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Development of a Gelatin Methacrylate and Human Testicular Extracellular Matrix Bioink for Engineering Bioartificial Testis Publisher



H Eyni HOSSEIN ; R Shabani RONAK ; N Baheiraei NAFISEH ; R Falak REZA ; S Dehghani SANAZ ; Hr Asgari Hamid REZA ; Sm Koruji Seyed MORTEZA
Authors

Source: Polymers for Advanced Technologies Published:2025


Abstract

Advancements in biomaterial engineering have enabled the creation of sophisticated in vitro culture platforms that closely mimic natural cellular microenvironments. This study focuses on designing an innovative bioprinted scaffold using a bioink composed of gelatin methacrylate (GelMA) and human testicular extracellular matrix (htECM) for constructing a bioartificial testis. Human testis was decellularized and confirmed using hematoxylin and eosin (H&E) staining and DNA quantification, while maintaining htECM components, as shown by Masson's trichrome, Alcian blue, and Orcein staining. This ECM was combined with GelMA to form a bioink for scaffold fabrication. Human spermatogonial stem cells (hSSCs) isolated using enzymatic digestion and confirmed by RT-PCR. Then, we evaluated the mechanical and biological properties of the scaffolds, focusing on surface morphology, degradation rate, swelling behavior, MTT assay, and biocompatibility. The decellularization protocol for testicular tissue was effective in eliminating cellular debris while conserving the ECM compounds. GelMA and GelMA/htECM scaffolds revealed a porous structure with suitable pore size and porosity, while the GelMA/htECM scaffold had more porosity (p < 0.05). The swelling ratio of the GelMA scaffold was enhanced by adding htECM, whereas its degradation rate decreased. The MTT assay showed that both scaffolds have no cytotoxic properties. Moreover, the GelMA/htECM scaffold displayed more cell attachment and better biocompatibility properties compared to the GelMA scaffold. These results highlight the potential of the GelMA/htECM scaffold as an advanced platform for in vitro spermatogenesis models, facilitating sperm production from hSSCs. This approach offers a promising tool for studying SSC behavior and developing personalized treatments for male infertility. © 2025 Elsevier B.V., All rights reserved.
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