Evaluation of the Effect of Platelet Rich Plasma on Wound Healing in the Tongue of Normal and Streptozotocin-induced Diabetic Albino Rats: Histological, Immunohistochemical, and Ultrastructural Study
DOI:
https://doi.org/10.3889/oamjms.2020.5366Keywords:
Platelet rich plasma, Wound healing, Diabetes, Rat, Tongue, p63, Vimentin, TEMAbstract
Background: Delayed healing of diabetic wounds has been well-documented. Currently, the use of platelet-rich plasma (PRP) has attracted great attention in many medical fields including wound healing.
Aim: Histological, immunohistochemical and ultrastructural evaluation of the effect of PRP on wound healing in the tongue of normal and Streptozotocin-induced diabetic albino rats.
Methodology: A total number of 108 adult male albino rats with average weight 200gm, were used in the study. The animals were classified into two main groups: non-diabetic and diabetic groups. Each group was further divided into three subgroups: non- treated wound, PRP-treatment before wound, and PRP-treatment after wound. Tongue specimens were dissected on postoperative days 1, 3, and 7. The specimens were examined histologically by H&E, immunohistochemically by p63 and vimentin, and ultra-structurally by TEM.
Results: The most accelerated wound healing was revealed in the subgroups treated with PRP before the wound, whether non-diabetic or diabetic, which occurred very early at the 3rd day postoperative in both cases. While complete wound healing was revealed at the 7th day postoperative in both the non-diabetic and diabetic subgroups treated with PRP after the wound, which was like the non-diabetic control subgroup. Whilst, the diabetic non-treated subgroup only showed partial wound healing at the 7th day postoperative.
Conclusion: A single injection of PRP could be used as a prophylactic to prevent expected impaired wound healing in diabetic oral mucosal wounds and to enhance wound healing in non-diabetic wounds. PRP could be used as a therapeutic to enhance wound healing in diabetic and non-diabetic oral mucosal wounds.
Key Words: platelet rich plasma, wound healing, diabetes, rat, tongue, p63, vimentin, TEM
BACKGROUND: Delayed healing of diabetic wounds has been well-documented. At present, the use of platelet-rich plasma (PRP) has attracted great attention in many medical fields including wound healing.
AIM: Histological, immunohistochemical, and ultrastructural evaluation of the effect of PRP on wound healing in the tongue of normal and streptozotocin-induced diabetic albino rats.
METHODOLOGY: A total number of 108 adult male albino rats with average weight 200 g were used in the study. The animals were classified into two main groups: Non-diabetic and diabetic groups. Each group was further divided into three subgroups: Non-treated wound, PRP-treatment before wound, and PRP-treatment after wound. Tongue specimens were dissected on post-operative days 1, 3, and 7. The specimens were examined histologically by H&E, immunohistochemically by p63 and vimentin, and ultrastructurally by TEM.
RESULTS: The most accelerated wound healing was revealed in the subgroups treated with PRP before the wound, whether non-diabetic or diabetic, which occurred very early at the 3rd day post-operative in both cases. While complete wound healing was revealed at the 7th day post-operative in both the non-diabetic and diabetic subgroups treated with PRP after the wound, which was like the non-diabetic control subgroup. While, the diabetic non-treated subgroup only showed partial wound healing at the 7th day post-operative.
CONCLUSION: A single injection of PRP could be used as a prophylactic to prevent expected impaired wound healing in diabetic oral mucosal wounds and to enhance wound healing in non-diabetic wounds. PRP could be used as a therapeutic to enhance wound healing in diabetic and non-diabetic oral mucosal wounds.
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References
Abd-Elmotelb MA. Morphometric, histological and immunohistochemical study of tongue epithelium in diabetic rats. Life Sci J. 2018;15:1-6.
Rabo A, Mohamed RA. Comparative study of the effect of Allium sativum (Garlic), Allium cepa (Onion) and insulin on the lingual papillae of streptozotocin induced diabetic albino rats. In: Oral Biology. Egypt: Ain Shams University, Faculty of Dentistry 2018. p. 150. https://doi.org/10.21608/ejh.2020.26939.1267
Tesseromatis C, Kotsiou A, Parara H, Vairaktaris E, Tsamouri M. Morphological changes of gingiva in streptozotocin diabetic rats. Int J Dent. 2009;2009:725628. https://doi. org/10.1155/2009/725628 PMid:20339569
Umasankar K, Balwin N, Backyavathy DM. Wound healing activity of topical mentha piperita and cymbopogan citratus essential oil on streptozotocin induced rats. Asian J Pharm Clin Res. 2013;6:180-3.
Venter NG, Marques RG, Dos Santos JS, Monte-Alto-Costa A. Use of platelet-rich plasma in deep second and third-degree burns. Burns. 2016;42:807-14. https://doi.org/10.1016/j. burns.2016.01.002 PMid:26822695
Graves DT, Liu R, Oates TW. Diabetes-enhanced inflammation and apoptosis impact on periodontal pathosis. Periodontol 2000. 2007;45(1):128-37. https://doi. org/10.1111/j.1600-0757.2007.00219.x PMid:17850453
Berdal M, Appelbom HI, Eikrem JH, Lund A, Zykova S, Busund LT, et al. Aminated β-1,3-d-glucan improves wound healing in diabetic db/db mice. Wound Repair Regen. 2007;15(6):825-32. https://doi.org/10.1111/j.1524-475x.2007.00286.x PMid:18028130
Thorne CH, Bartlett SP, Beasley RW, Aston SJ, Gurtner GC, Spear SL. Wound healing: Normal and abnormal. In: Grabb and Smith’s Plastic Surgery. Alphen aan den Rijn: Wolters Kluwer Health Adis (ESP); 2013.
Nanci A. Repair and regeneration of oral tissues. In: Ten Cate’s Oral Histology: Development, Structure, and Function. St. Louis, Missouri: Elsevier; 2018. p. 729-740.
Gazaerly HE, Elbardisey DM, Eltokhy HM, Teaama D. Effect of transforming growth factor beta 1 on wound healing in induced diabetic rats. Int J Health Sci. 2013;7(2):160-72. https://doi. org/10.12816/0006040 PMid:24421745
Setta HS, Elshahat A, Elsherbiny K, Massoud K, Safe I. Platelet-rich plasma versus platelet-poor plasma in the management of chronic diabetic foot ulcers: A comparative study. Int Wound J. 2011;8(3):307-12. https://doi. org/10.1111/j.1742-481x.2011.00797.x PMid:21470370
Marx RE. Platelet-rich plasma (PRP): What is PRP and what is not PRP? Implant Dent. 2001;10(4):225-8. https://doi. org/10.1097/00008505-200110000-00002 PMid:11813662
Kramer ME, Keaney TC. Systematic review of platelet-rich plasma (PRP) preparation and composition for the treatment of androgenetic alopecia. J Cosmet Dermatol. 2018;17(5):666-71. https://doi.org/10.1111/jocd.12679 PMid:29790267
Tian J, Cheng LH, Cui X, Lei XX, Tang JB, Cheng B. Application of standardized platelet-rich plasma in elderly patients with complex wounds. Wound Repair Regen. 2019;27(3):268-76. https://doi.org/10.1111/wrr.12702 PMid:30693614
Dhillon RS, Schwarz EM, Maloney MD. Platelet-rich plasma therapy future or trend? Arthritis Res Ther. 2012;14(4):219. https://doi.org/10.1186/ar3914 PMid:22894643
Textor J. Platelet-rich plasma (PRP) as a therapeutic agent: Platelet biology, growth factors and a review of the literature. In: Lana JF, Santana MH, Belangero WD, Luzo AC, editors. Platelet-Rich Plasma Regenerative Medicine: Sports Medicine, Orthopedic, and Recovery of Musculoskeletal Injuries. Berlin, Germany: Springer Science and Business Media; 2014. p. 61-94. https://doi.org/10.1007/978-3-642-40117-6_2
Davydova L, Tkach G, Tymoshenko A, Moskalenko A, Sikora V, Kyptenko L, et al. Anatomical and morphological aspects of papillae, epithelium, muscles, and glands of rats’ tongue: Light, scanning, and transmission electron microscopic study. Int Med Appl Sci. 2017;9(3):168-77. https://doi. org/10.1556/1646.9.2017.21 PMid:29201443
Goździewska-Harłajczuk K, Klećkowska-Nawrot J, Barszcz K, Marycz K, Nawara T, Modlińska K, et al. Biological aspects of the tongue morphology of wild-captive WWCPS rats: A histological, histochemical and ultrastructural study. Anat Sci Int. 2018;93(4):514-32. https://doi.org/10.1007/s12565-018-0445-y PMid:29948977
Noszczyk B, Majewski S. p63 Expression during normal cutaneous wound healing in humans. Plast Reconstr Surg 2001;108:1242-7; discussion 1248. https://doi. org/10.1097/00006534-200110000-00023 PMid:11604626
Fuyuhiro Y, Yashiro M, Noda S, Kashiwagi S, Matsuoka J, Doi Y, et al. Clinical significance of vimentin-positive gastric cancer cells. Anticancer Res 2010;30(12):5239-43. PMid:21187520
Robinson-Bennett B, Han A. Handbook of Immunohistochemistry and in situ Hybridization of Human Carcinomas. Amsterdam, Netherlands: Elsevier Science; 2006.
Choudhary P, Choudhary OP. Uses of transmission electron microscope in microscopy and its advantages and disadvantages. Int J Curr Microbiol Appl Sci. 2018;7(5):743-7. https://doi.org/10.20546/ijcmas.2018.705.090
Williams DB, Carter CB. The transmission electron microscope. In: McNamara A, editor. Transmission Electron Microscopy a Textbook for Materials Science. United States: Springer; 2009. p. 3-22. https://doi.org/10.1007/978-0-387-76501-3_1
Middleton KK, Barro V, Muller B, Terada S, Fu FH. Evaluation of the effects of platelet-rich plasma (PRP) therapy involved in the healing of sports-related soft tissue injuries. Iowa Orthop J. 2012;32:150-63. PMid:23576936
Tandon PN, Mahajan RC, Anand N, Basu SK, Ganguly NK, Kamboj VP, et al. Guidelines for Care and Use of Animals in Scientific Research. New Delhi: Indian National Science Academy; 2000.
Bhattacharya S, Aggarwal R, Singh VP, Ramachandran S, Datta M. Downregulation of miRNAs during delayed wound healing in diabetes: Role of dicer. Mol Med. 2015;21:847-60. https://doi.org/10.2119/molmed.2014.00186 PMid:26602065
Ebaid H, Ahmed OM, Mahmoud AM, Ahmed RR. Limiting prolonged inflammation during proliferation and remodeling phases of wound healing in streptozotocin-induced diabetic rats supplemented with camel undenatured whey protein. BMC Immunol. 2013;14:31. https://doi.org/10.1186/1471-2172-14-31 PMid:23883360
Dhurat R, Sukesh MS. Principles and methods of preparation of platelet-rich plasma: A review and author’s perspective. J Cutan Aesthet Surg. 2014;7:189-97. https://doi. org/10.4103/0974-2077.150734 PMid:25722595
Elsaadany B, El Kholy S, El Rouby D, Rashed L, Shouman T. Effect of transplantation of bone marrow derived mesenchymal stem cells and platelets rich plasma on experimental model of radiation induced oral mucosal injury in albino rats. Int J Dent. 2017;2017:8634540. https://doi.org/10.1155/2017/8634540 PMid:28337218
Drury RA, Wallington EA. Carleton’s Histological Techniques. 5th ed. New York: Oxford University Press; 1980. p. 195. https://doi. org/10.1038/modpathol.2011.89
Martin SE, Temm CJ, Goheen MP, Ulbright TM, Hattab EM. Cytoplasmic p63 immunohistochemistry is a useful marker for muscle differentiation: An immunohistochemical and immunoelectron microscopic study. Mod Pathol. 2011;24:1320-6. PMid:21623385
Kurihara K, Isobe T, Yamamoto G, Tanaka Y, Katakura A, Tachikawa T. Expression of BMI1 and ZEB1 in epithelial-mesenchymal transition of tongue squamous cell carcinoma. Oncol Rep. 2015;34(2):771-8. https://doi.org/10.3892/or.2015.4032 PMid:26043676
Bozzola JJ, Russell LD. Electron Microscopy: Principles and Techniques for Biologists. 2nd ed. Boston: Jones and Bartlett Publishers International; 1999.
Sciubba JJ, Waterhouse JP, Meyer J. A fine structural comparison of the healing of incisional wounds of mucosa and skin. J Oral Pathol Med 1978;7(4):214-27. https://doi. org/10.1111/j.1600-0714.1978.tb01596.x PMid:99502
De Masi EC, Campos AC, De Masi FD, Ratti MA, Ike IS, De Masi RD. The influence of growth factors on skin wound healing in rats. Braz J Otorhinolaryngol. 2016;82(5):512-21. https://doi. org/10.1016/j.bjorl.2015.09.011 PMid:26832633
Duymus ME, et al. Comparison of the effects of platelet rich plasma prepared in various forms on the healing of dermal wounds in rats. Wounds. 2016;28(3):99-108.
Wong JW, Gallant-Behm C, Wiebe C, Mak K, Hart DA, Larjava H, et al. Wound healing in oral mucosa results in reduced scar formation as compared with skin: Evidence from the red Duroc pig model and humans. Wound Repair Regeneration. 2009;17(5):717-29. https://doi.org/10.1111/j.1524-475x.2009.00531.x PMid:19769724
Galkowska H, Wojewodzka U, Olszewski WL. Chemokines, cytokines, and growth factors in keratinocytes and dermal endothelial cells in the margin of chronic diabetic foot ulcers. Wound Repair Regen. 2006;14(5):558-65. https://doi. org/10.1111/j.1743-6109.2006.00155.x
Abiko Y, Selimovic D. The mechanism of protracted wound healing on oral mucosa in diabetes. Review. Bos J Basic Med Sci. 2010;10(3):188-91. https://doi.org/10.17305/bjbms.2010.2683
Kidman K. Tissue repair and regeneration: The effects of diabetes on wound healing. Diabetic Foot J. 2008;11(2):73-80.
Dionyssiou D, Demiri E, Foroglou P, Cheva A, Saratzis N, Aivazidis C, et al. The effectiveness of intralesional injection of platelet-rich plasma in accelerating the healing of chronic ulcers: An experimental and clinical study. Int Wound J. 2013;10(4):397- 406. https://doi.org/10.1111/j.1742-481x.2012.00996. PMid:22672105
Pastar I, Stojadinovic O, Yin NC, Ramirez H, Nusbaum AG, Sawaya A, et al. Epithelialization in wound healing: A comprehensive review. Adv Wound Care. 2014;3(7):445-64. PMid:25032064
Rashed FM, GabAllah OM, AbuAli SY, Shredah MT. The effect of using bone marrow mesenchymal stem cells versus platelet rich plasma on the healing of induced oral ulcer in albino rats. Int J Stem Cells. 2019;12(1):95-106. https://doi.org/10.15283/ijsc18074 PMid:30836730
Di Como CJ, Urist MJ, Babayan I, Drobnjak M, Hedvat CV, Teruya-Feldstein J, et al. p63 expression profiles in human normal and tumor tissues. Clin Cancer Res. 2002;8(2):494-501. PMid:11839669
Häkkinen L, Larjava H, Koivisto L. Granulation tissue formation and remodeling. Endod Top. 2012;24(1):94-129. https://doi. org/10.1111/etp.12008
Gurtner GC, Werner S, Barrandon Y, Longaker MT. Wound repair and regeneration. Nature. 2008;453(7193):314-21. https://doi.org/10.1038/nature07039
Kumar GS. Oral mucous membrane. In: Orban’s Oral Histology and Embryology. India: Elsevier; 2015. p. 194-240.
Farahat A, Salah HE, Al-Shraim M. Evaluation of the clinical and histopathological effect of Platelet rich plasma on chronic wound healing. Int Res J Basic Clin Stud. 2014;2(6):55-61.
Gonzalez AC, Costa TF, de Araújo Andrade Z, Medrado AR. Wound healing a literature review. Anais Bras Dermatol. 2016;91:614- 20. https://doi.org/10.1590/abd1806-4841.20164741 PMid:27828635
Eming SA, Krieg T, Davidson JM. Inflammation in wound repair: Molecular and cellular mechanisms. J Investig Dermatol. 2007;127(3):514-25. https://doi.org/10.1038/sj.jid.5700701 PMid:17299434
Italiano JE Jr., Richardson JL, Patel-Hett S, Battinelli E, Zaslavsky A, Short S, et al. Angiogenesis is regulated by a novel mechanism: Pro and antiangiogenic proteins are organized into separate platelet alpha granules and differentially released. Blood. 2008;111(3):1227-33. https://doi.org/10.1182/ blood-2007-09-113837
Jee CH, Eom NY, Jang HM, Jung HW, Choi ES, Won JH, et al. Effect of autologous platelet-rich plasma application on cutaneous wound healing in dogs. J Vet Sci. 2016;17(1):79-87.
Molina-Miñano F, López-Jornet P, Camacho-Alonso F, Vicente- Ortega V, et al. The use of plasma rich in growth factors on wound healing in the skin: Experimental study in rabbits. Int Wound J. 2009;6(2):145-8. https://doi.org/10.1111/j.1742-481x.2009.00592.x PMid:19432664
Marchetti C, Farina A, Cornaglia AI. Microscopic, immunocytochemical, and ultrastructural properties of peri-implant mucosa in humans. J Periodontol. 2002;73:555-63. https://doi.org/10.1902/jop.2002.73.5.555 PMid:12027260
Ostvar O, Shadvar S, Yahaghi E, Azma K, Fayyaz AF, Ahmadi K, et al. Effect of platelet-rich plasma on the healing of cutaneous defects exposed to acute to chronic wounds: A clinico-histopathologic study in rabbits. Diagn Pathol. 2015;10(85):1-6. https://doi.org/10.1186/s13000-015-0327-8 PMid:27802803
Yu H, Yuan L, Xu M, Zhang Z, Duan H. Sphingosine kinase 1 improves cutaneous wound healing in diabetic rats. Injury. 2014;45:1054-8.
Velander P, Theopold C, Hirsch T, Bleiziffer O, Zuhaili B, Fossum M, et al. Impaired wound healing in an acute diabetic pig model and the effects of local hyperglycemia. Wound Repair Regen. 2008;16(2):288-93. https://doi. org/10.1111/j.1524-475x.2008.00367.x PMid:18318812
Shubin AV, Demidyuk IV, Komissarov AA, Rafieva LM, Kostrov SV. Cytoplasmic vacuolization in cell death and survival. Oncotarget. 2016;7(34):55863-89. https://doi.org/10.18632/oncotarget.10150 PMid:27331412
Henics T, Wheatley DN. Cytoplasmic vacuolation, adaptation and cell death: A view on new perspectives and features. Biol Cell. 1999;91(7):485-98. https://doi.org/10.1016/ s0248-4900(00)88205-2 PMid:10572624
Leoni G, Neumann PA, Sumagin R, Denning TL, Nusrat A. Wound repair: Role of immune-epithelial interactions. Mucosal Immunol. 2015;8(5):959-68. PMid:26174765
Qing C. The molecular biology in wound healing and non-healing wound. Chin J Traumatol. 2017;20(4):189-93. https:// doi.org/10.1016/j.cjtee.2017.06.001 PMid:28712679
Li H, Hamza T, Tidwell JE, Clovis N, Li B. Unique antimicrobial effects of platelet-rich plasma and its efficacy as a prophylaxis to prevent implant-associated spinal infection. Adv Healthc Mater. 2013;2(9):1277-84. https://doi.org/10.1002/adhm.201200465 PMid:23447088
El Backly R, Ulivi V, Tonachini L, Cancedda R, Descalzi F, Mastrogiacomo M. Platelet lysate induces in vitro wound healing of human keratinocytes associated with a strong proinflammatory response. Tissue Eng Part A. 2011;17:1787- 800. https://doi.org/10.1089/ten.tea.2010.0729 PMid:21385008
Koh TJ, DiPietro LA. Inflammation and wound healing: The role of the macrophage. Exp Rev Mol Med. 2011;13:e23-3. PMid:21740602
Marx RE. Platelet-rich plasma: Evidence to support its use. J Oral Maxillofac Surg. 2004;62(4):489-96. https://doi.org/10.1016/j. joms.2003.12.003 PMid:15085519
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