| Sign up

PDF (30.8 MB)

Cite

EndNote(RIS) BibTeX

Collect

Collect

Submit Manuscript

Show Outline

Figures (7)

Scheme 1

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Research Article | Open Access

Macro-to-nano dynamic hydrogels in sustained transdermal therapy of diffuse ocular surface malignancies

Jie Chen^{¹^,²^,^§}, Yirong Zeng^{³^,^§}, Hao Tian^{¹^,²^,^§}, Qixiao Guan^{³^,^§}, Ziyue Huang^{¹^,²}, Hongjing Dou^³(), Shiqiong Xu^{¹^,²}(), Renbing Jia^{¹^,²}()

1Department of Ophthalmology, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China

2Shanghai Key Laboratory of Orbital Diseases and Ocular Oncology, Shanghai 200011, China

3The State Key Laboratory of Metal Matrix Composites, School of Materials Science and Engineering, Shanghai Jiao Tong University, Shanghai 200240, China

^§ Jie Chen, Yirong Zeng, Hao Tian, and Qixiao Guan contributed equally to this work.

Show Author Information

Graphical Abstract

View original image Download original image

BRAF/MEKi dual-targeted liposome-doped dynamic hydrogels (B/MLDDHs) serve as a model to treat conjunctival melanoma (CoM). With favorable efficacy and biocompatibility, B/MLDDHs offer a versatile and expandable approach for ocular surface drug delivery applicable to a broader spectrum of ocular surface disorders.

Abstract

Conjunctival melanoma (CοΜ) is of high malignancy that diffusely impacts the ocular surface, which is challenging to surgically remove and pose a life-threatening risk. As one of the most suitable treatment options, drug dropping drugs to the eye surface, however, is limited in its clinical application for CoM treatment due to low drug utilization. Using BRAF/MEK inhibitors (BRAF/MEKi) as model drugs, this study proposes a long-lasting eyedrop based on BRAF/MEK dual-targeted liposome-doped dynamic hydrogels with drugs (B/MLDDHs), which can overcome the tear flushing to succeed in effective ocular surface drug delivery. B/MLDDHs encapsulate drugs within lipid bilayers, enhancing drug bioavailability and biocompatibility. The aldehyde dextran and protonated chitosan-based injectable hydrogel system facilitates drug retention on the ocular surface exerting a sustained liposome release, which significantly enhances the drug bioavailability. In therapeutics, this study demonstrates significantly enhanced in vivo therapeutic effect on orthotopic mouse model of CoM. Given their ability to fulfill permeable and long-term release of hydrophobic drugs, B/MLDDHs may serve as a promising platform for non-invasive delivery of various drug molecules, thereby improving therapeutic outcomes for CoM.

Keywords

hydrogels targeted therapy conjunctival melanoma ocular surface malignancies

Electronic Supplementary Material

Download File(s)

7187_ESM.pdf (1.3 MB)

References

[1]

Khidr, L.; Chen, P. L. RB, the conductor that orchestrates life, death and differentiation. Oncogene 2006, 25, 5210–5219.

Crossref Google Scholar

[2]

Hu, D. N.; Yu, G. P.; McCormick, S. A.; Finger, P. T. Population-based incidence of conjunctival melanoma in various races and ethnic groups and comparison with other melanomas. Am. J. Ophthalmol. 2008, 145, 418–423.E1.

Crossref Google Scholar

[3]

Grimes, J. M.; Shah, N. V.; Samie, F. H.; Carvajal, R. D.; Marr, B. P. conjunctival melanoma: Current treatments and future options. Am. J. Clin. Dermatol. 2020, 21, 371–381.

Crossref Google Scholar

[4]

Rossi, E.; Schinzari, G.; Maiorano, B. A.; Pagliara, M. M.; Di Stefani, A.; Bria, E.; Peris, K.; Blasi, M. A.; Tortora, G. Conjunctival melanoma: Genetic and epigenetic insights of a distinct type of melanoma. Int. J. Mol. Sci. 2019, 20, 5447.

Crossref Google Scholar

[5]

Mikkelsen, L. H. Molecular biology in conjunctival melanoma and the relationship to mucosal melanoma. Acta Ophthalmol 2020, 98 Suppl 115, 1–27.

Crossref Google Scholar

[6]

Mikkelsen, L. H.; Larsen, A. C.; von Buchwald, C.; Drzewiecki, K. T.; Prause, J. U.; Heegaard, S. Mucosal malignant melanoma-a clinical, oncological, pathological and genetic survey. APMIS 2016, 124, 475–486.

Crossref Google Scholar

[7]

Rodrigues, M.; de Koning, L.; Coupland, S. E.; Jochemsen, A. G.; Marais, R.; Stern, M. H.; Valente, A.; Barnhill, R.; Cassoux, N.; Evans, A. et al. So close, yet so far: Discrepancies between uveal and other melanomas. A position paper from UM cure 2020. Cancers (Basel) 2019, 11, 1032.

Crossref Google Scholar

[8]

Jia, S. C.; Zhu, T. Y.; Shi, H. H.; Zong, C. Y.; Bao, Y. Y.; Wen, X. Y.; Ge, S. F.; Ruan, J.; Xu, S. Q.; Jia, R. B. et al. American joint committee on cancer tumor staging system predicts the outcome and metastasis pattern in conjunctival melanoma. Ophthalmology 2022, 129, 771–780.

Crossref Google Scholar

[9]

Larsen, A. C.; lDahl, C.; Dahmcke, C. M.; Lade-Keller, J.; Siersma, V. D.; Toft, P. B.; Coupland, S. E.; Prause, J. U.; Guldberg, P.; Heegaard, S. et al. BRAF mutations in conjunctival melanoma: Investigation of incidence, clinicopathological features, prognosis and paired premalignant lesions. Acta Ophthalmol. 2016, 94, 463–470.

Crossref Google Scholar

[10]

Finger, P. T.; Pavlick, A. C. Checkpoint inhibition immunotherapy for advanced local and systemic conjunctival melanoma: A clinical case series. J. Immunother. Cancer 2019, 7, 83.

Crossref Google Scholar

[11]

Subbiah, V.; Baik, C.; Kirkwood, J. M. Clinical development of BRAF plus MEK inhibitor combinations. Trends Cancer 2020, 6, 797–810.

Crossref Google Scholar

[12]

Wu, Y.; Li, X.; Fu, X. Y.; Huang, X. M.; Zhang, S. R.; Zhao, N.; Ma, X. W.; Saiding, Q.; Yang, M.; Tao, W. et al. Innovative nanotechnology in drug delivery systems for advanced treatment of posterior segment ocular diseases. Adv. Sci. (Weinh) 2024, 11, e2403399.

Crossref Google Scholar

[13]

Wu, Y. M.; Liu, Y. Y.; Li, X. Y.; Kebebe, D.; Zhang, B.; Ren, J.; Lu, J.; Li, J. W.; Du, S. Y.; Liu, Z. D. Research progress of in-situ gelling ophthalmic drug delivery system. Asian J. Pharm. Sci. 2019, 14, 1–15.

Crossref Google Scholar

[14]

Arıcı, M. K.; Arıcı, D. S.; Topalkara, A.; Güler, C. Adverse effects of topical antiglaucoma drugs on the ocular surface. Clin. Exp. Ophthalmol. 2000, 28, 113–117.

Crossref Google Scholar

[15]

Gholizadeh, S.; Wang, Z. Q.; Chen, X.; Dana, R.; Annabi, N. Advanced nanodelivery platforms for topical ophthalmic drug delivery. Drug Discov. Today 2021, 26, 1437–1449.

Crossref Google Scholar

[16]

Zahir-Jouzdani, F.; Khonsari, F.; Soleimani, M.; Mahbod, M.; Arefian, E.; Heydari, M.; Shahhosseini, S.; Dinarvand, R.; Atyabi, F. Nanostructured lipid carriers containing rapamycin for prevention of corneal fibroblasts proliferation and haze propagation after burn injuries: In vitro and in vivo. J. Cell. Physiol. 2019, 234, 4702–4712.

Crossref Google Scholar

[17]

Wang, X. Y.; Chen, L. B.; Wang, X. L.; Zhang, M. M.; Yang, F. M.; Wu, F.; Liu, J. Y.; Lu, L.; Pang, Y. Long-acting protective ocular surface by instilling adhesive dual-antiviral nanoparticles. Adv. Healthc. Mater. 2022, 11, e2200283.

Crossref Google Scholar

[18]

Allen, T. M.; Cullis, P. R. Liposomal drug delivery systems: From concept to clinical applications. Adv. Drug Deliv. Rev. 2013, 65, 36–48.

Crossref Google Scholar

[19]

Blanco, E.; Shen, H. F.; Ferrari, M. Principles of nanoparticle design for overcoming biological barriers to drug delivery. Nat. Biotechnol. 2015, 33, 941–951.

Crossref Google Scholar

[20]

Filipczak, N.; Pan, J. Y.; Yalamarty, S. S. K.; Torchilin, V. P. Recent advancements in liposome technology. Adv. Drug Deliv. Rev. 2020, 156, 4–22.

Crossref Google Scholar

[21]

Shi, J. J.; Kantoff, P. W.; Wooster, R.; Farokhzad, O. C. Cancer nanomedicine: Progress, challenges and opportunities. Nat. Rev. Cancer 2017, 17, 20–37.

Crossref Google Scholar

[22]

van der Koog, L.; Gandek, T. B.; Nagelkerke, A. Liposomes and extracellular vesicles as drug delivery systems: A comparison of composition, pharmacokinetics, and functionalization. Adv. Healthc. Mater. 2022, 11, e2100639.

Crossref Google Scholar

[23]

Battaglia, L.; Gallarate, M. Lipid nanoparticles: State of the art, new preparation methods and challenges in drug delivery. Expert Opin. Drug Deliv. 2012, 9, 497–508.

Crossref Google Scholar

[24]

Wang, Y.; Hu, Y. H.; An, J. Y.; Zhang, H.; Liu, X.; Li, X. R.; Zhang, Z. Z.; Zhang, X. M. Liposome-based permeable eyedrops for effective posterior segment drug delivery. Adv. Funct. Mater. 2024, 34, 2403142.

Crossref Google Scholar

[25]

Li, X. Y.; Gong, J. P. Design principles for strong and tough hydrogels. Nat. Rev. Mater. 2024, 9, 380–398.

Crossref Google Scholar

[26]

Wang, J.; Su, W.; Zhang, T. T.; Zhang, S. S.; Lei, H. W.; Ma, F. D.; Shi, M. N.; Shi, W. J.; Xie, X. D.; Di, C. X. Aberrant cyclin D1 splicing in cancer: From molecular mechanism to therapeutic modulation. Cell Death Dis. 2023, 14, 244.

Crossref Google Scholar

[27]

Feng, W. B.; Huang, W. J.; Chen, J.; Qiao, C. Y.; Liu, D. F.; Ji, X. Y.; Xie, M.; Zhang, T. Y.; Wang, Y. J.; Sun, M. Y. CXCL12-mediated HOXB5 overexpression facilitates colorectal cancer metastasis through transactivating CXCR4 and ITGB3. Theranostics 2021, 11, 2612–2633.

Crossref Google Scholar

[28]

Hamidi, H.; Ivaska, J. Every step of the way: Integrins in cancer progression and metastasis. Nat. Rev. Cancer 2018, 18, 533–548.

Crossref Google Scholar

[29]

Yousefi, H.; Vatanmakanian, M.; Mahdiannasser, M.; Mashouri, L.; Alahari, N. V.; Monjezi, M. R.; Ilbeigi, S.; Alahari, S. K. Understanding the role of integrins in breast cancer invasion, metastasis, angiogenesis, and drug resistance. Oncogene 2021, 40, 1043–1063.

Crossref Google Scholar

[30]

Zheng, Z. Q.; Li, Z. X.; Zhou, G. Q.; Lin, L.; Zhang, L. L.; Lv, J. W.; Huang, X. D.; Liu, R. Q.; Chen, F. P.; He, X. J. et al. Long noncoding RNA FAM225A promotes nasopharyngeal carcinoma tumorigenesis and metastasis by acting as ceRNA to sponge miR-590-3p/miR-1275 and upregulate ITGB3. Cancer Res. 2019, 79, 4612–4626.

Crossref Google Scholar

[31]

Yuan, J. M.; Dong, X. D.; Yap, J.; Hu, J. C. The MAPK and AMPK signalings: Interplay and implication in targeted cancer therapy. J. Hematol. Oncol. 2020, 13, 113.

Crossref Google Scholar

[32]

Chang, L.; Jung, N. Y.; Atari, A.; Rodriguez, D. J.; Kesar, D.; Song, T. Y.; Rees, M. G.; Ronan, M.; Li, R. T.; Ruiz, P. et al. Systematic profiling of conditional pathway activation identifies context-dependent synthetic lethalities. Nat. Genet. 2023, 55, 1709–1720.

Crossref Google Scholar

[33]

Yang, R. H.; Tang, S. C.; Xie, X. L.; Jin, C. F.; Tong, Y. H.; Huang, W. J.; Zan, X. J. Enhanced ocular delivery of beva via ultra-small polymeric micelles for noninvasive anti-VEGF therapy. Adv. Sci. (Weinh) 2024, 36, e2314126.

Crossref Google Scholar

[34]

Chen, L. B.; Wu, F.; Pang, Y.; Yan, D.; Zhang, S. Y.; Chen, F. J.; Wu, N. X.; Gong, D. N.; Liu, J. Y.; Fu, Y. et al. Therapeutic nanocoating of ocular surface. Nano Today 2021, 41, 101309.

Crossref Google Scholar

[35]

Chen, L. B.; Yan, D.; Wu, N. X.; Yao, Q. K.; Sun, H.; Pang, Y.; Fu, Y. Injectable bio-responsive hydrogel for therapy of inflammation related eyelid diseases. Bioact. Mater. 2021, 6, 3062–3073.

Crossref Google Scholar

[36]

Geroski, D. H.; Edelhauser, H. F. Transscleral drug delivery for posterior segment disease. Adv. Drug Deliv. Rev. 2001, 52, 37–48.

Crossref Google Scholar

[37]

Gumbiner, B. Structure, biochemistry, and assembly of epithelial tight junctions. Am. J. Physiol. 1987, 253, C749–C758.

Crossref Google Scholar

[38]

Peng, C.; Kuang, L. J.; Zhao, J. Y.; Ross, A. E.; Wang, Z. Q.; Ciolino, J. B. Bibliometric and visualized analysis of ocular drug delivery from 2001 to 2020. J. Control. Release 2022, 345, 625–645.

Crossref Google Scholar

[39]

Wang, K.; Jiang, L.; Zhong, Y. Y.; Zhang, Y.; Yin, Q. C.; Li, S.; Zhang, X. B.; Han, H. J.; Yao, K. Ferrostatin-1-loaded liposome for treatment of corneal alkali burn via targeting ferroptosis. Bioeng. Transl. Med. 2022, 7, e10276.

Crossref Google Scholar

[40]

Yang, J.; Bahreman, A.; Daudey, G.; Bussmann, J.; Olsthoorn, R. C. L.; Kros, A. Drug delivery via cell membrane fusion using lipopeptide modified liposomes. ACS Cent. Sci. 2016, 2, 621–630.

Crossref Google Scholar

[41]

de Salamanca, A. E.; Diebold, Y.; Calonge, M.; García-Vazquez, C.; Callejo, S.; Vila, A.; Alonso, M. J. Chitosan nanoparticles as a potential drug delivery system for the ocular surface: Toxicity, uptake mechanism and in vivo tolerance. Invest. Ophthalmol. Vis. Sci. 2006, 47, 1416–1425.

Crossref Google Scholar

[42]

Wang, Y. Y.; Zhou, L.; Fang, L.; Cao, F. Multifunctional carboxymethyl chitosan derivatives-layered double hydroxide hybrid nanocomposites for efficient drug delivery to the posterior segment of the eye. Acta Biomater. 2020, 104, 104–114.

Crossref Google Scholar

[43]

Li, Y.; Liu, S. Y.; Zhang, J. J.; Wang, Y. M.; Lu, H. J.; Zhang, Y. X.; Song, G. Z.; Niu, F. H.; Shen, Y. F.; Midgley, A. C. et al. Elastic porous microspheres/extracellular matrix hydrogel injectable composites releasing dual bio-factors enable tissue regeneration. Nat Commun. 2024, 15, 1377.

Crossref Google Scholar

[44]

Chen, S. L.; Luo, Y.; He, Y.; Li, M.; Liu, Y. J.; Zhou, X. S.; Hou, J. W.; Zhou, S. B. In-situ-sprayed therapeutic hydrogel for oxygen-actuated Janus regulation of postsurgical tumor recurrence/metastasis and wound healing. Nat. Commun. 2024, 15, 814.

Crossref Google Scholar

[45]

Geng, S. Z.; Guo, P. K.; Wang, J.; Zhang, Y. Y.; Shi, Y. R.; Li, X. L.; Cao, M. N.; Song, Y. T.; Zhang, H. L.; Zhang, Z. Z. et al. Ultrasensitive optical detection and elimination of residual microtumors with a postoperative implantable hydrogel sensor for preventing cancer recurrence. Adv Mater. 2024, 36, 2307923.

Crossref Google Scholar

[46]

Wang, D. Y.; Liu, J. W.; Duan, J.; Yi, H.; Liu, J. J.; Song, H. W.; Zhang, Z. Z.; Shi, J. J.; Zhang, K. X. Enrichment and sensing tumor cells by embedded immunomodulatory DNA hydrogel to inhibit postoperative tumor recurrence. Nat. Commun. 2023, 14, 4511.

Crossref Google Scholar

[47]

Nam, S.; Mooney, D. Polymeric tissue adhesives. Chem. Rev. 2021, 121, 11336–11384.

Crossref Google Scholar

Volume 18 Issue 3,
March 2025

Article number: 94907187

DOI: 10.26599/NR.2025.94907187

Cite this article:

Chen J, Zeng Y, Tian H, et al. Macro-to-nano dynamic hydrogels in sustained transdermal therapy of diffuse ocular surface malignancies. Nano Research, 2025, 18(3): 94907187. https://doi.org/10.26599/NR.2025.94907187

Scheme 1Sketch map of construction of B/MLDDHs and how they efficiently prevent the proliferation and migration of CoM, leading to a boosted therapeutic effect. (a) The construction of B/MLDDHs and the detailed illustration of slow-release B/MLDDHs for treating CoM. (b) The molecular mechanisms by which B/MLDDHs boost active internalization through endocytosis and inhibits tumor proliferation and migration by regulating CCND1 and ITGβ3.
Figure 1BRAF/MEKi are identified as a promising therapeutic strategy in CoM cell line. (a) and (b) CCK8 assay showed good safety profiles in normal cells and significant cytotoxic effects on tumor cells, (a) melanocyte cell lines (PIG1) and (b) human CoM cell lines (CM2005.1). (c) Colony formation assay to evaluate the proliferation of tumor cells with monotherapy or combined BRAF/MEKi. (d) Wound healing assay to assess the migration rates of tumor cells treated with either monotherapy or a combination of BRAF/MEK inhibitors. (e) Statistical results of migration rates of tumor cells, **p < 0.01, ****p < 0.0001.
Figure 2BRAF/MEKi impede cell cycle and cell adhesion, thereby significantly curtailing the proliferation and migration of CoM. (a) GO enrichment of different expression genes in D&T group compared to Control group, and fold change > 1.5 were included. (b) KEGG analysis of different expression genes between D&T group and Control group. (c) Volcano plot of different expression genes between D&T group and NC group. (d) and (e) GSEA analysis performed in cell cycle and focal adhesion pathway. (f) and (g) Integrative Genomics Viewer (IGV) tracks for CCND1 and ITGβ3 from RNA-seq data in Control and D&T groups. Data were collected from three independent biological replicates.
Figure 3Characterizations of BRAF/MEKi dual-targeted Liposome(B/ML) and BRAF/MEKi dual-targeted liposome-doped dynamic hydrogels (B/MLDDHs). (a) Malvern particle size analyzer to measure B/ML particle size. (b) Cryogenic TEM image of B/ML, scale bar: 100 nm. (c) and (d) Rheology test scanned by time and frequency to assess self-healing and hydrogel characters. (e) and (f) Cyro-SEM images showing the microstructures of B/MLDDHs with varient magnifications. (e) Scale bar: 50 μm, (f) Scale bar: 2 μm. (g) Hoechst staining in CoM cell lines treated with liposomes labeled with Cy3 dye (LIPO group) or PBS-based Cy3 dye (Control group) exhibiting equal fluorescence intensity for 24 h, scale bar: 50 μm.
Figure 4In-vitro cytotoxicity and specificity of BRAF/MEKi dual-targeted Liposome(B/ML) in CoM cells. (a) Colony formation assay to evaluate the proliferation of CM2005.1 treated with D&T or B/ML. (b) and (c) Representative flow cytometry images of Annexin V/PI-stained cell apoptosis assay and cell cycle assay following 48-h treatment with either 5 nM DMSO, 5 nM Dabrafenib & 5 nM Trametinib or 0.5 μg/mL Β/ΜL (equipotent D&T concentration nearly 5 nM Dabrafenib & 5 nM Trametinib). (d) Western blot to perform the expression of CCND1, ITGβ3, and factors in the MAPK pathway of CM2005.1 cells treated by either 5 nM DMSO, 5 nM Dabrafenib & 5 nM Trametinib or 0.5 μg/mL Β/ΜL for 48 h. (e) Co-culture of normal melanocytes and CoM cells. PIG1 (green) cells were respectively co-cultured with CM2005.1 (red). PBS, 5 nM Dabrafenib & 5 nM Trametinib, 0.5 μg/mL Β/ΜL, Gel, Gel embedded 5 nM Dabrafenib & 5 nM Trametinib, B/ML embedded with 0.5 μg/mL Β/ΜL were administered in each group separately. Scale bar: 50 μm.
Figure 5In vivo biocompatibility and ocular retention rate assessment of B/MLDDHs. (a) Under physiological conditions, the in vivo biocompatibility of PBS-eyedrop-treated and 125 μg/mL B/MLDDHs-eyedrop-treated groups were assessed on day 6 using slit-lamp examination, fluorescein sodium staining, and H&E staining, scale bar: 50 μm for H&E staining images. (b) Quantification (n = 3) of the central corneal thickness after 5-day-topical administration of PBS or B/MLDDHs. (c) and (d) The IVIS imaging and quantification (n = 3) of fluorescence signal at different time points under different treatments, results were presented as the mean ± SD, ***p < 0.001.
Figure 6Antitumor efficacy of B/MLDDHs in CoM orthotopic model. (a) Schematic illustration of CoM orthotopic model and treatment. (b) Timeline of the in vivo experiments in CoM orthotopic model. (c) The digital image of orthotopic tumor burden from different group. One mouse of D&T group died before reaching the endpoint of treatment. (d)–(f) Quantification data of tumor size, relative tumor size, and tumor weight from different group, *p < 0.05, **p < 0.01. (g) H&E and IHC analysis of ITGβ3/CCND1/Ki67 protein expressions in CoM mouse model after different treatments, scale bar: 200 μm.

About Us

Learn about Open Access

Tsinghua University Press

Publish with Us

Peer Review Policy

Copyright and Licensing

Article Processing Charge

Contact Us

Journal Collaboration: Yao Meng (Ms.)✉️ +86-10-83470574

Technical Support: Kuo Zhao (Mr.)✉️ +86-10-83470507

Media Contact: Hao Jin (Mr.)✉️ +86-10-83470559

Address: Floor 6, Tower B, Xueyan Building, Shuangqing Road, Haidian District, Beijing 100084, China.

SciOpen——中国科技期刊卓越行动计划支持项目

Copyright © 2025 Tsinghua University Press Ltd.

京ICP备 10035462号-42 京公网安备11010802044758号