Evaluation of platelet-rich plasma on hair regrowth and lesional T-cell cytokine expression in alopecia areata: A randomized observer-blinded, placebo-controlled, split-head pilot study.

BACKGROUND: Platelet-rich plasma has shown some promise in the treatment of alopecia areata. OBJECTIVE: To evaluate the effect of platelet-rich plasma on hair regrowth and lesional T-cell cytokine expression in alopecia areata. METHODS: This was a randomized, placebo-controlled, split-head study involving 27 patients with alopecia areata (Severity of Alopecia Tool score ≥25%). Alopecia patches on either side of the scalp were randomized to receive 3 intradermal injections of platelet-rich plasma or normal saline at monthly intervals and evaluated 3 months after the last session. Lesional T-cell cytokine messenger RNA expression was compared pre- and posttreatment in the platelet-rich plasma-treated sites. RESULTS: The mean Severity of Alopecia Tool score did not change significantly compared with baseline with either platelet-rich plasma or placebo injections at any visit; however, the mean percentage reduction in the score in the platelet-rich plasma arm was more than in the placebo arm (9.05% ± 36.48% vs 4.99% ± 33.88%; P = .049) at final assessment. The mean interferon gamma (P = .001) and interleukin 17 cytokine (P = .009) messenger RNA expression decreased, whereas the mean interleukin 10 (P = .049) and FOXP3 (P = .011) messenger RNA expression increased significantly after platelet-rich plasma treatment. LIMITATIONS: Small sample size and a relatively short follow-up. CONCLUSION: Platelet-rich plasma was found to have limited efficacy in alopecia areata. However, it may play a role in restoring immune balance in the alopecic patches.

Effect of Q-switched Nd-YAG laser on the clinical, pigmentary, and immunological markers in patients with lichen planus pigmentosus: A pilot study.

The persistent discoloration in lichen planus pigmentosus (LPP) is secondary to persistent melanophages in the superficial dermis in these patients. We evaluated the effect of Q-switched Nd-YAG laser on the clinical, pigmentary, and immunological markers in patients with LPP. Nine females with a clinical diagnosis of LPP were included in the study. After six sessions of laser with toning protocol, performed over a representative area of 5 × 5 cm2 at 2-weekly intervals, the mean clinical improvement as per the physician assessment was 25.7% (10-40%). There was no significant reduction in melanin and erythema index. On expression analysis using quantitative-polymerase chain reaction, the reduction in tyrosinase (p = 0.03) was statistically significant, though, the reduction in other pigment and immunological markers was not significant. The immunohistochemistry quantification data of corresponding proteins also did not show any significant difference. Post inflammatory hypopigmentation was noted in one patient. Q-switched Nd-YAG laser toning protocol resulted in modest clinical and histological improvement in patients of LPP.

Transethnic analysis of psoriasis susceptibility in South Asians and Europeans enhances fine-mapping in the MHC and genomewide.

Because transethnic analysis may facilitate prioritization of causal genetic variants, we performed a genomewide association study (GWAS) of psoriasis in South Asians (SAS), consisting of 2,590 cases and 1,720 controls. Comparison with our existing European-origin (EUR) GWAS showed that effect sizes of known psoriasis signals were highly correlated in SAS and EUR (Spearman ρ = 0.78; p < 2 × 10-14). Transethnic meta-analysis identified two non-MHC psoriasis loci (1p36.22 and 1q24.2) not previously identified in EUR, which may have regulatory roles. For these two loci, the transethnic GWAS provided higher genetic resolution and reduced the number of potential causal variants compared to using the EUR sample alone. We then explored multiple strategies to develop reference panels for accurately imputing MHC genotypes in both SAS and EUR populations and conducted a fine-mapping of MHC psoriasis associations in SAS and the largest such effort for EUR. HLA-C*06 was the top-ranking MHC locus in both populations but was even more prominent in SAS based on odds ratio, disease liability, model fit and predictive power. Transethnic modeling also substantially boosted the probability that the HLA-C*06 protein variant is causal. Secondary MHC signals included coding variants of HLA-C and HLA-B, but also potential regulatory variants of these two genes as well as HLA-A and several HLA class II genes, with effects on both chromatin accessibility and gene expression. This study highlights the shared genetic basis of psoriasis in SAS and EUR populations and the value of transethnic meta-analysis for discovery and fine-mapping of susceptibility loci.

Comparison of Platelet-Rich Plasma Prepared Using Two Methods: Manual Double Spin Method versus a Commercially Available Automated Device.

BACKGROUND: In the absence of a standard protocol, several methods and devices have been used for preparing platelet-rich plasma (PRP) with varying platelet concentrations. METHODS: Venous blood sample from 20 patients was used for preparing PRP using two methods: a manual double-spin method (1st spin at 160 g × 10 min, 2nd spin at 400 g × 10 min), and using a commercially available automated device (DrPRP-Kit®, REMI Laboratory Instruments). Platelet, erythrocyte, and total leukocyte counts were calculated for each PRP sample and compared. RESULTS: Platelet count in the PRP prepared with the manual double-spin method (PRPm, 12.51 ± 5.89 × 105/µL) as well as with the automated device (PRPa, 7.25 ± 4.74 × 105/µL) had significantly higher mean platelet count than whole blood (2.58 ± 0.81 × 105/µL, P < 0.001). The mean platelet count in PRPm was statistically significantly higher than PRPa (P < 0.001). The platelet capture efficiency of the manual method (mean 47.11%, median 41.75%) was statistically significantly higher than that of the automated device (mean 31.89%, 29.51%, P = 0.012). Platelet counts in both PRPs were variable, but the counts were more dispersed in PRPa(coefficient of variation 65%) as compared to PRPm(coefficient of variation 47%). CONCLUSION: The manual double-spin method had a higher platelet capture efficiency resulting in a higher platelet concentration as compared to the automated device. Though there was a significant interindividual variation in the platelet yield in the PRPs produced by both methods, results were more consistent with the manual method.

Use of platelet-rich plasma to suspend noncultured epidermal cell suspension improves repigmentation after autologous transplantation in stable vitiligo: a double-blind randomized controlled trial.

BACKGROUND: Noncultured epidermal cell suspension (NCES) is an effective surgical modality for stable vitiligo which involves transplantation of the basal layer of epidermal cells onto the dermabraded vitiliginous patch. Platelet-rich plasma (PRP) has growth factors which may stimulate melanocyte migration and proliferation of keratinocytes and fibroblasts. The objective of this study was to compare the extent of repigmentation achieved by transplantation of NCES suspended in PRP with that of NCES suspended in phosphate buffered saline (PBS). METHODS: Twenty-one patients of stable vitiligo with at least two lesions of comparable size were included. The two vitiligo patches were randomized to receive NCES suspended in PRP or PBS. Postoperatively after 1 week, patients were given heliotherapy for 15 minutes daily. RESULTS: At 6 months follow-up, mean repigmentation by area method in PRP arm was 75.6 ± 30% SD and in non-PRP arm was 65 ± 34% SD (P = 0.0036). Patient satisfaction by visual analogue scale at 6 months also showed better results in PRP arm (P = 0.001). Assessment by three independent observers showed better repigmentation in PRP side both at 3 and 6 months. CONCLUSIONS: Suspending NCES in PRP can result in significantly greater mean repigmentation and patient satisfaction than suspending in PBS.

Safety and efficacy of autologous noncultured dermal cell suspension transplantation in the treatment of localized facial volume loss: A pilot study.

BACKGROUND: Available options for correction of facial volume loss, such as synthetic fillers, autologous fat and cultured fibroblasts, have limitations viz. temporary effect and high cost. AIM: To assess the use of a novel technique, autologous non-cultured dermal cell suspension transplantation, for correction of localized facial volume loss due to inflammatory pathologies. METHODS: It was a pilot study conducted in the Dermatology Outpatient Department, All India Institute of Medical Sciences (AIIMS), New Delhi, India. Autologous non-cultured dermal cell suspension was transplanted in a total of 10 patients, out of which 5 had predominantly dermal loss and the rest had predominantly lipoatrophy. The donor tissue from the gluteal region was digested into a single cell suspension using collagenase-1 and injected into the recipient area. The outcome was assessed subjectively by patients and investigators and objectively using ultrasonography. Cell count, viability testing and measurement of mesenchymal stem cells were also done. RESULTS: On assessment of patients, the median improvement in the predominantly dermal atrophy group at 3 and 6 months was 70% (range: 10-90%) and 80% (range: 0-90%), respectively, and in the predominantly lipoatrophy group, 0% (range: 0-40) and 0% (range: 0-50), respectively. Mean thickness of dermis + subcutis at the baseline was 1.835 mm (range: 0.89-6.04 mm), which increased to 2.912 mm (range: 0.88-7.07 mm, P = 0.03) at 6 months. LIMITATIONS: Our pilot study has some limitations such as small sample size and heterogeneity of the recruited patients. CONCLUSIONS: Autologous non-cultured dermal cell suspension transplantation appears to be safe and effective in localized facial dermal defects because of inflammatory pathologies, but not effective in deeper defects.

Role of lupus anticoagulants in immediate acting inhibitor positivity in congenital haemophilia A patients.

OBJECTIVES: Presence of lupus anticoagulants (LA) in haemophilia and their interference in coagulation assays is well-known. Factor VIII (FVIII) inhibitors are generally time and temperature dependent whereas LAs are immediate acting inhibitors (IAIs). The present study reports the challenges in laboratory detection of both progressive and non-progressive, specific FVIII inhibitors in the presence of LA. METHODS: From 2012 through 2015, 4900 HA patients were screened for inhibitors. APTT based inhibitor screening tests and Nijmegen-modified Bethesda assay (NBA) were done in all samples. LA test and FVIII inhibitors by ELISA were done in patients with IAIs. RESULTS: Out of 451 patients positive for inhibitors in the initial screening tests, classical and progressive FVIII inhibitors were observed in 398 patients while 53 had IAIs showing no/partial correction in 1:1 mixtures of NPP and patient plasma. In 27 patients, both FVIII and FIX activity levels were <1%, resulting in difficulty in diagnosis. In 48 HA patients with IAIs, 42 were LA positive. 4 patients were found to have only LA with false positive results in NBA while 38 had a combination of LA and FVIII inhibitors. Six patients were LA negative and had only FVIII IAIs. Five (62.5%) of 8 HA patients initiated on immune tolerance induction (ITI) also were positive for IAIs. CONCLUSION: The findings emphasizes the presence of specific FVIII inhibitors in congenital HA with absence of time dependent inactivation kinetics in a small proportion of cases. ELISA or chromogenic assays along with LA testing can offer accurate laboratory diagnosis in patients with coexisting LA.

Participation of T cell immunoglobulin and mucin domain-3 (TIM-3) and its ligand (galectin-9) in the pathogenesis of active generalized vitiligo.

Vitiligo is a depigmentary disease where melanocytes of the basal layer of epidermis are selectively destroyed by immune-cell-mediated cytotoxicity. The T cell immunoglobulin- and mucin-domain-containing molecules (TIMs) are involved in immune regulation, and their participation is not known in vitiligo. The present study revealed significant increase in the percentage of CD3+CD4+TIM3+ T cells (P < 0.05) in peripheral blood and was positively correlated with percentage body surface area involvement in aGV group. Further, increased expression of TIM-3 and its ligand galectin-9 (Gal-9) mRNA was found in peripheral blood and lesional/perilesional skin of active generalized vitiligo (aGV) compared with controls. Characteristic migration pattern of TIM-3-positive immune cells in lesional (near/in the epidermis) and perilesional (towards epidermis) skin section suggested that TIM-3+ immune cells may be involved in melanocyte destruction. Further, investigation is required to understand the role of TIM-3/Gal-9 signalling pathways in aGV and it can be targeted in the management of vitiligo.

The Epidemiology of FVIII Inhibitors in Indian Haemophilia A Patients.

A serious complication of replacement therapy in patients with bleeding disorders is the development of 'inhibitors', particularly FVIII inhibitors in haemophilia A patients. This leads to an increase in the management cost, morbidity and mortality, especially post-operatively. The mechanism of FVIII inhibitor development is quite complex and it is difficult to predict inhibitor development, but a prompt and accurate diagnosis is critical as early therapy can save lives. The aim of this study was to screen patients with bleeding disorders in India for inhibitors, and to analyse and compare the prevalence of inhibitors in different regions in India. Patient details were recorded and blood samples were collected in sodium citrate vacutainers from 1,505 patients with bleeding disorders, in different cities in India. Coagulation and inhibitor screening assays were performed, followed by the Bethesda assay in inhibitor positive samples to quantify the FVIII inhibitor titre. Out of the 1,505 samples analysed, 1,285 were Haemophilia A patients, out of which 78 (6.07 %) were positive for 'FVIII Inhibitors'. The highest incidence of FVIII Inhibitors was seen in South India (13.04 %). The highest incidence of 20.99 % was observed in Chennai, followed by Hyderabad (13.33 %), Jammu (9.90 %) and Guwahati (8.51 %), respectively, with respect to the samples analysed. The other regions showed an inhibitor incidence <8 %. The incidence of inhibitors in haemophilia A patients is different in different regions of India; this may be due to the intensity of treatment, type of product or the genetic characteristics of these patients.