{"doi":"10.1167/tvst.14.11.10","title":"Quantitative Measurements of Stromal and Epithelial Riboflavin With a Novel Transepithelial High-Concentration Riboflavin Soak-and-Rinse Protocol","abstract":"Purpose: To enrich stromal riboflavin concentration and reduce epithelial riboflavin in transepithelial corneal collagen crosslinking (CXL). Methods: Ex vivo experiments on rabbit corneas were performed. The control group followed the standard (epi-off) Dresden CXL: a 30-minute epi-off application of 0.1% riboflavin and 20% dextran. The transepithelial riboflavin solutions consisted of various riboflavin concentrations in 1% hydroxypropyl methylcellulose in 0.45% saline, with or without 0.01% benzalkonium chloride (BAK). The novel soak-and-rinse protocol consists of 0.8% riboflavin with 0.01% BAK and 1% hydroxypropyl methylcellulose (hypotonic) applied for 20 minutes, followed by a 10-minute saline rinse. Stromal and epithelial thicknesses were measured by optical coherence tomography; riboflavin concentrations were quantified by spectrophotometry on 3-mm stromal buttons and epithelial eluates. Statistical analysis employed one-way analysis of variance, linear regression, and one-tailed unpaired t-tests. Results: The 20-minute soak increased stromal riboflavin compared to the 10-minute soak. A 76% higher stromal concentration was achieved by adding BAK to the transepithelial 0.8% riboflavin solution (P < 0.05). The 10-minute rinse achieved a Dresden-equivalent stromal riboflavin level and reduced epithelial riboflavin by 5.9-fold compared to a 20-second rinse (P < 0.0001). Conclusions: Stromal riboflavin concentrations equivalent to those achieved with the epi-off protocol can be achieved by transepithelial application of the novel high-concentration riboflavin formulation. The additional 10-minute rinse effectively reduced epithelial riboflavin levels, facilitating the delivery of ultraviolet light and oxygen into the stroma during CXL. Translational Relevance: The novel transepithelial riboflavin soak-and-rinse protocol may potentially enhance the efficacy of transepithelial CXL by reducing epithelial consumption of ultraviolet light and oxygen and increasing the stromal CXL reaction.","journal":"Translational Vision Science & Technology","year":2025,"id":580886,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":0,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9604,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1447732,"name":"Elizabeth White","orcid":"0000-0001-5090-1729","position":1,"is_corresponding":false},{"id":465027,"name":"Reid L. Wilson","orcid":"0000-0002-4240-2361","position":2,"is_corresponding":false},{"id":644499,"name":"Shuibin Ni","orcid":"0000-0003-3146-5006","position":3,"is_corresponding":false},{"id":525574,"name":"Yifan Jian","orcid":"0000-0001-7988-7385","position":4,"is_corresponding":false},{"id":254821,"name":"Ted S. Acott","orcid":"0000-0002-1445-6012","position":5,"is_corresponding":false},{"id":1155318,"name":"Yan Li","orcid":"0000-0001-9670-6339","position":6,"is_corresponding":false},{"id":418220,"name":"David Huang","orcid":"0000-0003-2592-8393","position":7,"is_corresponding":false},{"id":1492296,"name":"Omkar C Thaware","orcid":null,"position":0,"is_corresponding":true}],"reference_count":25,"raw_metadata":null,"created_at":"2026-07-19T02:58:43.046112Z","pmid":"41222196","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}