Retinoic acid-releasing scaffold based on chitosan hydrogel and testis decellular plates.

Introduction: The use of releasing scaffolds is promising for testes tissue engineering. Chitosan (CS) is a natural biopolymer extensively used as a delivery system. The decellularized testis provides a structure resembling natural extracellular matrix (ECM). All-trans retinoic acid (atRA) is an important factor for spermatogonia differentiation, meiosis completion, and mature sperm release. In this study, thermosensitive CS/βGP hydrogel was served as a novel atRA-releasing support for testis decellular plates (TDPs).

Methods: The CS/βGP hydrogel was evaluated for gelation time, morphology, wettability, cytocompatibility, and atRA-releasing behavior. Mouse testes were treated with 1% SDS and evaluated for decellularization efficacy through morphological assessments, DNA content assays, and DAPI staining. TDPs were obtained from the decellularized testes and placed on an atRA-releasing CS/βGP hydrogel support.

Results: The CS/βGP hydrogels were prepared with different formulations. It was found that increasing the βGP concentration significantly decreased the gelation time. The addition of atRA did not considerably affect the hydrophilicity of hydrogel. The in vitro release studies showed a sustained atRA release behavior, although an initial low burst release was recorded. Also, increasing the amount of atRA led to a decrease in the rate of drug release. The decellularization procedure successfully removed cells while preserving the ECM. The atRA-releasing CS-TDP scaffold was found to be non-toxic with good biocompatibility.

Conclusion: Results showed that the novel atRA-releasing CS-TDP scaffold can sustainably deliver atRA to the culture system and create a cytocompatible environment for testicular cells. Therefore, this scaffold may be useful in developing new tissue engineering approaches for various types of male infertility diseases.