral actions such as antimicrobial, immunostimulating,antihypoxic effects, and activates antioxidant enzymes andangiogenesis. The aim of this study is to investigate the effect ofgaseous ozone on bone healing in diabetic rat calvarial defectstreated with autogenous bone graft, morphometrically andimmunohistochemically.Material and Methods: Diabetes was induced and critical sizedefects were created on rats. Study groups: 1-Empty defect(Control, n=14) group, 2-Autograft (AG, n=14) group, 3-Emptydefect+ozone therapy (Control+Ozone, n=14) group, 4-Autograft+ozone application (AG+Ozone, n=14) group. Gaseousozone was applied on the operation day and the following 2 weeksdaily (140ppm @ 2L/d, 2.24 mg). Total bone area was measured.Osteocalcin and Bone morphogenic protein-2 protein expressionswere evaluated.Results: Control and Control+Ozone groups had no osteoclastand residuel lacunae during the study . Osteoblasts in AG+Ozonegroup were higher than AG group at 4th week (p>0.05).AG+Ozone group had more total bone area than AG group at 4thweek. AG+Ozone group revealed more BMP-2 immune positivitycompared to the other groups. Osteocalcin immune positivity inAG groups was higher than those of the Control groups.Conclusion: Within the limitations of this study, gaseous ozoneapplication decreased osteoclast number and increasedosteoblast number and bone regeneration, especially, in earlystages of bone regeneration in diabetic rats

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1. Gurav AN. Management of diabolical diabetes mellitus and periodontitis nexus: Are we doing enough? World J Diabetes. 2016; 7(4): 50-66.
2. Jiao H, Xiao E, Graves DT. Diabetes and Its Effect on Bone and Fracture Healing. Curr Osteoporos Rep. 2015; 13(5): 327-35.
3. Daugela P, Cicciu M, Saulacic N. Surgical Regenerative Treatments for Peri-Implantitis: Metaanalysis of Recent Findings in a Systematic Literature Review. J Oral Maxillofac Res. 2016; 7(3): e15.
4. Dimitriou R, Jones E, McGonagle D, Giannoudis PV. Bone regeneration: current concepts and future directions. BMC Med. 2011; 9: 66.
5. Yin W, Qi X, Zhang Y, Sheng J, Xu Z, Tao S, et al. Advantages of pure platelet-rich plasma compared with leukocyte- and platelet-rich plasma in promoting repair of bone defects. J Transl Med. 2016; 14(1): 73.
6. Peng W, Kim IK, Cho HY, Pae SP, Jung BS, Cho HW, et al. Assessment of the autogenous bone graft for sinus elevation. J Korean Assoc Oral Maxillofac Surg. 2013; 39(6): 274-82.
7. Keskiner I, Alkan A, Acikgoz G, Arpak N, Kaplan S, Arslan H. Platelet-rich plasma and autogenous bone graft combined with guided tissue regeneration in periodontal fenestration defects in dogs. Int J Periodontics Restorative Dent. 2014; 34(6): e112-20.
8. Kokdere NN, Baykul T, Findik Y. The use of plateletrich fibrin (PRF) and PRF-mixed particulated autogenous bone graft in the treatment of bone defects: An experimental and histomorphometrical study. Dent Res J (Isfahan). 2015; 12(5): 418-24.
9. Toker H, Ozdemir H, Ozer H, Eren K. Alendronate enhances osseous healing in a rat calvarial defect model. Arch Oral Biol. 2012; 57(11): 1545-50.
10.Ozdemir H, Toker H, Balci H, Ozer H. Effect of ozone therapy on autogenous bone graft healing in calvarial defects: a histologic and histometric study in rats. J Periodontal Res. 2013; 48(6): 722-6.
11.Abelev B, Adam J, Adamova D, Adare AM, Aggarwal MM, Aglieri Rinella G, et al. J/psi suppression at forward rapidity in Pb-Pb collisions at radicals(NN) = 2.76 TeV. Phys Rev Lett. 2012; 109(7): 072301.
12.Bocci VA. Scientific and medical aspects of ozone therapy. State of the art. Arch Med Res. 2006; 37(4): 425-35.
13.Huth KC, Jakob FM, Saugel B, Cappello C, Paschos E, Hollweck R, et al. Effect of ozone on oral cells compared with established antimicrobials. Eur J Oral Sci. 2006; 114(5): 435-40.
14.Nagayoshi M, Fukuizumi T, Kitamura C, Yano J, Terashita M, Nishihara T. Efficacy of ozone on survival and permeability of oral microorganisms. Oral Microbiol Immunol. 2004; 19(4): 240-6.15.Huth KC, Quirling M, Lenzke S, Paschos E, Kamereck K, Brand K, et al. Effectiveness of ozone against periodontal pathogenic microorganisms. Eur J Oral Sci. 2011; 119(3): 204-10.
16.Kazancioglu HO, Ezirganli S, Aydin MS. Effects of laser and ozone therapies on bone healing in the calvarial defects. J Craniofac Surg. 2013; 24(6): 2141-6.
17.Kazancioglu HO, Kurklu E, Ezirganli S. Effects of ozone therapy on pain, swelling, and trismus following third molar surgery. Int J Oral Maxillofac Surg. 2014; 43(5): 644-8.
18.Erdemci F, Gunaydin Y, Sencimen M, Bassorgun I, Ozler M, Oter S, et al. Histomorphometric evaluation of the effect of systemic and topical ozone on alveolar bone healing following tooth extraction in rats. Int J Oral Maxillofac Surg. 2014; 43(6): 777-83.
19.Lektemur Alpan A, Toker H, Ozer H. Ozone Therapy Enhances Osseous Healing In Diabetic Rats With Calvarial Defect: A Morphometric and Immunohistochemical Study. J Periodontol. 2016: 1-15.
20.Deliberador TM, Giovanini AF, Lopes TR, Zielak JC, Moro A, Baratto Filho F, et al. Immunoexpression of PPAR-gamma and osteocalcin proteins for bone repair of critical-size defects treated with fragmented autogenous abdominal adipose tissue graft. Braz Dent J. 2014; 25(3): 179-85.
21.Kumar V, Mahdi F, Khanna AK, Singh R, Chander R, Saxena JK, et al. Antidyslipidemic and Antioxidant Activities of Hibiscus rosa sinensis Root Extract in Alloxan Induced Diabetic Rats. Indian J Clin Biochem. 2013; 28(1): 46-50.
22.Mohammadi Sartang M, Mazloomi SM, Tanideh N, Rezaian Zadeh A. The Effects of Probiotic Soymilk Fortified with Omega-3 on Blood Glucose, Lipid Profile, Haematological and Oxidative Stress, and Inflammatory Parameters in Streptozotocin Nicotinamide-Induced Diabetic Rats. J Diabetes Res. 2015; 2015: 696372.
23.Islam MS, Loots du T. Experimental rodent models of type 2 diabetes: a review. Methods Find Exp Clin Pharmacol. 2009; 31(4): 249-61.
24.Srinivas NR. Strategies for preclinical pharmacokinetic investigation in streptozotocininduced diabetes mellitus (DMIS) and alloxaninduced diabetes mellitus (DMIA) rat models: case studies and perspectives. Eur J Drug Metab Pharmacokinet. 2015; 40(1): 1-12.
25.Frode TS, Medeiros YS. Animal models to test drugs with potential antidiabetic activity. J Ethnopharmacol. 2008; 115(2): 173-83
26.Ishibashi T, Tanaka K, Taniguchi Y. Platelet aggregation and coagulation in the pathogenesis of diabetic retinopathy in rats. Diabetes. 1981; 30(7): 601-6.
27.Valentovic MA, Alejandro N, Betts Carpenter A, Brown PI, Ramos K. Streptozotocin (STZ) diabetes enhances benzo(alpha)pyrene induced renal injury in Sprague Dawley rats. Toxicol Lett. 2006; 164(3): 214-20.
28.Lei YC, Hwang JS, Chan CC, Lee CT, Cheng TJ. Enhanced oxidative stress and endothelial dysfunction in streptozotocin-diabetic rats exposed to fine particles. Environ Res. 2005; 99(3): 335-43.
29.Schmitz JP, Hollinger JO. The critical size defect as an experimental model for craniomandibulofacial nonunions. Clin Orthop Relat Res. 1986(205): 299- 308.
30.Freeman E, Turnbull RS. The value of osseous coagulum as a graft material. J Periodontal Res. 1973; 8(4): 229-36.
31.Petridis X, Diamanti E, Trigas G, Kalyvas D, Kitraki E. Bone regeneration in critical-size calvarial defects using human dental pulp cells in an extracellular matrix-based scaffold. J Craniomaxillofac Surg. 2015; 43(4): 483-90.
32.Vo TN, Ekenseair AK, Spicer PP, Watson BM, Tzouanas SN, Roh TT, et al. In vitro and in vivo evaluation of self-mineralization and biocompatibility of injectable, dual-gelling hydrogels for bone tissue engineering. J Control Release. 2015; 205: 25-34.
33.Abreu TC, Lima RP, Souza VS, Campos Junior O, Albuquerque AV, Aguiar JL, et al. The biopolymer sugarcane as filling material of critical defects in rats. Acta Cir Bras. 2016; 31(1): 53-8.
34.Gorla LF, Spin-Neto R, Boos FB, Pereira Rdos S, Garcia-Junior IR, Hochuli-Vieira E. Use of autogenous bone and beta-tricalcium phosphate in maxillary sinus lifting: a prospective, randomized, volumetric computed tomography study. Int J Oral Maxillofac Surg. 2015; 44(12): 1486-91.
35.Shyng YC, Devlin H, Sloan P. The effect of streptozotocin-induced experimental diabetes mellitus on calvarial defect healing and bone turnover in the rat. Int J Oral Maxillofac Surg. 2001; 30(1): 70-4.
36.Pacios S, Kang J, Galicia J, Gluck K, Patel H, Ovaydi-Mandel A, et al. Diabetes aggravates periodontitis by limiting repair through enhanced inflammation. FASEB J. 2012; 26(4): 1423-30.
37.Lu H, Kraut D, Gerstenfeld LC, Graves DT. Diabetes interferes with the bone formation by affecting the expression of transcription factors that regulate osteoblast differentiation. Endocrinology. 2003; 144(1): 346-52
38.Kan B, Sencimen M, Bayar GR, Korkusuz P, Coskun AT, Korkmaz A, et al. Histomorphometric and microtomographic evaluation of the effects of hyperbaric oxygen and systemic ozone, used alone and in combination, on calvarial defect healing in rats. J Oral Maxillofac Surg. 2015; 73(6): 1231 e1-10.
39.Salazar VS, Gamer LW, Rosen V. BMP signalling in skeletal development, disease and repair. Nat Rev Endocrinol. 2016; 12(4): 203-21.
40.Pacios S, Andriankaja O, Kang J, Alnammary M, Bae J, de Brito Bezerra B, et al. Bacterial infection increases periodontal bone loss in diabetic rats through enhanced apoptosis. Am J Pathol. 2013; 183(6): 1928-35.
41.Sarkar PD, Choudhury AB. Relationships between serum osteocalcin levels versus blood glucose, insulin resistance and markers of systemic inflammation in central Indian type 2 diabetic patients. Eur Rev Med Pharmacol Sci. 2013; 17(12): 1631-5.
42.Zanatta LC, Boguszewski CL, Borba VZ, Kulak CA. Osteocalcin, energy and glucose metabolism. Arq Bras Endocrinol Metabol. 2014; 58(5): 444-51.
43.Starup-Linde J, Vestergaard P. Biochemical bone turnover markers in diabetes mellitus - A systematic review. Bone. 2015.
44.Roszer T. Inflammation as death or life signal in diabetic fracture healing. Inflamm Res. 2011; 60(1): 3-10.