CO₂ fractional laser-assisted transdermal delivery of silk nanofiber carriers in a rabbit ear hypertrophic scar model

Abstract

Background: Hypertrophic scars are skin fibrotic diseases, characterized by fibroblast hyperproliferation and excessive accumulation of extracellular matrix. However, topical drug application for hypertrophic scars are unsatisfactory. The purpose of this study was to explore the permeability of silk nanofiber hydrogels (SNFs) loaded with rhodamine 6G (R6G) and rhodamine 110 (R110) mediated by CO₂ fractional laser irradiation into hypertrophic scar tissues. Methods: In this work, R6G and R110 were chosen as hydrophilic and hydrophobic model molecules. They were loaded inside SNFs. In vivo rabbit ear hypertrophic scars were treated with CO₂ fractional laser irradiation and then R6G/R110-laden SNFs were applied to the scars to evaluate their synergetic effect on drug penetration efficiency. Their permeability was quantified by fluorescence intensity and measured by confocal laser scanning microscopy on days 1, 3, 5 and 7. More specifically, the thermal coagulation zone (CZ) and its surrounding area (peri-CZ) caused by the thermal coagulation of the laser were discussed separately. Results: Our data indicated that the SNFs promoted the penetration of R6G but not that of R110 in the peri-CZ on day 1 when combined with laser irradiation. Interestingly, both R6G and R110 were abundant in the CZ and remained stable on days 1, 3 and 5. Moreover, rapid re-epithelialization hindered the long-term permeability of both drugs. Conclusion: Combining CO₂ fractional laser irradiation with SNF drug delivery could improve the efficiency of hydrophilic drug delivery within 24 h before total re-epithelialization.

Keywords

CO₂ fractional laser / Hypertrophic scar / Silk nanofiber hydrogels / Transdermal delivery / Laser-assisted drug delivery

References

[1] Ogawa R.Keloid and hypertrophic scars are the result of chronic inflammation in the reticular dermis. Int J Mol Sci. 2017; 18: 606.

[2] Do NN, Eming SA. Skin fibrosis: models and mechanisms. Curr Res Transl Med. 2016; 64: 185-93.

[3] Finnerty CC, Jeschke MG, Branski LK, Barret JP, Dziewulski P, Herndon DN. Hypertrophic scarring: the greatest unmet challenge after burn injury. Lancet. 2016; 388: 1427-36.

[4] Sen CK, Gordillo GM, Roy S, Kirsner R, Lambert L, Hunt TK, et al. Human skin wounds: a major and snowballing threat to public health and the economy. Wound Repair Regen. 2009; 17: 763-71.

[5] Ward RE, Sklar LR, Eisen DB. Surgical and noninvasive modalities for scar revision. Dermatol Clin. 2019; 37: 375-86.

[6] Berman B, Maderal A, Raphael B. Keloids and hypertrophic scars: pathophysiology, classification, and treatment. Dermatol Surg. 2017; 43: S3-18.

[7] Zouboulis CC, Wild T, Zouboulis VA, Har-Shai Y. Intralesional cryosurgery of keloids: required treatment hold time. Br J Dermatol. 2021; 184: 173-5.

[8] Lee Peng G, Kerolus JL. Management of Surgical Scars. Facial Plast Surg Clin North Am. 2019; 27: 513-7.

[9] Chen Z, Chen Z, Pang R, Wei Z, Zhang H, Liu W, et al. The effect of botulinum toxin injection dose on the appearance of surgical scar. Sci Rep. 2021; 11: 13670.

[10] Hietanen KE, Järvinen TA, Huhtala H, Tolonen TT, Kuokkanen HO, Kaartinen IS. Treatment of keloid scars with intralesional triamcinolone and 5-fluorouracil injections-a randomized controlled trial. J Plast Reconstr Aesthet Surg. 2019; 72: 4-11.

[11] Prausnitz MR, Langer R. Transdermal drug delivery. Nat Biotechnol. 2008; 26: 1261-8.

[12] Karppinen SM, Heljasvaara R, Gullberg D, Tasanen K, Pihlajaniemi T. Toward understanding scarless skin wound healing and pathological scarring. F1000Res. 2019; 8: 787.

[13] Ali FR, Al-Niaimi F. Treatment of nonmelanoma skin cancers using laser-assisted drug delivery. Dermatol Surg. 2018; 44: 310.

[14] Alsalhi W, Alalola A, Randolph M, Gwillim E, Tosti A. Novel drug delivery approaches for the management of hair loss. Expert Opin Drug Deliv. 2020; 17: 287-95.

[15] Hsiao CY, Yang SC, Alalaiwe A, Fang JY. Laser ablation and topical drug delivery: a review of recent advances. Expert Opin Drug Deliv. 2019; 16: 937-52.

[16] Alegre-Sanchez A, Jimenez-Gomez N, Boixeda P. Laser-assisted drug delivery. Actas Dermo-Sifilogr. 2018; 109: 858-67.

[17] Huang C, Li P, Wang B, Deng Y, Li J, Mao M, et al. Multi-factors associated with efficacy and adverse events of fractional erbium:YAG laser-assisted delivery of topical betamethasone for stable vitiligo: a retrospective analysis. Lasers Surg Med. 2020; 52: 590-6.

[18] Badawi AM, Osman MA. Fractional erbium-doped yttrium aluminum garnet laser-assisted drug delivery of hydroquinone in the treatment of melasma. Clin Cosmet Investig Dermatol. 2018;Volume 11: 13-20.

[19] Al Janahi S, Lee M, Lam C, Chung HJ. Laser-assisted drug delivery in the treatment of keloids: a case of extensive refractory keloids successfully treated with fractional carbon dioxide laser followed by topical application and intralesional injection of steroid suspension. JAAD case reports. 2019; 5: 840-3.

[20] Todorova K, Mandinova A. Novel approaches for managing aged skin and nonmelanoma skin cancer. Adv Drug Deliv Rev. 2020; 153: 18-27.

[21] Miletta N, Siwy K, Hivnor C, Clark J, Shofner J, Zurakowski D, et al. Fractional ablative laser therapy is an effective treatment for hypertrophic burn scars: a prospective study of objective and subjective outcomes. Ann Surg. 2021; 274: e574-e80.

[22] Banzhaf CA, Lin LL, Dang N, Freeman M, Haedersdal M, Prow TW. The fractional laser-induced coagulation zone characterized over time by laser scanning confocal microscopy-a proof of concept study. Lasers Surg Med. 2018; 50: 70-7.

[23] Banzhaf CA, Ortner VK, Philipsen PA, Haedersdal M. The ablative fractional coagulation zone influences skin fluorescence intensities of topically applied test molecules-an in vitro study with fluorescence microscopy and fluorescence confocal microscopy. Lasers Surg Med. 2019; 51: 68-78.

[24] Wenande E, Olesen UH, Nielsen MM, Janfelt C, Hansen SH, Anderson RR, et al. Fractional laser-assisted topical delivery leads to enhanced, accelerated and deeper cutaneous 5-fluorouracil uptake. Expert Opin Drug Deliv. 2017; 14: 307-17.

[25] Banzhaf CA, Thaysen-Petersen D, Bay C, Philipsen PA, Mogensen M, Prow T, et al. Fractional laser-assisted drug uptake: impact of time-related topical application to achieve enhanced delivery. Lasers Surg Med. 2017; 49: 348-54.

[26] Zhang Z, Chen J, Huang J, Wo Y, Zhang Y, Chen X. Experimental study of 5-fluorouracil encapsulated Ethosomes combined with CO2 fractional laser to treat hypertrophic scar. Nanoscale Res Lett. 2018; 13: 26.

[27] Krishnan V, Mitragotri S. Nanoparticles for topical drug delivery: potential for skin cancer treatment. Adv Drug Deliv Rev. 2020; 153: 87-108.

[28] Farokhi M, Mottaghitalab F, Reis RL, Ramakrishna S, Kundu SC. Functionalized silk fibroin nanofibers as drug carriers: advantages and challenges. J Control Release. 2020; 321: 324-47.

[29] Liu L, Ding Z, Yang Y, Zhang Z, Lu Q, Kaplan DL. Asiaticoside-laden silk nanofiber hydrogels to regulate inflammation and angiogenesis for scarless skin regeneration. Biomater Sci. 2021; 9: 5227-36.

[30] Ding Z, Zhang Y, Guo P, Duan T, Cheng W, Guo Y, et al. Injectable Desferrioxamine-laden silk nanofiber hydrogels for accelerating diabetic wound healing. ACS Biomater Sci Eng. 2021; 7: 1147-58.

[31] Zhang X, Zhang Z, Xiao L, Ding Z, He J, Lu G, et al. Natural nanofiber shuttles for transporting hydrophobic cargo into aqueous solutions. Biomacromolecules. 2020; 21: 1022-30.

[32] Bai S, Zhang X, Lu Q, Sheng W, Liu L, Dong B, et al. Reversible hydrogel-solution system of silk with high beta-sheet content. Biomacromolecules. 2014; 15: 3044-51.

[33] Taudorf EH, Haak CS, Erlendsson AM, Philipsen PA, Anderson RR, Paasch U, et al. Fractional ablative erbium YAG laser: histological characterization of relationships between laser settings and micropore dimensions. Lasers Surg Med. 2014; 46: 281-9.

[34] Wenande E, Tam J, Bhayana B, Schlosser SK, Ishak E, Farinelli WA, et al. Laser-assisted delivery of synergistic combination chemotherapy in in vivo skin. J Control Release. 2018; 275: 242-53.

[35] Wenande E, Anderson RR, Haedersdal M. Fundamentals of fractional laser-assisted drug delivery: an in-depth guide to experimental methodology and data interpretation. Adv Drug Deliv Rev. 2020; 153: 169-84.

[36] Yu J, Kalaria DR, Kalia YN. Erbium:YAG fractional laser ablation for the percutaneous delivery of intact functional therapeutic antibodies. J Control Release. 2011; 156: 53-9.

[37] Takeuchi I, Shimamura Y, Kakami Y, Kameda T, Hattori K, Miura S, et al. Transdermal delivery of 40-nm silk fibroin nanoparticles. Colloids Surf B Biointerfaces. 2019; 175: 564-8.

[38] Xiao L, Ding Z, Zhang X, Wang X, Lu Q, Kaplan DL. Silk Nanocarrier size optimization for enhanced tumor cell penetration and cytotoxicity in vitro. ACS Biomater Sci Eng. 2022; 8: 140-50.

[39] Wenande E, Olesen UH, Boesen MR, Persson DP, Lerche CM, Stürup S, et al. Laser-assisted delivery enhances topical uptake of the anticancer agent cisplatin. Drug Deliv. 2018; 25: 1877-85.

[40] Taudorf EH. Laser-assisted delivery of topical methotrexate-in vitro investigations. Dan Med J. 2016; 63: B5254.

[41] Olesen UH, Clergeaud G, Lerche CM, Andresen TL, Haedersdal M. Topical delivery of vismodegib using ablative fractional laser and micro-emulsion formulation in vitro. Lasers Surg Med. 2019; 51: 79-87.

[42] Banzhaf CA, Wind BS, Mogensen M, Meesters AA, Paasch U, Wolkerstorfer A, et al. Spatiotemporal closure of fractional laser-ablated channels imaged by optical coherence tomography and reflectance confocal microscopy. Lasers Surg Med. 2016; 48: 157-65.

[43] Haak CS, Hannibal J, Paasch U, Anderson RR, Haedersdal M. Laser-induced thermal coagulation enhances skin uptake of topically applied compounds. Lasers Surg Med. 2017; 49: 582-91.

[44] Manuskiatti W, Kaewkes A, Yan C, Ng JN, Glahn JZ, Wanitphakdeedecha R. Hypertrophic scar outcomes in fractional laser monotherapy versus fractional laser-assisted topical corticosteroid delivery: a randomized clinical trial. Acta Derm Venereol. 2021; 101: adv00416.

[45] Sabry HH, Abdel Rahman SH, Hussein MS, Sanad RR, Abd El Azez TA. The efficacy of combining fractional carbon dioxide laser with verapamil hydrochloride or 5-fluorouracil in the treatment of hypertrophic scars and keloids: a clinical and Immunohistochemical study. Dermatol Surg. 2019; 45: 536-46.

[46] Mao KL, Fan ZL, Yuan JD, Chen PP, Yang JJ, Xu J, et al. Skin-penetrating polymeric nanoparticles incorporated in silk fibroin hydrogel for topical delivery of curcumin to improve its therapeutic effect on psoriasis mouse model. Colloids Surf B Biointerfaces. 2017; 160: 704-14.