title: The viability of mouse spermatogonial germ cells on a novel scaffold, containing human serum albumin and calcium phosphate nanoparticles.
abstract: BACKGROUND
  In spermatogenesis, spermatogonial cells differentiate to the haploid gametes. It has been shown that spermatogenesis can be done at in vitro condition. In vitro spermatogenesis may provide an open window to treat male infertility.
  
  
  OBJECTIVE
  The aim of this study was to evaluate the effects of a novel scaffold containing human serum albumin (HSA)/tri calcium phosphate nanoparticles (TCP NPs) on the mouse spermatogonial cell line (SCL).
  
  
  METHODS
  First, TCP NPs were synthesized by reaction of calcium nitrate and diammonium phosphate at pH 13. Then, serial concentrations of TCP NPs were separately added to 500 mg/mL HSA, and incubated in the 100(o)C water for 30 min. In the next step, each scaffold was cut (2×2mm), placed into sterile well of microplate, and then incubated for 1, 2, and 3 days at 37(o)C with mouse SCL. After incubation, the cytotoxicity of the scaffolds was evaluated by different tests including 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) assay, lactate dehydrogenase (LDH) assay, vital staining, and cell counting. On the other hand, the release of TCP NPs and HSA from the scaffolds was measured.
  
  
  RESULTS
  Based on microscopic observation, the size of cavities for all scaffolds was near 200-500 µm, and the size of TCP NPs was near 50-100 nm. All toxicity tests showed that the increase of TCP concentration in the scaffold did not affect mouse SCL. It means that the percentage of cell viability, LDH release, vital cells, and cell quantity was 85%, 105%, 90%, and 110%, respectively. But, the increase of incubation time led to increase of LDH release (up to 115%) and cell count (up to 115%). Also, little decrease of cell viability and vital cells was seen when incubation time was increased. Here, no release of TCP NPs and HSA was seen after increase of TCP concentration and incubation time.
  
  
  CONCLUSIONS
  It can be concluded that the increase of TCP concentration in HSA/ TCP NPs scaffold does not lead to cytotoxicity. On the other hand, the increase of incubation time leads to increase of mouse SCL cell death. In this study, it was found that TCP NPs and HSA could not release from the scaffolds. In future, both proliferation and differentiation of mouse SCL on HSA/TCP NPs scaffold must be checked over more wide incubation times.
authors.affiliation: Department of Biology, East Tehran Branch, Islamic Azad University, Tehran, Iran.
authors.forename: Mona
authors.initials: M
authors.lastname: Yadegar
authors.affiliation: Department of Laboratory Sciences, School of Paramedicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
authors.forename: Seyed Hossein
authors.initials: SH
authors.lastname: Hekmatimoghaddam
authors.affiliation: Department of Biology, Ashkezar Branch, Islamic Azad University, Ashkezar, Iran.
authors.forename: Saeide
authors.initials: S
authors.lastname: Nezami Saridar
authors.affiliation: Department of Biology, Ashkezar Branch, Islamic Azad University, Ashkezar, Iran.
authors.forename: Ali
authors.initials: A
authors.lastname: Jebali
keywords: Calcium phosphate nanoparticles; Cytotoxic effect; Human serum albumin; Scaffold; Spermatogonial cell line