P-018 Structural and biocompatibility analysis of gold nanoparticle integrated polymeric scaffolds for infertility diagnosis
Authors: Andone Rotaru B A , Girlovanu M , Vulpoi A , Botiz I , Boca S 
Published: June 2023

Abstract

Study question
Can infertility diagnosis and sperm selection be improved by using bio- nanotechnologies?

Summary answer
Gold nanoparticle integrated biopolymeric scaffolds show good biocompatibility when interacting with human sperm cells, being suitable to be used as cell sorting and biosensing platforms.

What is known already
Microfluidic platforms are used in sperm selection for increasing the in-vitro fertilization success but they are limited by the exclusive analysis of sperm cell motility. The integration of microfluidic platforms with nanomaterials such as nanopatterned substrates can greatly enhance the selectivity and the sensitivity of the detection. The accuracy can be further improved by employing the intrinsic properties of gold nanoparticles (optical properties, high biocompatibility, surface chemistry favoring the conjugation of specific biomolecules), which makes them suitable for the successful development of precise and reliable detection assays, so far used to a limited extent in infertility research and clinical approaches.

Study design, size, duration
Concentration, motility and morphology of spermatozoa were analyzed in semen samples collected for spermogram. The normozoospermic and teratozoospermic samples were selected and prepared by density gradient centrifugation. The sediment obtained contained 100% motile spermatozoa, which were incubated on the thin films for 24h. After 24h, the probe from each incubation dish was collected and the survival rate was analyzed using the Makler chamber.

Participants/materials, setting, methods
Thin films of atactic polystyrene, chitosan and mixtures of each of the two polymers with gold nanoparticles were used to produce scaffolds of microchannels. The substrates were morphologically characterized by optical microscopy, scanning transmission electron microscopy and atomic force microscopy. Normozoospermic and teratozoospermic sperm samples were collected and incubated on the scaffolds. After 24h incubation, the viability was assessed by counting the motile spermatozoa under microscopic observation. Statistical analysis was performed using two-way ANOVA test.

Main results and the role of chance
Analysis of the sperm cell viability incubated on the polymeric scaffolds revealed good biocompatibility with higher survival rate in the case of chitosan-gold nanoparticle integrated samples. In multiple comparisons of normozoospermic samples, the survival rates of the sample incubated on the chitosan thin films to the control sample were 18% to 80% (-62%) (p = 0.0143), 16.66% to 45% (-28.34%) (p = 0.0143), 0% to 15.38% (-15.38%) (p = 0.0239), 4.76% to 55.35% (-50.59%) (p = 0.0239). In multiple comparisons of teratozoospermic samples, the survival rates of the sample incubated on the chitosan thin films to the control sample were 14.67% to 41% (-26.33%) (p = 0.0143), 0% to 44.82% (-33.82%) (p = 0.0239). For polystyrene, the viability was 0% which excludes its further use in infertility-related assays due to its high cytotoxicity. Overall, the colloidal gold nanoparticles integrated into the polymeric matrix improved the biocompatibility of the films. For example, citrate gold nanoparticles of spherical shape and sizes of about 20 nm increased the survival rate of normozoospermic samples, while PEGylation of nanoparticles showed further improvement of the biocompatibility. The influence of the particle type, composition and coating material onto the spermatozoa viability is currently under investigation for optimizing the effectiveness of the polymeric scaffolds.

Limitations, reasons for caution
NA

Wider implications of the findings
Due to the unique properties of gold nanoparticles, the nanoparticle-polymer scaffold systems have the potential to become the foundation of a precise biosensor that can enhance the accuracy of the current approaches in infertility assessment.

Trial registration number
Not applicable