Spermatogonia stem cells (SSCs) are a unique cell population maintaining male spermatogenesis during life, through their potential for proliferation and differentiation. The application of silicon nanoparticles (SNs) and hyaluronic acid (HA) to induce the differentiation of SSCs seems promising. Herein, we investigate the effect of SN and HA scaffolds on the progression of SSCs spermatogenesis in mice. Initially SSCs were isolated from healthy immature mice and cultured on prepared scaffolds (HA, SN, and HA/SN) in a 3D culture system. Then viability of SSCs cultured on scaffolds was examined using MTT assay and Acridine Orange staining. Then SSCs cultured on scaffolds were transplanted into epididymal adipose tissue (EAT) in mature mice and the result was studied by H&E and IHC staining 8 weeks after transplantation. MTT and Acridine Orange analysis revealed that among three different scaffolds HA/SN based scaffold causes considerable toxicity on SSCs (P < 0.05) while H&E staining showed that culture of SSCs on HA, SN, and HA/SN scaffolds has a positive effect on the progression of SSCs spermatogenesis after transplantation into EAT. IHC staining identified TP1, TEKT1, and PLZF as crucial biomarkers in the spermatogenesis development of SSCs transplanted to EAT. According to the presence of these biomarkers in different experimental groups, we found the most spermatogenesis development in SSCs cultured on HA/SN scaffold (PLZF, P < 0.01) (TEKT1, P < 0.01) (TP1, P < 0.001). Our study showed that, although the cytotoxic effect of the HA/SN scaffold decreases the viability rate of SSCs; however, SSCs that survive on HA/SN scaffold showed more ability to progress in spermatogenesis after transplantation into EAT.

精原干细胞（SSC）是一种独特的细胞群，通过其增殖和分化的潜力维持男性一生中的精子发生。应用硅纳米粒子（SNs）和透明质酸（HA）来诱导SSCs的分化似乎很有前景。在此，我们研究了 SN 和 HA 支架对小鼠 SSC 精子发生进展的影响。最初，从健康的未成熟小鼠中分离 SSC，并在 3D 培养系统中在准备好的支架（HA、SN 和 HA/SN）上培养。然后使用 MTT 测定和吖啶橙染色检查支架上培养的 SSC 的活力。然后将在支架上培养的SSC移植到成熟小鼠的附睾脂肪组织(EAT)中，并在移植后8周通过H&E和IHC染色研究结果。MTT和吖啶橙分析显示，在三种不同的支架中，基于HA/SN的支架对SSCs造成相当大的毒性（P  < 0.05），而H&E染色显示，在HA、SN和HA/SN支架上培养SSCs对SSCs的毒性有积极作用。 SSCs 移植到 EAT 后精子发生的进展。IHC 染色鉴定出 TP1、TEKT1 和 PLZF 是移植到 EAT 的 SSC 精子发生发育中的关键生物标志物。根据不同实验组中这些生物标志物的存在情况，我们发现在HA/SN支架上培养的SSC中精子发生发育最多（PLZF，P  < 0.01）（TEKT1，P  < 0.01）（TP1，P  < 0.001）。我们的研究表明，虽然HA/SN支架的细胞毒性作用降低了SSC的存活率；然而，在 HA/SN 支架上存活的 SSC 在移植到 EAT 后表现出更强的精子发生能力。