Generation of Retinal Pigmented Epithelium-Like Cells from Pigmented Spheres Differentiated from Bone Marrow Stromal Cell-Derived Neurospheres

Hamid Aboutaleb Kadkhodaeian,Taki Tiraihi,Hamid Ahmadieh,Hossein Ziaei,Narsis Daftarian,Taher Taheri 한국조직공학과 재생의학회 2019 조직공학과 재생의학 Vol.16 No.3

BACKGROUND: Retinal degeneration causes blindness, and cell replacement is a potential therapy. The purpose of this study is to formation of pigmented neurospheres in a simple medium, low-cost, high-performance manner over a short period of time while expressing markers of RPE cells and the activation of specific genes of the pigment cells. Also, these neurospheres have the ability to produce a monolayer of retinal pigment epithelium-like cells (RPELC) with the ability of photoreceptor outer segment phagocytosis. METHODS: BMSC were isolated from pigmented hooded male rats and were immunoreactive to BMSC markers, then converted into neurospheres, differentiated into pigmented spheres (PS), and characterized using Retinal pigment epithelium-specific 65 kDa protein (RPE65), Retinaldehyde-binding protein 1 (CRALBP) and orthodenticle homeobox 2 (OTX2) markers by immunocytochemistry, RT-PCR and RT-qPCR. The PS were harvested into RPELC. The functionality of RPELC was evaluated by phagocytosis of fluorescein-labeled photoreceptor outer segment. RESULTS: The BMSC immunophenotype was confirmed by immunostained for fibronectin, CD90, CD166 and CD44. These cells differentiated into osteogenic and lipogenic cells. The generated neurospheres were immunoreactive to nestin and stemness genes. The PS after 7–14 days were positive for RPE65 (92.76–100%), CRALBP (95.21–100%) and OTX2 (94.88–100%), and after 30 days RT-PCR, qPCR revealed increasing in gene expression. The PS formed a single layer of RPELC after cultivation and phagocyte photoreceptor outer segments. CONCLUSION: Bone marrow stromal stem cells can differentiate into functional retinal pigmented epithelium cells in a simple, low-cost, high-performancemanner over a short period of time. These cells due to expressing theRPELCgenes andmarkers can be used in cell replacement therapy for degenerative diseases including age-relatedmacular degeneration as well as retinitis pigmentosa.

Predictors and Outcomes of Vitrectomy and Silicone Oil Injection in Advanced Diabetic Retinopathy

Alireza Ramezani,Hamid Ahmadieh,Amin Rozegar,Masoud Soheilian,Morteza Entezari,Siamak Moradian,Mohammad H Dehghan,Homayoun Nikkhah,Mehdi Yaseri 대한안과학회 2017 Korean Journal of Ophthalmology Vol.31 No.3

Purpose: To evaluate visual and anatomical results and identify factors that influence vitrectomy and silicone oil(SO) injection outcomes in proliferative diabetic retinopathy (PDR). Methods: This retrospective study included 236 eyes with PDR that were undergoing vitrectomy and SO injectionwith >3-month follow-up. The primary outcomes were final best-corrected visual acuity (BCVA) and retinalattachment rate. Results: At the final visit (mean, 88 ± 58 weeks), complete, partial, and no retinal attachment were observed in86.9%, 10.6%, and 2.5% of patients, respectively. A total of 155 eyes had experienced SO removal, while 81had SO in place. The mean initial BCVA was 1.9 ± 0.7 logarithm of the minimum angle of resolution (logMAR)and significantly improved to 1.7 ± 0.8 logMAR (p = 0.001). Initial macular detachment (adjusted odds ratio[AOR], 0.25), development of iatrogenic break (AOR, 0.25), and use of heavy SO (AOR, 0.13) were independentlyassociated with a lower risk of final retinal attachment, and SO removal was associated with a higherincidence (AOR, 7.55). Better baseline BCVA was associated with a higher risk of final BCVA ≥20 / 200. Conclusions: Despite an encouraging outcome based on anatomical data in advanced PDR treated with vitrectomyand SO, the functional prognosis was not satisfying for patients. Eyes with better vision at baseline hada more favorable prognosis, whereas eyes with initial macular detachment, intraoperative iatrogenic break, orheavy SO showed more unfavorable outcomes. In selected cases, extending the time of SO use did not worsenthe prognosis.

Intravitreal Triamcinolone Reinjection for Refractory Diabetic Macular Edema

( Alireza Ramezani ),( Hamid Ahmadieh ),( Homa Tabatabaei ) 대한안과학회 2006 Korean Journal of Ophthalmology Vol.20 No.3

Purpose: To evaluate the effect of intravitreal triamcinolone acetonide (IVT) reinjection on clinical and optical coherence tomographic features in refractory diabetic macular edema. Methods: In a prospective interventional case series, all IVT treated patients enrolled in a previous clinical trial were recalled to have a new ophthalmologic examination and optical coherence tomography (OCT) performed. Eyes found suitable for reinjection received 4 mg IVT. Complete clinical examination and OCT were repeated at 2 and 4 months post-injection. The changes were statistically analyzed using a paired t test and were compared to the results of the first injections. Results: Of all returning patients, 12 cases with complete records were considered candidates for reinjection. Visual acuity (VA) changes were not significant after the first and second interventions, although there was a relative improvement (0.14 logMAR) 2 months after the first injection. Reductions of central macular thickness (CMT) were 43±69 μm, and 40±69 μm after the first injection and 27±48 μm, 49±58 μm after the reinjection at 2 & 4 months, respectively. None of the mentioned changes was significant. After the second injection, however, intraocular pressure was elevated at both 2 & 4 months (3.6 & 2.4 mmHg respectively, P<0.05). Two months after the first administration, intraocular pressure was found to be raised significantly (5.58 mmHg, P=0.001). Conclusions: The transient beneficial effects of IVT on diabetic macular edema are not repeated with second injections. However, IVT-related ocular hypertension is more persistent after reinjection. Purpose: To evaluate the effect of intravitreal triamcinolone acetonide (IVT) reinjection on clinical and optical coherence tomographic features in refractory diabetic macular edema. Methods: In a prospective interventional case series, all IVT treated patients enrolled in a previous clinical trial were recalled to have a new ophthalmologic examination and optical coherence tomography (OCT) performed. Eyes found suitable for reinjection received 4 mg IVT. Complete clinical examination and OCT were repeated at 2 and 4 months post-injection. The changes were statistically analyzed using a paired t test and were compared to the results of the first injections. Results: Of all returning patients, 12 cases with complete records were considered candidates for reinjection. Visual acuity (VA) changes were not significant after the first and second interventions, although there was a relative improvement (0.14 logMAR) 2 months after the first injection. Reductions of central macular thickness (CMT) were 43±69 μm, and 40±69 μm after the first injection and 27±48 μm, 49±58 μm after the reinjection at 2 & 4 months, respectively. None of the mentioned changes was significant. After the second injection, however, intraocular pressure was elevated at both 2 & 4 months (3.6 & 2.4 mmHg respectively, P<0.05). Two months after the first administration, intraocular pressure was found to be raised significantly (5.58 mmHg, P=0.001). Conclusions: The transient beneficial effects of IVT on diabetic macular edema are not repeated with second injections. However, IVT-related ocular hypertension is more persistent after reinjection.

Diabetic Macular Edema Before and After Intravitreal Triamcinolone Injection

( Alireza Ramezani ),( Homa Tabatabaie ),( Hamid Ahmadieh ) 대한안과학회 2007 Korean Journal of Ophthalmology Vol.21 No.2

Purpose: To compare intravitreal triamcinolone acetonide (IVT) versus natural course in refractory diabetic macular edema. Methods: In a prospective interventional case series, twenty five eyes with refractory DME which had been allocated to the sham group of a previous clinical trial underwent new examination and optical coherence tomography about 9 months after their first enrollment. Twenty eyes that met the inclusion criteria, visual acuity (VA) < 20/50 and central macular thickness (CMT) > 200 μm, were treated by 4 mg IVT. Evaluations were repeated at 2 and 4 months post-injection to imitate the similar examination intervals after sham injection. Corrected visual acuity and macular thickness changes following IVT were compared to the corresponding changes after sham injection (the natural course). Results: Visual acuity changes within and between each period were not statistically significant. Visual acuity decreased 0.08 & 0.09 logMAR by 2 months and 0.06 & 0.04 logMAR by 4 months after sham and IVT injections, respectively. The changes of macular thickness after IVT and sham intervention were not meaningful either. However, the difference between thickness changes by 4 months (52±50 μm increase after sham vs. 262±115 μm reduction after IVT) was significant (P=0.014). Conclusions: Concerning macular thickness, IVT has beneficial effect on refractory diabetic macular edema as opposed to observation. However, considering visual acuity, it does not induce significant difference in comparison to the natural course of the disease. Korean Journal of Ophthalmology 21(2):95-99, 2007