Target area treatment ratio of varied lesions in the cultured pure melanocyte transplantation repigmentation of vitiligo: A retrospective study.

Autologous cultured pure melanocyte transplantation (CMT) can be utilized to treat stable vitiligo cases, but clinical data are insufficient to improve its efficacy. To evaluate the influence of various factors on the therapeutic effect of CMT, this single-center retrospective study enrolled stable vitiligo patients who underwent CMT between 2009 and 2020. Univariate and multivariable analysis were used to determine the factors affecting the outcome of repigmentation. The study included 491 patients with long-term follow-up data (6-120 months). It was found that 69.7% of patients achieved an excellent re-color effect and 18.4% achieved a good re-color effect. There were statistically significant differences in pigmentation between patients with stable disease course, vitiligo type, and lesion site. Overall, a significant positive correlation between the target area treatment ratio of varied lesions and the percentage of repigmentation was found. CMT is effective and well tolerated in the treatment of stable vitiligo. Various factors, especially the target area treatment ratio of varied lesions, should be carefully assessed before using CMT. As the target area treatment ratio of varied lesions could further improve the post-operative repigmentation other than type of vitiligo. This clinic trial was approved by Hangzhou Third People's Hospital (number 2023KA015, national clinical record number MR-33-23-034502).

The Treatment of Refractory Vitiligo With Autologous Cultured Epithelium Grafting: A Real-World Retrospective Cohort Study.

BACKGROUND: Surgical intervention is the main therapy for refractory vitiligo. We developed a modified autologous cultured epithelial grafting (ACEG) technique for vitiligo treatment. Between January 2015 and June 2019, a total of 726 patients with vitiligo underwent ACEG in China, with patient characteristics and clinical factors being meticulously documented. Using a generalized linear mixed model, we were able to assess the association between these characteristics and the repigmentation rate. RESULTS: ACEG demonstrated a total efficacy rate of 82.81% (1754/2118) in treating 726 patients, with a higher repigmentation rate of 64.87% compared to conventional surgery at 52.69%. Notably, ACEG showed a better response in treating segmental vitiligo, lesions on lower limbs, age ≤ 18, and stable period > 3 years. A keratinocyte:melanocyte ratio below 25 was found to be advantageous too. Single-cell RNA sequencing analysis revealed an increase in melanocyte count and 2 subclusters of keratinocytes after ACEG, which remained higher in repigmented sites even after 1 year. CONCLUSIONS: ACEG is a promising therapy for refractory vitiligo. Patient age, clinical type, lesion site, and stability before surgery influence repigmentation in ACEG. The mechanism of repigmentation after ACEG treatment is likely not confined to the restoration of melanocyte populations. It may also involve an increase in the number of keratinocytes that support melanocyte function within the affected area. These keratinocytes may aid the post-transplant survival and function of melanocytes by secreting cytokines and extracellular matrix components. TRIAL REGISTRATION: registered with Chictr.org.cn (ChiCTR2100051405).

Melanocyte transplantation to skin prepared by controlled PUVA-induced sunburn-like blistering for vitiligo treatment - A pilot clinical trial.

Vitiligo is a common acquired disease of pigment loss. In lesions recalcitrant to non-invasive treatment, transplantation of cultured autologous melanocytes is an emerging choice. Conventionally, the recipient site is often prepared by laser-mediated or mechanical dermabrasion. Such preparation procedures have disadvantages including prolonged transplantation duration, long period for reepithelialization and potential scarring. We propose a method of preparing recipient sites by psoralen and controlled ultraviolet A (PUVA)-induced blistering followed by transplanting suspended melanocytes. We introduced this method in 10 patients with segmental vitiligo on their recipient site 3 to 5 days before transplantation and blistering developed in 2 to 3 days afterwards. On the day of transplantation, the blister roof could be peeled off easily without bleeding and the recipient site preparation could be completed in 20 min. The recipient site became reepithelialized within 1 week. Progressive repigmentation was observed for up to 6 months, with an average of 65.06% repigmentation in the recipient site without scarring at the end of follow-up. Hence, preparation of the recipient site by controlled PUVA-induced sunburn-like blistering can potentially facilitate melanocyte transplantation and prevent scarring.

Autologous cell transplant as a treatment for stable segmental vitiligo: a systematic review.

OBJECTIVE: This systematic review provides a comprehensive analysis of the efficacy of autologous cell transplant as a therapeutic approach for stable segmental vitiligo. Vitiligo poses significant challenges for healthcare professionals in terms of treatment selection. Autologous cell transplant has emerged as a promising modality for managing vitiligo, with cultured and noncultured transplants being considered when determining the patient's treatment approach. There is little knowledge and literature on the subject, so we analyze the different studies. METHOD: Using online medical literature databases and the PRISMA guidelines, six out of 60 articles met the acceptance criteria to be analyzed, emphasizing the lack of current literature on this subject. RESULTS: Autologous cell transplant achieves excellent pigmentation rates for many body parts. We found that cultivated cells had better results than noncultivated ones. Both types of treatments could pigment 80% or more where needed. CONCLUSION: This review highlights the importance of autologous cell transplant as a new and reliable tool for the treatment of stable segmental vitiligo, cultured transplants being the most effective. By employing autologous cell transplant, the repigmentation rate is notably high and consistently achievable. Although its cost and logistical complexities hinder the current accessibility to this therapy, efforts are being made to enhance its availability, and its scope is expected to expand further. More studies are needed to understand this therapy method in other kinds of vitiligo.

Safety and Efficacy of Autologous Melanocyte/Keratinocyte Transplantation in Patients with Refractory Stable Vitiligo.

BACKGROUND: Vitiligo is a common depigmentation skin disease associated with significant psychosocial morbidity and profound effect on the quality of life. The treatment of vitiligo is still a major challenge in the field of dermatology. Currently, topical steroids, calcineurin inhibitors, ultraviolet phototherapy, surgery, and cultured and non-cultured epidermal melanocyte transplantation are used for the treatment of vitiligo. However, the effectiveness of these treatment modalities is limited by the lack of response, long-term treatment periods, high cost, and inevitable adverse effects. OBJECTIVES: In this study, we aimed to evaluate the efficacy of intraepidermal injection of autologous non-cultured melanocytes and keratinocytes as an alternative therapy for the refractory and stable (RS) vitiligo. METHODS: The treatment procedure was performed on thirty-nine RS vitiligo patients. The autologous skin grafts obtained from the buttock area and epidermis were separated from dermis using dispase. Single-cell autologous melanocytes and keratinocytes were prepared from the epidermis by trypsin/ethylene diamine tetra acetic acid and injected at the concentration of 100-400 × 103 cells/cm2, intra-epidermally to the selected vitiligo lesions. Vitiligo re-pigmentation was monitored employing photography. Photographs were taken prior to and 2, 4, and 6 months after the cell transplantation. Improvement of the skin depigmentation was classified as follows: <25% as minimal response, 26-50% as moderate response, 51-75% as good response, and finally 76-100% as excellent response. RESULTS: Cell infusion appeared to be safe as none of the patients exhibited any adverse effects. At the end of the sixth month follow-up period, of the treated patients, 12.8% demonstrated an excellent response, 36% exhibited a good response, and 51.2% showed a moderate to minimal response to the administered therapy. Obtained significant p value for Wilcoxon test over the checkpoints at 2nd, 4th, and 6th month (p = 0.03, 0.04, and 0.039, respectively) post-cell transplantation confirmed notable growing trend in the re-pigmentation. CONCLUSION: Our findings provide a strong support for the therapeutic efficacy of autologous non-cultured melanocytes and keratinocytes in patients with RS vitiligo.

Recipient-to-Donor Ratios in the Surgical Treatment of Vitiligo, Leukoderma, and Piebaldism: A Retrospective Review.

BACKGROUND: The autologous noncultured melanocyte keratinocyte transplant procedure (MKTP) has emerged as a popular grafting technique with proven efficacy for achieving repigmentation. However, there remains no consensus regarding the optimal recipient-to-donor (RD) ratio required to achieve acceptable repigmentation. In this retrospective cohort study of 120 patients, we sought to examine whether expansion ratios impact the repigmentation success rates following MKTP. RESULTS: A total of 69 patients (mean [SD] age was 32.4 [14.3] years, mean follow-up was 30.4 [22.5] months, 63.8% were male; 55% were dark-skinned individuals [Fitzpatrick IV-VI]) were included. The mean percent change in the Vitiligo Area Scoring Index (VASI) was 80.2 (±23.7; RD of 7.3) in patients with focal/segmental vitiligo (SV), 58.3 (±33.0; RD of 8.2) in those with non-segmental vitiligo (NSV), and 51.8 (±33.6; RD of 3.7) in those with leukoderma and piebaldism. Focal/SV was positively associated with a higher percent change in VASI (parameter estimate: 22.6, p value <0.005). In the SV/focal group, non-white patients had a higher RD ratio compared to White individuals (8.2 ± 3.4 vs. 6.0 ± 3.1, respectively, p value = 0.035). DISCUSSION: In our study, we found that patients with SV were significantly more likely to achieve higher repigmentation rates compared to those with NSV. Although repigmentation rates were higher in the low expansion ratio group than in the high expansion ratio group, we did not observe a significant difference between the two groups. CONCLUSION: MKTP is an effective therapy for restoring repigmentation in patients with stable vitiligo. Therapeutic response of vitiligo to MKTP appears to be influenced by the type of vitiligo, rather than a specific RD ratio.

Dermabrasion Versus Microneedling in Transplantation of Autologous Noncultured Melanocyte-Keratinocyte Cell Suspension in Patients With Vitiligo.

BACKGROUND: Autologous noncultured melanocyte-keratinocyte transplantation is one of the procedures used to treat stable vitiligo with varying reported results. Recipient site preparation is one of the variables that could affect repigmentation outcomes. OBJECTIVE: To assess the effectiveness of transplanting autologous melanocyte-keratinocyte suspension in patients with stable vitiligo and to compare recipient site preparation using dermabrasion versus microneedling. METHODS: From March 2020 to September 2022, this randomized comparative study included 40 patients with 40 stable vitiligo lesions managed by suspension transplants of melanocytes. Patients were divided into 2 groups: group A, where the recipient site was prepared using dermabrasion, and group B, which was done by microneedling. The assessment was performed 3 months after the treatment based on the degree of repigmentation (excellent, ≥90%; good, 50%-89%; fair, 20%-49%; and poor response, <20%). RESULTS: Both modalities resulted in effective repigmentation, but the dermabrasion group showed a statistically significant improvement and a satisfactory repigmentation rate. CONCLUSION: Autologous melanocyte transplantation is a safe and effective treatment method for stable vitiligo lesions that have not responded to other therapies. When compared with microneedling, dermabrasion produced better outcomes for recipient site preparation.

Transplantation of autologous epidermal melanocyte-keratinocyte cells suspension for cell therapy of vitiligo: A clinical evaluation and biometric assessment.

INTRODUCTION: Among several surgical treatments, the use of transplantation of epidermal cultured melanocytes or melanocytes-keratinocytes cell suspension has gained many researchers and dermatologists' attention as a new technique for the treatment of vitiligo. The present study aimed to transplant autologous epidermal melanocytes-keratinocytes cell suspension for the treatment of vitiligo. METHODS: In this study, 15 volunteer patients aged between 18 and 45 years old were studied. The autologous melanocytes-keratinocytes cell suspension was then transplanted to the region after dermabrasion. The included patients were evaluated by VisioFace, MPA9, and Skin Scanner-DUB once before and 1, 2, and 6 months after the transplantation, while the extents of stainability and changes in the transplanted region were recorded. RESULTS: The color contrast between the lesion and normal skin significantly decreased after 1, 2, and 6 months of the melanocytes transplantation compared with the pre-procedure (13.8 ± 0.45 before vs. 12.9 ± 0.43, 12.2 ± 0.45, and 10.2 ± 0.34 at months 1, 2, and 6, p < 0.001). Furthermore, melanin index significantly increased six months after cell transplantation compared to the pretreatment (168.3 ± 4.22 vs. 130.5 ± 3.98, p < 0.001). CONCLUSION: Transplantation of melanocytes cells with dermabrasion can be effective on vitiligo improvement, so it is recommended.

Combination of epidermal keratinocyte-melanocyte cells suspension and microneedling: Safe surgical approach in vitiligo.

INTRODUCTION: Vitiligo is a skin disease that is associated with impaired skin immune systems and pigment degradation in skin melanocyte cells. Despite the significant impact of the disease on the quality of life of patients, treatment of the disease using an effective method such as the transplantation of uncultivated melanocytes was considered by researchers around the world. The goal of this research was to use microneedling to transplant epidermal keratinocyte-melanocyte cells suspension for the treatment of vitiligo patients. METHODS: In this study, 15 male and female vitiligo in face region patients aged 18-45 years were studied. In this study, melanocyte-keratinocyte cells suspension was sprayed. Patients underwent microneedling treatment after spraying the cells. Before and after transplantation, patients were biometrically examined, and the quantity of pigmentation and changes in the transplanted region were documented. Statistical software was used to examine the results. RESULTS: The color difference between the lesion area and normal skin in one, two, and six months after treatment with cell suspension was significantly reduced compared with before treatment (by 48.95%). Moreover, the amount of melanin was significantly increased 6 months after treatment compared with before treatment (129.8 ± 4.16 vs. 195.2 ± 3.54, p = 0.000). A significant decrease in skin brightness in the skin of the lesion area was observed compared with normal skin, 6 months after treatment compared with before treatment (43.7 ± 1.44 vs. 27.9 ± 1.24, p = 0.000). CONCLUSION: Epidermal keratinocyte-melanocyte cells suspension in combination with microneedling could be considered as an effective treatment of vitiligo.

Efficacy and safety of traditional and surgical treatment modalities in segmental vitiligo: A review article.

BACKGROUND: Segmental vitiligo (SV) is a distinctive variant of vitiligo that is usually resistant to traditional treatments. Therefore, surgical therapies are mainstay of treatment in this type of vitiligo. To date, there is no review article or systematic review that evaluates specifically efficacy of treatment modalities on SV. AIM: To evaluate current evidence regarding efficacy and safety of traditional and surgical treatment modalities in SV. METHODS: We conducted a search in PubMed, Embase, Web of science, and Google Scholar for key words of "vitiligo" AND "segmental" AND "treatment" OR "therapy" OR "surgical treatments" OR "medical treatments" OR "laser" OR "phototherapy". Inclusion criteria were English literature that investigated efficacy of different treatments on three or more cases on SV from January 2000 until July 2021. RESULTS: A total of thirty-four articles were selected for detailed assessment. Different treatment modalities include medical treatment [systemic corticosteroid (SCS), topical CS (TCS), and topical calcineurin inhibitors (TCI)], phototherapy [narrow band-ultraviolet B (NB-UVB), psoralen and UVA (PUVA) and psoralen+solar exposure (PUVASOL)], laser/lights [helium-neon and Excimer laser/light (EL)] and surgical treatments [punch graft (PG), follicular graft, suction blister epidermal grafting (SBEG), spilt-thickness skin graft (STSG), and cultured/non-cultured-melanocytes-keratinocytes transplantation (MKTP)]. CONCLUSION: There were few randomized controlled trials (RCT) evaluating the efficacy of treatments in SV. Therefore, future high quality studies are required for better assessment of various treatment modalities in SV. Results of current evidence indicate resistance of SV to traditional therapies unless in patients with short duration of vitiligo. Therefore, surgical interventions are the first-line of treatment in refractory cases, long-standing disease, or presence of leukotrichia in depigmented patches.

Fractional versus full ablative CO2 laser in recipient site of non-cultured melanocytes and keratinocyte transplantation in treatment of vitiligo.

BACKGROUND: Melanocyte-keratinocyte transplant procedure (MKTP) or non-cultured epidermal cell suspension transplantation is a very popular surgical modality for treating stable vitiligo. The recipient-site preparation is one potential determinant in the repigmentation outcomes. AIM: To assess the efficacy of fractional CO2 (FCO2 ) laser in recipient-site preparation before MKTP and comparing it to the frequently used full surface laser ablation. METHODS: This randomized comparative trial included 19 patients with 40 stable vitiligo lesions. In each patient, the treated sites were randomly categorized into two groups according to the recipient-site ablation (either fractional or full ablative CO2 laser). Assessment of repigmentation was performed six months after the procedure. RESULTS: Both modalities achieved successful repigmentation of a median of 80% and 77.5% for fractional and full ablation groups, respectively, with a non-statistically significant difference between them. The median of VASI change percent was -73% and -71% with fractional and full surface ablation, respectively. CONCLUSIONS: FCO2 laser ablation is effective for recipient-site preparation before cell suspension transplantation as well as the full ablative CO2 laser.

CXCL9 as a key biomarker of vitiligo activity and prediction of the success of cultured melanocyte transplantation.

This study aimed to investigate the potential biomarkers of vitiligo by evaluating the disease activity and curative effect of autologous cultured pure melanocyte transplantation (CMT) on patients. Altogether, 36patients with stable vitiligo were treated with CMT. Blister fluid samples were collected from patients with stable vitiligo. Patients with active vitiligo were matched with healthy controls. The chemokine levels in the serum and blister fluid samples were measured using Luminex. The curative effect on patients with stable vitiligo was evaluated 6 months after treatment. Treatment responses were defined according to the extent of repigmentation as effective (if 50% or more repigmentation was achieved) or ineffective (if less than 50% or worse repigmentation was achieved). Patients received re-transplantation if the initial treatment was ineffective. The levels of C-X-C motif chemokine ligand (CXCL)9 and CXCL10 in blister fluid samples were significantly lower in stable patients than in active participants. Receiver operating characteristic analysis revealed that the levels of CXCL9 and CXCL10 were sensitive and specific in diagnosing active vitiligo. Further, 65.6% (21/32) of patients who received CMT had effective treatment responses. The high CXCL9 level in the blister fluid was a significant predictor of ineffective treatment responses. The treatment response was significantly enhanced after treatment. Four patients with ineffective treatment responses received anti-inflammatory treatment and re-transplantation. The CXCL9 and CXCL10 levels in the blister fluid were related to the presence of active vitiligo. Also, the CXCL9 level was a predictor of the effectiveness of CMT in treating vitiligo.

A retrospective study of long term follow-up of 2283 vitiligo patients treated by autologous, non-cultured melanocyte-keratinocyte transplantation.

BACKGROUND: Autologous non-cultured melanocyte-keratinocyte transplantation (MKTP) can be used to treat stable vitiligo cases, but there were insufficient clinical data to evaluate its safety and efficacy. OBJECTIVE: To assess the influence of various factors on the therapeutic outcome of MKTP. METHOD: The single-center retrospective study included stable vitiligo patients who underwent MKTP between June 2009 and June 2018. Univariate and/or multivariable analysis were used to determine the factors affecting the outcome of repigmentation. RESULT: The study comprised 2283 patients who had long-term follow-up data (12-108months). Excellent repigmentation was achieved in 400/606 (66%),788/1341 (58.8%),437/684 (63.9%),18/24 (75%) patients with segmental vitiligo, pre-MKTP phototherapy, younger than 24 years, the lesion on the perineum and scrotum, respectively. However, the patients with a positive family history, Koebner phenomenon responded worse(χ2=29.417, P<0.001; χ2=107.397, P<0.001; respectively). Overall, a significant positive correlation between duration of stability and percentage of repigmentation was found (χ2=42.053, P<0. 001). CONCLUSION: MKTP is efficient and well tolerated for stable vitiligo treatment. Various factors such as duration of disease stability, vitiligo type, family history, site of lesion should be carefully assessed before using MKTP, as it would further improve the post-operative repigmentation.

A decellularized human corneal scaffold for anterior corneal surface reconstruction.

Allogenic transplants of the cornea are prone to rejection, especially in repetitive transplantation and in scarred or highly vascularized recipient sites. Patients with these ailments would particularly benefit from the possibility to use non-immunogenic decellularized tissue scaffolds for transplantation, which may be repopulated by host cells in situ or in vitro. So, the aim of this study was to develop a fast and efficient decellularization method for creating a human corneal extracellular matrix scaffold suitable for repopulation with human cells from the corneal limbus. To decellularize human donor corneas, sodium deoxycholate, deoxyribonuclease I, and dextran were assessed to remove cells and nuclei and to control tissue swelling, respectively. We evaluated the decellularization effects on the ultrastructure, optical, mechanical, and biological properties of the human cornea. Scaffold recellularization was studied using primary human limbal epithelial cells, stromal cells, and melanocytes in vitro and a lamellar transplantation approach ex vivo. Our data strongly suggest that this approach allowed the effective removal of cellular and nuclear material in a very short period of time while preserving extracellular matrix proteins, glycosaminoglycans, tissue structure, and optical transmission properties. In vitro recellularization demonstrated good biocompatibility of the decellularized human cornea and ex vivo transplantation revealed complete epithelialization and stromal repopulation from the host tissue. Thus, the generated decellularized human corneal scaffold could be a promising biological material for anterior corneal reconstruction in the treatment of corneal defects.