Topical Tacrolimus as an adjunct to Conventional Therapy for Stromal Herpetic Keratitis: a Randomized Clinical Trial

Purpose: This study investigates the effects of 0.05% topical tacrolimus as an adjunct therapy for patients with non-necrotizing herpetic stromal keratitis (HSK). Methods: Patients with non-necrotizing HSK, referred to the Cornea Clinic at Hospital in Rasht, Iran, between September 2016 and February 2018, were randomly assigned to two groups. The case group (N = 25) and the control group (N = 25) received conventional treatment with systemic acyclovir and topical prednisolone. The case group (N = 25) additionally received 0.05% tacrolimus eye drops four times a day for one month. Complete ocular examinations, including best-corrected visual acuity (BCVA) assessment, intraocular pressure (IOP) measurement, slit lamp biomicroscopy, and photo slit lamp imaging, were performed before treatment, and 3, 7, 14, 21, and 28 days after the intervention. Results: The mean age of the patients was 46.2 ± 12.9 years, and 70% of the patients were male. There was no difference between the groups in terms of age, sex, and baseline ocular measurements (P > 0.05). The case group had a lower mean logarithm of the minimum angle of resolution (LogMAR) for BCVA, lower grading scores, and steeper decreasing trends for corneal haziness, edema, neovascularization, and epitheliopathy compared to the control group after the second week (P < 0.05), while IOP remained unchanged between groups (P > 0.05). Conclusion: The addition of 0.05% topical tacrolimus enhances visual acuity and reduces corneal inflammation, neovascularization, and scarring; thus, it can used as an appropriate adjunct treatment for patients with HSK

Acute myocarditis and acute myopathy as the first manifestations of COVID-19; a case report

Coronavirus disease 2019 (COVID-19) mainly manifests with flu-like and respiratory symptoms such as fever, chill, myalgia, cough, dyspnea and in severe cases, it leads to acute respiratory distress syndrome and respiratory failure. However, there is evidence of extra-pulmonary involvements in patients with COVID-19. Some case reports and studies have reported severe and life-threatening complications related to COVID-19 such as cardiovascular complications (acute heart failure, myocarditis, acute coronary syndrome, thromboembolic events) and neuromuscular complications (stroke, transient ischemic attack, myositis, myopathy, Guillain-Barre syndrome). Here, we report a 51-year-old woman without a previous history of cardiovascular disease or neuromuscular disease referred to the emergency department of our hospital with new onset severe respiratory distress and progressive symmetric quadriparesis. We concluded that, the patient was infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and we therefore have encountered acute myocarditis and acute myopathy due to COVID-19 disease. In the intensive care unit (ICU), the patient was treated with oxygen therapy without mechanical ventilation, dexamethasone, intravenous human immunoglobulin (IVIG), beta interferon and remdesivir. The clinical feature, cardiac, respiratory, neuromuscular and hemodynamic parameters improved clearly five days after taking above mentioned treatments. The troponin, N-terminal pro-B type natriuretic peptide (NT-proBNP), creatine phosphokinase (CPK), returned to normal values. Following improvement of cardiac and neurologic problems, the patient was transferred from ICU to general ward and then after 10 days, she was discharged with oral anticoagulant, anti-platelet, low-dose of corticosteroids and other conservative treatments

Treatment of Hypertrophic Scar in Human with Autologous Transplantation of Cultured Keratinocytes and Fibroblasts along with Fibrin Glue

Objective: Hypertrophic scar involves excessive amounts of collagen in dermal layer and may be painful. Nowadays, we can’t be sure about effectiveness of procedure for hypertrophic scar management. The application of stem cells with natural scaffold has been the best option for treatment of burn wounds and skin defect, in recent decades. Fibrin glue (FG) was among the first of the natural biomaterials applied to enhance skin deformity in burn patients. This study aimed to identify an efficient, minimally invasive and economical transplantation procedure using novel FG from human cord blood for treatment of hypertrophic scar and regulation collagen synthesis. Materials and Methods: In this case series study, eight patients were selected with hypertrophic scar due to full-thickness burns. Human keratinocytes and fibroblasts derived from adult skin donors were isolated and cultured. They were tested for the expression of cytokeratin 14 and vimentin using immunocytochemistry. FG was prepared from pooled cord blood. Hypertrophic scars were extensively excised then grafted by simply placing the sheet of FG containing autologous fibroblast and keratinocytes. Histological analyses were performed using Hematoxylin and eosin (H&E) and Masson’s Trichrome (MT) staining of the biopsies after 8 weeks. Results: Cultured keratinocytes showed a high level of cytokeratin 14 expression and also fibroblasts showed a high level of vimentin. Histological analyses of skin biopsies after 8 weeks of transplantation revealed re-epithelialization with reduction of hypertrophic scars in 2 patients. Conclusion: These results suggest may be the use of FG from cord blood, which is not more efficient than previous biological transporters and increasing hypertrophic scar relapse, but could lead to decrease pain rate

Long-Term Follow-up of Autologous Fibroblast Transplantation for Facial Contour Deformities, A Non-Randomized Phase IIa Clinical Trial

Objective: Recently, the promising potential of fibroblast transplantation has become a novel modality for skin rejuvenation. We investigated the long-term safety and efficacy of autologous fibroblast transplantation for participants with mild to severe facial contour deformities. Materials and Methods: In this open-label, single-arm phase IIa clinical trial, a total of 57 participants with wrinkles (n=37, 132 treatment sites) or acne scars (n=20, 36 treatment sites) who had an evaluator’s assessment score of at least 2 out 7 (based on a standard photo-guide scoring) received 3 injections of autologous cultured fibroblasts administered at 4-6 week intervals. Efficacy evaluations were performed at 2, 6, 12, and 24 months after the final injection based on evaluator and patient’s assessment scores. Results: Our study showed a mean improvement of 2 scores in the wrinkle and acne scar treatment sites. At sixth months after transplantation, 90.1% of the wrinkle sites and 86.1% of the acne scar sites showed at least a one grade improvement on evaluator assessments. We also observed at least a 2-grade improvement in 56.1% of the wrinkle sites and 63.9% of the acne scar sites. A total of 70.5% of wrinkle sites and 72.2% of acne scar sites were scored as good or excellent on patient assessments. The efficacy outcomes remained stable up to 24-month. We did not observe any serious adverse events during the study. Conclusion: These results have shown that autologous fibroblast transplantation could be a promising remodeling modality with long-term corrective ability and minimal adverse events (Registration Number: NCT01115634)

Defining microRNA signatures of hair follicular stem and progenitor cells in healthy and androgenic alopecia patients

[Background]: The exact pathogenic mechanism causes hair miniaturization during androgenic alopecia (AGA) has not been delineated. Recent evidence has shown a role for non-coding regulatory RNAs, such as microRNAs (miRNAs), in skin and hair disease. There is no reported information about the role of miRNAs in hair epithelial cells of AGA.[Objectives]: To investigate the roles of miRNAs affecting AGA in normal and patient’s epithelial hair cells.[Methods]: Normal follicular stem and progenitor cells, as well as follicular patient’s stem cells, were sorted from hair follicles, and a miRNA q-PCR profiling to compare the expression of 748 miRNA (miRs) in sorted cells were performed. Further, we examined the putative functional implication of the most differentially regulated miRNA (miR-324-3p) in differentiation, proliferation and migration of cultured keratinocytes by qRT-PCR, immunofluorescence, and scratch assay. To explore the mechanisms underlying the effects of miR-324-3p, we used specific chemical inhibitors targeting pathways influenced by miR-324-3p.[Result]: We provide a comprehensive assessment of the "miRNome" of normal and AGA follicular stem and progenitor cells. Differentially regulated miRNA signatures highlight several miRNA candidates including miRNA-324-3p as mis regulated in patient’s stem cells. We find that miR-324-3p promotes differentiation and migration of cultured keratinocytes likely through the regulation of mitogen-activated protein kinase (MAPK) and transforming growth factor (TGF)-β signaling. Importantly, pharmacological inhibition of the TGF-β signaling pathway using Alk5i promotes hair shaft elongation in an organ-culture system.[Conclusion]: Together, we offer a platform for understanding miRNA dynamic regulation in follicular stem and progenitor cells in baldness and highlight miR-324-3p as a promising target for its treatment.This study was funded by a grant provided from Royan Institute and Disease Models & Mechanisms Travelling Fellowship by Biologists Company.Peer reviewe

Virus-Specific T Cells: Promising Adoptive T Cell Therapy Against Infectious Diseases Following Hematopoietic Stem Cell Transplantation

Hematopoietic stem cell transplantation (HSCT) is a life-saving therapy for various hematologic disorders. Due to the bone marrow suppression and its long recovery period, secondary infections, like cytomegalovirus (CMV), Epstein-Bar virus (EBV), and adenovirus (AdV), are the leading causes of morbidity and mortality in HSCT cases. Drug resistance to the antiviral pharmacotherapies makes researchers develop adoptive T cell therapies like virus-specific T cell therapy. These studies have faced major challenges such as finding the most effective T cell expansion methods, isolating the expected subtype, defining the functionality of the end-cell population, product quality control, and clinical complications after the injection. This review discusses the viral infections after HSCT, T cells characteristics during chronic viral infection, application of virus-specific T cells (VSTs) for refractory infections, standard methods for producing VSTs and their limitation, clinical experiences on VSTs, focusing on outcomes and side effects that can be helpful in decision-making for patients and further researches

Subcutaneous Injection of Allogeneic Adipose-Derived Mesenchymal Stromal Cells in Psoriasis Plaques: Clinical Trial Phase I

Objective: Mesenchymal stromal cells (MSCs) play immunomodulatory role in various autoimmune diseases. Previouspre-clinical and clinical studies have shown that MSCs could be a therapeutic modality for psoriasis. However, themechanisms of treatment and its possible side effects are under investigation. In this study, the safety and probableefficacy of injecting allogeneic adipose-derived mesenchymal stromal cells (ADSCs) in psoriatic patients were evaluated.Materials and Methods: In this phase I clinical study with six months of follow-up, total number of 1×106 or 3×106cells/cm2 of ADSCs were injected into the subcutaneous tissue of each plaque as a single dose in three males and twofemales (3M/2F) with a mean age of 32.8 ± 8.18. The primary outcome was safety. Changes in clinical and histologicalindexes, the number of B and T lymphocytes in local and peripheral blood, and serum levels of inflammatory cytokineswere assessed. Paired t test was used to compare variables at two time points (baseline and six months after injection)and repeated measures ANOVA test was utilized for variables at three time points in follow-up visits.Results: No major adverse effects such as burning, pain, itching, or any systemic side effects were observed followingADSCs injection, and the lesions showed slight to considerable improvement after injection. The mRNA expressionlevels of pro-inflammatory factors were reduced in the dermis of the patients after injection. The increased expressionlevel of Foxp3 transcription factor in the patient blood samples suggested modulation of inflammation after ADMSCsadministration. Six months after the intervention, no major side effects were reported, but skin thickness, erythema, andscaling of the plaques, as well as the PASI score, were decreased in majority of patients.Conclusion: Our study suggested that ADSC injection could be considered as a safe and effective therapeuticapproach for psoriatic plaques (registration number: IRCT20080728001031N24)

Regenerative Medicine and Cell Therapy

XIV, 318 p.online resource

Methods for Isolation of Bone Marrow Stem Cells: Comparative Analysis

During the past decade, regenerative medicine has emerged as a key technology in thenext generation of medical care, and cell therapy and organ repair using stem cells havebecome very attractive options for regenerative medicine. The application of stem cells inregenerative medicine has required modified methods for isolation. Furthermore, the processof cell separation plays an important role in cell therapy and regenerative medicineusing stem cells. So, in this review, we compare different methods for the separation ofcells from bone marrow for transplantation to humans, with emphasis on the advantagesand disadvantages of each method

Investigation the effect of copper nanoparticles on the toxicity and migration of keratinocyte cells

Background: Re-epithelialization has an important role in skin wound healing. Delays in re-epithelialization are more likely to create the chronic wound. Impaired wound healing leads to a large burden of morbidity and mortality. Current treatments based on the use of autografts, allografts and xenografts, suffer from limitations such as, quantity of donor skin available, donor-site infection, potential risk of disease transmission and rejection of the graft. Given this problems, nanomaterial such as copper nanoparticles has attracted considerable research interest because of their high surface area to volume ratio, high stability, clinical safety, and antibacterial effects. Epithelialization involves keratinocyte migration and proliferation to the wound site. Therefore, this study was conducted to investigate the effect of copper nanoparticles on keratinocyte cell migration and proliferation. Methods: This experimental study was performed in Royan Institute, Tehran in 2016. In this study we investigated the effect of copper nanoparticles on viability, migration and proliferation of keratinocyte cells. Cultured human foreskin Keratinocyte cells were exposed to various concentration (1, 10 and 100 µmol) and sizes (40 and 80 nm) of copper nanoparticles for 24, 48 and 72 hours. The copper nanoparticles toxicity was examined by MTS assay. Cell migration has also been investigated with the Scratch assay. Results: The results showed that the 1, 10 and 100 µmol concentrations of 40 and 80 nm copper nanoparticles were not toxic for cultured human foreskin keratinocyte cells after 24h. It was also found keratinocyte cell proliferation was increased by 1 µmol concentration of 80 nm copper nanoparticles after 72h. The results of the Scratch assay showed that the 1 µmol concentration of 80 nm copper nanoparticles significantly (P<0.05) increased keratinocyte cell migration compared to deionized water as of control group after 24h. Conclusion: It seems the 1 µmol concentration of 80 nm copper nanoparticle could stimulate keratinocyte cell migration and proliferation. However, in vivo studies conducted on animal model wound healing subjects are needed for determining re-epithelialization