J Korean Med Sci.  2017 Jan;32(1):13-21. 10.3346/jkms.2017.32.1.13.

Effects of Platelet-Rich Plasma on Kidney Regeneration in Gentamicin-Induced Nephrotoxicity

• Moghadam A
• Khozani TT
• Mafi A
• Namavar MR
• Dehghani F

• KMID: 2417579
• DOI: http://doi.org/10.3346/jkms.2017.32.1.13

Abstract

Platelet-rich plasma (PRP) as a source of growth factors may induce tissue repairing and improve fibrosis. This study aimed to assess the effects of PRP on kidney regeneration and fibrosis in gentamicin (GM)-induced nephrotoxicity rat model by stereological study. Thirty-two male rats were selected. Nephrotoxicity was induced in animals by administration of GM (80 mg/kg/daily, intraperitoneally [IP], 8 day) and animals were treated by PRP (100 µL, intra-cortical injection using surgical microscopy, single dose). Blood samples were collected for determine blood urea nitrogen (BUN) and creatinine (Cr) before and after PRP therapy. At the end of experiment, right kidneys were sectioned by Isotropic Uniform Random (IUR) method and stained with H & E and Masson's Trichrome. The stereological methods were used for estimating the changes in different structures of kidney. PRP increased the number of epithelial cells in convoluted tubules, and decreased the volume of connective tissue, renal corpuscles and glomeruli in GM-treated animals (P < 0.05). Our findings indicate that PRP had beneficial effects on proliferation of epithelial cells in convoluted tubules and ameliorated GM-induced fibrosis.

Keyword

Fibrosis; Kidney; Regeneration

MeSH Terms

Animals
Blood Urea Nitrogen
Connective Tissue
Creatinine
Epithelial Cells
Fibrosis
Gentamicins
Humans
Intercellular Signaling Peptides and Proteins
Kidney*
Male
Methods
Microscopy
Models, Animal
Platelet-Rich Plasma*
Rats
Regeneration*
Creatinine
Gentamicins
Intercellular Signaling Peptides and Proteins

Figure

• 

  Fig. 1 (A) A grid of points on the images for estimation of the volume density of the interstitial connective tissue stained by Masson Trichrome. (B) The length density of the cortical convoluted tubules is estimated by an unbiased counting frame on the images stained by H & E.

• 

  Fig. 2 H & E staining of rat kidney sections at (A) control group; (B) GM group: dilatation (◄), cellular debris (*), inflammation (↓); (C) GM+PRP group: debris clearing (*) and regenerating tubular epithelium (◄); (D) GM+NS group: necrosis with cellular debris (*). GM = gentamicin, PRP = platelet-rich plasma, NS = normal saline.

• 

  Fig. 3 Masson Trichrome staining of cortex (A-D) and medulla (E-H) of rat kidney sections at different experimental groups. (A and E: control group), (B and F: GM group), (C and G: GM+PRP group), and (D and H: GM+NS group). GM = gentamicin, PRP = platelet-rich plasma, NS = normal saline.

• 

  Fig. 4 Effect of platelet-rich plasma on the number of convoluted tubule epithelial cells (A), kidney cortex volume (B), connective tissue volume (C), mean volume-weighted glomerulus volume (D), and mean volume-weighted renal corpuscle volume (E) in GM-induced rats. GM = gentamicin, PRP = platelet-rich plasma, NS = normal saline.

Reference

1. Fortier LA, Barker JU, Strauss EJ, McCarrel TM, Cole BJ. The role of growth factors in cartilage repair. Clin Orthop Relat Res. 2011; 469:2706–2715.

2. Lyras DN, Kazakos K, Verettas D, Polychronidis A, Tryfonidis M, Botaitis S, Agrogiannis G, Simopoulos C, Kokka A, Patsouris E. The influence of platelet-rich plasma on angiogenesis during the early phase of tendon healing. Foot Ankle Int. 2009; 30:1101–1106.

3. Kark LR, Karp JM, Davies JE. Platelet releasate increases the proliferation and migration of bone marrow-derived cells cultured under osteogenic conditions. Clin Oral Implants Res. 2006; 17:321–327.

4. Debus ES, Schmidt K, Geiger D, Dietz UA, Thiede A. Growth factors for preventing amputation in delayed wound healing. Kongressbd Dtsch Ges Chir Kongr. 2001; 118:829–833.

5. Peerbooms JC, Sluimer J, Bruijn DJ, Gosens T. Positive effect of an autologous platelet concentrate in lateral epicondylitis in a double-blind randomized controlled trial: platelet-rich plasma versus corticosteroid injection with a 1-year follow-up. Am J Sports Med. 2010; 38:255–262.

6. Randelli PS, Arrigoni P, Cabitza P, Volpi P, Maffulli N. Autologous platelet rich plasma for arthroscopic rotator cuff repair. A pilot study. Disabil Rehabil. 2008; 30:1584–1589.

7. Norman J, Tsau YK, Bacay A, Fine LG. Epidermal growth factor accelerates functional recovery from ischaemic acute tubular necrosis in the rat: role of the epidermal growth factor receptor. Clin Sci (Lond). 1990; 78:445–450.

8. Matsumoto K, Nakamura T. Hepatocyte growth factor: renotropic role and potential therapeutics for renal diseases. Kidney Int. 2001; 59:2023–2038.

9. Bledsoe G, Crickman S, Mao J, Xia CF, Murakami H, Chao L, Chao J. Kallikrein/kinin protects against gentamicin-induced nephrotoxicity by inhibition of inflammation and apoptosis. Nephrol Dial Transplant. 2006; 21:624–633.

10. Abdelsameea AA, Mohamed AM, Amer MG, Attia SM. Cilostazol attenuates gentamicin-induced nephrotoxicity in rats. Exp Toxicol Pathol. 2016; 68:247–253.

11. Franco D, Franco T, Schettino AM, Filho JM, Vendramin FS. Protocol for obtaining platelet-rich plasma (PRP), platelet-poor plasma (PPP), and thrombin for autologous use. Aesthetic Plast Surg. 2012; 36:1254–1259.

12. Scherle W. A simple method for volumetry of organs in quantitative stereology. Mikroskopie. 1970; 26:57–60.

13. Nyengaard JR. Stereologic methods and their application in kidney research. J Am Soc Nephrol. 1999; 10:1100–1123.

14. Gamble M. The hematoxylins and eosin. In : Bancroft JD, Gamble M, editors. Theory and Practice of Histological Techniques. 6th ed. Philadelphia, PA: Churchill livingstone/Elsevier;2008. p. 121–126.

15. Weibel ER. Stereological Methods. London: Academic Press;1979.

16. Maldonado PD, Barrera D, Rivero I, Mata R, Medina-Campos ON, Hernández-Pando R, Pedraza-Chaverrí J. Antioxidant S-allylcysteine prevents gentamicin-induced oxidative stress and renal damage. Free Radic Biol Med. 2003; 35:317–324.

17. Choi KH, Kim TI, Chong DL, Lee HY, Han DS. Gentamicin induced apoptosis of renal tubular epithelial (LLC-PK1) cells. Korean J Intern Med. 2000; 15:218–223.

18. Quiros Y, Vicente-Vicente L, Morales AI, López-Novoa JM, López-Hernández FJ. An integrative overview on the mechanisms underlying the renal tubular cytotoxicity of gentamicin. Toxicol Sci. 2011; 119:245–256.

19. Abdeen A, Sonoda H, El-Shawarby R, Takahashi S, Ikeda M. Urinary excretion pattern of exosomal aquaporin-2 in rats that received gentamicin. Am J Physiol Renal Physiol. 2014; 307:F1227–37.

20. Böttinger EP, Bitzer M. TGF-beta signaling in renal disease. J Am Soc Nephrol. 2002; 13:2600–2610.

21. Kim J, Shon E, Kim CS, Kim JS. Renal podocyte injury in a rat model of type 2 diabetes is prevented by metformin. Exp Diabetes Res. 2012; 2012:210821.

22. Rodriguez-Barbero A, L’Azou B, Cambar J, López-Novoa JM. Potential use of isolated glomeruli and cultured mesangial cells as in vitro models to assess nephrotoxicity. Cell Biol Toxicol. 2000; 16:145–153.

23. Hom DB, Linzie BM, Huang TC. The healing effects of autologous platelet gel on acute human skin wounds. Arch Facial Plast Surg. 2007; 9:174–183.

24. Lin JJ, Cybulsky AV, Goodyer PR, Fine RN, Kaskel FJ. Insulin-like growth factor-1 enhances epidermal growth factor receptor activation and renal tubular cell regeneration in postischemic acute renal failure. J Lab Clin Med. 1995; 125:724–733.

25. Kempczinski RF, Caulfield JB. A light and electron microscopic study of renal tubular regeneration. Nephron. 1968; 5:249–264.

26. Yang J, Dai C, Liu Y. Hepatocyte growth factor suppresses renal interstitial myofibroblast activation and intercepts Smad signal transduction. Am J Pathol. 2003; 163:621–632.

27. Yang J, Liu Y. Blockage of tubular epithelial to myofibroblast transition by hepatocyte growth factor prevents renal interstitial fibrosis. J Am Soc Nephrol. 2002; 13:96–107.

28. Brown R, Ollerstam A, Johansson B, Skøtt O, Gebre-Medhin S, Fredholm B, Persson AE. Abolished tubuloglomerular feedback and increased plasma renin in adenosine A1 receptor-deficient mice. Am J Physiol Regul Integr Comp Physiol. 2001; 281:R1362–7.