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.

<h4>Aims and objectives</h4>The aim of this study is to examine the efficacy and safety of various regenerative medicine treatments, such as cell therapy, platelet-rich plasma (PRP), plasma-poor platelet (PPP), plasma-rich fibrin (PRF), mesenchymal stem cells, stromal vascular fraction (SVF), exosomes, adipose-derived stem cells (ADSC), and stem cell-conditioned media (SC-CM), for treating vitiligo.<h4>Method</h4>We conducted a thorough search of major databases such as PubMed, Scopus, and Web of Science, and selected 48 articles based on specific criteria. We used EndNote X8 and Google Sheets to review and extract data from the articles. After analyzing the studies, we categorized them accordingly.<h4>Results</h4>This systematic review analyzed 48 articles involving 2186 patients with vitiligo to assess the effectiveness of regenerative medicine treatments. Key findings revealed that methods such as autologous non-cultured melanocyte-keratinocyte transplantation and platelet-rich plasma (PRP) injection exhibited significant repigmentation, particularly when combined with modalities like NB-UVB phototherapy and laser treatments. Notably, the autologous melanocyte-keratinocyte transplantation achieved over 50% repigmentation within 9 months, while PRP demonstrated an average repigmentation of 58.7%, especially effective with CO₂ laser treatment. Hair follicle-derived cell transplantation also showed impressive response rates, achieving good to excellent results in up to 93.8% of patients. Side effects were noted in 21 of 28 studies, primarily involving pain, with no serious adverse events reported. The risk of bias assessment indicated that 37.21% of studies were low risk, while 48.84% had high risks overall. These findings suggest that while regenerative medicine holds promise for vitiligo treatment, further clinical trials are necessary to explore additional methods like stromal vascular fraction and exosomes.<h4>Conclusion</h4>We have concluded that regenerative medicine plays an effective role in the treatment of vitiligo lesions. Furthermore, this treatment method is safe and does not cause serious complications. It can be used alone or in combination with other methods for treating vitiligo. To advance the treatment of vitiligo, we recommend conducting clinical trials on the unexplored branches of regenerative medicine.

Abstract BackgroundMelanocytes have been cultivated from the outer root sheath of hair follicles for two decades. So far, these isolation and culturing procedures have been carried out from human and mouse follicles. In this study, we have translated the established procedure for obtaining melanocytes from stem cells and precursors of human hair follicle to a rabbit species in order to isolate and cultivate rabbit melanocytes from whisker follicle outer root sheath (ORS), hereby named rMORS, and compare them to rabbit epidermal rabbit melanocytes (rEMs).ResultsThe rMORS were isolated and cultured by extracting whisker follicles from enzymatically digested skin and submitting them to the air-liquid interface hypoxic conditions. The cells were allowed to migrate from the follicle ORS onto the nylon mesh of Transwell inserts, collected and subcultured in melanocyte culture medium for 11 passages. From early passages on, the rMORS cells displayed typical melanocyte characteristics comparable to those of rEM used as experimental control. Melanocyte features were assessed on morphological level by the means of microscopy, functional and gene expression level. Functionality was qualitatively assessed by the Von Kossa and Nile Blue staining of melanin and by biochemically quantifying melanin content and Tyrosinase activity. Gene expression of neuroectodermal and melanocytic lineage markers NES, PAX3, MITF, TYR, CKIT and PMEL was determined by the means of qRT-PCR. ConclusionsWe concluded that the method of isolating and culturing rMORS efficiently and reproducibly corresponded to the prior art method established in human follicle, yielding melanocytes comparable to the rabbit epidermal melanocytes in all assessed features. This method is not crucial in terms of regenerative therapy, it rather paves way for a non-invasive ex vivo sampling of hair and it may give way to deeper insights into rabbit follicle biology or to postulating useful species-specific in vivo experimental models.

rate in the early Anagen II stage and continue through the Anagen III stage of the hair growth cycle. After harvesting, melanocytes were either grown in vitro using enriched keratinocyte S-FM culture for 2 weeks or transplanted immediately. Among the cultured melanocytes, a pigmented area was identified around the implanted hair follicle in the matrix before transplantation (Fig 1 ). This area corresponds to an area of potential melanocyte migration from the hair follicle bulb. We could see the pigmented area around the base of cultured hair follicles on both day 7 and day 14. After the transplantation, the dark-brown-colored area (arrow) is still visible around the transplanted hair follicle (Fig 2 ). Figure 2. Open in a new tab Hair follicles grown in vitro. Hair follicle stalks demonstrate darker pigmentation at after 2 weeks of incubation in enriched keratinocyte serum-free media. When hair follicles were transplanted immediately after harvest and allowed to heal, we were also able to identify the pigmented area 2 weeks posttransplantation. This pigmented area is similar to that seen among cultured melanocytes. We could see a dark-brown area around the transplanted hair follicles, whereas no pigmentation was observed in the nontransplanted wound. Over time, as the surgical wounds healed, this dark-brown color became more pronounced (Fig 3 ). Figure 3. Open in a new tab Healing wounds 2 weeks postoperatively. Melanocyte-transplanted wounds (transplantation site) (right) demonstrate central areas of pigmentation, whereas nontransplanted wounds contain no pigmentation (control site) (left). After complete healing (4 weeks posttransplantation), central areas of pigmentation observed in hair follicle transplanted wounds appeared even darker and thicker than their counterparts at 2 weeks posttransplantation (Fig 4 ). Despite marked wound contraction, a rim of nonpigmented tissue visibly demarcated the central area of hair follicle melanocyte pigmentation from the rim