Therapeutic Efficacy of 308-nm Excimer Laser Combined with Matrine in Vitiligo by Modulation of Serum Immunoglobulins and Inflammatory Cytokines

Summary

The combined use of a 308-nm excimer laser and oral matrine markedly enhanced repigmentation in vitiligo by promoting melanocyte regeneration and restoring immune balance. This dual-modality therapy reduced pro-inflammatory cytokines, normalized immunoglobulins, elevated IL-10, and achieved superior clinical efficacy with excellent safety and patient tolerance.

Abstract

Vitiligo is a chronic autoimmune depigmenting disorder characterized by the loss of functional melanocytes and aberrant immune activation. Despite advances in phototherapy, clinical responses remain inconsistent, largely due to persistent inflammatory activity and inadequate immune regulation. This study investigated the therapeutic efficacy and immunologic mechanisms of combining 308-nm excimer laser phototherapy with oral matrine, a natural alkaloid possessing anti-inflammatory and immunoregulatory properties. A total of 232 patients with non-segmental vitiligo were prospectively assigned to receive either excimer laser monotherapy or combination therapy for 12 weeks. Clinical improvement was evaluated using the Vitiligo Area Scoring Index (VASI), and serum immunoglobulins and cytokines were analyzed to assess systemic immune modulation. Both regimens produced significant repigmentation, yet the combination therapy achieved a more pronounced VASI reduction (from 9.72 ± 3.24 at baseline to 5.76 ± 2.18 after treatment) than the excimer laser alone (from 9.68 ± 3.37 to 6.63 ± 2.34, p < 0.001). The proportion of patients demonstrating excellent or good repigmentation was notably higher with the combined regimen (76.7%) compared with monotherapy (58.7%). In parallel, serum IgG levels declined markedly (13.72 ± 2.64 g/L to 11.85 ± 2.18 g/L, p < 0.001), whereas IgA levels rose modestly (2.07 ± 0.68 g/L to 2.23 ± 0.61 g/L, p = 0.041). Pro-inflammatory cytokines including IFN-γ and IL-17 were significantly reduced (both p < 0.001), while anti-inflammatory IL-10 increased from 8.3 ± 3.2 to 12.9 ± 3.5 pg/mL (p < 0.001). VASI improvement correlated inversely with IFN-γ (r = –0.482, p < 0.05) and IL-17 (r = –0.451, p < 0.001) changes, and positively with IL-10 elevation (r = 0.503, p < 0.001). These findings demonstrate that matrine enhances excimer laser efficacy by promoting melanocyte regeneration and restoring immune equilibrium, establishing a biomarker-based dual-modality strategy for precise and durable management of vitiligo.

Introduction

Vitiligo is a chronic, acquired depigmenting skin disorder characterized by the selective destruction or dysfunction of epidermal melanocytes, leading to the development of well-defined white macules of variable size and distribution. It affects approximately 0.5%–2% of the global population, without gender or racial predilection, and often presents before the age of 30^1,2. Although the disease is not life-threatening, it imposes a substantial psychosocial and emotional burden, significantly impairing self-esteem, social functioning, and quality of life³. The pathogenesis of vitil....

Protocol

This study was conducted in accordance with the ethical principles of the Declaration of Helsinki and the institutional regulations governing human research. The study was conducted in accordance with the ethical principles of the Declaration of Helsinki and approved by the Ethics Committee of Xingtai People’s Hospital, Department of Dermatology (Approval ID: 2021[029]). All participants provided written informed consent before participation.

Patient selection
A total of 232 patients clinically diagnosed with vitiligo were enrolled over a 3-year period from January 2021 to December 2023. All participants were di....

Results

Baseline clinical characteristics demonstrated comparable demographic and disease profiles between groups
A total of 232 patients with non-segmental vitiligo were enrolled and evenly allocated to the excimer group (n = 116) or the combination therapy group receiving laser plus oral matrine (n = 116). Baseline demographic and clinical parameters, including age (35.8 ± 6.0 vs. 35.4 ± 5.6 years), sex distribution (male/female: 59/57 vs. 59/57), disease duration (4.8 ± 1.0 vs. .......

Discussion

This study introduces a combined therapeutic approach integrating a 308-nm excimer laser with oral matrine for the treatment of non-segmental vitiligo. The central proposition is that dual modulation—local photobiological stimulation of melanocytes coupled with systemic immunoregulation—can achieve more robust and sustained repigmentation than the excimer group. The findings support this hypothesis: the combination therapy produced earlier perifollicular pigmentation, greater VASI reduction, and higher patien.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
308-nm XeCl Excimer Laser System	STRATA Skin Sciences (XTRAC)	Velocity 400 (Model)	Targeted 308-nm UVB laser for vitiligo lesions; verify model at your site; or equivalent device
–80 °C Ultra-Low Freezer	Thermo Scientific	Forma 900 Series (Model 980)	Long-term serum storage
96-Well ELISA Plates (High-Bind)	Corning Costar	9018	If using in-house coating or single-plex workflows
Adjustable Medical Chair	Midmark	641 Procedure Chair	Patient positioning for consistent irradiation distance
Aloe-based Soothing Gel	3M	9236	Thin post-irradiation layer for erythema relief; or equivalent medical gel
Amber Glass Bottle (500 mL)	Fisherbrand	02-897-2J	Light-protected storage at 4 °C for matrine solution
Analytical Balance (0.1 mg)	METTLER TOLEDO	ME104E	Gravimetric prep and QC
Automated Immunonephelometer	Siemens Healthineers	BN ProSpec System	Quantifies IgG/IgA/IgM per manufacturer’s IFU
Barcode Labels (Cryo-safe)	Diversified Biotech	CRY-1000	Sample tracking and chain-of-custody
Biotinylated Detection Antibody Set	MSD	Included with panel	Matched to capture antibodies
Broad-spectrum Sunscreen SPF ≥30	Neutrogena	070501017052 (Ultra Sheer SPF 50)	Post-session photoprotection; outpatient counseling item
Calibrators for Immunoglobulins	Siemens Healthineers	N Protein Standard SL	Traceable calibration for nephelometry
Control Sera (High/Low) for Ig Assays	Siemens Healthineers	N/T Protein Controls	Daily QC of nephelometric runs
Cryogenic Vials, 0.5 mL	Thermo Scientific Nunc	375418	Serum aliquots; external thread; DNase/RNase-free
Dermatoscope	DermLite	DL4	Polarized handheld dermatoscope for lesion inspection
Dermoscopy/Photography Scale Card	3Gen	Scale/Color Reference Card	Ensures color consistency across timepoints
Digital Camera for Standardized Images	Canon	EOS 80D	Fixed distance 60 cm; exposure 1/60 s; ISO 200; tripod recommended
Fixed-Angle Refrigerated Centrifuge	Eppendorf	5810 R	800 × g, 10 min, 4 °C serum separation
HPLC Column (C18, 4.6×250 mm, 5 µm)	Agilent	ZORBAX Eclipse Plus C18, 959990-902	Routine alkaloid separation; or equivalent
HPLC System	Agilent	1260 Infinity II (G7111B)	Purity verification and batch standardization of matrine
Image Analysis / Measurement	ImageJ (NIH)	1.54 (RRID:SCR_003070)	Optional lesion area quantification from photographs
Lint-free Medical Gauze (Sterile)	Johnson & Johnson	546112	Drying treatment area post-cleansing
Matrine (≥98%, HPLC)	MedChemExpress	HY-N0156	Oral solution preparation; verify local supplier and pharmacopeia compliance
Microplate Reader (450 nm)	BioTek (Agilent)	Synergy H1	Absorbance at 450 nm; pathlength correction
Microplate Washer	BioTek (Agilent)	ELx405	Automated wash cycles; reduces variability
Multiplex Human Cytokine Panel (Custom 6-plex)	Meso Scale Discovery (MSD)	U-PLEX Platform (K15067L)	Includes IFN-γ, TNF-α, IL-1β, IL-6, IL-17A, IL-10; or equivalent
PBS Tablets	Gibco	18912-014	PBS preparation for washes/buffers
Plate Sealer Films	Bio-Rad	MSB1001	Prevents evaporation during incubations
Protective UV Goggles (operator)	Uvex (Honeywell)	S0360X Ultra-Spec 2000	Operator eye protection; use dedicated pair
Protective UV Goggles (patient)	Uvex (Honeywell)	S0360X Ultra-Spec 2000	UV-blocking eyewear for patient safety during irradiation
Serum Separation Tubes (SST II Advance)	BD Vacutainer	367955	Fasting blood collection at baseline and week 12
Statistical Software	IBM	SPSS Statistics v26.0 (RRID:SCR_002865)	Primary data analysis per protocol
Sterile Physiological Saline (0.9%)	Baxter	2B1324 (500 mL)	Lesion cleansing prior to each laser session
Stop Solution (2 M H2SO4)	Sigma-Aldrich	258105	Stops TMB reaction
Streptavidin-HRP	Thermo Scientific	N100	Detection reagent; kit-compatible
TMB Substrate (ELISA)	Thermo Scientific	34028	Chromogenic development; 10 min in dark
Tripod with Distance Marker	Manfrotto	MKCOMPACTLT-BK	Maintains constant camera-to-subject distance
Tween-20 (for PBST 0.05%)	Sigma-Aldrich	P1379	Wash buffer surfactant
UV Radiometer / Power Meter	International Light Technologies	ILT2400	For daily output verification and MED testing; with SED240/NS254 detector or equivalent
UV-Blocking Opaque Drape/Sheet	Thorlabs	BK5 Blackout Fabric	Covers surrounding normal skin to avoid unintended exposure; or equivalent
VASI Scoring Sheets (Printed)	In-house	NA-VASI-2025	Standardized clinical scoring forms; archive with subject ID
Wood’s Lamp (UV-A, 365 nm)	Spectroline	ENF-260C	Clinical diagnosis and lesion edge assessment
Wood’s Lamp Shield Hood	DermLite	DL Hood	Improves contrast under UV-A exam