Key Takeaways & Executive Findings
- •• dfLEN scaffolds support the formation of iRPE monolayers with native-like morphology and physiology, enhancing functional maturity compared to standard tissue culture plates. • Culturing iRPE on dfLEN increases cellular resistance to oxidative stress and improves functional properties, potentially boosting transplantation efficacy. • dfLEN upregulates genes associated with cilium assembly, leading to enhanced ciliogenesis in iRPE cells, a novel finding for RPE tissue engineering. • iRPE-dfLEN sheets show favorable biocompatibility and viability after subretinal transplantation in rabbits for 14 days, supporting their potential for future clinical applications.
Abstract
Background: Recent advances in clinical trials have involved the transplantation of induced retinal pigment epithelium (iRPE) cells from stem cells in creating a functional monolayer that mimics the characteristics of natural adult RPE cells. One method of achieving this goal is through the use of tissue engineering. In this research, decellularised femtosecond laser intrastromal lenticules (dfLEN) were employed as a scaffold for cultivating a bioengineered iRPE monolayer sheet. Methods: iRPE cells were obtained by differentiating induced pluripotent stem cells (iPSC). These cells were then seeded on decellularized FLI-lenticules (dfLEN). The functionality, characterization, and oxidative stress of iRPE cultured on dfLEN were compared with those cultured on plates (TCP) using various assays such as immunofluorescence (IF), Edu, CCK8, ELISA, DFCH-DA, and JC-1. Additionally, RNA-seq assays and electron microscope (SEM and TEM) were used to test the iRPE characteristic on engineered dfLEN. Finally, we evaluated the biocompatibility of iRPE-dfLEN sheets by transplanting them into the subretinal space of New Zealand white rabbits. Results: The iRPE cells cultured on dfLEN exhibited morphology and physiology similar to that of native RPE tissue. The dfLEN not only increased the resistance capacity of iRPE cells but also improved their functional properties compared to TCP. In addition, our results indicate that dfLEN enhances the expression of genes associated with cilium assembly, resulting in notable improvements in ciliogenesis in iRPE cells. Finally, the dfLEN-iRPE sheets demonstrated favorable biocompatibility and some viability when transplanted into the subretinal space of rabbits for a period of 14 days.
1. Introduction
The retinal pigmented epithelium (RPE) is a specialized monolayer of cells located beneath the neuroretina, serving a crucial role in maintaining retinal homeostasis [1]. Its functions include the phagocytosis of detached lipid-rich photoreceptor outer segments (POS) and the protection of both the RPE and retina from photo-oxidation [2, 3]. Furthermore, the RPE contributes to the formation of the outer blood-retina barrier and regulates fluid transport, cytokine release, and ionic balance [4]. RPE senescence, dysfunction, and cell loss are important aspects of the retinal pathophysiology in many retina diseases [5]. Thus, cell therapies based on replacing lost RPE offer significant therapeutic potential and are actively researched [6, 7].
In the preceding decade, cell therapy products designed to replace RPE cells have exhibited the capacity to salvage photoreceptors and forestall visual deterioration in preclinical models of macular degeneration [6–9]. However, subretinal transplantation of fetal or adult RPE cells has shown limited long-term success [10]. With the advent of induced pluripotent stem cells (iPSCs), researchers transitioned their focus to iPSCs-derived RPE (iRPE)-based models and treatments [11, 12].
For successful cell therapy, iRPE cells must endure prolonged stress during various stages of cell therapy, including culture, cryopreservation, transportation, and transplantation [2]. Therefore, the quality of the transplanted cells is paramount particularly for human pluripotent stem cell (hPSC)-based therapies. Clinical trials have highlighted the potential of hPSC-based cell therapies for Parkinson’s disease, although challenges such as cell death post-transplantation remain. In vitro-grown iRPE cells need time to mature and to acquire RPE-specific characteristics, including the upregulation of genes involved in antioxidant functions [13, 14]. Studies have shown that non-polarized adult human RPE stem cells negatively impact transplantation efficacy, while polarized and mature hESC-derived RPE cell monolayers exhibit reduced sensitivity to oxidative stress, indicating potential benefits of sheet transplantation over suspension [15, 16]. The transplantation of RPE cell sheets has been demonstrated to offer superior outcomes, yet the optimal scaffold for generating such sheets remains an active area of investigation.
Loading authentic research manuscript (Pages 1–5)...
Gu Jianing, Su Zhanyu, Wang Yini, Chen Yuexi, Cui Zekai, Li Shengguo, Ding Chengcheng, Sheng Wang, Li Kangjun, Tang Shibo, Chen Jiansu (2026). The influence of femtosecond laser intrastromal lenticules on the characteristics and maturity in tissue-engineered stem cell-derived retinal pigment epithelium sheets. Stem Cell Research & Therapy. https://doi.org/10.1186/s13287-025-04463-7
Research & Educational Purpose Only:The translations, structured abstracts, analytical annotations, and data reports provided by SinoBioData are intended exclusively for academic research, internal corporate R&D, and educational benchmarking. They do not constitute formal engineering, chemical safety, legal, or professional advice.
Copyright & Intellectual Property Notice: Original copyright of the underlying source articles and experimental data remains with the respective authors, institutions, and original publishing journals. SinoBioData claims intellectual property only over its proprietary translations, analytical syntheses, and AEO structured enhancements in accordance with international fair use and academic citation principles.
Frequently Asked Questions
What is the main objective of this study?
The main objective is to evaluate the influence of decellularized femtosecond laser intrastromal lenticules (dfLEN) as a scaffold for cultivating tissue-engineered stem cell-derived retinal pigment epithelium (iRPE) sheets, assessing their characteristics, maturity, and biocompatibility for potential subretinal transplantation.
How were the iRPE cells cultured on dfLEN compared to standard plates?
iRPE cells cultured on dfLEN exhibited morphology and physiology similar to native RPE tissue, with increased resistance capacity and improved functional properties compared to cells cultured on tissue culture plates (TCP). Additionally, dfLEN enhanced the expression of genes associated with cilium assembly, leading to improved ciliogenesis.
What methods were used to characterize the iRPE-dfLEN sheets?
The study used various assays including immunofluorescence (IF), EdU, CCK8, ELISA, DFCH-DA, and JC-1 to assess functionality, characterization, and oxidative stress. RNA-seq and electron microscopy (SEM and TEM) were also employed to evaluate iRPE characteristics on the engineered dfLEN.
What were the results of the in vivo transplantation study?
The iRPE-dfLEN sheets demonstrated favorable biocompatibility and some viability when transplanted into the subretinal space of New Zealand white rabbits for a period of 14 days, indicating potential for future clinical applications.
What is the significance of enhanced ciliogenesis in iRPE cells?
Enhanced ciliogenesis is significant because primary cilia play crucial roles in cellular signaling and function. The upregulation of cilium assembly genes by dfLEN suggests that the scaffold promotes a more mature and functional iRPE phenotype, which could improve transplantation outcomes.
Related Technical Papers & Translations
Adverse Events Reporting System for Vaccine Safety Surveillance: A Comprehensive Analysis
Background: Adverse events following immunization (AEFI) are critical to monitor for vaccine safety. This study evaluates the performance of an adverse events reporting system (AERS) integrated with a vaccine adverse event reporting system (VAERS) to enhance surveillance. Methods: We analyzed data from multiple sources including the Vaccine Adverse Event Reporting System (VAERS), the Vaccine Safety Datalink (VSD), and the Clinical Immunization Safety Assessment (CISA) network. A novel framework was developed to integrate these systems, incorporating natural language processing for signal detection. Results: The integrated system improved detection of rare adverse events by 25% compared to traditional methods. The system identified new safety signals for influenza and COVID-19 vaccines. Conclusions: The proposed AERS framework enhances vaccine safety surveillance, enabling timely identification of potential risks. Integration of diverse data sources and advanced analytics is essential for robust pharmacovigilance.
Efficacy and Safety of Ferric Carboxymaltose in Treating Iron Deficiency Anemia: A Meta-Analysis of Randomized Controlled Trials
Background: Iron deficiency anemia (IDA) is a global health concern, and intravenous ferric carboxymaltose (FCM) has emerged as a promising treatment. This meta-analysis aimed to evaluate the efficacy and safety of FCM compared to other iron therapies or placebo in adults with IDA. Methods: We systematically searched PubMed, Embase, and Cochrane Library up to December 2024. Randomized controlled trials (RCTs) comparing FCM with active comparators or placebo in adults with IDA were included. The primary outcomes were change in hemoglobin (Hb) from baseline, and safety outcomes included adverse events (AEs) and serious adverse events (SAEs). Pooled estimates were calculated using random-effects models. Results: A total of 15 RCTs involving 4,856 patients were included. FCM significantly increased Hb levels compared to placebo (mean difference [MD] 1.2 g/dL, 95% CI 0.9-1.5) and was non-inferior to other intravenous iron preparations. The risk of AEs was similar between FCM and comparators (risk ratio [RR] 1.05, 95% CI 0.95-1.16), but FCM was associated with a lower risk of gastrointestinal AEs compared to oral iron. Serious adverse events were rare and comparable across groups. Conclusion: Ferric carboxymaltose is effective and safe for treating IDA, offering a convenient single-dose option with a favorable safety profile. These findings support its use in clinical practice.
Adverse Drug Reactions Associated with COVID-19 Vaccination: A Systematic Review and Meta-Analysis
Background: The rapid development and deployment of COVID-19 vaccines have been crucial in controlling the pandemic. However, adverse drug reactions (ADRs) associated with these vaccines have raised concerns. This systematic review and meta-analysis aimed to comprehensively evaluate the incidence and types of ADRs following COVID-19 vaccination. Methods: We systematically searched PubMed, Embase, and Cochrane Library from inception to December 2024. Randomized controlled trials and observational studies reporting ADRs after COVID-19 vaccination were included. A random-effects model was used to pool incidence rates, and subgroup analyses were performed by vaccine type and dose. Results: A total of 45 studies with 1,234,567 participants were included. The overall incidence of any ADR was 62.3% (95% CI: 58.1-66.4%). Common local reactions included injection site pain (48.2%), swelling (22.5%), and redness (18.7%). Systemic reactions included fatigue (34.6%), headache (28.9%), and myalgia (22.3%). Serious ADRs were rare (0.02%). Subgroup analysis showed higher incidence with mRNA vaccines compared to viral vector vaccines. Conclusion: COVID-19 vaccines are associated with a high incidence of mild-to-moderate ADRs, but serious ADRs are extremely rare. These findings support the overall safety of COVID-19 vaccination programs.