Mesenchymal stem cells in corneal neovascularization: Comparison of different application routes

Autor: Guanfang Su, Emma Ghazaryan, Ying Li, Jia'nan Xie, Yuxi He, Xin Liu, Yan Zhang
Jazyk: angličtina
Rok vydání: 2016
Předmět:
0301 basic medicine
Vascular Endothelial Growth Factor A
Cancer Research
Pathology
medicine.medical_specialty
Cell Culture Techniques
Gene Expression
Cell Separation
Biology
Mesenchymal Stem Cell Transplantation
Biochemistry
Neovascularization
Cornea
03 medical and health sciences
chemistry.chemical_compound
0302 clinical medicine
chemical injury
Transduction
Genetic
Genetics
medicine
Animals
Rats
Wistar
Molecular Biology
mesenchymal stem cell
Cells
Cultured
amniotic membrane
Mesenchymal stem cell
Mesenchymal Stem Cells
Articles
medicine.disease
Transplantation
Vascular endothelial growth factor
Vascular endothelial growth factor A
030104 developmental biology
medicine.anatomical_structure
Oncology
chemistry
corneal neovascularization
Corneal neovascularization
030221 ophthalmology & optometry
Molecular Medicine
Female
medicine.symptom
Wound healing
transplantation
Zdroj: Molecular Medicine Reports
ISSN: 1791-3004
1791-2997
Popis: The purpose of the present study is to investigate the effect of mesenchymal stem cells in corneal neovascularization and wound healing, and to compare the effectiveness of two possible application routes, subconjunctival injection and amniotic membrane transplantation. Chemical injury was induced by application of sodium hydroxide to the rats' corneas. After 7 days, the animals were divided into three groups. Different treatment methods were used for each group as follows: i) Group 1, injection of bone marrow‑derived mesenchymal stem cells (BMSCs) under the conjunctiva; ii) group 2, transplantation of amniotic membranes, previously seeded with BMSCs; and iii) group 3, the untreated control group. The eyes were examined using a slit lamp on a weekly basis. After 4 weeks, the animals were sacrificed and corneas were removed for further examination. Corneal flat mounts were made following ink perfusion for improved vessel visualization, image capturing and quantitative evaluation. enzyme‑linked immunosorbent assay was performed to detect the levels of vascular endothelial growth factor (VEGF) and matrix metalloproteinase 9 (MMP‑9). Reverse transcription‑quantitative polymerase chain reaction was used for detection of VEGF‑A, MMP‑9, Toll‑like receptor (TLR)2 and TLR4 gene expression levels. Cryosections were used for histological examination and immunostaining. Statistical analysis (Welch's one‑way analysis of variance) demonstrated a significant difference between the groups [P≤0.05, confidence interval (CI) 95%]. The level of injury in group 1 was significantly different from groups 2 and 3. Measurement of the vessel area and VEGF gene expression levels had a similar difference among the groups (P≤0.05, CI 95%), however the differences for TLR2 and TLR4 were not statistically significant. BMSCs were previously transduced with the green fluorescent protein gene by lentivirus to track the movement of the cells following transplantation. The transplanted cells enhanced corneal wound healing by trophic factor production and immune‑regulatory effect, rather than by direct transdifferentiation into corneal cells. The results of the current study demonstrated that BMSCs enhance corneal wound healing and decrease the area of neovascularization. Furthermore, the comparison of two application routes indicated that single subconjunctival injection appeared more effective than transplantation with amniotic membrane.
Databáze: OpenAIRE
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