Stem cell-based therapy represents a promising approach for the treatment of numerous currently uncurable diseases. However, wider application of this therapy is still bound by various limitations. To increase the effectiveness of cell therapy, a combined application of stem cells with various types of chemicals or agents, which could support the immunoregulatory and therapeutic properties of stem cells, has been proposed and tested. One prospective approach is offered by the co-application of mesenchymal stem cells (MSCs), which have potent immunomodulatory and regenerative properties, and selected metal nanoparticles (NPs) which have been used in various fields of medicine for their immunomodulatory, anti-oxidant and antibacterial properties. It has been shown that the main mechanism of the therapeutic action of MSCs is the production of immunomodulatory molecules and growth factors, and that the secretory activity of MSCs can be modified by different types of NPs. For this purpose, metal NPs are extremely useful. They possess unique characteristics and can influence the growth and repair of tissues, exert strong antimicrobial activity and serve as nanocarriers. Thus, treatment based on the simultaneous application of MSCs and selected NPs combines the therapeutic effects of MSCs and impacts of NPs on applied MSCs, and on the cells and tissues of the recipient. In this review we outline the current state of studies combining the administration of MSCs and the application of metal NPs, with a focus on perspectives to use such treatment for corneal and retinal injuries and diseases.

• Keywords
• combined application, mesenchymal stem cells, metal nanoparticles, ocular disorders, therapeutic effect,
• MeSH
• Combined Modality Therapy methods   MeSH
• Metal Nanoparticles * chemistry   therapeutic use   MeSH
• Humans MeSH
• Mesenchymal Stem Cells * cytology   MeSH
• Eye Diseases * therapy   MeSH
• Mesenchymal Stem Cell Transplantation * methods   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Review MeSH

The human amniotic membrane (HAM) is widely used for its wound healing effect in clinical practice, as a feeder for the cell cultivation, or a source of cells to be used in cell therapy. The aim of this study was to find effective and safe enzymatic HAM de-epithelialization method leading to harvesting of both denuded undamaged HAM and viable human amniotic epithelial cells (hAECs). The efficiency of de-epithelialization using TrypLE Express, trypsin/ ethylenediaminetetraacetic (EDTA), and thermolysin was monitored by hematoxylin and eosin staining and by the measurement of DNA concentration. The cell viability was determined by trypan blue staining. Scanning electron microscopy and immunodetection of collagen type IV and laminin α5 chain were used to check the basement membrane integrity. De-epithelialized hAECs were cultured and their stemness properties and proliferation potential was assessed after each passage. The HAM was successfully de-epithelialized using all three types of reagents, but morphological changes in basement membrane and stroma were observed after the thermolysin application. About 60% of cells remained viable using trypsin/EDTA, approximately 6% using TrypLE Express, and all cells were lethally damaged after thermolysin application. The hAECs isolated using trypsin/EDTA were successfully cultured up to the 5th passage with increasing proliferation potential and decreased stem cell markers expression (NANOG, SOX2) in prolonged cell culture. Trypsin/EDTA technique was the most efficient for obtaining both undamaged denuded HAM and viable hAECs for consequent culture.

• MeSH
• Amnion cytology   metabolism   pathology   MeSH
• DNA analysis   isolation & purification   MeSH
• Edetic Acid chemistry   MeSH
• Epithelial Cells cytology   metabolism   pathology   MeSH
• Collagen Type IV metabolism   MeSH
• Cells, Cultured MeSH
• Laminin metabolism   MeSH
• Humans MeSH
• Microscopy, Electron, Scanning MeSH
• Nanog Homeobox Protein metabolism   MeSH
• Cell Proliferation MeSH
• Re-Epithelialization MeSH
• SOXB1 Transcription Factors metabolism   MeSH
• Trypsin metabolism   MeSH
• Cell Survival MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• DNA MeSH
• Edetic Acid MeSH
• Collagen Type IV MeSH
• laminin alpha5 MeSH Browser
• Laminin MeSH
• Nanog Homeobox Protein MeSH
• SOXB1 Transcription Factors MeSH
• Trypsin MeSH

Retinal degenerative diseases, which include age-related macular degeneration, retinitis pigmentosa, diabetic retinopathy, and glaucoma, mostly affect the elderly population and are the most common cause of decreased quality of vision or even blindness. So far, there is no satisfactory treatment protocol to prevent, stop, or cure these disorders. A great hope and promise for patients suffering from retinal diseases is represented by stem cell-based therapy that could replace diseased or missing retinal cells and support regeneration. In this respect, mesenchymal stem cells (MSCs) that can be obtained from the particular patient and used as autologous cells have turned out to be a promising stem cell type for treatment. Here we show that MSCs can differentiate into cells expressing markers of retinal cells, inhibit production of pro-inflammatory cytokines by retinal tissue, and produce a number of growth and neuroprotective factors for retinal regeneration. All of these properties make MSCs a prospective cell type for cell-based therapy of age-related retinal degenerative diseases.

• Keywords
• age-related retinal degenerative diseases, mesenchymal stem cells, stem cell therapy,
• MeSH
• Cell Differentiation genetics   physiology   MeSH
• Retinal Degeneration metabolism   therapy   MeSH
• Mesenchymal Stem Cells cytology   metabolism   MeSH
• Mice, Inbred BALB C MeSH
• Mice MeSH
• Retinal Diseases metabolism   therapy   MeSH
• Prospective Studies MeSH
• Stem Cell Transplantation methods   MeSH
• Animals MeSH
• Check Tag
• Mice MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

PURPOSE: The present study aims to prepare poly(D,L-lactic acid) (PLA) nanofibers loaded by the immunosuppressant cyclosporine A (CsA, 10 wt%). Amphiphilic poly(ethylene glycol)s (PEG) additives were used to modify the hydrophobic drug release kinetics. METHODS: Four types of CsA-loaded PLA nanofibrous carriers varying in the presence and molecular weight (MW) of PEG (6, 20 and 35 kDa) were prepared by needleless electrospinning. The samples were extracted for 144 h in phosphate buffer saline or tissue culture medium. A newly developed and validated LC-MS/MS method was utilized to quantify the amount of released CsA from the carriers. In vitro cell experiments were used to evaluate biological activity. RESULTS: Nanofibers containing 15 wt% of PEG showed improved drug release characteristics; significantly higher release rates were achieved in initial part of experiment (24 h). The highest released doses of CsA were obtained from the nanofibers with PEG of the lowest MW (6 kDa). In vitro experiments on ConA-stimulated spleen cells revealed the biological activity of the released CsA for the whole study period of 144 h and nanofibers containing PEG with the lowest MW exhibited the highest impact (inhibition). CONCLUSIONS: The addition of PEG of a particular MW enables to control CsA release from PLA nanofibrous carriers. The biological activity of CsA-loaded PLA nanofibers with PEG persists even after 144 h of previous extraction. Prepared materials are promising for local immunosuppression in various medical applications.

• Keywords
• LC-MS/MS, cyclosporine A, drug release kinetics, poly(D,L-lactic acid) nanofibers, poly(ethylene glycol),
• MeSH
• Cell Line MeSH
• Cyclosporine administration & dosage   chemistry   MeSH
• Hydrophobic and Hydrophilic Interactions MeSH
• Immunosuppressive Agents administration & dosage   chemistry   MeSH
• Kinetics MeSH
• Culture Media MeSH
• Humans MeSH
• Nanofibers chemistry   MeSH
• Drug Carriers MeSH
• Polyesters chemistry   MeSH
• Polyethylene Glycols chemistry   MeSH
• Surface Properties MeSH
• Spleen cytology   MeSH
• Tissue Culture Techniques MeSH
• Drug Liberation MeSH
• Particle Size MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Cyclosporine MeSH
• Immunosuppressive Agents MeSH
• Culture Media MeSH
• Drug Carriers MeSH
• poly(lactide) MeSH Browser
• Polyesters MeSH
• Polyethylene Glycols MeSH

The aim of this study was to examine whether mesenchymal stem cells (MSCs) and/or corneal limbal epithelial stem cells (LSCs) influence restoration of an antioxidant protective mechanism in the corneal epithelium and renewal of corneal optical properties changed after alkali burns. The injured rabbit corneas (with 0.25 N NaOH) were untreated or treated with nanofiber scaffolds free of stem cells, with nanofiber scaffolds seeded with bone marrow MSCs (BM-MSCs), with adipose tissue MSCs (Ad-MSCs), or with LSCs. On day 15 following the injury, after BM-MSCs or LSCs nanofiber treatment (less after Ad-MSCs treatment) the expression of antioxidant enzymes was restored in the regenerated corneal epithelium and the expressions of matrix metalloproteinase 9 (MMP9), inducible nitric oxide synthase (iNOS), α-smooth muscle actin (α-SMA), transforming growth factor-β1 (TGF-β1), and vascular endothelial factor (VEGF) were low. The central corneal thickness (taken as an index of corneal hydration) increased after the injury and returned to levels before the injury. In injured untreated corneas the epithelium was absent and numerous cells revealed the expressions of iNOS, MMP9, α-SMA, TGF-β1, and VEGF. In conclusion, stem cell treatment accelerated regeneration of the corneal epithelium, restored the antioxidant protective mechanism, and renewed corneal optical properties.

• MeSH
• Alkalies MeSH
• Antioxidants therapeutic use   MeSH
• Cell Differentiation drug effects   MeSH
• Burns, Chemical enzymology   genetics   pathology   therapy   MeSH
• Immunohistochemistry MeSH
• Rabbits MeSH
• Limbus Corneae cytology   MeSH
• Matrix Metalloproteinase 9 metabolism   MeSH
• Mesenchymal Stem Cells cytology   drug effects   MeSH
• Protective Agents pharmacology   therapeutic use   MeSH
• Corneal Pachymetry MeSH
• Gene Expression Regulation drug effects   MeSH
• Epithelium, Corneal pathology   MeSH
• Superoxide Dismutase metabolism   MeSH
• Nitric Oxide Synthase Type II metabolism   MeSH
• Transforming Growth Factor beta genetics   metabolism   MeSH
• Mesenchymal Stem Cell Transplantation * MeSH
• Adipocytes cytology   drug effects   MeSH
• Vascular Endothelial Growth Factor A metabolism   MeSH
• Corneal Opacity complications   therapy   MeSH
• Animals MeSH
• Check Tag
• Rabbits MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Retracted Publication MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Alkalies MeSH
• Antioxidants MeSH
• Matrix Metalloproteinase 9 MeSH
• Protective Agents MeSH
• Superoxide Dismutase MeSH
• Nitric Oxide Synthase Type II MeSH
• Transforming Growth Factor beta MeSH
• Vascular Endothelial Growth Factor A MeSH

UNLABELLED: Stem cell-based therapy has become an attractive and promising approach for the treatment of severe injuries or thus-far incurable diseases. However, the use of stem cells is often limited by a shortage of available tissue-specific stem cells; therefore, other sources of stem cells are being investigated and tested. In this respect, mesenchymal stromal/stem cells (MSCs) have proven to be a promising stem cell type. In the present study, we prepared MSCs from bone marrow (BM-MSCs) or adipose tissue (Ad-MSCs) as well as limbal epithelial stem cells (LSCs), and their growth, differentiation, and secretory properties were compared. The cells were grown on nanofiber scaffolds and transferred onto the alkali-injured eye in a rabbit model, and their therapeutic potential was characterized. We found that BM-MSCs and tissue-specific LSCs had similar therapeutic effects. Clinical characterization of the healing process, as well as the evaluation of corneal thickness, re-epithelialization, neovascularization, and the suppression of a local inflammatory reaction, were comparable in the BM-MSC- and LSC-treated eyes, but results were significantly better than in injured, untreated eyes or in eyes treated with a nanofiber scaffold alone or with a nanofiber scaffold seeded with Ad-MSCs. Taken together, the results show that BM-MSCs' therapeutic effect on healing of injured corneal surface is comparable to that of tissue-specific LSCs. We suggest that BM-MSCs can be used for ocular surface regeneration in cases when autologous LSCs are absent or difficult to obtain. SIGNIFICANCE: Damage of ocular surface represents one of the most common causes of impaired vision or even blindness. Cell therapy, based on transplantation of stem cells, is an optimal treatment. However, if limbal stem cells (LSCs) are not available, other sources of stem cells are tested. Mesenchymal stem cells (MSCs) are a convenient type of cell for stem cell therapy. The therapeutic potential of LSCs and MSCs was compared in an experimental model of corneal injury, and healing was observed following chemical injury. MSCs and tissue-specific LSCs had similar therapeutic effects. The results suggest that bone marrow-derived MSCs can be used for ocular surface regeneration in cases when autologous LSCs are absent or difficult to obtain.

• Keywords
• Alkali-injured ocular surface, Corneal regeneration, Limbal stem cells, Mesenchymal stem cells, Stem cell-based therapy,
• MeSH
• Biomarkers metabolism   MeSH
• Cell- and Tissue-Based Therapy methods   MeSH
• Cell Differentiation MeSH
• Bone Marrow Cells cytology   physiology   MeSH
• Burns, Chemical pathology   therapy   MeSH
• Epithelial Cells cytology   physiology   transplantation   MeSH
• Gene Expression MeSH
• Neovascularization, Physiologic MeSH
• Rabbits MeSH
• Limbus Corneae blood supply   injuries   MeSH
• Mesenchymal Stem Cells cytology   physiology   MeSH
• Primary Cell Culture MeSH
• Cell Proliferation MeSH
• Re-Epithelialization physiology   MeSH
• Epithelium, Corneal blood supply   injuries   MeSH
• Tissue Scaffolds MeSH
• Mesenchymal Stem Cell Transplantation * MeSH
• Adipose Tissue cytology   physiology   MeSH
• Adipocytes cytology   physiology   MeSH
• Animals MeSH
• Check Tag
• Rabbits MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Comparative Study MeSH
• Names of Substances
• Biomarkers MeSH

Ocular surface defects represent one of the most common causes of impaired vision or even blindness. For treatment, keratoplasty represents the first choice. However, if corneal defects are more extensive and associated with a limbal stem cell (LSC) deficiency, corneal transplantation is not a sufficient therapeutic procedure and only viable approach to treatment is the transplantation of LSCs. When the LSC deficiency is a bilateral disorder, autologous LSCs are not available. The use of allogeneic LSCs requires strong immunosuppression, which leads to side-effects, and the treatment is not always effective. The alternative and perspective approach to the treatment of severe ocular surface injuries and LSC deficiency is offered by the transplantation of autologous mesenchymal stem cells (MSCs). These cells can be obtained from the bone marrow or adipose tissue of the particular patient, grow well in vitro and can be transferred, using an appropriate scaffold, onto the damaged ocular surface. Here they exert beneficial effects by possible direct differentiation into corneal epithelial cells, by immunomodulatory effects and by the production of numerous trophic and growth factors. Recent experiments utilizing the therapeutic properties of MSCs in animal models with a mechanically or chemically injured ocular surface have yielded promising results and demonstrated significant corneal regeneration, improved corneal transparency and a rapid healing process associated with the restoration of vision. The use of autologous MSCs thus represents a promising therapeutic approach and offers hope for patients with severe ocular surface injuries and LSC deficiency.

• MeSH
• Transplantation, Autologous MeSH
• Models, Biological MeSH
• Cell Differentiation MeSH
• Bone Marrow Cells cytology   metabolism   MeSH
• Antigens, CD metabolism   MeSH
• Cells, Cultured MeSH
• Humans MeSH
• Mesenchymal Stem Cells cytology   metabolism   MeSH
• Intercellular Signaling Peptides and Proteins metabolism   MeSH
• Nanofibers * MeSH
• Corneal Diseases surgery   MeSH
• Cell Movement MeSH
• Stem Cell Transplantation methods   MeSH
• Mesenchymal Stem Cell Transplantation methods   MeSH
• Adipose Tissue cytology   metabolism   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Antigens, CD MeSH
• Intercellular Signaling Peptides and Proteins MeSH

Limbal stem cells (LSC), which reside in the basal layer of the limbus, are thought to be responsible for corneal epithelial healing after injury. When the cornea is damaged, LSC start to proliferate, differentiate, and migrate to the site of injury. To characterize the signaling molecules ensuring communication between the cornea and LSC, we established a mouse model of mechanical corneal damage. The central cornea or limbal tissue was excised at different time intervals after injury, and the expression of genes in the explants was determined. It was observed that a number of genes for growth and differentiation factors were significantly upregulated in the cornea rapidly after injury. The ability of these factors to regulate the differentiation and proliferation of limbal cells was tested. It was found that the insulin-like growth factor-I (IGF-I), which is rapidly overexpressed after injury, enhances the expression of IGF receptor in limbal cells and induces the differentiation of LSC into cells expressing the corneal cell marker, cytokeratin K12, without any effect on limbal cell proliferation. In contrast, the epidermal growth factor (EGF) and fibroblast growth factor-β (FGF-β), which are also produced by the damaged corneal epithelium, supported limbal cell proliferation without any effect on their differentiation. Other factors did not affect limbal cell differentiation or proliferation. Thus, IGF-I was identified as the main factor stimulating the expression of IGF receptors in limbal cells and inducing the differentiation of LSC into cells expressing corneal epithelial cell markers. The proliferation of these cells was supported by EGF and FGF.

• MeSH
• Cell Differentiation genetics   MeSH
• Epidermal Growth Factor biosynthesis   metabolism   MeSH
• Fibroblast Growth Factors biosynthesis   metabolism   MeSH
• Wound Healing physiology   MeSH
• Insulin-Like Growth Factor I biosynthesis   metabolism   MeSH
• Keratin-12 biosynthesis   MeSH
• Stem Cells metabolism   MeSH
• Limbus Corneae cytology   metabolism   MeSH
• Mice, Inbred BALB C MeSH
• Mice MeSH
• Cell Proliferation MeSH
• Receptor, IGF Type 1 biosynthesis   metabolism   MeSH
• Epithelium, Corneal * cytology   injuries   metabolism   MeSH
• Signal Transduction MeSH
• Gene Expression Profiling MeSH
• Up-Regulation MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Mice MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Epidermal Growth Factor MeSH
• Fibroblast Growth Factors MeSH
• Insulin-Like Growth Factor I MeSH
• Keratin-12 MeSH
• Receptor, IGF Type 1 MeSH

Polyvinyl alcohol nanofibers incorporating the wide spectrum antibiotic gentamicin were prepared by Nanospider™ needleless technology. A polyvinyl alcohol layer, serving as a drug reservoir, was covered from both sides by polyurethane layers of various thicknesses. The multilayered structure of the nanofibers was observed using scanning electron microscopy, the porosity was characterized by mercury porosimetry, and nitrogen adsorption/desorption measurements were used to determine specific surface areas. The stability of the gentamicin released from the electrospun layers was proved by high-performance liquid chromatography (HPLC) and inhibition of bacterial growth. Drug release was investigated using in vitro experiments with HPLC/MS quantification, while the antimicrobial efficacy was evaluated on Gram-positive Staphylococcus aureus and Gram-negative Pseudomonas aeruginosa. Both experiments proved that the released gentamicin retained its activity and showed that the retention of the drug in the nanofibers was prolonged with the increasing thickness of the covering layers.

• Keywords
• drug release, electrospinning, gentamicin, morphology, multilayered structure, nanofibers,
• MeSH
• Anti-Bacterial Agents administration & dosage   chemistry   MeSH
• Diffusion MeSH
• Electrochemistry methods   MeSH
• Gentamicins administration & dosage   MeSH
• Gram-Positive Bacteria drug effects   physiology   MeSH
• Delayed-Action Preparations administration & dosage   chemistry   MeSH
• Nanocapsules chemistry   ultrastructure   MeSH
• Rotation MeSH
• Materials Testing MeSH
• Particle Size MeSH
• Cell Survival drug effects   MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Anti-Bacterial Agents MeSH
• Gentamicins MeSH
• Delayed-Action Preparations MeSH
• Nanocapsules MeSH

Bone marrow-derived mesenchymal stem cells (MSCs) modulate immune response and can produce significant levels of transforming growth factor-β (TGF-β) and interleukin-6 (IL-6). These 2 cytokines represent the key factors that reciprocally regulate the development and polarization of naive T-cells into regulatory T-cell (Treg) population or proinflammatory T helper 17 (Th17) cells. In the present study we demonstrate that MSCs and their products effectively regulate expression of transcription factors Foxp3 and RORγt and control the development of Tregs and Th17 cells in a population of alloantigen-activated mouse spleen cells or purified CD4(+)CD25(-) T-cells. The immunomodulatory effects of MSCs were more pronounced when these cells were stimulated to secrete TGF-β alone or TGF-β together with IL-6. Unstimulated MSCs produce TGF-β, but not IL-6, and the production of TGF-β can be further enhanced by the anti-inflammatory cytokines IL-10 or TGF-β. In the presence of proinflammatory cytokines, MSCs secrete significant levels of IL-6, in addition to a spontaneous production of TGF-β. MSCs producing TGF-β induced preferentially expression of Foxp3 and activation of Tregs, whereas MSC supernatants containing TGF-β together with IL-6 supported RORγt expression and development of Th17 cells. The effects of MSC supernatants were blocked by the inclusion of neutralization monoclonal antibody anti-TGF-β or anti-IL-6 into the culture system. The results showed that MSCs represent important players that reciprocally regulate the development and differentiation of uncommitted naive T-cells into anti-inflammatory Foxp3(+) Tregs or proinflammatory RORγt(+) Th17 cell population and thereby can modulate autoimmune, immunopathological, and transplantation reactions.

• MeSH
• Cell Differentiation immunology   MeSH
• Cell Culture Techniques MeSH
• Cytokines biosynthesis   MeSH
• Immunity MeSH
• Culture Media, Conditioned pharmacology   MeSH
• Mesenchymal Stem Cells physiology   MeSH
• Mice MeSH
• Paracrine Communication immunology   MeSH
• T-Lymphocytes, Regulatory immunology   MeSH
• T-Lymphocytes immunology   MeSH
• Inflammation immunology   MeSH
• Animals MeSH
• Check Tag
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Cytokines MeSH
• Culture Media, Conditioned MeSH