Golden Perspective: Application of Laser‐Generated Gold Nanoparticle Conjugates in Reproductive Biology

Abstract

Contents The current demand for female calves has grown rapidly and controlling the sex of offspring provides an economically flexible management for the livestock producer. The only functioning method of efficiently producing separate populations of X and Y sperm in mammals is based on relative DNA differentiation by high‐speed flow cytometry. In this context, gold nanoparticles conjugated to sex chromosome‐specific moieties display promising application as novel fluorophor‐alternative for the high‐throughput screening, since they feature no photo bleaching, high quantum yield, good biocompatibility and the possibility of non‐destructive membrane penetration. Especially, gold nanoparticles fabricated by pulsed laser ablation are in the recent focus of interest, due to excellent biocompatibility, fabrication‐dependent, tuneable particle size as well as surface charge and ease of (bio)‐functionalization with a remarkably strong ligand binding. For the purpose of our studies functionalized gold nanoparticles may be used as novel markers for sex‐sorting of mammalian sperm and, depending on the selected probe, also for the selection of sperm with heritable DNA‐sequences interesting for animal breeding.

Nanobiotechnology in reproduction – pros and cons. A review

“…Moreover, NPs, as an alternative to traditional fluorophores, are used for spermatozoa sorting by flow cytometry and fluorescent-activated cell sorting (Chattopadhyay et al, 2010). Several publications report using AuNPs covalently bonded with membrane-penetration agents for sorting populations of gametes by recognizing and binding with specific DNA sequences at the Y-chromosome (Barchanski et al, 2011;Rath et al, 2013) without affecting fertilization potential (Feugang et al, 2012). Improvement of this technique will allow not only broader use of sex-sorted spermatozoa, mainly in breeding livestock (Rath et al, 2013), but also development of bioimaging systems in the future (Barchanski et al, 2011).…”

“…Application of noble-metal NPs may impact sperm vitality parameters like motility, morphology and membrane integrity, resulting in impairment of male gametes, which become incapable of fertilization. These effects were observed after incubation of spermatozoa with extremely high concentrations of ligand-free AuNPs and AgNPs (Barchanski et al, 2011;Moretti et al, 2013). This cytotoxic consequence was manifested by failure of chromatin to decondense and in the nucleus structure (Zakhidov et al, 2010).…”

Evaluation of pulsed laser ablation in liquids generated gold nanoparticles as novel transfection tools: efficiency and cytotoxicity

“…In general, the generation of PLAL-AuNP is carried out by focusing femtosecond laser pulses through a lens on a solid pure gold plate located in a vessel with ddH 2 O [11]. The laser irradiation induced photomechanical effects lead to the direct formation PLAL-AuNP from the gold target [11]. These generated, positively charged, pure PLAL-AuNPs were shown to easily adsorb biomolecules such as unmodified, negatively charged DNA oligonucleotides [12].…”

“…This method has been shown to be a potent tool to obtain highly pure colloidal particles avoiding chemical surface modifications making these particles interesting for biomedical applications [9,10]. In general, the generation of PLAL-AuNP is carried out by focusing femtosecond laser pulses through a lens on a solid pure gold plate located in a vessel with ddH 2 O [11]. The laser irradiation induced photomechanical effects lead to the direct formation PLAL-AuNP from the gold target [11].…”

Nanomedicine and mammalian sperm: Lessons from the porcine model

“…57 58 2. Nanotechnology as a versatile tool for targeted biological delivery 59 Nanotechnology is an innovative multifaceted discipline, combining the aspects of physics, 60 chemistry, biology, mathematics, engineering and computer sciences, which designs and 61 investigates the properties of small-scale (in general, 1-100nm) structures for the variety of 62 industrial, consumer and scientific applications. In biomedicine, nanotechnology allows us to 63 engineer versatile nano-sized platforms that are comparable in size to biological molecules and 64 intracellular organelles.…”