Sperm DNA fragmentation induced by cryopreservation: new insights and effect of a natural extract from Opuntia ficus-indica.
OBJECTIVE
To analyze the effect of cryopreservation on sperm DNA fragmentation (SDF) in two cytometric sperm populations, PI(brighter) and PI(dimmer), and to test the effects of Opuntia ficus-indica (OFI) extracts, which contain antioxidants and flavanoids, and of resveratrol on cryopreservation of human semen.


METHODS
In vitro prospective study.


METHODS
Institutional study.


METHODS
Twenty-one normozoospermic men undergoing semen analysis for couple infertility.


METHODS
Cryopreservation using the routine method in the presence of OFI extracts or resveratrol.


METHODS
Measurement of SDF by TUNEL/PI flow cytometric method to evaluate sperm motility (by automated motion analysis, CASA system) and viability (by eosin/nigrosin staining) in the two populations of sperm PI(br) and PI(dim).


RESULTS
Cryopreservation induced an increase of SDF only in the PI(br) sperm population. The increase was negatively dependent on the basal values of SDF in the same population. Addition of OFI extracts and resveratrol to the cryopreservation medium slightly but statistically significantly reduced SDF in the PI(br) population without affecting the deleterious effect of cryopreservation on sperm motion parameters or viability.


CONCLUSIONS
The increase of SDF in the PI(br) population, which is unrelated to semen quality, suggests that caution must be taken in using cryopreserved semen, as morphologically normal and motile sperm may be damaged. The addition of substances with multifunctional properties such as OFI extracts to cryopreservation medium is only slightly effective in preventing the dramatic effects on SDF.
Department of Clinical Physiopathology, University of Florence, Florence, Italy.
Mehrdad
M
Meamar
Nassira
N
Zribi
Marta
M
Cambi
Lara
L
Tamburrino
Sara
S
Marchiani
Erminio
E
Filimberti
Maria Grazia
MG
Fino
Annibale
A
Biggeri
Yves
Y
Menezo
Gianni
G
Forti
Elisabetta
E
Baldi
Monica
M
Muratori
D015925:Cryopreservation / Q000379:methods*; D053938:DNA Fragmentation / Q000187:drug effects*; D006801:Humans; D007248:Infertility, Male / Q000175:diagnosis / Q000188:drug therapy; D008297:Male; D029747:Opuntia; D010936:Plant Extracts / Q000302:isolation & purification / Q000494:pharmacology* / Q000627:therapeutic use; D011446:Prospective Studies; D013094:Spermatozoa / Q000187:drug effects* / Q000473:pathology / Q000502:physiology
D010936:Plant Extracts