Keratinocyte Transplantation Research Articles

Chemotherapy-Mediated Complications of Wound Healing: An Understudied Side Effect.

Significance: Chemotherapy is a primary method to treat cancer. While chemotherapeutic drugs are designed to target rapidly dividing cancer cells, they can also affect other cell types. In the case of dermal cells and macrophages involved in wound healing, cytotoxicity often leads to the development of chronic wounds. The situation becomes even more severe when chemotherapy is combined with surgical tumor excision. Recent Advances: Despite its significant impact on patients' recovery from surgery, the issue of delayed wound healing in individuals undergoing chemotherapy remains inadequately explored. Critical Issues: This review aims to analyze the harmful impact of chemotherapy on wound healing. The analysis showed that chemotherapy drugs could inhibit cellular metabolism, cell division, and angiogenesis and lead to nerve damage. They impede the migration of cells into the wound and reduce the production of extracellular matrix. At the molecular level, they interfere with replication, transcription, translation, and cell signaling. This work reviews skin problems that patients may experience during and after chemotherapy and demonstrates insights into the cellular and molecular mechanisms of these pathologies. Future Directions: In the future, the problem of impaired wound healing in patients treated with chemotherapy may be addressed by cell therapies like autologous keratinocyte transplantation, which has already proved effective in this case. Epigenetic intervention to mitigate the side effects of chemotherapy is also worth considering, but epigenetic consequences of chemotherapy on skin cells are largely unknown and should be investigated.

Lesional CD8+ T-Cell Number Predicts Surgical Outcomes of Melanocyte–Keratinocyte Transplantation Surgery for Vitiligo

The melanocyte-keratinocyte transplantation procedure (MKTP) treats stable and recalcitrant vitiligo. Despite careful selection of candidates based on clinical stability, the success of the procedure is unpredictable. The aim of our study was to define the immunological profile of stable vitiligo lesions undergoing MKTP and correlate them with clinical outcomes. We included 20 MKTP candidates with vitiligo and a patient with piebaldism as a control. Prior to MKTP, T-cell subsets and chemokines in the recipient skin were measured by flow cytometry and ELISA. During MKTP, melanocytes in the donor skin were quantified by flow cytometry. After MKTP, patients were followed for 12 months and repigmentation was assessed clinically and by ImageJ analysis of clinical photographs. Baseline immunologic biomarkers, duration of clinical stability, and transplanted melanocyte number were correlated to postsurgical repigmentation scores. CD8+ T cells were elevated in 43% of the clinically stable vitiligo lesions. CD8+ T-cell number negatively correlated with postsurgical repigmentation scores (r=-0.635, P= 0.002). Duration of clinical stability, skin chemokines, and transplanted melanocyte number did not influence postsurgical repigmentation. This study demonstrates that CD8+ T-cell number correlates negatively with success of postsurgical repigmentation and can be a biomarker to identify ideal surgical candidates.

Risk Factors for Infection After Noncultured Melanocyte Keratinocyte Transplantation for Vitiligo.

Noncultured autologous melanocyte keratinocyte transplantation is considered a safe and effective treatment option in stable vitiligo. Factors associated with risk of infection are still poorly explored. To search for factors associated with the risk of infection after noncultured autologous melanocyte keratinocyte transplantation (MKTP). This was a retrospective multicentric study including all patients with vitiligo who had undergone noncultured autologous MKTP between January 2010 and December 2020. Data included age, sex, site, and size of the treated area, recipient area preparation method, and antibiotic prescription preceding the procedure. Univariate and multivariate analyses to search for factors associated with infection after MKTP were conducted. A total of 672 patients were included. Infection was present in 39 of the patients (6%) (95% confidence interval [CI]: 4.2%-7.7%). The following factors were independently associated with higher rate of infection: cryotherapy for recipient area preparation (OR 19.76, 95% CI: 3.21-121.74) and treated lesions on the trunk (OR 2.67, 95% CI: 1.21-5.90), lower extremity (OR 5.99, 95% CI: 2.49-14.40), and foot (OR 13.15, 95% CI: 4.37-39.62). Infection after noncultured autologous MKTP is not uncommon. Cryotherapy for recipient area preparation and lesions on the trunk, lower extremity, or foot was independently associated with an increased risk of infection.

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

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. 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. 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. 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. FCO2 laser ablation is effective for recipient-site preparation before cell suspension transplantation as well as the full ablative CO2 laser.

Effect of melanocyte keratinocyte transplantation procedure, using ablative CO2 laser and followed by NB-UVB phototherapy, on the expression of E-Cadherin and matrix metalloproteinase-2 in acral vitiligo: an immunohistochemical study

Background Melanocyte keratinocyte transplantation procedure (MKTP) is a multi-procedural intervention that could hypothetically alter the environment surrounding grafted epidermal cells, helping melanocytes’ migration and adherence to keratinocytes in their basal position. Objective To evaluate the effect of MKTP in acral vitiligo skin prepared by total ablative CO2 laser resurfacing and followed by narrow-band ultraviolet B (NB-UVB) phototherapy on matrix metalloproteinase-2 (MMP2) and E-Cadherin expression and the reflection of these changes on repigmentation. Patients and methods Acral lesions in 20 stable nonsegmental vitiligo patients were prepared by full CO2 ablation down to the papillary dermis followed by MKTP and NB-UVB sessions. Two 4 mm punch biopsies were taken: the first at baseline and the second after the onset of repigmentation or after 6 months of phototherapy if no repigmentation occurred. Immunohistochemical staining for evaluation of E-Cadherin and MMP2 expression was performed. Results Eight patients (40%) achieved repigmentation ranging from 10 to 90% with a median of 62.5%. E-Cadherin intensity was significantly increased after treatment (P Conclusion MKTP with recipient site prepared by full CO2 laser ablation followed by NB-UVB phototherapy increased the expression of E-Cadherin and MMP2 in acral vitiliginous skin. The increase of dermal MMP2 could play a favorable role in repigmentation.

Recipient site preparation by cryoblebbing in melanocyte keratinocyte transplantation procedure over the fingers in vitiligo: A pilot study.

Surgical treatment of vitiligo lesions over the fingers has poor outcome. In this intra-patient comparative study, 12 patients with stable non-segmental vitiligo (NSV) affecting the middle three fingers of one hand were included. Three variations were used in treatment of finger vitiligo lesions: minipuch grafting, melanocytes keratinocyte transplantation procedure (MKTP) preceded by cryoblebbing or full CO2 laser resurfacing of the recipient site. Liquid nitrogen was used to create blebs in one finger 24 hours before therapy. On the following day, the second finger was treated by minipunch grafting and the third finger was resurfaced by CO2 laser. A suspension was prepared and 0.1 mL was injected into each cryobleb. It was also applied to the resurfaced skin. All patients underwent topical PUVA therapy and were followed-up for 12 months. Ten cases with 52 lesions completed the follow-up period. About 4/18 lesions treated by cryoblebbing followed by MKTP showed ≥75% repigmentation while only 1/17 lesions treated by laser resurfacing + MKTP and 1/17 lesions treated by minipunch grafting showed 30% and 10% repigmentation, respectively. No complications occurred in MKTP treated lesions. Cryoblebbing of the recipient site seems to improve the outcome of MKTP in lesions over the fingers in stable NSV.

Outcomes of autologous non-cultured melanocyte keratinocyte transplantation in vitiligo and nevus depigmentosus

Background Melanocyte-keratinocyte transplantation procedure (MKTP) is an effective surgical technique for restoring skin pigmentation in all types of vitiligo and leukoderma patients who are unresponsive to medical and/or phototherapy treatment. Data specific to the outcomes of MKTP among Thai vitiligo and nevus depigmentosus patients are currently scarce. Objectives To evaluate the efficacy and safety of MKTP in patients with vitiligo or nevus depigmentosus at the short-term (≤6 months) and long-term (≥12 months) follow-up. Materials and methods A retrospective review of the medical records of vitiligo or nevus depigmentosus patients who underwent MKTP at the Department of Dermatology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand during 2016 to 2019 was conducted. Repigmentation outcomes were evaluated by Vitiligo Area Scoring Index (VASI). Results Twenty-five patients had 27 MKTP surgeries on 32 anatomically-based lesions. The mean age was 32.4 years, the mean age at onset was 25.5 years, and 19 patients were male. Segmental vitiligo, non-segmental vitiligo, and nevus depigmentosus had significantly improved VASI scores at the short-term follow-up (−74.2% ± 23.2%, −100%, and −62.5% ± 17.6%, respectively) and the long-term follow-up (−81% ± 27.7%, −95.0% ± 7.0%, and −83.3% ± 14.4%, respectively). Conclusion MKTP is a safe and effective method for treating refractory vitiligo and nevus depigmentosus in Thai patients.

Needling as a Method to Highlight the Margins of a Vitiligo Patch During Melanocyte-Keratinocyte Transplantation.

Needling as a Method to Highlight the Margins of a Vitiligo Patch During Melanocyte-Keratinocyte Transplantation.

Feasibility Study to Evaluate RECELL and Melanocyte Keratinocyte Transplantation Procedure for Repigmentation of Stable Vitiligo Lesions

Feasibility Study to Evaluate RECELL and Melanocyte Keratinocyte Transplantation Procedure for Repigmentation of Stable Vitiligo Lesions

Poly(D,L-lactide)/PEG blend films for keratinocyte cultivation and skin reconstruction

The objective of this study was to develop a novel porous thin poly(D,L-lactide) (PLA) film as a tissue-engineering scaffold for keratinocytes used for the replacement of damaged skin. Poly(D,l-lactic acid)/poly(ethylene glycol) (PEG: Mw 6000 or 15 000) blend films were formed by a spin coating technique. The properties and structures of these blend films were investigated. PDLA/PEG (6000) blend films were modified by microfibrillar collagen after water incubation to increase hydrophilicity and improve keratinocyte adhesion. Primary keratinocytes were seeded on PLA films, cultivated for 9 d and transplanted to rats with a model skin defect wound. The wound’s healing after keratinocyte transplantation was assayed with histological and immunochemical methods. It was found that skin damage recovery was the most effective after transplantation of keratinocytes on porous PLA film modified with collagen.

The Nurse’s Role in Melanocyte–Keratinocyte Transplantation Procedure

Journal of the Dermatology Nurses’ Association ():, September 5, 2019. | DOI: 10.1097/JDN.0000000000000485

Six-year follow-up of vitiligo patients successfully treated with autologous non-cultured melanocyte-keratinocyte transplantation.

Although autologous non-cultured melanocyte-keratinocyte transplantation is a treatment option for stable vitiligo, there is lack of long-term maintenance data for this specific treatment. To search for factors associated with long-term maintenance of patients with stable vitiligo successfully treated with melanocyte-keratinocyte transplantation. This was a single-centre retrospective study including stable vitiligo patients who underwent successful melanocyte-keratinocyte transplantation in the National Center for Vitiligo, Riyadh, Saudi Arabia, between 1 January 2004 and 30 June 2015. Cox proportional hazard model was used to estimate factors associated with relapse at 6 years of followup. Co-variates included, gender, type of vitiligo, age at vitiligo onset, age at surgical procedure, disease duration, disease stability, affected body surface area, treated surface area, fingertip involvement, type of recipient area treatment and recurrence defined as the onset of new lesions on previously untreated areas. The risk of developing relapse defined as re-appearance of more than 10% depigmentation in a previously treated and repigmented site was considered as the main outcome. In total, 602 patients were included in the study of whom 410 (67%) were women. Mean age was 24.25 years [4.0-67.0]. Affected body surface area of less than 1% (adjusted HR = 0.37; P = 0.04) and mechanical dermabrasion (adjusted HR = 0.26; P = 0.03) were independently associated with lower rates of relapse. On the contrary, non-segmental type of vitiligo (adjusted HR = 2.11; P = 0.03) and fingertip involvement (adjusted HR = 3.75; P = 0.01) were independently associated with higher rates of relapse. Criteria for selecting patients with stable vitiligo for surgery should include careful assessment of vitiligo type including body surface area of vitiligo and involvement of fingertip before undergoing surgical procedure.

Does the transplantation of keratinocytes really reduce the risk of death? Survival analysis of patients hospitalized at the Dr Stanisław Sakiel Centre for Burns Treatment in 2008–2015

Keratinocyte transplantation is an adjuvant procedure in the extensive burn therapy method. However, it must be taken into consideration that clinical results of keratinocyte transplantation are ambiguous and progress achieved in this method is still being verified, especially due to the high cost of cultured epithelial autograft (CEA) transplants. The aim of this study was to verify the impact of cultured keratinocyte application on patients' survival. This study included a group of patients with the highest chance for a successful outcome of the therapy and excluded patients with no compelling reason to apply for such an expensive therapy. This study included all the patients with burns diagnosed between January 1, 2008 and January 1, 2016, who were treated with cultured skin cells. Patients' age and gender, percentage of total body surface area (TBSA) affected, percentage of burn depth of the 3rd/4th degree, number of days between admission and surgery, and need for rehabilitation were analyzed. The cultured cell application did not significantly affect the risk of death (p > 0.05). Keratinocytes should be applied as an adjunctive method for the treatment of burns with at least 40% TBSA affected, but with a maximal burn depth of the 2nd degree. In the group of patients below 50 years of age, a higher number of transplants with a cell population above 20 million/mL and a significantly lower mortality rate were observed, which means that in the mentioned age group, this graft was more effective. It has been suggested that patients older than 50 years of age with burns deeper than of the 2nd degree should be treated with more advanced methods like, e.g., the application of stem cells.

Autologous non-cultured melanocyte-keratinocyte transplantation in the treatment of vitiligo: patient selection and perspectives.

Autologous non-cultured melanocyte–keratinocyte transplantation procedure (MKTP) is one of the simplest cellular grafting techniques. Various modifications were done over the years to make the technique easier and more economical which led to its great popularity among dermatologists. Proper patient selection and good technical skills are essential for achieving success with this technique. In this review, different patient-related and procedure-related factors that affect the outcome are discussed. This review may guide dermatologists to select suitable candidates, and explains what to expect in each case and indicates different techniques which can be used. The expected complications and stability of acquired pigmentation, which are an essential part of the pretreatment patient counseling, are also discussed.

1292 Comparative study of the immunological profile in stable segmental and non segmental vitiligo patients undergoing melanocyte keratinocyte transplantation

1292 Comparative study of the immunological profile in stable segmental and non segmental vitiligo patients undergoing melanocyte keratinocyte transplantation

Novel Technique for Retention of Cell Suspension in Melanocyte–Keratinocyte Transplantation Procedure Using Glued Nylon Suture or Mesh

Novel Technique for Retention of Cell Suspension in Melanocyte–Keratinocyte Transplantation Procedure Using Glued Nylon Suture or Mesh

Advances in keratinocyte delivery in burn wound care.

This review gives an updated overview on keratinocyte transplantation in burn wounds concentrating on application methods and future therapeutic cell delivery options with a special interest in hydrogels and spray devices for cell delivery.To achieve faster re-epithelialisation of burn wounds, the original autologous keratinocyte culture and transplantation technique was introduced over 3 decades ago. Application types of keratinocytes transplantation have improved from cell sheets to single-cell solutions delivered with a spray system. However, further enhancement of cell culture, cell viability and function in vivo, cell carrier and cell delivery systems remain themes of interest.Hydrogels such as chitosan, alginate, fibrin and collagen are frequently used in burn wound care and have advantageous characteristics as cell carriers.Future approaches of keratinocyte transplantation involve spray devices, but optimisation of application technique and carrier type is necessary.

Modeling AEC—New approaches to study rare genetic disorders

Ankyloblepharon-ectodermal defects-cleft lip/palate (AEC) syndrome is a rare monogenetic disorder that is characterized by severe abnormalities in ectoderm-derived tissues, such as skin and its appendages. A major cause of morbidity among affected infants is severe and chronic skin erosions. Currently, supportive care is the only available treatment option for AEC patients. Mutations in TP63, a gene that encodes key regulators of epidermal development, are the genetic cause of AEC. However, it is currently not clear how mutations in TP63 lead to the various defects seen in the patients' skin. In this review, we will discuss current knowledge of the AEC disease mechanism obtained by studying patient tissue and genetically engineered mouse models designed to mimic aspects of the disorder. We will then focus on new approaches to model AEC, including the use of patient cells and stem cell technology to replicate the disease in a human tissue culture model. The latter approach will advance our understanding of the disease and will allow for the development of new in vitro systems to identify drugs for the treatment of skin erosions in AEC patients. Further, the use of stem cell technology, in particular induced pluripotent stem cells (iPSC), will enable researchers to develop new therapeutic approaches to treat the disease using the patient's own cells (autologous keratinocyte transplantation) after correction of the disease-causing mutations.

Foreskin-isolated keratinocytes provide successful extemporaneous autologous paediatric skin grafts

Severe burns in children are conventionally treated with split-thickness skin autografts or epidermal sheets. However, neither early complete healing nor quality of epithelialization is satisfactory. An alternative approach is to graft isolated keratinocytes. We evaluated paediatric foreskin and auricular skin as donor sources, autologous keratinocyte transplantation, and compared the graft efficiency to the in vitro capacities of isolated keratinocytes to divide and reconstitute epidermal tissue. Keratinocytes were isolated from surgical samples by enzymatic digestion. Living cell recovery, in vitro proliferation and epidermal reconstruction capacities were evaluated. Differentiation status was analysed, using qRT-PCR and immunolabelling. Eleven children were grafted with foreskin-derived (boys) or auricular (girls) keratinocyte suspensions dripped onto deep severe burns. The aesthetic and functional quality of epithelialization was monitored in a standardized way. Foreskin keratinocyte graft in male children provides for the re-epithelialization of partial deep severe burns and accelerates wound healing, thus allowing successful wound closure, and improves the quality of scars. In accordance, in vitro studies have revealed a high yield of living keratinocyte recovery from foreskin and their potential in terms of regeneration and differentiation. We report a successful method for grafting paediatric males presenting large severe burns through direct spreading of autologous foreskin keratinocytes. This alternative method is easy to implement, improves the quality of skin and minimizes associated donor site morbidity. In vitro studies have highlighted the potential of foreskin tissue for graft applications and could help in tissue selection with the prospect of grafting burns for girls.

Treatment of post-burn leucoderma with non-cultured melanocyte–keratinocyte transplantation (MKTP)

Introduction Deep burns often result in hypo-pigmentation, referred to as leucoderma which has a similar psychosocial impact on the patients as that of vitiligo. Several tissue grafting methods have been reported to treat post-burn leucoderma. Methods A simple method consisting of harvesting a donor skin sample, preparation of an epidermal cell suspension and the transplantation of the cell suspension on to a dermabraded recipient area, was performed in a clean procedure room. No special laboratory set-up was used for the cell separation procedure. Patients were treated with 18 sessions of excimer laser starting 1 month post-operatively, to hasten the repigmentation. Observation Of the 10 patients treated with MKTP, 3 were lost to follow-up. The remaining 7 patients showed repigmentation ranging from 90% to 100% with good color matching. Conclusion MKTP is an effective method to treat post-burn leucoderma. No special precautions are required to treat any anatomical site or uneven scarred surface.