Outer Root Sheath Cell Suspension Research Articles

Treatment of stable vitiligo using cultured and non-cultured autologous melanocytes from hair follicle outer root sheath cell suspension

Background Several surgical treatment methods have been proposed for the treatment of stable vitiligo, but they remain inadequate in terms of patient satisfaction. Objective To assess the efficacy and safety of a modified autologous cultured hair follicle outer root sheath (HF-ORS) cell suspension transplantation in the treatment of stable vitiligo lesions, and to compare it with transplantation of autologous non-cultured HF-ORS cell suspension for the same patient. Patients and methods Twenty-four patients were enrolled in this prospective controlled study. Hairs were epilated, and HF-ORS cell suspensions were prepared using two methods for the same patient. The recipient site was prepared by CO2 laser resurfacing. Assessment of cellular activity and viability included cellular melanin content determined by enzyme-linked immunosorbent assay and measurement of the fold change in premelanosome (Pmel-17) gene expression. The patients were followed-up for 6 months postprocedure. Results Nineteen patients completed the study. The results demonstrated that the cultured method showed superior repigmentation response in 15 (79%) patients, three (16%) responded equally, and one (5%) was more tolerant of the noncultured method. Conclusion Cultured autologous HF-ORS melanocytes show superior repigmentation response, increased melanin content, and upregulated gene expression of Pmel-17 than that of noncultured ones. However, the study also demonstrated that both methods are potential options for the treatment of stable vitiligo.

A Prospective Observational Comparative Study of Novel Autologous Negative Pressure Epidermal Harvesting System (ANPEHS or EHS) and Suction Blister Grafting (SBG) in Treatment of Stable Vitiligo.

Background:Vitiligo surgery has evolved a long way from punch skin grafts to the epidermal cell suspension and latest to the extracted hair follicle outer root sheath cell suspension transplantation (EHFORSCS). In the latest development, the novel technique autologous negative pressure epidermal harvesting system (ANPEHS) seems to be a good addition to the prevailing therapy in vitiligo.Aims and Objectives:The aim of this work was to study and compare novel ANPEHS and suction blister grafting (SBG) in the treatment of stable vitiligo.Materials and Methods:This was a prospective, single-centered, observational, open-labeled comparative study of the rate and final extent of repigmentation in ANPEHS or EHS and SBG in the management of stable vitiligo. The patients in this study were drawn from the outpatient Department of Dermatology of a Tertiary Care Hospital of the Armed Forces from July 1 2015 to December 31, 2016. A total of 40 patients with at least two comparable depigmented patches of 6 months’ stability were recruited. Both procedures were performed in the same patient on the same day. Informed consents were taken from all the patients. Each patient in the study had undergone a detailed clinical, general physical, systemic, and a dermatological examination. Clinical photographs had been taken before and after grafting, monthly for the first 6 months then bimonthly for the next 6 months. Donor site was preferably inner aspect of thigh.Statistical Analysis Used:“Chi-square test” and “statistical significance” (P value) methods.Results:Of total 80 patches, excellent results were seen in 82.9% patches by using the EHS method and 80% excellent results by using the SBG method. Similarly, very good results were seen in 2.9% patches and good results in 5.7% patches by using both methods. 11.4% patches showed poor results by using the SBG method and 8.6% patches showed poor results by using the EHS method.Conclusions:The EHS method is a simple, painless, less time-consuming, expensive but effective technique to produce homogeneous repigmentation without any donor site anesthesia and complication.

Noncultured Extracted Hair Follicle Outer Root Sheath Cell Suspension versus Noncultured Epidermal Cell Suspension in the Treatment of Stable Vitiligo.

Background:Various treatment modalities exist for vitiligo, yet none of them are curative. Vitiligo is still considered a challenging disease to manage. Surgical treatment offers an excellent option for patients with stable vitiligo, especially those who fail to respond to medical treatment. Cell suspension techniques carry the advantage of covering large surface areas.Objective:To compare noncultured extracted hair follicle outer root sheath cell suspension (NCORSHFS) and noncultured epidermal cell suspension (NCES) in producing repigmentation.Subjects and Methods:Twenty patients were randomly allocated into two groups. They were objectively evaluated for the extent of repigmentation (after 1, 2, and 3 months), complications, cosmetic outcome, and satisfaction.Results:In NCORSHFS group, 10% showed excellent pigmentation, 20% showed good pigmentation, 50% fair, and 20% poor pigmentation. In NCES group, 10% showed excellent pigmentation, 10% good pigmentation, 40% fair, and 40% poor pigmentation. This difference was not statistically significant. Excellent color match was observed in 80% of NCORSHFS and in 70% of NCES. Donor area complications were absent in NCORSHFS group, whereas in NCES, mild scarring in 20% and hyperpigmentation in 40% of patients was observed. The difference in patients’ satisfaction between the two groups was not statistically significant.Conclusion:Both NCORSHFS and NCES are effective in producing good repigmentation with perfect color match and patients’ satisfaction. NCORSHFS has no donor area complications because it is a scarless procedure.

Study of efficacy and safety of noncultured, extracted follicular outer root sheath cell suspension transplantation in the management of stable vitiligo.

Noncultured, extracted follicular outer root sheath suspension (NC-EHF-ORS-CS) is a recently introduced technique for the treatment of stable vitiligo. To study the clinical efficacy of this technique and to determine the viability and cell composition of the suspension. Twenty-five patients with stable vitiligo were included in this prospective study. Fifty follicles were extracted from occipital scalp and were incubated with trypsin-ethylenediaminetetraacetic acid to separate outer root sheath cells. The cell suspension was filtered and centrifuged to obtain a cell pellet, which was resuspended and applied to the dermabraded recipient area. Cell viability of the suspension was assessed using trypan blue staining, and markers of keratinocyte stem cells (CD200) and melanocytes (S100) were evaluated using flow cytometry and immunocytochemistry, respectively. At 6 months, the mean (±SD) repigmentation was 52 ± 25.1%, and >75% repigmentation was seen in 8/25 (32%) patients. Mean percentage cell viability of the suspension was 80 ± 17.2% with a mean concentration of CD200 + and S100 + cells being 7.91 ± 8.68% and 9.93 ± 1.22% (n = 3), respectively. Recipient site infection was seen in 4 of 25 (16%) patients and a color mismatch in 11 of 25 (44%) patients. NC-EHF-ORS-CS is a useful minimally invasive therapy for vitiligo.

Comparative Study of Efficacy of Epidermal Melanocyte Transfer Versus Hair Follicular Melanocyte Transfer in Stable Vitiligo.

Background:Vitiligo surgery has come up a long way from punch skin grafts to epidermal cell suspension and latest to the extracted hair follicle outer root sheath cell suspension (EHFORSCS) transplantation. The progressive development from one technique to the other is always on a quest for the best. In the latest development, EHFORSCS, which is an enriched source of follicular inactive melanocyte (melanocyte stem cells), seems to be a good addition to the prevailing cell-based therapies for vitiligo. However, it needs to be explored further in larger, clinical trials.Methodology:A total of 11 patients with sixty stable vitiligo sites attending dermatology outpatient department were included for the open-labeled, prospective, comparative study. The sites were sequentially distributed into two groups of thirty each. Sites of one group were subjected to epidermal melanocyte transfer (EMT) and the others to hair follicular melanocyte transfer (HFMT). Response to treatment was evaluated on the basis of degree of repigmentation; final evaluation of area of involvement was done after completion of 6 months.Results:At the end of 6 months, repigmentation >90% was observed in 83.33% patches of EMT group and 43.33% in HFMT group. Repigmentation >75% was observed in 90% of patches in Group A and 43.34% of patches in Group B, respectively. There was statistically significant difference in the overall pigmentation between these two groups.Conclusion:Both noncultured autologous epidermal cell suspension transfer and noncultured EHFORSCS transfer are safe and effective surgical modalities in the management of stable vitiligo though EMT has shown a better response in the present study. Outer root sheath cell suspension transfer is a novel, minimally invasive technique in its nascent stage in the surgical management of vitiligo which requires further larger clinical trials for evaluation of its efficacy.

A Study of Noncultured Extracted Hair Follicle Outer Root Sheath Cell Suspension for Transplantation in Vitiligo.

Context:Vitiligo surgeries have come a long way from tissue grafts to cultured and non cultured cell transplantation. Extracted hair follicle outer root sheath cell transplantation (EHF ORS) suspension is more enriched with melanocyte. In a hair bulb, there is one melanocyte for every five keratinocytes which is much higher than the epidermal melanin unit.Aims:To analyse the effectiveness of cultured EHF ORS and to perform objective evaluation based on clinical improvement & photographic evidence. To observe any untoward events or side effects.Settings and Design:The study was open and uncontrolled. All the patients were screened at preliminary visit. Reviews were done every two weeks. The endpoint selected was six months post procedure.Materials and Methods:Twenty five patients of stable Vitiligo were included in the study and follicular unit were harvested by Follicular Unit Extraction method. Outer root sheath cells were extracted by trypsinization. The solution was transplanted over dermabraded recipient site. Pressure dressing was given. Patients were followed up regularly.Statistical Analysis Used:Descriptive Statistics, Chi-Square.Results:Mean ± SD repigmentation was 80.15% ± 22.9% with excellent repigmentation (90-100%) in 60% of patients.Conclusions:This method is safe, effective, and simpler than the other methods involving cell culturing and requiring a laboratory set-up but selection of patients is crucial for the success of the outcome.

Comparison between autologous noncultured extracted hair follicle outer root sheath cell suspension and autologous noncultured epidermal cell suspension in the treatment of stable vitiligo: a randomized study

Vitiligo is an acquired disorder of pigmentation caused by loss of epidermal melanocytes. Autologous noncultured epidermal cell suspension (NCES) and autologous noncultured extracted hair follicle outer root sheath cell suspension (NCORSHFS) are important surgical modalities for the treatment of stable vitiligo. To compare NCES and NCORSHFS for producing repigmentation in stable vitiligo. We randomized 30 patients with 47 stable vitiligo lesions into two groups. Patients in group 1 were treated with NCES, and those in group 2 with NCORSHFS. They were evaluated 16 weeks postsurgery for the extent of repigmentation, colour match, change in Dermatology Life Quality Index (DLQI) score and patient satisfaction. The extent of repigmentation was excellent (90-100% repigmentation) in 83% of lesions in the NCES group and 65% of lesions in the NCORSHFS group (P = 0·154). Repigmentation ≥ 75% (good repigmentation) was observed in 92% of lesions in the NCES group and 78% of lesions in the NCORSHFS group (P = 0·425). There was a significant improvement in DLQI score in both the groups, but the mean decrease among groups did not differ significantly (P = 0·244). However, patients in the NCES group were significantly more satisfied than the patients in the NCORSHFS group. No significant difference was seen in colour match and pattern of repigmentation. Adverse effects were minimal. Both NCES and NCORSHFS are safe and effective techniques with comparable efficacy. To the best of our knowledge, this is the first study directly comparing two different cellular techniques.

Extracted hair follicle outer root sheath cell suspension for pigment cell restoration in vitiligo

Vitiligo surgery has come up a long way from punch skin grafts to epidermal cell suspension and latest to the extracted hair follicle outer root sheath cell suspension (EHF-ORS-CS) transplantation. The progressive development from one technique to the other is always in a quest for the best. In the latest development- EHF-ORS-CS, which is an enriched source of follicular inactive melanocyte (melanocyte stem cells), seems to be a good addition to the prevailing cell-based therapies for vitiligo; however, need to be explored further in larger, and preferably randomized blinded studies. This review discusses the principle, technical details, and stem cell composition of hair follicular outer root sheath cell suspension.

Depletion of CD200+ hair follicle stem cells in human prematurely gray hair follicles

Introduction:Melanocyte stem cells (MelSCs) are known to be depleted in gray hair follicles. Hair follicle stem cells (HFSCs) are important for maintenance of stemness of MelSCs.Methods:We compared the proportion of CD200+ (Cluster of Differentiation 200 positive) stem cells in the outer root sheath cell suspension of gray and pigmented hair follicles of three patients with the premature graying of hair. In addition, explants culture for HFSCs was also carried out from gray and pigmented hair follicles. Cultured HFSCs were also differentiated into melanocytes.Results:The mean ± SD CD200+ HFSCs population were 9.4 ± 1.4% and 3.5 ± 0.5% for pigmented and gray hair follicles, respectively (P = 0.002). In explants culture, the growth of HFSCs from the gray hair follicle stopped at around day 20–22, whereas the growth of the cells from the pigmented follicle continued.Conclusion:CD200+ HFSCs are depleted in prematurely gray hair in the humans. CD200+ hair follicle stem cell yield is poorer in gray hair explant culture than pigmented hair explant culture.

Noncultured extracted hair follicle outer root sheath cell suspension for transplantation in vitiligo

Current noncultured cell-based transplantation therapies for vitiligo largely involve shave skin biopsy for preparation of noncultured melanocyte suspension. As the overall proportion of melanocytes is low in the epidermis, these techniques require basal cell layer enrichment, which adds additional steps. We tried follicular unit extraction (FUE) to harvest hair follicles as a source of melanocytes. To evaluate the efficacy of a novel surgical method for vitiligo: noncultured extracted hair follicular outer root sheath (ORS) cell suspension transplantation. Fourteen patients with vitiligo, stable for at least 3 months, were included in this prospective study. Fifteen to 25 hair follicles were extracted from occipital scalp using the FUE method. Hair follicles were incubated with trypsin-ethylenediamine tetraacetic acid solution at 37°C for 90 min to separate ORS cells. The cell suspension was filtered through a 70-μm cell strainer, then centrifuged for 5 min at 1000 r.p.m. to obtain a cell pellet. The pellet was resuspended and applied to the dermabraded recipient area and dressed. The mean ± SD repigmentation was 65·7 ± 36·7%. Overall, nine of 14 patients achieved > 75% repigmentation. Mean percentage repigmentation was significantly higher in patients with ≥ 1 year stability than those with < 1 year stability (P = 0·02). Extracted hair follicular ORS cell suspension can be a useful simplified transplantation method for vitiligo. The transplantation procedure should be reserved for patients with vitiligo stable for at least 1 year. A larger study is needed for further evaluation.