In a groundbreaking advance poised to redefine the landscape of regenerative medicine and women’s health, researchers have unveiled an optimized method for creating human ovarian scaffolds using supercritical carbon dioxide (scCO₂). This innovative protocol stands to revolutionize tissue engineering by providing a more efficient, biocompatible, and structurally intact extracellular matrix (ECM) scaffold derived from human ovarian tissue. The implications for treating primary ovarian insufficiency and other estrogen deficiency-related conditions could be profound, offering new hope to millions of women worldwide.
Primary ovarian insufficiency (POI), a condition affecting approximately 1 to 3 percent of women under 40, precipitates premature menopause and a cascade of hormonal deficiencies that trigger a host of biologically and psychologically challenging symptoms. As life expectancy rises globally, the need for effective ovarian tissue replacement strategies grows exponentially. Conventional hormone replacement therapies, while helpful, fail to fully replicate the complex endocrine and paracrine environment of the natural ovary. The vision of bioengineering a functional ovary hinges on accurately mimicking the ovarian niche—a matrix critical for supporting folliculogenesis, hormonal synthesis, and overall ovarian physiology.
Key to this vision is the development of decellularized extracellular matrix scaffolds that preserve the native architecture and biochemical cues of ovarian tissue. Decellularization involves removing cellular components while retaining the ECM framework, which houses proteins, glycosaminoglycans (GAGs), and other factors essential for cell migration, differentiation, and survival. Achieving this delicate balance has historically been marred by protocols that either inadequately remove cells or excessively damage ECM components, rendering scaffolds unsuitable for clinical use.
Enter the application of supercritical carbon dioxide—a state of CO₂ that possesses unique physical properties bridging the characteristics of liquids and gases, including low viscosity and high diffusivity. This makes scCO₂ an exceptionally promising agent for decellularization tasks. The recent study harnessed these properties to develop a novel protocol aimed at optimizing ovarian scaffold preparation in terms of efficacy, structural integrity, and cytocompatibility.
The researchers meticulously examined the effects of pressure variations within the scCO₂ environment, setting experiments at 200 and 300 bar while keeping temperature fixed at 40°C for 1.5 hours. Their experimental design also introduced a two-pronged approach to augment the decellularization process: the use of 70% ethanol as a co-solvent within the scCO₂ medium and a pretreatment stage utilizing 1% sodium dodecyl sulfate (SDS) for four hours before the scCO₂ application. SDS, a well-known detergent, was employed for its robust cell lysis capabilities, aiming to facilitate more thorough removal of cellular materials without compromising ECM integrity.
Through a comprehensive suite of analytical techniques, including DNA quantification and histological staining such as hematoxylin and eosin (H&E), the team confirmed the effective removal of cellular components. Notably, scanning electron microscopy (SEM) revealed that the three-dimensional microarchitecture of the ECM remained remarkably well-preserved under optimal protocol conditions. This is critical because the physical scaffold acts as a structural and biochemical venue for future cell seeding and tissue regeneration.
Preserving the glycosaminoglycan content proved essential, as these molecules regulate cellular behavior through signaling pathways and contribute to the mechanical properties of tissue. Using a dimethyl methylene blue assay, the study demonstrated that the GAG concentration was maintained at levels conducive to biological functionality. This retention further endorses the protocol’s potential in creating a supportive microenvironment that could better facilitate ovarian cell integration and function.
Cytocompatibility tests, notably the MTT assay, further validated the scaffold’s suitability for cellular colonization and proliferation. Scaffolds prepared with the 1% SDS pretreatment followed by scCO₂ at 200 bar did not exhibit cytotoxic effects, underscoring the biocompatibility of this preparation method. This represents a key milestone in potential clinical translation, as any implanted scaffold must promote cell viability and avoid eliciting detrimental immune responses.
Compared to traditional decellularization methods, which often involve harsh detergents and prolonged exposure times that risk ECM degradation, this scCO₂-based approach offers a safer, faster, and more environmentally sound alternative. The supercritical fluid technique minimizes chemical residue and mechanical damage, contributing to the scaffold’s robustness and functionality.
The success of this optimized protocol marks a significant stride toward practical applications in ovarian tissue engineering. Bioengineered ovarian scaffolds could one day be implanted to restore endocrine function, support follicle maturation, and potentially preserve fertility in women facing premature ovarian failure or those undergoing gonadotoxic treatments like chemotherapy.
Moreover, the scalability and reproducibility of this supercritical carbon dioxide-assisted decellularization process suggest it could be adapted to other tissue types requiring engineered scaffolds, thereby broadening its utility in regenerative medicine.
While these findings represent promising preliminary data, further research involving in vivo studies, functional assessments of reseeded scaffolds, and long-term biocompatibility evaluations remains essential before clinical adoption. Additionally, integration of vascularization strategies will be indispensable for enhancing graft survival and function post-transplantation.
In summary, this pioneering study leverages the unique physicochemical properties of supercritical carbon dioxide combined with targeted SDS pretreatment to engineer ovarian scaffolds that faithfully preserve extracellular matrix composition and architecture, while ensuring cytocompatibility. This innovation significantly advances the field of female reproductive tissue engineering and opens new horizons for therapeutic interventions aimed at combating ovarian insufficiency and associated hormonal deficits.
Subject of Research:
Article Title: Preparation and characterization of human decellularized ovarian scaffold based on supercritical carbon dioxide protocol
Article References: Hosseinpour, F., Zeinolabedini Hezave, A., Talaei-Khozani, T. et al. Preparation and characterization of human decellularized ovarian scaffold based on supercritical carbon dioxide protocol. BioMed Eng OnLine 24, 59 (2025). https://doi.org/10.1186/s12938-025-01392-7
Image Credits: AI Generated
DOI: https://doi.org/10.1186/s12938-025-01392-7
