This work was financially supported by the Key Project of Medical Research Jointly from Chongqing Health Commission and Chongqing Science and Technology Bureau (2022ZDXM037), the Key Projects of Scientific Research Fund of Traditional Chinese Medicine of Wuhan Health Commission (WZ22A03), and Natural Science Foundation of Chongqing (cstc2020jcyj-msxmX0592).

All research procedures involving human participants comply with the ethical standards of the Ethics Committee of Chongqing Hospital of Traditional Chinese Medicine and the principles of the Declaration of Helsinki (1964) and its subsequent amendments and have received approval for the study from the aforementioned Ethics Committee. Image data collection was performed with both verbal and written consent from the patients, and photographs taken for teaching purposes as a routine part of the examination were obtained using the Multispectral Dermoscope.
1. Patient selection
<ol>
	<li>Set the following inclusion criteria5:
	<ol>
		<li>Select patients based on the presence of facial light to dark brown, well-defined patches, typically with symmetrical distribution, absence of inflammation, and scaling.</li>
		<li>Select patients based on the absence of significant subjective symptoms such as itching, burning, and pain.</li>
		<li>Select patients considering that melasma predominantly occurs in females, primarily after puberty.</li>
		<li>While selecting patients, consider seasonal variations in the condition-often worse in summer and milder in winter.</li>
		<li>Exclude patients if other conditions (such as nevus fuscoceruleus zygomaticus, Riehl&#39;s melanosis, and pigmentary photodermatitis) that may cause hyperpigmentation are present.</li>
	</ol>
	</li>
</ol>
2. Preparation for roller microneedle treatment
<ol>
	<li>Preoperative communication
	<ol>
		<li>Understand the patient&#39;s treatment goals and cosmetic aspirations and gather complete medical history, including medication history, allergies, contraindications, and past cosmetic treatments.</li>
		<li>Inform the patients about the treatment&#39;s effects, steps, potential complications, recovery time, duration, and costs. Discuss specific treatment plans and alternative options to ensure all patient queries are addressed, helping them establish realistic expectations and achieve higher satisfaction.</li>
		<li>Ask patients to sign an informed consent form.</li>
	</ol>
	</li>
	<li>Environmental requirements
	<ol>
		<li>Set aside a quiet, well-lit treatment room for the operation of the roller microneedle and disinfect it for 30 min with ultraviolet light before the procedure.</li>
		<li>Ask the patients to wear shoe covers and hats upon entering the treatment room.</li>
	</ol>
	</li>
	<li>Preparation of the necessary items and medication for treatment
	<ol>
		<li>Check and prepare all the materials listed in the Table of Materials.</li>
	</ol>
	</li>
	<li>Makeup removal and cleansing
	<ol>
		<li>If the treatment area was covered with makeup, thoroughly remove it with makeup remover before cleansing.</li>
		<li>For non-makeup wearers, clean the treatment area with a gentle cleansing product.</li>
	</ol>
	</li>
	<li>Multispectral dermatoscopy recording 
	NOTE: The Multispectral dermoscope uses different light sources to capture facial ultra-high pixel images.
	<ol>
		<li>Prior to treatments and after three treatments, use the Multispectral dermoscope to take photos of the patient&#39;s front, left (45&#176;), and right (45&#176;) from three different angles and record the values of patients&#39; UV-shallow and -deep spots and brown spots</li>
	</ol>
	</li>
	<li>Application of topical anesthesia
	<ol>
		<li>Apply 5% lidocaine cream evenly to the treated area, with a total dosage of 30-40 g, and wrap it with cling film to enhance the anesthesia effect for 40-60 min. 
		NOTE: Some patients may exhibit symptoms of allergy to the topical anesthetic. Patients may experience severe itching, burning, or stinging at the site of application. If such reactions occur, promptly remove the anesthesia and provide appropriate symptomatic treatment.</li>
		<li>Before the next treatment, ensure proper moisturization of the skin in the anesthesia application area.</li>
	</ol>
	</li>
	<li>Preparation of microneedle solution
	<ol>
		<li>Extract 5 mL of 5% tranexamic acid solution using a syringe and set it aside for use.</li>
	</ol>
	</li>
	<li>Preparation of microneedle instruments
	<ol>
		<li>Select a 0.5 mm roller microneedle (Figure 1) and soak it in 0.1% benzalkonium bromide solution for approximately 20 min to disinfect it.</li>
		<li>Rinse the microneedle 3x with physiological saline to remove the disinfectant. 
		&#8203;NOTE: Avoid cross-contamination by using a single roller microneedle for each patient to prevent unnecessary damage to the skin due to blunt needles and to prevent the spread of diseases between patients.</li>
	</ol>
	</li>
	<li>Removal of topical anesthesia
	<ol>
		<li>After a sufficient duration of topical anesthesia application, remove the cling film and scrape off the anesthetic with a spatula. Use gauze soaked in 0.9% saline solution to wipe off any remaining anesthetic from the face.</li>
	</ol>
	</li>
	<li>Disinfection
	<ol>
		<li>Expose the treatment area, wrap the hair with a disinfectant towel when treating the face, and disinfect the facial skin with a 0.1% benzalkonium bromide cotton ball 3x from the top to the hairline edge, to the lower jaw edge, and chin, following the principle of &#34;bottom-up, inward-outward&#34; to prevent infection.</li>
	</ol>
	</li>
</ol>
3. Roller microneedle procedure
<ol>
	<li>Have the therapist wash their hands and wear a hat, mask, and sterile gloves.</li>
	<li>After informing the patient about beginning the procedure, instruct the patient to close their eyes and treat in the order of left cheek, right cheek, forehead, jaw, upper lip, and nose. Inject an appropriate amount of tranexamic acid solution onto the skin using a syringe, apply evenly by hand, and then rinse the roller microneedle with physiological saline, roll it in a short distance along the horizontal, vertical, and oblique directions in the treatment area.</li>
	<li>Gradually perform the treatment in different areas, applying the solution while rolling to enhance absorption. Repeat 3-5x in one area, preferably with skin flushing and no obvious bleeding. Apply force evenly, neither too heavy nor too light, and do not drag a microneedle on the skin.</li>
</ol>
4. Efficacy evaluation
<ol>
	<li>Use a combination of roller microneedles and topical tranexamic acid solution 3x for the treatment of melasma, once every 4 weeks, as a course of treatment.</li>
	<li>Use the multispectral dermatoscope before and after treatment, 3x to observe the efficacy according to the values of the patients&#39; UV shallow and deep spots and brown spots. The smaller the values, the lighter the melasma.</li>
	<li>Statistically analyze the changes in the UV-shallow and -deep spots and brown spots before and after treatment.</li>
</ol>

Enhancing Melasma Treatment with Tranexamic Acid and Microneedling Therapy

<ol>
	<li>Passeron, T., Picardo, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Melasma,+a+photoaging+disorder.">Melasma, a photoaging disorder.</a> Pigment Cell &amp; Melanoma Research. 31 (4), 461-465 (2018).</li><li>Pigment Disease Group. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dermatovenereology+Professional+Committee+of+Chinese+Society+of+Integrative+Medicine,+Vitiligo+Research+Center+of+the+Dermatology+and+Venereal+Disease+Branch+of+the+Chinese+Medical+Association,+Pigment+Disease+Working+Group+of+the+Dermatology+and+Venereal+Disease+Branch+of+the+Chinese+Medical+Association,+Chinese+Medical+Doctor+Association.+Consensus+on+diagnosis+and+treatment+of+melasma+in+China+(2021+vision).">Dermatovenereology Professional Committee of Chinese Society of Integrative Medicine, Vitiligo Research Center of the Dermatology and Venereal Disease Branch of the Chinese Medical Association, Pigment Disease Working Group of the Dermatology and Venereal Disease Branch of the Chinese Medical Association, Chinese Medical Doctor Association. Consensus on diagnosis and treatment of melasma in China (2021 vision).</a> Chinese Journal Dermatology. 54 (2), 110-115 (2021).</li><li>Vineet, K. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microneedle+arrays+for+cutaneous+and+transcutaneous+drug+delivery,+disease+diagnosis,+and+cosmetic+aid.">Microneedle arrays for cutaneous and transcutaneous drug delivery, disease diagnosis, and cosmetic aid.</a> Journal of Drug Delivery Science and Technology. 79, 104058 (2023).</li><li>Skin Laser Medical Aesthetics Group, Dermatology and Venereology Branch, Chinese Medical Association, Skin Laser Technology Application Research Center of the Chinese Medical Association. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chinese+dermatologists'+consensus+on+laser+and+IPLS+treatment+and+skin+repairment+of+melasma.">Chinese dermatologists' consensus on laser and IPLS treatment and skin repairment of melasma.</a> Journal Practical Dermatology. 13 (2), 65-69 (2020).</li><li>Pigment Disease Group. Dermatovenereology Professional Committee of Chinese Society of Integrative Medicine. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+diagnosis+and+efficacy+criteria+for+melasma+(revised+in+2003).">Clinical diagnosis and efficacy criteria for melasma (revised in 2003).</a> Chinese Journal of Dermatology Venereology Integrative Traditional and Western Medicine. 3 (1), 66 (2004).</li><li>McKesey, J., Tovar-Garza, A., Pandya, A. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Melasma+treatment:+an+evidence-based+review.">Melasma treatment: an evidence-based review.</a> American Journal of Clinical Dermatology. 21 (2), 173-225 (2020).</li><li>Budamakuntla, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+randomised,+open-label,+comparative+study+of+tranexamic+acid+microinjections+and+tranexamic+acid+with+microneedling+in+patients+with+melasma.">A randomised, open-label, comparative study of tranexamic acid microinjections and tranexamic acid with microneedling in patients with melasma.</a> Journal of Cutaneous and Aesthetic Surgery. 6 (3), 139-143 (2013).</li><li>Cohen, B. E., Elbuluk, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microneedling+in+skin+of+color:+a+review+of+uses+and+efficacy.">Microneedling in skin of color: a review of uses and efficacy.</a> Journal of the American Academy of Dermatology. 74 (2), 348-355 (2016).</li><li>Lee, H. C., Thng, T. G. S., Goh, C. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oral+tranexamic+acid+(TA)+in+the+treatment+of+melasma:+a+retrospective+analysis.">Oral tranexamic acid (TA) in the treatment of melasma: a retrospective analysis.</a> Journal of the American Academy of Dermatology. 75 (2), 385-392 (2016).</li><li>Banihashemi, M., Zabolinejad, N., Jaafari, M. R., Salehi, M., Jabari, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+therapeutic+effects+of+liposomal+tranexamic+acid+and+conventional+hydroquinone+on+melasma.">Comparison of therapeutic effects of liposomal tranexamic acid and conventional hydroquinone on melasma.</a> Journal of Cosmetic Dermatology. 14 (3), 174-177 (2015).</li><li>Ebrahimi, B., Naeini, F. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Topical+tranexamic+acid+as+a+promising+treatment+for+melasma.">Topical tranexamic acid as a promising treatment for melasma.</a> Journal of Research in Medical Sciences. 19 (8), 753-757 (2014).</li><li>Na, J. I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+tranexamic+acid+on+melasma:+a+clinical+trial+with+histological+evaluation.">Effect of tranexamic acid on melasma: a clinical trial with histological evaluation.</a> Journal of the European Academy of Dermatology and Venereology. 27 (8), 1035-1039 (2013).</li><li>Kim, E. H., Kim, Y. C., Lee, E. S., Kang, H. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+vascular+characteristics+of+melasma.">The vascular characteristics of melasma.</a> Journal of Dermatological Science. 46 (2), 111-116 (2007).</li><li>Yuan, C., Wang, X. M., Yang, L. J., Wu, P. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tranexamic+acid+accelerates+skin+barrier+recovery+and+upregulates+occluding+in+damaged+skin.">Tranexamic acid accelerates skin barrier recovery and upregulates occluding in damaged skin.</a> International Journal of Dermatology. 53 (8), 959-965 (2014).</li><li>Tohgasaki, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enolase-1+expression+in+the+stratum+corneum+is+elevated+with+parakeratosis+of+atopic+dermatitis+and+disrupts+the+cellular+tight+junction+barrier+in+keratinocytes.">Enolase-1 expression in the stratum corneum is elevated with parakeratosis of atopic dermatitis and disrupts the cellular tight junction barrier in keratinocytes.</a> International Journal of Cosmetic Science. 40 (2), 178-186 (2018).</li><li>Lima, E. V. A., Lima, M. M. D. A., Paix&#227;o, M. P., Miot, H. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+the+effects+of+skin+microneedling+as+adjuvant+therapy+for+facial+melasma:+a+pilot+study.">Assessment of the effects of skin microneedling as adjuvant therapy for facial melasma: a pilot study.</a> BMC Dermatology. 17 (1), 14 (2017).</li><li>Saleh, F., Abdel-Azim, E., Ragaie, M., Guendy, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Topical+tranexamic+acid+with+microneedling+versus+microneedling+alone+in+treatment+of+melasma:+clinical,+histopathologic,+and+immunohistochemical+study.">Topical tranexamic acid with microneedling versus microneedling alone in treatment of melasma: clinical, histopathologic, and immunohistochemical study.</a> Journal of Egypt Women's Dermatologic Society. 16 (2), 89 (2019).</li><li>Bailey, A. J. M., Li, H. O., Tan, M. G., Cheng, W., Dover, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microneedling+as+an+adjuvant+to+topical+therapies+for+melasma:+A+systematic+review+and+meta-analysis.">Microneedling as an adjuvant to topical therapies for melasma: A systematic review and meta-analysis.</a> Journal of the American Academy of Dermatology. 86 (4), 797-810 (2022).</li></ol>

The authors have no conflicts of interest to declare.

The pathogenesis of melasma is complex, and treatment methods include systemic medication, topical depigmenting agents, chemical peels, and laser therapy. Unfortunately, no single treatment is universally effective for all patients, and satisfactory results cannot be always guaranteed6. Phototherapy shows a decreasing trend in treating melasma, and some patients experience worsening of melasma after phototherapy. This is due to an increase in individuals with sensitive skin and impaired skin barri...

Melasma, also known as chloasma, is a chronic and acquired facial hyperpigmentation skin condition. Clinically, it presents as symmetrical, light to dark brown patches with unclear borders, distributed on the cheeks, forehead, and jawline. The incidence rate among Asian women of childbearing age can be as high as 30%1. The disease is easily diagnosed but difficult to cure. The pathogenesis of melasma is not yet fully understood, but three main factors contribute to its development: genetic susceptibility, exposure to sunlight, and hormonal changes. Increased melanin synthesis, vascular proliferation at the site of skin lesions, inflammatory reactions, and impaired skin barrier function all play a role in the occurrence of melasma2.
Currently, treatment options for melasma include laser therapy and medications, among which, transdermal administration is a widely used method. However, the main challenge lies in the barrier function of the stratum corneum, which hinders drug penetration. Microneedle therapy creates multiple micro-channels in the skin within a short period, disrupting the stratum corneum, inducing collagen production, and increasing skin permeability. This approach holds promise in allowing drugs to diffuse into the skin and then rapidly reseal, thus avoiding the introduction of toxic or irritating substances3.
Commonly used microneedle tools include the electronic water light injection microneedle, roller microneedle, electrically assisted nanocrystalline microneedle, plum blossom microneedle, and single needle microneedle4. The roller microneedle is characterized by less trauma, good curative effect, convenient operation, and can be combined with some other drugs to treat melasma. It has a good therapeutic prospect for melasma and is worth more promotion and use in clinical practice. This article introduces the operation and precautions of roller microneedles combined with tranexamic acid in the treatment of melasma and demonstrates the efficacy of roller microneedle combined with tranexamic acid in the treatment of melasma through clinical cases.

<table><tbody><tr><td>5 mL syringe</td><td></td><td></td><td></td></tr><tr><td>Benzalkonium bromide solution</td><td>Chongqing Traditional Chinese Medicine Hospital</td><td>H20050531</td><td>0.1%</td></tr><tr><td>cling film</td><td></td><td></td><td></td></tr><tr><td>Compound lidocaine cream</td><td>Beijing Ziguang Pharmaceutical Co., Ltd.</td><td>H20063466</td><td>5.0%</td></tr><tr><td>disinfectant towel</td><td></td><td></td><td></td></tr><tr><td>disposable curved disc</td><td></td><td></td><td></td></tr><tr><td>Multispectral Dermoscope</td><td>CBS Medical Skin Analysis, Wuhan, China</td><td>CBS-2022</td><td></td></tr><tr><td>Roller microneedle</td><td>Guangzhou Yuanxiang Biotechnology Co., Ltd</td><td>221001</td><td>(model: 0.22 x 0.5 mm, Figure 1)</td></tr><tr><td>Sodium chloride solution</td><td>Hubei Tiansheng Pharmaceutical Co., Ltd.</td><td>H42021838</td><td>0.9%</td></tr><tr><td>sterile cotton swab</td><td></td><td></td><td></td></tr><tr><td>sterile gauze</td><td></td><td></td><td></td></tr><tr><td>sterile gloves</td><td></td><td></td><td></td></tr><tr><td>tongue depressor</td><td></td><td></td><td></td></tr><tr><td>Tranexamic acid solution</td><td>Shanghai Hyundai Hasen (Shangqiu) Pharmaceutical Co., Ltd.</td><td>H20184047</td><td>5.0%</td></tr><tr><td>treatment vehicle</td><td></td><td></td><td></td></tr></tbody></table>

roller microneedle combined with tranexamic acid solution in treating melasma

Melasma, a common, acquired facial pigmentation skin disorder, presents a straightforward clinical diagnosis but poses challenges in terms of effective management. The precise underlying causes of melasma remain elusive, and the current therapeutic approaches predominantly encompass pharmaceutical and laser interventions, with limited efficacy. Transdermal administration stands as a prevalent treatment method for melasma, often facilitated by the application of microneedles. Among these, tranexamic acid emerges as a frequently employed therapeutic agent. A subset of microneedles, known as roller microneedles, plays a significant role in this approach by delicately puncturing the epidermis with multiple fine needles, synergizing with drug delivery. This methodology not only enhances drug absorption but also augments treatment efficacy while minimizing tissue trauma. These attributes forecast promising avenues for the treatment of melasma. This article primarily introduces the combination of roller microneedle and tranexamic acid solution in the treatment of melasma and demonstrates the efficacy of roller microneedle and tranexamic acid solution in the treatment of melasma through clinical cases.

Case 1, a 57-year-old female with a medical history of 10 years, was selected for treatment and efficacy evaluation. The efficacy of the combination of roller microneedle and tranexamic acid solution was evaluated before treatment and after three treatment sessions (Figure 2, Figure 3, and Figure 4) and observed from different angles using a multispectral dermatoscope. A reduction of 35.91 mm2, 163.32 mm2, and ...

Watch this Scientific Journal Video about Roller Microneedle Combined with Tranexamic Acid Solution in Treating Melasma at JoVE.com

Author Spotlight: Non-Surgical Treatment of Melasma&#8211; Microneedling with Tranexamic Acid

This article presents a protocol to conduct the treatment of melasma by using roller microneedles in combination with tranexamic acid solution and evaluate the efficacy.

Roller Microneedle Combined with Tranexamic Acid Solution in Treating Melasma

Melasma, a common, acquired facial pigmentation skin disorder, presents a straightforward clinical diagnosis but poses challenges in terms of effective ...

roller-microneedle-combined-with-tranexamic-acid-solution-treating

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

791 Views.  Chongqing Hospital of Traditional Chinese Medicine. This article presents a protocol to conduct the treatment of melasma by using roller microneedles in combination with tranexamic acid solution and evaluate the efficacy.

Video: Author Spotlight: Non-Surgical Treatment of Melasma– Microneedling with Tranexamic Acid

Spatial and Temporal Control of Murine Melanoma Initiation from Mutant Melanocyte Stem Cells

Cutaneous melanoma is well known as the most aggressive skin cancer. Although the risk factors and major genetic alterations continue to be documented with increasing depth, the incidence rate of cutaneous melanoma has shown a rapid and continuous increase during recent decades. In order to find effective preventative methods, it is important to understand the early steps of melanoma initiation in the skin. Previous data has demonstrated that follicular melanocyte stem cells (MCSCs) in the adult skin tissues can act as melanoma cells of origin when expressing oncogenic mutations and genetic alterations. Tumorigenesis arising from melanoma-prone MCSCs can be induced when MCSCs transition from a quiescent to active state. This transition in melanoma-prone MCSCs can be promoted by the modulation of either hair follicle stem cells&#39; activity state or through extrinsic environmental factors such as ultraviolet-B (UV-B). These factors can be artificially manipulated in the laboratory by chemical depilation, which causes transition of hair follicle stem cells and MCSCs from a quiescent to active state, and by UV-B exposure using a benchtop light. These methods provide successful spatial and temporal control of cutaneous melanoma initiation in the murine dorsal skin. Therefore, these in vivo model systems will be valuable to define the early steps of cutaneous melanoma initiation and could be used to test potential methods for tumor prevention.

The following procedure describes a method for spatial and temporal control of melanocytic tumor initiation in murine dorsal skin, using a genetically engineered mouse model. This protocol describes macroscopic as well as microscopic cutaneous melanoma initiation.

Cutaneous melanoma is well known as the most aggressive skin cancer. Although the risk factors and major genetic alterations continue to be documented ...

Cancer Research

Pharmacologic Induction of Epidermal Melanin and Protection Against Sunburn in a Humanized Mouse Model

Fairness of skin, UV sensitivity and skin cancer risk all correlate with the physiologic function of the melanocortin 1 receptor, a Gs-coupled signaling protein found on the surface of melanocytes. Mc1r stimulates adenylyl cyclase and cAMP production which, in turn, up-regulates melanocytic production of melanin in the skin. In order to study the mechanisms by which Mc1r signaling protects the skin against UV injury, this study relies on a mouse model with "humanized skin" based on epidermal expression of stem cell factor (Scf). K14-Scf transgenic mice retain melanocytes in the epidermis and therefore have the ability to deposit melanin in the epidermis. In this animal model, wild type Mc1r status results in robust deposition of black eumelanin pigment and a UV-protected phenotype. In contrast, K14-Scf animals with defective Mc1r signaling ability exhibit a red/blonde pigmentation, very little eumelanin in the skin and a UV-sensitive phenotype. Reasoning that eumelanin deposition might be enhanced by topical agents that mimic Mc1r signaling, we found that direct application of forskolin extract to the skin of Mc1r-defective fair-skinned mice resulted in robust eumelanin induction and UV protection 1. Here we describe the method for preparing and applying a forskolin-containing natural root extract to K14-Scf fair-skinned mice and report a method for measuring UV sensitivity by determining minimal erythematous dose (MED). Using this animal model, it is possible to study how epidermal cAMP induction and melanization of the skin affect physiologic responses to UV exposure.

Epidermal melanin is induced by topical application of forskolin in a murine model of the fair-skinned UV-sensitive human. Pharmacologic manipulation of cAMP levels in the skin and epidermal darkening strongly protect against UV-mediated inflammation (sunburn) as measured by the minimum erythematous dose (MED) assay.

Fairness of skin, UV sensitivity and skin cancer  risk all correlate with the physiologic function of the melanocortin 1  receptor, a Gs-coupled signaling ...

Polymeric Microneedle Array Fabrication by Photolithography

This manuscript describes the fabrication of polymeric microneedle (MN) arrays by photolithography. It involves a simple mold-free process by using a photomask consisting of embedded micro-lenses. Embedded micro-lenses were found to influence MN geometry (sharpness). Robust MN arrays with tip diameters ranging between 41.5 &#181;m &#177; 8.4 &#181;m and 71.6 &#181;m &#177; 13.7 &#181;m, with two different lengths (1,336 &#181;m &#177; 193 &#181;m and 957 &#181;m &#177; 171 &#181;m) were fabricated. These MN arrays may provide potential applications in delivery of low molecular and macromolecular therapeutic agents through skin.

Here, we present a protocol describing a mold-free fabrication process of the polymeric microneedles by photolithography.

This manuscript describes the fabrication of polymeric microneedle (MN) arrays by photolithography. It involves a simple mold-free process by using a ...

Bioengineering

Tumor Treating Field Therapy in Combination with Bevacizumab for the Treatment of Recurrent Glioblastoma

A novel device that employs TTF therapy has recently been developed and is currently in use for the treatment of recurrent glioblastoma (rGBM). It was FDA approved in April 2011 for the treatment of patients 22 years or older with rGBM. The device delivers alternating electric fields and is programmed to ensure maximal tumor cell kill1.
Glioblastoma is the most common type of glioma and has an estimated incidence of approximately 10,000 new cases per year in the United States alone2. This tumor is particularly resistant to treatment and is uniformly fatal especially in the recurrent setting3-5. Prior to the approval of the TTF System, the only FDA approved treatment for rGBM was bevacizumab6. Bevacizumab is a humanized monoclonal antibody targeted against the vascular endothelial growth factor (VEGF) protein that drives tumor angiogenesis7. By blocking the VEGF pathway, bevacizumab can result in a significant radiographic response (pseudoresponse), improve progression free survival and reduce corticosteroid requirements in rGBM patients8,9. Bevacizumab however failed to prolong overall survival in a recent phase III trial26. A pivotal phase III trial (EF-11) demonstrated comparable overall survival between physicians&#8217; choice chemotherapy and TTF Therapy but better quality of life were observed in the TTF arm10.
There is currently an unmet need to develop novel approaches designed to prolong overall survival and/or improve quality of life in this unfortunate patient population. One appealing approach would be to combine the two currently approved treatment modalities namely bevacizumab and TTF Therapy. These two treatments are currently approved as monotherapy11,12, but their combination has never been evaluated in a clinical trial. We have developed an approach for combining those two treatment modalities and treated 2 rGBM patients. Here we describe a detailed methodology outlining this novel treatment protocol and present representative data from one of the treated patients.

A novel methodology that is employed for the treatment of recurrent glioblastomas is described. This treatment approach employs the application of alternating electric tumor treating fields (TTFields), known as TTF Therapy in combination with bevacizumab, a targeted agent that is currently FDA approved as monotherapy.

A novel device that employs TTF therapy has recently been developed and is currently in use for the treatment of recurrent glioblastoma (rGBM). It was FDA ...

Apparatus for Harvesting Tissue Microcolumns

This manuscript describes the production process for a laboratory apparatus, made from off-the-shelf components, that can be used to collect microcolumns of full-thickness skin tissue. The small size of the microcolumns allows donor sites to heal quickly without causing donor site scarring, while harvesting full-thickness tissue enables the incorporation of all cellular and extracellular components of skin tissue, including those associated with deeper dermal regions and the adnexal skin structures, which have yet to be successfully reproduced using conventional tissue engineering techniques. The microcolumns can be applied directly into skin wounds to augment healing, or they can be used as the autologous cell/tissue source for other tissue engineering approaches. The harvesting needles are made by modifying standard hypodermic needles, and they can be used alone for harvesting small amounts of tissue or coupled with a simple suction-based collection system (also made from commonly available laboratory supplies) for high-volume harvesting to facilitate studies in large animal models.

Here we describe a protocol for producing harvesting needles that can be used to collect full-thickness skin tissue without causing donor site scarring. The needles can be combined with a simple collection system to achieve high-volume harvesting.

This manuscript describes the production process for a laboratory apparatus, made from off-the-shelf components, that can be used to collect microcolumns ...

Intense Pulsed Light for the Treatment of Dry Eye Owing to Meibomian Gland Dysfunction

Dry eye disease (DED) is an increasingly common condition and one of the most common complaints of patients. The vast majority of DED is caused by the so-called &#34;evaporative&#34; subtype, that is mainly caused by meibomian gland dysfunction (MGD). Intense pulsed light (IPL) devices employ high intensity pulses of polychromatic lights with a broad range of wavelength (515-1200 nm). IPL treatment has been utilized for years in the field of dermatology, and then its use was applied to ophthalmology for the treatment of MGD. Recently, a new device employing IPL was specifically designed for the periocular application. This procedure determines the thermal selective coagulation and ablation of superficial blood vessels and telangiectasias of the eyelids skin, reducing the release of inflammatory mediators and tear cytokines levels, and improving meibomian glands outflow. IPL treatment is noninvasive and easy to perform, lasts for only a few minutes and can be conducted in an office setting. In the present study, 19 patients underwent 3 sessions of IPL treatment. After treatment, both mean noninvasive break-up time and lipid layer thickness grade significantly increased, as a result of an improvement of tear film stability and quality, respectively. Conversely, no statistically significant changes were found for meibomian gland loss and tear osmolarity. Furthermore, the vast majority of the treated patients (17/19; 89.5% of the total) perceived an improvement of their ocular discomfort symptoms after IPL treatment. Although IPL treatment provides an improvement of both ocular surface parameters and ocular discomfort symptoms after one cycle of three sessions, regular repeated treatments are usually required to maintain the persistence over the time of its beneficial effects.

Dry eye disease is an increasingly common condition, which strongly impair patients&#39; life quality. Recently, a new device employing intense pulsed light, specifically designed for the periocular area, has been shown to improve tear film stability and ocular discomfort symptoms in dry eye disease owing to meibomian gland dysfunction.

Dry eye disease (DED) is an increasingly common condition and one of the most common complaints of patients. The vast majority of DED is caused by the ...

Microneedle Patches for Sustained Delivery of Exosomes in Biomedical Applications

Fabrication and Characterization of Microneedle Patches for Loading and Delivery of Exosomes

Exosomes, as emerging &#34;next-generation&#34; biotherapeutics and drug delivery vectors, hold immense potential in diverse biomedical fields, ranging from drug delivery and regenerative medicine to disease diagnosis and tumor immunotherapy. However, the rapid clearance by traditional bolus injection and poor stability of exosomes restrict their clinical application. Microneedles serve as a solution that prolongs the residence time of exosomes at the administration site, thereby maintaining the drug concentration and facilitating sustained therapeutic effects. In addition, microneedles also possess the ability to maintain the stability of bioactive substances. Therefore, we introduce a microneedle patch for loading and delivering exosomes and share the methods, including isolation of exosomes, fabrication, and characterization of exosome-loaded microneedle patches. The microneedle patches were fabricated using trehalose and hyaluronic acid as the tip materials and polyvinylpyrrolidone as the backing material through a two-step casting method. The microneedles demonstrated robust mechanical strength, with tips able to withstand 2 N. Pig skin was used to simulate human skin, and the tips of microneedles completely melted within 60 s after skin puncture. The exosomes released from the microneedles exhibited morphology, particle size, marker proteins, and biological functions comparable to those of fresh exosomes, enabling dendritic cells uptake and promoting their maturation.

Author Spotlight: Innovative Microneedle-Based Strategies for Enhanced Exosome Delivery and Stability

Exosomes possess significant clinical potential, but their practical application is limited due to easy in vivo clearance and poor stability. Microneedles present a solution by enabling localized delivery by puncturing physiological barriers and dry-state preservation, thereby addressing the limitations of exosome administration and expanding their clinical utility.

Exosomes, as emerging &#34;next-generation&#34; biotherapeutics and drug delivery vectors, hold immense potential in diverse biomedical fields, ranging ...

Stimulation of Stem Cell Niches and Tissue Regeneration in Mouse Skin by Switchable Protoporphyrin IX-Dependent Photogeneration of Reactive Oxygen Species In Situ

Here, we describe a protocol to induce switchable in vivo photogeneration of endogenous reactive oxygen species (ROS) in mouse skin. This transient production of ROS in situ efficiently activates cell proliferation in stem cell niches and stimulates tissue regeneration as strongly manifested through the acceleration of burn healing and hair follicle growth processes. The protocol is based on a regulatable photodynamic treatment that treats the tissue with precursors of the endogenous photosensitizer protoporphyrin IX and further irradiates the tissue with red light under tightly controlled physicochemical parameters. Overall, this protocol constitutes an interesting experimental tool to analyze ROS biology.

The aim of this protocol is to induce transient in vivo production of nonlethal levels of reactive oxygen species (ROS) in mouse skin, further promoting physiological responses in the tissue.

Here, we describe a protocol to induce switchable in vivo photogeneration of endogenous reactive oxygen species (ROS) in mouse skin. This transient ...

Biology

An Efficient Modified Method for Treating Wasting Marmoset Syndrome

Modification of the Treatment Methods for Wasting Marmoset Syndrome with Tranexamic Acid and Supportive Measures

Wasting marmoset syndrome (WMS), a serious disease in captive common marmoset (Callithrix jacchus) colonies, is associated with a high mortality rate. The specific cause of WMS is still unclear and there are few effective treatments. Previously, we had reported a tranexamic acid therapy with supportive measures as a useful treatment for WMS. In the present study, we describe the modified method: a combination of 0.1 mL of 5% tranexamic acid subcutaneously five times per week, 2.0 mL of amino acid formulation intravenously three times per week, 5.0 mL of Ringer's lactate with 0.1 mL of a vitamin formulation subcutaneously three times per week, and oral administration of 0.1 mL of an iron formulation five times per week. We also describe how to administer the solution intravenously via the saphenous vein with a tip of restraining the animal, as well as the detailed methods for oral and subcutaneous administration. The modified methods have comparable efficiency to the original WMS treatment method.

Author Spotlight: Developing a Safer and More Efficient Treatment Protocol for Wasting Marmoset Syndrome (WMS)

Here, we describe the modified treatment methods for wasting marmoset syndrome (WMS, also known as inflammatory bowel disease (IBD)-like disease) with tranexamic acid. We also present how to administer the therapeutic agents orally, subcutaneously, and intravenously.

Wasting marmoset syndrome (WMS), a serious disease in captive common marmoset (Callithrix jacchus) colonies, is associated with a high mortality rate. The ...

Minimizing Pigmentation in Laser Treatment Using Salicylic Acid

Enhancement of Facial Rejuvenation Through a Combination of 1565 nm Non-Ablative Fractional Laser with 30% Supramolecular Salicylic Acid

The 1565 nm non-ablative fractional laser can effectively stimulate collagen regeneration, achieving the effect of skin rejuvenation and anti-aging. In recent years, it has been widely used in dermatology, especially in facial rejuvenation, treatment of stretch marks, and acne scar treatment, with significant results. However, after the 1565 nm non-ablative fractional laser treatment, patients may experience short-term erythema and edema. The inflammation after treatment may also cause pigmentation, making it difficult for some patients to accept. This article introduces a treatment method of 1565 nm non-ablative fractional laser combined with 30% super molecular salicylic acid for facial rejuvenation, effectively reducing posttreatment erythema and post-inflammatory pigmentation. The method includes specific treatment steps and the respective endpoint reactions of each step. The method starts by selecting appropriate energy parameters for the 1565 nm non-ablative fractional laser treatment, followed immediately by the application of 30% supramolecular salicylic acid. The treatment should be stopped when a frosty reaction occurs. The purpose of this project is to provide patients with a safer new method of facial rejuvenation treatment.

Author Spotlight: Advancing Facial Rejuvenation Therapy with Post-Laser Salicylic Acid Application

The purpose of this project is to effectively reduce the occurrence of adverse reactions such as erythema and pigmentation after non-ablative fractional laser treatment by combining 1565 non-ablative fractional laser with 30% super molecular salicylic acid, providing patients with a safer new method of facial rejuvenation treatment.

The 1565 nm non-ablative fractional laser can effectively stimulate collagen regeneration, achieving the effect of skin rejuvenation and anti-aging. In ...