The aim of this study was to investigate the efficiency of low-level laser therapy (LLLT) to recovery testicular degeneration in rams. In the first study, rams were induced to testicular degeneration by scrotal insulation, and then, they were treated using LLLT at 28 J/cm2 (INS28) or 56 J/cm2 (INS56) energy densities. Sperm kinetics, morphology, and membranes integrity as well as proportion of lumen area in seminiferous tubule were assessed. In the second study, rams were submitted or not to scrotal insulation and treated or not by the best protocol of LLLT defined by experiment 1 (INS28). In this study were evaluated sperm kinetics, morphology, membranes integrity, ROS production, and DNA integrity. Testosterone serum concentration and proportion of lumen area in seminiferous tubule were also analyzed. Insulation was effective in promoting sperm injuries in both experiments. Biostimulatory effect was observed in experiment 1: INS28 presented smaller proportion of lumen area (P = 0.0001) and less degeneration degree (P = 0.0002). However, in experiment 2, there was no difference between the groups (P = 0.17). In addition, LLLT did not improve sperm quality, and there was a decreasing for total and progressive motility (P = 0.02) and integrity of sperm membranes (P = 0.01) in LLLT-treated groups. Moreover, testosterone concentration was not improved by LLLT (P = 0.37). Stimulation of aerobic phosphorylation by LLLT may have led to a deregulated increase in ROS leading to sperm damages. Thus, LLLT at energy of 28 J/cm2 (808 nm of wavelength and 30 mW of power output) can induce sperm damages and increase the quantity of cells in seminiferous tubule in rams.
Biostimulation Sperm Spermatozoa Proliferation Male Infertility
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The authors thank Dr. Fábio Pogliani and Dr. Fábio Sellera for assistance to laser therapy protocols, Dr. Ricardo Strefezi for assistance in histopathology images evaluations, and Dr. José Antunes Rodrigues, technicians Marina Holanda, and Rogério Azevedo for assistance to testosterone assay. They also thank Dr. Eduardo Harry Birgel Junior, Dr. Daniela Becker Birgel, Mr. João Carlos Pinto de Campos, Mr. Márcio Donizete De Carli, and Mr. José Maria Bernardi for assistance to the animals.
Compliance with ethical standards
This work was supported by the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP, processes numbers 2011/16744-3, 2012/00040-0, 2012/15087-1, 2013/15745-1).
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1.Laboratory of Teaching and Research in Pathology of Reproduction, Center of Biotechnology in Animal Reproduction, Department of Animal Reproduction, School of Veterinary Medicine and Animal ScienceUniversity of Sao Paulo (USP)PirassunungaBrazil
2.Laboratory of Semen Biotechnology and Andrology, Center of Biotechnology in Animal Reproduction, Department of Animal Reproduction, School of Veterinary Medicine and Animal ScienceUniversity of Sao Paulo (USP)PirassunungaBrazil
3.Laboratory of Andrology and Embryo Technology, Department of Animal Reproduction, School of Veterinary Medicine and Animal ScienceUniversity of Sao Paulo (USP)PirassunungaBrazil
4.Laboratory of Neuroendocrinology and Reproduction, Department of Physiology – Faculty of MedicineUniversity of Sao Paulo (USP)Ribeirao PretoBrazil