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Effect of low intensity pulse ultrasound on the neutrophil function and epithelial thickness after skin excision in diabetes mellitus type 2 rats’ model (an immunohistochemistry study on matrix metalloproteinase-9)

  • Titis Laraswati ,
  • Cahya Y. Hasan ,
  • Bambang Dwirahardjo ,


Objective: To determine the effect of Low Intensity Pulse Ultrasound (LIPUS) on the neutrophil function and epithelial thickness after skin excision in diabetes mellitus type 2 rats’ model (an Immunohistochemistry Study on Matrix Metalloproteinase-9).Material and Methods: Thirty-six Sprague dawley rats were divided into six groups: DM2, DM4, DM7, DML2, DML4, and DML7, each of which consist of 6 rats. Rats that matched the inclusion criteria were made into diabetes mellitus type 2 by injecting Nicotinamide and Streptozotocin. In all rat’s excision wound was made with punch biopsy in the back area. DML group received LIPUS therapy in the wound area (frequency: 3 MHz, intensity: 0.5 W/cm2, duty cycle: 20%, duration: 3 minutes every day) for two days (DML2), four days (DML4) and seven days (DML7). In the wound tissue an immunohistochemical examination was performed with Polyclonal Antibody MMP-9 to observe MMP-9 expression and and Periodic Acid-Schiff staining to observe epithelial thickness.Results: Two-Way ANOVA and Post-Hoc LSD test showed lower level of MMP-9 expressions in the DML group than in the DM group, whereas the epithelial thickness in the DML group was found higher than in the DM group. The Pearson test showed a correlation between the level of MMP-9 expressions and epithelial thickness (p=0.000; r=-0.785).Conclusion: LIPUS therapy reduced MMP-9 expression and increases epithelial thickness.


  1. Sjamsuhidajat R, De-Jong W. Surgical Textbook. 3rd ed. Penerbit Buku Kedokteran EGC; Jakarta: 2010. p. 67. (In Indonesia)
  2. Valero AM, Garcia JCF, Ballester AH, et al. Effects of diabetes on the osseointegration of dental implants. Med Oral Patol Oral Cir Bucal 2007;12: 38-43.
  3. Luo HR and Loison F. Constitutive neutrophil apoptosis: mechanisms and regulation. Am J Hematol 2008;83: 288-295.
  4. Wilgus TA, Roy S, Jodi C, et al. Neutrophils and wound repair: positive actions and negative reactions. Adv Wound Care 2013;2: 379-388.
  5. Blakytny R, Jude E. The Molecular biology of chronic wounds and delayed healing in diabetes. Diabet Med 2006;23: 594-608.
  6. Volmer TM, Lobmann R. Neuropathy and diabetic foot syndrome. Int J Mol Sci 2016;17: 1-11.
  7. Tuhin RH, Begum MM, Rahman S, et al. Wound healing effect of euphorbia hirta linn. (euphorbiaceae) in alloxan induced diabetic rats. BMC Complement and Alternative Medicine 2017;17: 1-14.
  8. Kintoko, Karimatulhajj H, Elfasyari TY, et al. Effect of diabetes condition on topical treatment of binahong leaf fraction in wound healing process. Trad Med J 2017;22: 103-110.
  9. Loureiro TCA, Munhoz CD, Martins JO, et al. Neutrophil function and metabolism in individuals with diabetes mellitus. Braz J Med Biol Res 2007;40: 1037-1044.
  10. Pithon-Curi TC, Schumacher RI, Freitas JJ, et al. Glutamine delays spontaneous apoptosis in neutrophils. Am J Physiol Cell Physiol, 2003; 284: C1355-C1361.
  11. Pastar I, Stojadinovic O, Yin NC, et al. Epithelialization in wound healing: a comprehensive review. Adv Wound Care 2013;3: 445-464.
  12. Nooh N, Graves DT. Healing is delayed in oral compared to dermal excisional wounds. J Periodontol 2003;74: 242-246.
  13. Rousseau G, Blouin A, Monchalin JP. Non-contact photo acoustic tomography and ultrasonography for tissue imaging. Biomed Opt Expres 2012;3: 16-25.
  14. Lou S, Liv H, Li Z, et al. The effects of low intensity pulsed ultrasound on fresh fracture. Medicine 2017;96: e8181.
  15. Da-Silva-Junior EM, Mesquita-Ferrari RA, Franca CM, et al. Modulating effect of low intensity pulsed ultrasound on the phenotype of inflammatory cells. Biomed Pharmacother 2017;96: 1-7.
  16. Leng X, Shang J, Gao D, et al. Low intensity pulsed ultrasound promotes proliferation and migration of hacat keratinocytes through the PI3K/AKT and JNK Pathways. Braz J Med Biol Res 2018;51: e7862.
  17. Iwanabe Y, Masaki C, Tamura A, et al. The effect of low intensity pulsed ultrasound on wound healing using scratch assay in epithelial cells. J Prostho Res 2016;60: 308-314.
  18. Shindo T, Ito K, Ogata T, et al. Low intensity pulsed ultrasound enhances angiogenesis and ameliorates left ventricular dysfunction in a mouse model of acute myocardial infarction. Arterioscler Thromb Vasc Biol 2015; 36:1220-1229.
  19. Piedade MC, Galhardo MS, Battlehner CN, et al. Effect of ultrasound therapy on the repair of gastrocnemius muscle injury in rats. J Ultrasonics 2008;48: 403-411.
  20. Hill GE, Fenwick S, Matthews BJ, et al. The effect of low intensity pulsed ultrasound on repair of epithelial cell monolayers in vitro. Ultrasound Med Biol 2005;1: 1701-1706.
  21. Tian S, Li M, Dong F, et al. The role of low intensity pulsed ultrasound on bone and soft tissue healing. Int J Clin Exp Med 2016;9: 12450-12456.
  22. Nakhaee A, Bokaeian M, Saravani M, et al. Attenuation of oxidative stress in streptozotocin-induced diabetic rats by eucalyptus globulus. Indian J Clin Biochem 2009;24: 419-425.
  23. Kiran G, Nandini CD, Ramesh HP, et al. Progression of early phasediabetic nephropathy in streptozotocin induced diabetic rats: evaluation of various kidney related parameters. Ind J Exp Biol 2012;50: 133-140.
  24. Zhou S, Bachem MG, Seufferlein T, et al. Low intensity pulsed ultrasound accelerates macrophage phagocytosis by a pathway that requires actin polymerization Rho Src/MAPKs activity. Cell Signal 2008;20: 695-704.
  25. Freitas LBS, Freitas TP, Rocha LG, et al. Effect of therapeutic pulsed ultrasound on parameters of oxidative. Cell Biol Int 2007;31: 482-488.
  26. Renno ACM, Toma RL, Feitosa SM, et al. Comparative effects of low intensity pulsed ultrasound and low level laser therapy on injured skeletal muscle. Photomed Laser Surg 2011;29: 5-10.
  27. Shu B, Yang Z, Li X, et al. Effect of different pulsed ultrasound on the restoration of rat skeletal muscle contusion. Cell Biochem Biophys 2012;62: 329-336.
  28. Zhou S, Schmelz A, Seufferlein T, et al. Molecular mechanisms of low intensity pulsed ultrasound in human skin fibroblasts. J Biol Chem 2004;279: 54463–54469.
  29. Nainggolan LI, Gunasagaran L. Prevalence of alveolar bone deffect pattern in periodontitis patients with diabetes mellitus using bitewing radiography. J Dentomaxillofac Sci 2018;3: 88-90.

How to Cite

Laraswati, T., Hasan, C. Y., & Dwirahardjo, B. (2021). Effect of low intensity pulse ultrasound on the neutrophil function and epithelial thickness after skin excision in diabetes mellitus type 2 rats’ model (an immunohistochemistry study on matrix metalloproteinase-9). Journal of Dentomaxillofacial Science, 6(1), 54–60.




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