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Keywords = amniotic epithelial cells

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24 pages, 2232 KB  
Article
Comparison of the Effects of Sorafenib and Stem Cell Secretome on HepG2 Cancer Cells with Respect to the Ras/Raf/MEK/ERK Pathway
by Aleksandra Gładyś, Aleksandra Skubis-Sikora, Bartosz Sikora, Kinga Pogoda-Mieszczak, Patrycja Wieczorek, Edyta Bogunia and Piotr Czekaj
J. Clin. Med. 2026, 15(17), 6738; https://doi.org/10.3390/jcm15176738 - 30 Aug 2026
Viewed by 268
Abstract
Background/Objective: Liver cancer is a global health challenge due to its resistance to most systemic therapies. Human mesenchymal and epithelial stem cells exhibit anti-proliferative and pro-apoptotic effects on some cancer cell lines, which can be used to support standard therapy. The aim of [...] Read more.
Background/Objective: Liver cancer is a global health challenge due to its resistance to most systemic therapies. Human mesenchymal and epithelial stem cells exhibit anti-proliferative and pro-apoptotic effects on some cancer cell lines, which can be used to support standard therapy. The aim of this study was to investigate the effects of conditioned media (CM) derived from mesenchymal adipose tissue-derived stem cells (hADSCs) and amniotic membrane-derived cells expressing both mesenchymal and epithelial markers (hACs) on HepG2 liver cancer cells in vitro and to identify possible similarities to the action of sorafenib, a standard drug in hepatocellular carcinoma (HCC) therapy. Methods: HepG2 cells were cultured with hADSC-derived CM (CM-hADSC) or hAC-derived CM (CM-hAC), either alone or with 7.5 μm sorafenib for 48 h. HepG2 cell viability and the expression of genes and/or proteins related to the apoptosis, cell-cycle, and Ras/Raf/MEK/ERK signaling pathway, were assessed. Results: The effect of sorafenib administration alone, consisting in reducing HepG2 cell viability, cell cycle inhibition, and reducing the expression of the alpha-fetoprotein (AFP) gene and most proteins related to the Ras/Raf/MEK/ERK pathway, was dominant over the effect of CM administered in combination with one of them. The effect of sorafenib on increasing the number of early apoptotic cells was not associated with increased Bax protein expression or with an increased proportion of cleaved forms of caspase-7 and caspase-9. The effect of CM-hAC was often different from that of CM-hADSC and consisted in enhancing the inhibitory effect of sorafenib at the cell cycle level and increasing the number of apoptotic cells and caspase mRNA expression, while activating the Ras/Raf/MEK/ERK signaling pathway and inducing AFP gene expression. Conclusions: Combined administration of sorafenib and some CM may produce the desired effects by enhancing the action of the drug. However, some of the effects of sorafenib and CM derived from different stem cell types were significantly different, which means that the role of stem cell secretome as a potential anticancer therapy requires further investigation, taking into account different types of cancer cell lines and a thorough analysis of the secretome composition. Full article
(This article belongs to the Section Oncology)
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19 pages, 3397 KB  
Article
Evaluation of Two Methods for Decellularizing Human Amniotic Membranes for Use in Tissue Engineering
by Elga Johanna Vargas, Carlos Domínguez-Paz and Lina Andrea Gómez
Methods Protoc. 2026, 9(5), 127; https://doi.org/10.3390/mps9050127 - 28 Aug 2026
Viewed by 271
Abstract
Background: The human amniotic membrane (hAM) is widely used in tissue engineering due to its low immunogenicity and extracellular matrix (ECM) rich in structural proteins; in this context, decellularization aims to remove cellular components while preserving tissue integrity, and this study aimed to [...] Read more.
Background: The human amniotic membrane (hAM) is widely used in tissue engineering due to its low immunogenicity and extracellular matrix (ECM) rich in structural proteins; in this context, decellularization aims to remove cellular components while preserving tissue integrity, and this study aimed to compare the effects of two enzymatic methods (trypsin and thermolysin). Methods: Human amniotic membranes obtained from placentas were divided into three groups: non-decellularized control, trypsin-treated, and thermolysin-treated. The membranes were treated with 0.25 trypsin-EDTA for 60 min at 37 °C, followed by incubation with Triton X-100 for 30 min at 37 °C, or with 125 µg/mL thermolysin for 9 min at 37 °C. Histological analysis with hematoxylin–eosin and semi-quantitative scoring assessed epithelial and stromal integrity, immunohistochemistry evaluated laminin α and fibronectin expression with quantitative image analysis, and uniaxial tensile testing measured mechanical properties; no preregistration numbers or animal models were reported. Results: Trypsin achieved more effective epithelial removal, whereas thermolysin caused greater stromal disruption; no statistically significant differences in laminin α or fibronectin expression were found among groups (p > 0.05), indicating preservation of key ECM components; both treatments reduced mechanical performance, with thermolysin decreasing early- and mid-path stiffness and trypsin reducing late-path stress and work density. Conclusion: Both enzymatic protocols preserved ECM composition but induced distinct structural and mechanical alterations, suggesting that method selection should balance efficient cell removal with preservation of structural and functional properties according to the intended application. Full article
(This article belongs to the Section Tissue Engineering and Organoids)
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19 pages, 4370 KB  
Article
Construction of a Feline Amniotic Membrane Scaffold Loaded with Limbal Stem Cell Grafts
by Can Huang, Xiaoyan Hu, Yunying Peng, Xueling Piao, Xuanrui Tao, Shihao Yang, Heng Yang, Shicheng Bi, Dezhi Zhang, Lijing Cao, Bin Wu, Ling Gan and Huihao Xu
Vet. Sci. 2026, 13(9), 853; https://doi.org/10.3390/vetsci13090853 - 23 Aug 2026
Viewed by 249
Abstract
Amniotic membranes are extensively used in feline corneal repair; however, challenges such as rapid degradation and difficulties with preservation across species limit their clinical use. The aim of this study was to develop a feline amniotic membrane-based graft with stem cell activity and [...] Read more.
Amniotic membranes are extensively used in feline corneal repair; however, challenges such as rapid degradation and difficulties with preservation across species limit their clinical use. The aim of this study was to develop a feline amniotic membrane-based graft with stem cell activity and provide experimental evidence for its potential use as a corneal epithelial transplant material in cats. FLSCs were first treated with different concentrations of epidermal growth factor (EGF), and 0.5 mg/L EGF was identified as the optimal concentration for promoting cellular activity. Under this condition, FLSCs exhibited enhanced proliferation and wound closure ability, with complete scratch closure observed approximately 24 h earlier than that in the control group. Growth curve analysis showed that FLSCs maintained stable proliferation characteristics, with a calculated population doubling time of approximately 25.2 h. Subsequently, FLSCs were seeded onto the lyophilized feline amniotic membrane scaffold. Light microscopy, scanning electron microscopy, and transmission electron microscopy demonstrated that FLSCs adhered to and proliferated on the scaffold, forming cell–matrix interaction structures, including hemidesmosome-like junctions. Immunofluorescence staining confirmed the expression of stem cell-associated markers ABCG2 and p63 after scaffold culture. These findings demonstrate the successful fabrication of a feline FLSC-loaded amniotic membrane construct in which FLSCs retained the expression of the examined limbal/stem cell-associated markers after scaffold culture. The successfully constructed FLSC-loaded feline amniotic membrane graft demonstrated considerable potential as a novel bioengineered tissue substitute for corneal surface transplantation in clinical applications. Full article
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21 pages, 5538 KB  
Article
Human Amniotic Epithelial Stem Cells and Osteoblast Cells Behavior on Collagen Membranes for Bone Guided Regeneration
by Antonio Pérez-Pérez, Javier Gil, Isabela Bueno-Bianchi, Loreto Monsalve-Guil, Iván Ortiz-Garcia, Alvaro Jiménez-Guerra, Enrique Núñez-Márquez, Eugenio Velasco-Ortega, José Luis Rondón Romero, Victor Sánchez-Margalet and Jesús Moreno-Muñoz
Int. J. Mol. Sci. 2026, 27(15), 6660; https://doi.org/10.3390/ijms27156660 - 26 Jul 2026
Viewed by 386
Abstract
Guided bone regeneration (GBR) heavily relies on barrier membranes, with collagen being the clinical standard material. Human amniotic epithelial cells (hAECs) represent a promising, non-controversial stem cell source with substantial osteogenic and immunomodulatory potential. This study aimed to comparatively evaluate the structural characteristics [...] Read more.
Guided bone regeneration (GBR) heavily relies on barrier membranes, with collagen being the clinical standard material. Human amniotic epithelial cells (hAECs) represent a promising, non-controversial stem cell source with substantial osteogenic and immunomodulatory potential. This study aimed to comparatively evaluate the structural characteristics of three commercial collagen membranes (Biocollagen®, Derma®, and VantyColl®) and their influence on the biological behavior, viability, and osteogenic differentiation of hAECs and hFOB 1.19 human fetal osteoblasts. The microarchitecture was assessed via scanning electron microscopy (SEM). Biological response was evaluated over 14 days, using MTT assays, calcium and phosphorus quantification, alkaline phosphatase (ALP) activity, and quantitative real-time PCR (qRT-PCR) for osteogenic markers (Runx2, Osterix, ALP, and OPN). SEM revealed a dense lamellar structure for Biocollagen®, a fibrillar and oriented architecture for Derma®, and a highly porous network for VantyColl®. Both cell types adhered to and proliferated on all membranes. Derma® provided the best long-term proliferative support for both lineages. Conversely, VantyColl® induced robust early osteoblastic differentiation, marked by exceptional upregulation of Osterix (24.93-fold) and Runx2 (2.64-fold), though it exhibited diminished long-term hAEC viability. Ultimately, collagen membrane microarchitecture dictates cell fate; dense fibrillar networks (Derma®) favor sustained growth and late matrix maturation (OPN), whereas high-porosity scaffolds (VantyColl®) amplify early osteoinductive cascades. Full article
(This article belongs to the Special Issue Advanced Biomaterials for Tissue Regeneration)
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19 pages, 9448 KB  
Article
Effects of Hydrodynamic Ozonated Water Processing on the Thermal Stability and Structural Integrity of the Human Amniotic Membrane
by Marcia Guelma Santos Belfort, Francisco Dimitre Rodrigo Pereira Santos, Maycon Crispim de Oliveira Carvalho, Aline Casarin dos Santos, Pedro Augusto Laurindo Igreja Marrafa, João Gomes de Oliveira Neto, Carlos José de Lima and Adriana Barrinha Fernandes
J. Funct. Biomater. 2026, 17(7), 352; https://doi.org/10.3390/jfb17070352 - 20 Jul 2026
Viewed by 487
Abstract
This study aimed to verify the morphology, biochemical composition, and thermal characterization of hydrated human amniotic membrane (HAM) processed in an ozonated water hydrodynamic system. This is an in vitro experimental study in which HAM samples were divided into two groups: in natura [...] Read more.
This study aimed to verify the morphology, biochemical composition, and thermal characterization of hydrated human amniotic membrane (HAM) processed in an ozonated water hydrodynamic system. This is an in vitro experimental study in which HAM samples were divided into two groups: in natura (IN) and ozonated (O3). Analyses were performed using histology, Fourier-transform infrared spectroscopy (FT-IR), thermogravimetric analysis (TGA/DTG), and differential scanning calorimetry (DSC/dDSC). Ozonation for 40 min preserved the biochemical integrity of HAM, maintaining the characteristic vibrational bands of Amides I, II, and III. Histological analysis showed morphological changes in epithelial cells, with partial removal in some regions, while the basement membrane and the scaffold remained preserved. Thermal analysis revealed that the in natura sample presented a bimodal dehydration profile, with a first event occurring between 60 and 65 °C associated with the evaporation of free or weakly bound water, and a second event peaking around 80 °C related to the removal of structural water. In contrast, the ozonated HAM exhibited a unimodal profile, with the mass loss peak shifted to approximately 70 °C. These findings were corroborated by DSC analysis, which showed a reduction in denaturation temperature from approximately 85 °C in the in natura sample to around 75 °C in the ozonated sample. The dDSC analysis confirmed the transition from a bimodal to a unimodal behavior after treatment, indicating a reduced energy barrier for protein denaturation and lower thermal stability of the collagen matrix. These results suggest that ozonation promotes alterations in the epithelial layer, which may favor the loss of both free and bound water. It is concluded that processing with ozonated water induces structural modifications, especially in the epithelial layer, and reduces the thermal stability of hydrated HAM without significantly altering the biochemical signature of collagen. This approach shows potential as an alternative method for membrane processing; however, functional evaluations are required to confirm its clinical applicability. Full article
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21 pages, 4793 KB  
Article
Establishment of In Ovo Salmonella Enteritidis Infection and Synbiotic Delivery Models in Chick Embryos and Their Effects on Early Gut Health
by Riliang Liu, Jiguang Wang, Jiying Dai, Yamei Wang, Weijiang Zheng and Wen Yao
Animals 2026, 16(12), 1863; https://doi.org/10.3390/ani16121863 - 17 Jun 2026
Viewed by 1273
Abstract
Early microbial exposure during embryogenesis may shape post-hatch gut development in poultry, yet the effects of sublethal pathogenic exposure and in ovo synbiotics remain unclear. This model-establishment study preliminarily established in ovo Salmonella Enteritidis (SE) infection and synbiotic (SYN) delivery models and evaluated [...] Read more.
Early microbial exposure during embryogenesis may shape post-hatch gut development in poultry, yet the effects of sublethal pathogenic exposure and in ovo synbiotics remain unclear. This model-establishment study preliminarily established in ovo Salmonella Enteritidis (SE) infection and synbiotic (SYN) delivery models and evaluated their effects on hatchability, cecal microbiota, intestinal morphology, epithelial turnover, and barrier function in newly hatched chicks. In one group, the air cells of specific pathogen-free White Leghorn eggs were injected with SE on embryonic day 12; in another group, a synbiotic consisting of Lactobacillus plantarum, Pediococcus acidilactici, and inulin was injected into the amniotic cavity on embryonic day 17.5. Sterile saline was injected as the vehicle-only procedural control at the corresponding time points and injection sites. Based on their impacts on hatchability, SE1-L and SYN-H were selected. SE1-L reduced cecal microbial diversity, expanded Proteobacteria and Escherichia–Shigella, increased ileal apoptosis and crypt depth, decreased the villus height-to-crypt depth ratio, downregulated jejunal tight-junction genes, upregulated ileal MYD88 and TNF-α, and increased plasma lipopolysaccharide and D-lactate. In contrast, SYN-H maintained hatchability, promoted early Pediococcus colonization, suppressed potential pathogens, increased ileal villus height and villus height-to-crypt depth ratio, enhanced proliferation, reduced apoptosis, and improved mucosal barrier-related indices. These findings provide preliminary evidence that embryonic SE infection and synbiotic delivery differentially influence early intestinal microbiota succession and gut development in newly hatched chicks. Full article
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15 pages, 4594 KB  
Article
Comparative Analysis of Ectodermal Marker Expression in Human Adipose-Derived Stem Cells and Amniotic Epithelial Cells Exposed to Ectoderm-Inducing Conditions
by Bartosz Sikora, Aleksandra Skubis-Sikora, Marcin Ciekalski, Patrycja Wieczorek, Agnieszka Prusek-Kucharek and Piotr Czekaj
Int. J. Mol. Sci. 2026, 27(11), 4976; https://doi.org/10.3390/ijms27114976 - 30 May 2026
Cited by 1 | Viewed by 387
Abstract
Nervous system and corneal disorders are major causes of permanent disability worldwide, largely due to the limited regenerative capacity of ectoderm-derived tissues. Therefore, the development of accessible and ethically acceptable cell-based therapies promoting the repair and regeneration of these tissues is of considerable [...] Read more.
Nervous system and corneal disorders are major causes of permanent disability worldwide, largely due to the limited regenerative capacity of ectoderm-derived tissues. Therefore, the development of accessible and ethically acceptable cell-based therapies promoting the repair and regeneration of these tissues is of considerable translational importance. In this study, we aimed to comparatively evaluate the ectodermal differentiation potential of human adipose-derived stem cells (ADSCs) and human amniotic epithelial cells (hAECs) in vitro, with hAECs serving as a reference cell population with established ectodermal plasticity. Primary ADSCs and hAECs were characterized phenotypically using flow cytometry and functional differentiation assays. Cells were subjected to a directed ectodermal differentiation protocol and assessed via morphological analysis, immunostaining for ectoderm-associated proteins, and RT-qPCR analysis of lineage-specific genes. ADSCs exhibited morphological changes following differentiation, including a more epithelial-like phenotype and an increased nucleus-to-cytoplasm ratio. Immunostaining revealed the induction of nestin and OTX2 expression after differentiation, which was particularly pronounced in ADSCs. Gene expression analysis demonstrated statistically significant upregulation of the ectoderm-related genes EN2, SOX1, and PAX6 exclusively in hAECs. Results suggest that in ADSCs the differentiation process was only partially activated. In conclusion, our findings further support the suitability of hAECs as a reference cell line for studies investigating ectodermal differentiation protocols, while also demonstrating that ADSCs exhibit a limited but detectable capacity for acquiring ectoderm-specific characteristics under defined in vitro culture conditions. Full article
(This article belongs to the Special Issue Latest Research on Mesenchymal Stem Cells (2nd Edition))
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13 pages, 1471 KB  
Case Report
Amniotic Membrane Transplantation Preserves Vision in Pediatric Recessive Dystrophic Epidermolysis Bullosa: Case Series
by Seika Den, Yukako Abukawa, Nanami Kishimoto, Ryuichi Shimada, Yuka Higashi, Kozue Kasai and Tadashi Nakano
J. Clin. Med. 2026, 15(9), 3503; https://doi.org/10.3390/jcm15093503 - 3 May 2026
Viewed by 707
Abstract
Background: Recessive dystrophic epidermolysis bullosa (RDEB) is a rare inherited disorder characterized by extreme epithelial fragility and progressive cicatrization, frequently leading to severe ocular surface disease and early visual impairment. Surgical interventions such as ocular surface reconstruction (OSR) in childhood are often [...] Read more.
Background: Recessive dystrophic epidermolysis bullosa (RDEB) is a rare inherited disorder characterized by extreme epithelial fragility and progressive cicatrization, frequently leading to severe ocular surface disease and early visual impairment. Surgical interventions such as ocular surface reconstruction (OSR) in childhood are often delayed because of anesthetic risks and concerns regarding recurrence. Consequently, the effectiveness of OSR, including amniotic membrane transplantation (AMT), and its impact on visual development remain poorly documented. Methods: We report a case series of two pediatric patients (three eyes) with genetically confirmed RDEB who underwent single-step OSR using AMT. Clinical outcomes, long-term visual acuity, perioperative management, and histopathological findings were evaluated. Results: Ocular manifestations included corneal epithelial damage, symblepharon, and pseudopterygium extending over the cornea. One patient underwent symblepharon lysis, superficial keratectomy, and AMT onto the bare sclera in the right eye at age 4 and in the left eye at age 8, both under intubated general anesthesia. The other patient underwent the same procedure in the right eye at age 6. Best spectacle-corrected visual acuity improved from ≤20/300 to 20/30 in all eyes, and pupillary zone clarity was maintained during the follow-up period (up to 6 years). Histopathology confirmed pseudopterygium with squamous metaplasia, goblet cell loss, and fibrovascular stroma. Safe general anesthesia was achieved through meticulous multidisciplinary perioperative planning involving anesthesiologists, dermatologists, and pediatricians. No systemic complications related to anesthesia or perioperative management were observed. Conclusions: Single-step OSR with on-lay AMT can restore and preserve visual function in pediatric RDEB. Early surgical intervention may prevent profound amblyopia and provide durable ocular surface stability. A multidisciplinary approach enables safe general anesthesia and perioperative management. Full article
(This article belongs to the Section Ophthalmology)
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41 pages, 3361 KB  
Systematic Review
A Systematic Review on Amnion as a Cell Delivery Scaffolding Material for Cartilage Regeneration in Pre-Clinical and Clinical Studies
by Shu-Yong Liow, Sik-Loo Tan, Alvin Jiunn-Hieng Lu, Kwong Weng Loh, Seow Hui Teo, Chan Young Lee, Le Wan, Azlina Amir Abbas and Kyung-Soon Park
Bioengineering 2026, 13(3), 357; https://doi.org/10.3390/bioengineering13030357 - 18 Mar 2026
Cited by 2 | Viewed by 1584
Abstract
Cartilage is an important yet vulnerable tissue with limited self-healing capacity, where damage often progresses to joint degeneration, which eventually leads to severe osteoarthritis (OA). Current tissue engineering strategies focus on biocompatible scaffolds for cartilage regeneration, particularly amnion (or amniotic membrane), emerging as [...] Read more.
Cartilage is an important yet vulnerable tissue with limited self-healing capacity, where damage often progresses to joint degeneration, which eventually leads to severe osteoarthritis (OA). Current tissue engineering strategies focus on biocompatible scaffolds for cartilage regeneration, particularly amnion (or amniotic membrane), emerging as a promising biomaterial due to its wide availability, low immunogenicity, and naturally derived microenvironment that is advantageous for cartilage regeneration. This systematic review aims to evaluate the existing evidence on the efficacy of amnion as a tissue scaffolding material for cartilage regeneration in both preclinical and clinical studies. Using terms such as “cartilage damage”, “cartilage injuries”, “amnion” and “amniotic membrane”, 19 relevant studies were identified across three major databases (PubMed, Scopus and Web of Science) until 25 December 2025. All preclinical and clinical studies that utilized amnion for cartilage repair or as cartilage tissue engineering scaffolding materials were included. Evidence quality was assessed using the OHAT and MINORS risk of bias tool. This study is prospectively registered in the PROSPERO database under the ID 1178444. The findings consistently indicate that amniotic scaffolds, regardless of processing methods or cell seeding, yield favorable outcomes without adverse effects across different species. In vitro analysis revealed that treatment groups with amnion show better cell attachment, viability, and proliferation, and higher content of cartilage-related markers expressed by the seeded cells, either chondrocyte, bone marrow-derived mesenchymal stem cells (MSCs), adipose tissue-derived MSCs, placenta-derived MSCs, umbilical cord-derived MSCs, amniotic MSCs or amniotic epithelial cells. In in vivo and ex vivo studies, amnion-treated groups demonstrated improved quality of the treated cartilage, with better integration, as indicated by higher histological scores and the presence of type II collagen (COL-II). There was an inconsistency in the reporting of cartilage defect dimensions in the in vivo models across the different studies. Nevertheless, the outcome measurements were consistently reported with histological analysis, with or without International Cartilage Repair Society (ICRS) scoring and immunohistochemistry (IHC) analysis, across the studies. Clinically, most subjects show improvement in the Knee Injury and Osteoarthritis Outcome Score (KOOS) Sports and Recreation score and KOOS Quality of Life score, as well as reduced Visual Analogue Scale (VAS) average and maximum pain scores. In conclusion, preclinical and clinical studies support amnion as an ideal scaffold material for cartilage tissue engineering and regeneration. Future research should focus on optimizing and standardizing amnion scaffold preparation at a production scale to facilitate the translation of these positive outcomes into clinical applications. This study is funded by the Ministry of Higher Education Malaysia via Prototype Research Grant Scheme (PRGS/1/2021/SKK01/UM/02/1) and UM International Collaboration Grant—2023 SATU Joint Research Scheme Program: ST007-2024. Full article
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24 pages, 385 KB  
Review
Autoimmune Ocular Surface Disorders: From Molecular Immunopathogenesis to Regenerative and Surgical Therapeutics
by Wojciech Luboń, Marta Świerczyńska, Katarzyna Jadczyk-Sorek and Dorota Wyględowska-Promieńska
Cells 2026, 15(4), 378; https://doi.org/10.3390/cells15040378 - 22 Feb 2026
Cited by 2 | Viewed by 1912
Abstract
Autoimmune ocular surface diseases represent a complex group of disorders in which systemic immune dysregulation triggers chronic inflammation, epithelial dysfunction, and progressive tissue fibrosis. Systemic lupus erythematosus, primary Sjögren’s syndrome, and ocular cicatricial pemphigoid are the principal entities linking systemic autoimmunity to ocular [...] Read more.
Autoimmune ocular surface diseases represent a complex group of disorders in which systemic immune dysregulation triggers chronic inflammation, epithelial dysfunction, and progressive tissue fibrosis. Systemic lupus erythematosus, primary Sjögren’s syndrome, and ocular cicatricial pemphigoid are the principal entities linking systemic autoimmunity to ocular surface pathology. These conditions share convergent mechanisms—including dysregulated cytokine signaling (IFN-I, IL-6, and IL-17), complement activation, and epithelial–mesenchymal transition—culminating in tear film instability and visual impairment. Recent advances in molecular immunology and omics profiling have elucidated disease-specific pathways and identified actionable therapeutic targets. Conventional immunosuppressants such as corticosteroids and cyclosporine remain fundamental, yet emerging biologics targeting BAFF, IFNAR, and JAK/STAT signaling—alongside regenerative strategies employing mesenchymal and induced pluripotent stem cells—are transforming disease management. Parallel innovations in amniotic membrane transplantation, keratoprosthesis, and bioengineered corneal scaffolds integrate structural reconstruction with immune modulation. Furthermore, the convergence of multi-omics analytics, artificial intelligence-assisted diagnostics, and microbiome-based immunomodulation heralds a new era of precision ophthalmology. This review synthesizes current molecular insights, clinical observations, and translational advances that collectively redefine autoimmune ocular surface diseases—from chronic inflammatory disorders into a targetable, regenerative, and potentially reversible spectrum of conditions. Full article
32 pages, 3948 KB  
Article
Immuno-Instructive 3D Tendon Biomimetic Scaffolds Functionalized with Amniotic Epithelial Stem Cell Secretome for Controlled Inflammation and Targeted Macrophage Polarization
by Mohammad El Khatib, Annunziata Mauro, Giuseppe Prencipe, Oriana Di Giacinto, Valeria Giovanna Festinese, Carola Agostinone, Maura Turriani, Paolo Berardinelli, Barbara Barboni and Valentina Russo
Int. J. Mol. Sci. 2026, 27(4), 2029; https://doi.org/10.3390/ijms27042029 - 20 Feb 2026
Viewed by 887
Abstract
Tendon healing is often hindered by unresolved inflammation and dysregulated immune responses, highlighting the need for innovative regenerative strategies. This study developed an immune-informed platform by functionalizing validated 3D tendon-mimetic poly(lactide-co-glycolide) (PLGA) scaffolds with immunomodulatory conditioned media (CM), referred to as CMINF [...] Read more.
Tendon healing is often hindered by unresolved inflammation and dysregulated immune responses, highlighting the need for innovative regenerative strategies. This study developed an immune-informed platform by functionalizing validated 3D tendon-mimetic poly(lactide-co-glycolide) (PLGA) scaffolds with immunomodulatory conditioned media (CM), referred to as CMINF to emphasize its anti-inflammatory and immunomodulatory properties, derived from ovine amniotic epithelial stem cells (AECs), offering a potential cell-free therapeutic solution. Three functionalization methods were compared: physical adsorption, and hydrochloric acid (HCl) or sodium hydroxide (NaOH) pre-treatments. FT-IR spectroscopy and protein adsorption analyses identified NaOH as the most effective method, enhancing retention and release of Amphiregulin (AREG), an AEC key immunomodulatory protein. Kinetic studies revealed a sustained, controlled release of AREG over 7 days (d) from CMINF-functionalized scaffolds (3D-CMINF), preserving bioactivity. Functionally, 3D-CMINF scaffolds significantly suppressed T-cell activation and peripheral blood mononuclear cell (PBMC) proliferation. The released CM from 3D-CMINF (CMR) exhibited time-dependent immunomodulatory effects: early T-cell inhibition (6–72 h) and delayed suppression of PBMC proliferation (48 h–7 d). Macrophage polarization analysis revealed a shift towards the pro-regenerative M2 phenotype, with increased expression of M2 over M1 markers in 3D-CMINF-adherent cells. Flow cytometry confirmed a preferential induction of regulatory M2b macrophages alongside reductions in pro-inflammatory M1 and pro-fibrotic M2a subsets. These results demonstrate that 3D-CMINF scaffolds can finely modulate immune responses, balancing inflammatory and reparative cues relevant to early tendon healing processes. This platform, integrating structural and immunomodulatory elements, presents a promising, cell-free, and translational immunoengineering strategy to control inflammation and support tendon repair. Full article
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14 pages, 278 KB  
Review
Cultivated Oral Mucosal Epithelial Transplantation for Limbal Stem Cell Deficiency: A Scoping Review of Indications, Platforms, Outcomes and Safety
by Konstantinos Papadopoulos, Mohamed Elalfy, Hasan Naveed, Sokratis Zormpas and Artemis Matsou
J. Clin. Med. 2026, 15(3), 1134; https://doi.org/10.3390/jcm15031134 - 1 Feb 2026
Viewed by 921
Abstract
Background: Cultivated oral mucosal epithelial transplantation (COMET/CAOMECS) is an autologous, immunosuppression-sparing option for ocular surface reconstruction in limbal stem cell deficiency (LSCD). After two decades, indications, platforms and outcome definitions vary, and COMET’s position relative to limbal-derived epithelium remains uncertain. Methods: We conducted [...] Read more.
Background: Cultivated oral mucosal epithelial transplantation (COMET/CAOMECS) is an autologous, immunosuppression-sparing option for ocular surface reconstruction in limbal stem cell deficiency (LSCD). After two decades, indications, platforms and outcome definitions vary, and COMET’s position relative to limbal-derived epithelium remains uncertain. Methods: We conducted a PRISMA-ScR scoping review of human clinical studies (PubMed, 2000–30 December 2025) with hand-searching and regulatory sources. Eligible reports included COMET/CAOMECS series and comparative cohorts (CLET/ACLET, SLET, KLAL/CLAL). The primary outcome was anatomical success (stable epithelialised cornea without recurrent persistent epithelial defect, progressive conjunctivalisation or uncontrolled neovascularisation at last assessment). Given heterogeneity in definitions and analytic frames (fixed-time vs. Kaplan–Meier [KM]), results were synthesised narratively by indication and platform. Results: Twenty-five reports (893 eyes; 821 patients) were included. Aetiologies were predominantly burns and SJS/TEN. Across amniotic membrane-based mixed-aetiology series, 12-month anatomical success clustered around 55–70%. Aggregated descriptively across COMET eyes, 211/467 (45%) had a stable surface at last follow-up. Epithelialisation was generally rapid in quiet AM-based reconstructions and slower with severe adnexal disease or carrier-free platforms. Mean BCVA improved from 1.8 ± 0.7 to 1.4 ± 0.7 logMAR (471 eyes); ≥2-line gains occurred in 308/471 (65.4%). A matched comparison suggested better 12-month survival, less neovascularisation and better BCVA with substrate-free versus AM-carried COMET; a biomaterial-/feeder-free platform reconstructed most eyes but failed more often with four-quadrant symblepharon. Observational comparative cohorts suggested higher surface survival and average visual gain with limbal-derived epithelium, at the cost of systemic immunosuppression. Conclusions: In appropriately selected bilateral LSCD, COMET offers immunosuppression-sparing reconstruction with moderate, durable surface stability and clinically meaningful visual gains when performed on a quiet, optimised surface. Platform refinements—particularly substrate-free constructs—and prospective, indication-defined comparative studies with harmonised outcomes are needed to define COMET’s role relative to limbal-derived epithelium. Full article
8 pages, 2417 KB  
Case Report
Amniotic Membrane-Assisted Corneal Transplantation in Ocular Perforation Due to GVHD: A Case Report
by Nicola Cardascia, Maria Gabriella La Tegola, Francesco D’Oria, Giacomo Boscia, Francesco Boscia and Giovanni Alessio
J. Clin. Med. 2026, 15(2), 548; https://doi.org/10.3390/jcm15020548 - 9 Jan 2026
Viewed by 930
Abstract
Background/Objectives: Ocular graft-versus-host disease (oGVHD) is a chronic, immune-mediated complication of allogeneic hematopoietic stem cell transplantation that can progress to corneal ulceration or perforation. These cases are often refractory to standard therapy and present a high risk of graft failure after keratoplasty. We [...] Read more.
Background/Objectives: Ocular graft-versus-host disease (oGVHD) is a chronic, immune-mediated complication of allogeneic hematopoietic stem cell transplantation that can progress to corneal ulceration or perforation. These cases are often refractory to standard therapy and present a high risk of graft failure after keratoplasty. We report a case of oGVHD-related corneal perforation successfully managed with a novel amniotic membrane-assisted “envelope” technique during corneal transplantation. Case Report: A 42-year-old man with chronic oGVHD and a full-thickness corneal perforation underwent urgent repair with a lamellar patch graft completely wrapped in cryopreserved amniotic membrane, followed by penetrating keratoplasty (PKP) using an amniotic membrane envelope surrounding the donor lenticule. Results: The amniotic membrane provided a 360° biological barrier that isolated graft antigens from the inflammatory environment while supporting epithelial healing and stromal remodeling. Despite recurrent inflammatory episodes and multiple procedures—including cataract extraction, pars plana vitrectomy, and multilayer amniotic membrane transplantation—the graft remained clear and stable at 12-month follow-up, achieving a best-corrected visual acuity of 20/40. Conclusions: The amniotic membrane envelope technique may represent a valuable adjunct in managing high-risk corneal perforations secondary to oGVHD. By combining immune modulation and regenerative support, this approach can enhance tectonic stability, reduce rejection risk, and promote durable surface recovery, potentially delaying or avoiding keratoprosthesis in refractory cases. Full article
(This article belongs to the Special Issue Diagnosis and Management of Corneal Diseases)
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16 pages, 1688 KB  
Article
Outcome of Allogeneic Penetrating Limbo-Keratoplasty: A Single-Center Retrospective Cohort Study
by Marie Ella Horstmann, Alexander K. Schuster, Norbert Pfeiffer and Joanna Wasielica-Poslednik
J. Clin. Med. 2025, 14(24), 8958; https://doi.org/10.3390/jcm14248958 - 18 Dec 2025
Viewed by 746
Abstract
Introduction: Allogeneic penetrating limbo-keratoplasty (limbo-PK) is one of the surgical methods for the treatment of limbal stem cell deficiency (LSCD). We report real-life results on different entities. Methods: Patients treated with limbo-PK at the Department of Ophthalmology of the University Medical Center [...] Read more.
Introduction: Allogeneic penetrating limbo-keratoplasty (limbo-PK) is one of the surgical methods for the treatment of limbal stem cell deficiency (LSCD). We report real-life results on different entities. Methods: Patients treated with limbo-PK at the Department of Ophthalmology of the University Medical Center Mainz were evaluated retrospectively. The primary endpoint was the epithelialization of the graft one year postoperatively. In addition, the postoperative best corrected visual acuity (BCVA), ocular concomitant diseases, drug treatment, and the need for further eye surgery postoperatively were examined. Results: We included 14 eyes of 13 patients (4 female) aged 59.8 ± 14.1 years who underwent limbo-PK between 2020 and 2024. Indications for limbo-PK included chemical burns (n = 4), blast injuries (n = 4), thermal burns (n = 2), trauma (n = 1) graft-versus-host disease (n = 1), and ectrodactyly-ectodermal dysplasia (EEC) (n = 1). The mean preoperative BCVA was 2.2 ± 0.6 logMAR (range: light perception to 0.7 logMAR). Four limbo-PK-grafts were HLA-typed. All limbo-PKs were combined with amniotic membrane transplantation; three with cataract surgery and one with tarsorrhaphy. Postoperatively, all patients received local immunosuppression, and 12 (85.7%) received additional systemic immunosuppression. At one-year follow-up mean BCVA increased to 1.0 ± 0.7 logMAR (range: 2.3 to 0.1, p-value = 0.03) and 11 of 14 eyes showed a functional graft with closed epithelium. In the further postoperative course, four patients needed a further Limbo-PK due to graft failure (n = 2), immune graft rejection after stopping local immunosuppressive therapy (n = 1) and perforation of the graft in a severe case of GvHd (n = 1). Conclusions: Limbo-PK is an effective surgical method for the treatment of LSCD. In our study cohort, we observed a significant improvement in mean BCVA one year postoperatively, with a functional, epithelialized graft achieved in 11 of 14 eyes. Full article
(This article belongs to the Section Ophthalmology)
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30 pages, 11166 KB  
Article
Potential Therapeutic Effects of Epithelial and Mesenchymal Stem Cell Secretome in Benzalkonium Chloride-Induced Limbal Stem Cell Dysfunction
by Agnieszka Prusek-Kucharek, Bartosz Sikora and Piotr Czekaj
Cells 2025, 14(22), 1790; https://doi.org/10.3390/cells14221790 - 14 Nov 2025
Cited by 1 | Viewed by 1440
Abstract
Dry Eye Disease (DED) is a multifactorial condition of the ocular surface, with one potential cause being damage from eye drops containing preservatives such as benzalkonium chloride (BAC). Current treatments for DED are unsatisfactory; therefore, it is worth exploring new therapies based on [...] Read more.
Dry Eye Disease (DED) is a multifactorial condition of the ocular surface, with one potential cause being damage from eye drops containing preservatives such as benzalkonium chloride (BAC). Current treatments for DED are unsatisfactory; therefore, it is worth exploring new therapies based on the secretome derived from stem cells. Human stem cells are important sources of growth factors and cytokines that promote tissue regeneration. The secretome of these cells can be obtained in vitro in conditioned medium (CM). The aim of the study was to evaluate the effect of CM derived from adipose-derived stem cells (hADSCs) and amniotic membrane-derived cells expressing mesenchymal and/or epithelial markers on limbal stem cells (LSCs) damaged by BAC, focusing on their regenerative potential. The study used two experimental models: the first focused on neutralizing the toxic effects of BAC when each CM was administered concurrently, and the second on the therapeutic effects of CM after prior cell damage by BAC. The effects of CM on LSCs were assessed, including apoptosis, cell cycle progression, proliferation, migration, and inflammation. CM from ADSCs and amniotic cells were shown to significantly reduce BAC-induced damage to LSCs. All tested CM promoted LSC regeneration, although their efficacy varied among treatments. The application of CM during BAC exposure yielded stronger and more consistent benefits than post-injury treatment. Full article
(This article belongs to the Section Cell and Gene Therapy)
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