Download fulltext
HTML
1.
Delrish E, Ghassemi F, Jabbarvand M, Lashay A, Atyabi F, Soleimani M, Dinarvand R. Biodistribution of Cy5-labeled Thiolated and Methylated Chitosan-Carboxymethyl Dextran Nanoparticles in an Animal Model of Retinoblastoma. JOVR [Internet]. 2022Jan.21 [cited 2024Apr.19];17(1):58–68. Available from: https://knepublishing.com/index.php/JOVR/article/view/10171
https://doi.org/10.18502/jovr.v17i1.10171
statistics
- 1055 Abstract Views
- 536 PDF Views
- 0 HTML Views
- Cited by 2 articles
authors
-
Elham Delrish
Elham Delrish
Translational Ophthalmology Research Centre, Farabi Eye Hospital, Tehran University of Medical Sciences, Tehran, Iran
-
Fariba Ghassemi
Fariba Ghassemi
Translational Ophthalmology Research Centre, Farabi Eye Hospital, Tehran University of Medical Sciences, Tehran, Iran
-
Mahmoud Jabbarvand
Mahmoud Jabbarvand
Translational Ophthalmology Research Centre, Farabi Eye Hospital, Tehran University of Medical Sciences, Tehran, Iran
-
Alireza Lashay
Alireza Lashay
Translational Ophthalmology Research Centre, Farabi Eye Hospital, Tehran University of Medical Sciences, Tehran, Iran
-
Fatemeh Atyabi
Fatemeh Atyabi
Nanotechnology Research Centre, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran
-
Masoud Soleimani
Masoud Soleimani
Department of Hematology, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran
-
Rassoul Dinarvand
Rassoul Dinarvand
Nanotechnology Research Centre, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran
Published Date
- Jan 21, 2022
Biodistribution of Cy5-labeled Thiolated and Methylated Chitosan-Carboxymethyl Dextran Nanoparticles in an Animal Model of Retinoblastoma
Abstract
Purpose: The use of more potent medicine for local chemotherapy of retinoblastoma in order to minimize local and systemic adverse effects is essential. The main goal of this investigation was to assess the biodistribution of thiolated and methylated chitosancarboxymethyl dextran nanoparticles (CMD-TCs-NPs and CMD-TMC-NPs) following intravitreal (IVT) injection into rat eyes with retinoblastoma.
Methods: An ionic gelation method was used to fabricate Cy5-labelled CMD-TCs-NPs and CMD-TMC-NPs. The NPs were characterized. Cellular internalization of Cy5-labelled NPs was investigated using confocal microscopy and the absorption of labeled NPs was quantified by flow cytometry in human retinoblastoma (Y79) cells. In addition, the Cy5-labeled distribution of nanoparticles in the posterior segment of the eye was histologically imaged by confocal microscopy after IVT injection of NPs into the eyes of rats with retinoblastoma.
Results: CMD-TCs-NPs and CMD-TMC-NPs showed a mean diameter of 34 ± 3.78 nm and 42 ± 4.23 nm and zeta potential of +11 ± 2.27 mV and +29 ± 4.31mV, respectively. The in vivo study of intraocular biodistribution of Cy5-labeled CMD-TCs-NPs and CMD-TMCNPs revealed that there is more affinity of CMD-TCs-NPs to the retina and retinoblastoma tumor after IVT administration while methylated chitosan nanoparticles are immobilized in the vitreous and are not able to reach the retina even after 24 hr.
Conclusion: The ionic gelation technique was efficient in synthesizing a biocompatible polymeric nanosystem for drug delivery into the posterior segment of the eye. The current study demonstrated increased ocular bioavailability of CMD-TCs-NPs relative to CMD-TMC-NPs in retinoblastoma induced rat eyes.
Keywords:
Biodistribution, Carboxymethyl Dextran, Chitosan, Cy5-Labeled, Nanoparticles, Retinoblastoma
References
1. Lawrence MS, Miller JW. Ocular tissue permeabilities. Int Ophthalmol Clin 2004;44:53–61.
2. Cheruvu NP, Amrite AC, Kompella UB. Effect of eye pigmentation on transscleral drug delivery. Invest Ophthalmol Vis Sci 2008;49:333–341.
3. Kennedy BG, Mangini NJ. P-glycoprotein expression in human retinal pigment epithelium. Mol Vis 2002;8:422– 430.
4. Le Goff MM, Bishop PN. Adult vitreous structure and postnatal changes. Eye 2008;22:1214–1222.
5. Mains J, Wilson CG. The vitreous humor as a barrier to nanoparticle distribution. J Ocul Pharmacol Ther 2013;29:143–150.
6. Pershing S, Bakri SJ, Moshfeghi DM. Ocular hypertension and intraocular pressure asymmetry after intravitreal injection of anti-vascular endothelial growth factor agents. Ophthalmic Surg Lasers Imaging Retina 2013;44:460–464.
7. Ghasemi Falavarjani K, Nguyen QD. Adverse events and complications associated with intravitreal injection of anti- VEGF agents: a review of literature. Eye 2013;27:787–794.
8. Kumar A, Sehra SV, Thirumalesh MB, Gogia V. Secondary rhegmatogenous retinal detachment following intravitreal bevacizumab in patients with vitreous hemorrhage or tractional retinal detachment secondary to Eales’ disease. Graefes Arch Clin Exp Ophthalmol 2012;250:685–690.
9. Al-Halafi AM. Nanocarriers of nanotechnology in retinal diseases. Saudi J Ophthalmol 2014;28:304–309.
10. Yasin MN, Svirskis D, Seyfoddin A, Rupenthal ID. Implants for drug delivery to the posterior segment of the eye: a focus on stimuli-responsive and tunable release systems. J Control Release 2014;196:208–221.
11. Rawas-Qalaji M, Williams CA. Advances in ocular drug delivery. Curr Eye Res 2012;37:345–356.
12. Rowe-Rendleman CL, Durazo SA, Kompella UB, Rittenhouse KD, Di Polo A, Weiner AL, et al. Drug and gene delivery to the back of the eye: from bench to bedside. Invest Ophthalmol Vis Sci 2014;55:2714.
13. Kritchenkov AS, Andranovits S, Skorik YA. Chitosan and its derivatives: vectors in gene therapy. Russ Chem Rev 2017;86:231–239.
14. Tiwari S, Bahadur P. Modified hyaluronic acid based materials for biomedical applications. Int J Biol Macromol 2019;121:556–571.
15. Poshina DN, Raik SV, Poshin AN, Skorik YA. Accessibility of chitin and chitosan in enzymatic hydrolysis: a review. Polym Degrad Stab 2018;156:269–278.
16. Berezin AS, Lomkova EA, Skorik YA. Chitosan conjugates with biologically active compounds: design strategies, properties, and targeted drug delivery. Russ Chem Bull 2012;61:781–795.
17. Pettignano A, Charlot A, Fleury E. Carboxyl-functionalized derivatives of carboxymethyl cellulose: towards advanced biomedical applications. Polym Rev 2019;59:510–560.
18. Fernando IPS, Kim D, Nah JW, Jeon YJ. Advances in functionalizing fucoidans and alginates (bio) polymers by structural modifications: a review. Chem Eng J 2019;355:33–48.
19. Siafaka PI, Titopoulou A, Koukaras EN, Kostoglou M, Koutris E, Karavas E, Bikiaris DN. Chitosan derivatives as effective nanocarriers for ocular release of timolol drug. Int J Pharm 2015;495:249–264.
20. Rassu G, Gavini E, Jonassen H, Zambito Y, Fogli S, Breschi MC, Giunchedi P. New chitosan derivatives for the preparation of rokitamycin loaded microspheres designed for ocular or nasal administration. J Pharm Sci 2009;98:4852–4865.
21. Bongiovì F, Di Prima G, Palumbo FS, Licciardi M, Pitarresi G, Giammona G. Hyaluronic acid-based micelles as ocular platform to modulate the loading, release, and corneal permeation of corticosteroids. Macromol Biosci 2017;17:1700261.
22. Zambito Y, Di Colo G. Thiolated quaternary ammoniumchitosan conjugates for enhanced precorneal retention, transcorneal permeation and intraocular absorption of dexamethasone. Eur J Pharm Biopharm 2010;75:194–199.
23. Hume LR, Lee HK, Benedetti L, Sanzgiri YD, Topp EM, Stella VJ. Ocular sustained delivery of prednisolone using hyaluronic acid benzyl ester films. Int J Pharm 1994;111:295–298.
24. Xu Q, Boylan NJ, Suk JS, Wang Y-Y, Nance EA, Yang J-C, et al. Nanoparticle diffusion in, and microrheology of, the bovine vitreous ex vivo. J Control Release 2013;167:76–84.
25. Koo H, Moon H, Han H, Na JH, Huh MS, Park JH, et al. The movement of self-assembled amphiphilic polymeric nanoparticles in the vitreous and retina after intravitreal injection. Biomaterials 2012;33:3485–3493.
26. Xu Q, Boylan NJ, Suk JS, Wang YY, Nance EA, Yang JC, et al. Nanoparticle diffusion in, and microrheology of, the bovine vitreous ex vivo. J Control Release 2013;167:76–84.
27. Koo H, Moon H, Han H, Na JH, Huh MS, Park JH, et al. The movement of self-assembled amphiphilic polymeric nanoparticles in the vitreous and retina after intravitreal injection. Biomaterials 2012;33:3485–3493.
28. Sieval AB, Thanou M, Kotze´ AF, Verhoef JC, Brussee J, Junginger HE. NMR preparation characterization of highly substituted N-trimethyl chitosan chloride. Carbohydr Polym 1998;36:157–165.
29. Snyman D, Hamman JH, Kotze JS, Rolling JE, Kotze AF. The relationship between the absulote molecular weight and the degree of quaternization of N-trimethy chitosan chloride. Carbohydr Polym 2002;50:145–150.
30. Margit DH, Constantia EK, Bernkop-Schnürch A. In vitro evaluation of the viscoelastic properties of chitosan– thioglycolic acid conjugates. Eur J Pharm Biopharm 2003;55:185–190.
31. Roldo M, Hornof M, Caliceti P, danBernkop-Schnurch A. Mucoadhesive thiolated chitosans as platforms for oral controlled drug delivery: synthesis an in vitro evaluation. Eur J Pharm Biopharm 2004;57:115–121.
32. Tekie FSM, Kiani M, Zakerian A, Pilevarian F, Assali A, Soleimani M, et al. Nano polyelectrolyte complexes of carboxymethyl dextran and chitosan to improve chitosan-mediated delivery of miR-145, Carbohydr Polym 2017;159:66–75.
33. Lanlan L, Jiawei G, Yuquan W, Xiaoxing X, Hui T, Jin L, et al. A broad-spectrum ROS-eliminating material for prevention of inflammation and drug-induced organ toxicity. Adv Sci 2018;5:1800781.
34. Zhigang X, Shiying L, Yuejun K, Mingfeng W. Glutathione- and pH-responsive nonporous silica prodrug nanoparticles for controlled release and cancer therapy. Nanoscale 2015;7:5859–5868.
35. Chevez-Barrios P, Hurwitz MY, Louie K, Marcus KT, Holcombe VN, Schafer P, et al. Metastatic and nonmetastatic models of retinoblastoma. Am J Pathol 2000;4:1405–1412.
36. De Britto D, Celi Goy R, CampanaFilho SP, Assis OBG. Quaternary salts of chitosan: history, antimicrobial features, and prospects. Int J Carbohydr Chem 2011;2011:312539.
37. Fang L, LijieW, Jingjing Z, Wenqiang T, Yuan Ch, Fang D, et al. Preparation and characterization of quaternized chitosan derivatives and assessment of their antioxidant activity. Molecules 2018;23:516.
38. Zhu X, Su M, Tang S, Wang L, Liang X, Meng F, et al. Synthesis of thiolated chitosan and preparation nanoparticles with sodium alginate for ocular drug delivery. Mol Vis 2012;18:1973–1982.
39. Maciel MVOB, Almeida AR, Machado MH, de MeloA PZ, da Rosa CG, de Freitas DZ, et al. Syzygium aromaticum L. (clove) essential oil as a reducing agent for the green synthesis of silver nanoparticles. Open J Appl Sci 2019;9:45–54.
40. Irimia T, Ghica M, Popa L, Anuta V, Arsene AL, Dinu-Pîrvu CE. Strategies for improving ocular drug bioavailability and corneal wound healing with chitosan-based delivery systems. Polymers 2018;10:1221.
41. Felt O, Furrer P, Mayer J, Plazonnet B, Buri P, Gurny R. Topical use of chitosan in ophthalmology: tolerance assessment and evaluation of precorneal retention. Int J Pharm 1999;180:185–193.
42. Bernkop-Schnurch A, Hornof M, Zoidl T. Thiolatedpolymers-thiomers: synthesis and in vitro evaluation of chitosan-2-iminothiolane conjugates. Int J Pharm 2003;260:229–237.
43. Bernkop-Schnürch A. Thiomers: a new generation of mucoadhesive polymers. Adv Drug Deliv Rev 2005;57:1569–1582.
44. Shin JM, Song SH, Vijayakameswara Rao N, Lee ES, Ko H, Park JH. A carboxymethyl dextran-based polymeric conjugate as the antigen carrier for cancer immunotherapy. Biomater Res 2018;22:21.
45. Vasic K, Knez Ž, Konstantinova EA, Kokorin AI, Gyergyek S, Leitgeb M. Structural and magnetic characteristics of carboxymethyl dextran coated magnetic nanoparticles: from characterization to immobilization application. React Funct Polym 2020;148:104481.
46. Kulkarni AD, Patel HM, Surana SJ, Vanjari YH, Belgamwar VS, Pardeshi CV. N,N,N-Trimethyl chitosan: an advanced polymer with myriad of opportunities in nanomedicine. Carbohydr Polym 2017;157:875–902.
47. Foroozandeh P, Abdul Aziz A. Insight into cellular uptake and intracellular trafficking of nanoparticles. Nanoscale 2018;13:339.
48. Käsdorf BT, Arends F, Lieleg O. Diffusion regulation in the vitreous humor. Biophys J 2015;109:2171–2181.
49. Pitkänen L, Ruponen M, Nieminen J, Urtti A. Vitreous is a barrier in nonviral gene transfer by cationic lipids and polymers. Pharm Res 2003;20:576–583.
50. Peeters L, Sanders NN, Braeckmans K, Boussery K, de Voorde JV, Smedt SCD, et al. Vitreous: a barrier to nonviral ocular gene Therapy. Invest Ophthalmol Vis Sci 2005;46:3553–3561.
51. Lee J, Goh U, Lee HJ, Kim J, Jeong M. Effective retinal penetration of lipophilic and lipid-conjugated hydrophilic agents delivered by engineered liposomes. Mol Pharm 2017;14:423–430.
52. Ravar F, Saadat E, Gholami M, Dehghankelishadi P, Mahdavi M, Azami S. Hyaluronic acid-coated liposomes for targeted delivery of paclitaxel, in-vitro characterization and in-vivo evaluation. J Control Release 2016;10:10–22.
53. Kompella UB, Amrite AC, Pacha Ravi R, Durazo SA. Nanomedicines for back of the eye drug delivery, gene delivery, and imaging. Prog Retin Eye Res 2013;36:172– 198.
2. Cheruvu NP, Amrite AC, Kompella UB. Effect of eye pigmentation on transscleral drug delivery. Invest Ophthalmol Vis Sci 2008;49:333–341.
3. Kennedy BG, Mangini NJ. P-glycoprotein expression in human retinal pigment epithelium. Mol Vis 2002;8:422– 430.
4. Le Goff MM, Bishop PN. Adult vitreous structure and postnatal changes. Eye 2008;22:1214–1222.
5. Mains J, Wilson CG. The vitreous humor as a barrier to nanoparticle distribution. J Ocul Pharmacol Ther 2013;29:143–150.
6. Pershing S, Bakri SJ, Moshfeghi DM. Ocular hypertension and intraocular pressure asymmetry after intravitreal injection of anti-vascular endothelial growth factor agents. Ophthalmic Surg Lasers Imaging Retina 2013;44:460–464.
7. Ghasemi Falavarjani K, Nguyen QD. Adverse events and complications associated with intravitreal injection of anti- VEGF agents: a review of literature. Eye 2013;27:787–794.
8. Kumar A, Sehra SV, Thirumalesh MB, Gogia V. Secondary rhegmatogenous retinal detachment following intravitreal bevacizumab in patients with vitreous hemorrhage or tractional retinal detachment secondary to Eales’ disease. Graefes Arch Clin Exp Ophthalmol 2012;250:685–690.
9. Al-Halafi AM. Nanocarriers of nanotechnology in retinal diseases. Saudi J Ophthalmol 2014;28:304–309.
10. Yasin MN, Svirskis D, Seyfoddin A, Rupenthal ID. Implants for drug delivery to the posterior segment of the eye: a focus on stimuli-responsive and tunable release systems. J Control Release 2014;196:208–221.
11. Rawas-Qalaji M, Williams CA. Advances in ocular drug delivery. Curr Eye Res 2012;37:345–356.
12. Rowe-Rendleman CL, Durazo SA, Kompella UB, Rittenhouse KD, Di Polo A, Weiner AL, et al. Drug and gene delivery to the back of the eye: from bench to bedside. Invest Ophthalmol Vis Sci 2014;55:2714.
13. Kritchenkov AS, Andranovits S, Skorik YA. Chitosan and its derivatives: vectors in gene therapy. Russ Chem Rev 2017;86:231–239.
14. Tiwari S, Bahadur P. Modified hyaluronic acid based materials for biomedical applications. Int J Biol Macromol 2019;121:556–571.
15. Poshina DN, Raik SV, Poshin AN, Skorik YA. Accessibility of chitin and chitosan in enzymatic hydrolysis: a review. Polym Degrad Stab 2018;156:269–278.
16. Berezin AS, Lomkova EA, Skorik YA. Chitosan conjugates with biologically active compounds: design strategies, properties, and targeted drug delivery. Russ Chem Bull 2012;61:781–795.
17. Pettignano A, Charlot A, Fleury E. Carboxyl-functionalized derivatives of carboxymethyl cellulose: towards advanced biomedical applications. Polym Rev 2019;59:510–560.
18. Fernando IPS, Kim D, Nah JW, Jeon YJ. Advances in functionalizing fucoidans and alginates (bio) polymers by structural modifications: a review. Chem Eng J 2019;355:33–48.
19. Siafaka PI, Titopoulou A, Koukaras EN, Kostoglou M, Koutris E, Karavas E, Bikiaris DN. Chitosan derivatives as effective nanocarriers for ocular release of timolol drug. Int J Pharm 2015;495:249–264.
20. Rassu G, Gavini E, Jonassen H, Zambito Y, Fogli S, Breschi MC, Giunchedi P. New chitosan derivatives for the preparation of rokitamycin loaded microspheres designed for ocular or nasal administration. J Pharm Sci 2009;98:4852–4865.
21. Bongiovì F, Di Prima G, Palumbo FS, Licciardi M, Pitarresi G, Giammona G. Hyaluronic acid-based micelles as ocular platform to modulate the loading, release, and corneal permeation of corticosteroids. Macromol Biosci 2017;17:1700261.
22. Zambito Y, Di Colo G. Thiolated quaternary ammoniumchitosan conjugates for enhanced precorneal retention, transcorneal permeation and intraocular absorption of dexamethasone. Eur J Pharm Biopharm 2010;75:194–199.
23. Hume LR, Lee HK, Benedetti L, Sanzgiri YD, Topp EM, Stella VJ. Ocular sustained delivery of prednisolone using hyaluronic acid benzyl ester films. Int J Pharm 1994;111:295–298.
24. Xu Q, Boylan NJ, Suk JS, Wang Y-Y, Nance EA, Yang J-C, et al. Nanoparticle diffusion in, and microrheology of, the bovine vitreous ex vivo. J Control Release 2013;167:76–84.
25. Koo H, Moon H, Han H, Na JH, Huh MS, Park JH, et al. The movement of self-assembled amphiphilic polymeric nanoparticles in the vitreous and retina after intravitreal injection. Biomaterials 2012;33:3485–3493.
26. Xu Q, Boylan NJ, Suk JS, Wang YY, Nance EA, Yang JC, et al. Nanoparticle diffusion in, and microrheology of, the bovine vitreous ex vivo. J Control Release 2013;167:76–84.
27. Koo H, Moon H, Han H, Na JH, Huh MS, Park JH, et al. The movement of self-assembled amphiphilic polymeric nanoparticles in the vitreous and retina after intravitreal injection. Biomaterials 2012;33:3485–3493.
28. Sieval AB, Thanou M, Kotze´ AF, Verhoef JC, Brussee J, Junginger HE. NMR preparation characterization of highly substituted N-trimethyl chitosan chloride. Carbohydr Polym 1998;36:157–165.
29. Snyman D, Hamman JH, Kotze JS, Rolling JE, Kotze AF. The relationship between the absulote molecular weight and the degree of quaternization of N-trimethy chitosan chloride. Carbohydr Polym 2002;50:145–150.
30. Margit DH, Constantia EK, Bernkop-Schnürch A. In vitro evaluation of the viscoelastic properties of chitosan– thioglycolic acid conjugates. Eur J Pharm Biopharm 2003;55:185–190.
31. Roldo M, Hornof M, Caliceti P, danBernkop-Schnurch A. Mucoadhesive thiolated chitosans as platforms for oral controlled drug delivery: synthesis an in vitro evaluation. Eur J Pharm Biopharm 2004;57:115–121.
32. Tekie FSM, Kiani M, Zakerian A, Pilevarian F, Assali A, Soleimani M, et al. Nano polyelectrolyte complexes of carboxymethyl dextran and chitosan to improve chitosan-mediated delivery of miR-145, Carbohydr Polym 2017;159:66–75.
33. Lanlan L, Jiawei G, Yuquan W, Xiaoxing X, Hui T, Jin L, et al. A broad-spectrum ROS-eliminating material for prevention of inflammation and drug-induced organ toxicity. Adv Sci 2018;5:1800781.
34. Zhigang X, Shiying L, Yuejun K, Mingfeng W. Glutathione- and pH-responsive nonporous silica prodrug nanoparticles for controlled release and cancer therapy. Nanoscale 2015;7:5859–5868.
35. Chevez-Barrios P, Hurwitz MY, Louie K, Marcus KT, Holcombe VN, Schafer P, et al. Metastatic and nonmetastatic models of retinoblastoma. Am J Pathol 2000;4:1405–1412.
36. De Britto D, Celi Goy R, CampanaFilho SP, Assis OBG. Quaternary salts of chitosan: history, antimicrobial features, and prospects. Int J Carbohydr Chem 2011;2011:312539.
37. Fang L, LijieW, Jingjing Z, Wenqiang T, Yuan Ch, Fang D, et al. Preparation and characterization of quaternized chitosan derivatives and assessment of their antioxidant activity. Molecules 2018;23:516.
38. Zhu X, Su M, Tang S, Wang L, Liang X, Meng F, et al. Synthesis of thiolated chitosan and preparation nanoparticles with sodium alginate for ocular drug delivery. Mol Vis 2012;18:1973–1982.
39. Maciel MVOB, Almeida AR, Machado MH, de MeloA PZ, da Rosa CG, de Freitas DZ, et al. Syzygium aromaticum L. (clove) essential oil as a reducing agent for the green synthesis of silver nanoparticles. Open J Appl Sci 2019;9:45–54.
40. Irimia T, Ghica M, Popa L, Anuta V, Arsene AL, Dinu-Pîrvu CE. Strategies for improving ocular drug bioavailability and corneal wound healing with chitosan-based delivery systems. Polymers 2018;10:1221.
41. Felt O, Furrer P, Mayer J, Plazonnet B, Buri P, Gurny R. Topical use of chitosan in ophthalmology: tolerance assessment and evaluation of precorneal retention. Int J Pharm 1999;180:185–193.
42. Bernkop-Schnurch A, Hornof M, Zoidl T. Thiolatedpolymers-thiomers: synthesis and in vitro evaluation of chitosan-2-iminothiolane conjugates. Int J Pharm 2003;260:229–237.
43. Bernkop-Schnürch A. Thiomers: a new generation of mucoadhesive polymers. Adv Drug Deliv Rev 2005;57:1569–1582.
44. Shin JM, Song SH, Vijayakameswara Rao N, Lee ES, Ko H, Park JH. A carboxymethyl dextran-based polymeric conjugate as the antigen carrier for cancer immunotherapy. Biomater Res 2018;22:21.
45. Vasic K, Knez Ž, Konstantinova EA, Kokorin AI, Gyergyek S, Leitgeb M. Structural and magnetic characteristics of carboxymethyl dextran coated magnetic nanoparticles: from characterization to immobilization application. React Funct Polym 2020;148:104481.
46. Kulkarni AD, Patel HM, Surana SJ, Vanjari YH, Belgamwar VS, Pardeshi CV. N,N,N-Trimethyl chitosan: an advanced polymer with myriad of opportunities in nanomedicine. Carbohydr Polym 2017;157:875–902.
47. Foroozandeh P, Abdul Aziz A. Insight into cellular uptake and intracellular trafficking of nanoparticles. Nanoscale 2018;13:339.
48. Käsdorf BT, Arends F, Lieleg O. Diffusion regulation in the vitreous humor. Biophys J 2015;109:2171–2181.
49. Pitkänen L, Ruponen M, Nieminen J, Urtti A. Vitreous is a barrier in nonviral gene transfer by cationic lipids and polymers. Pharm Res 2003;20:576–583.
50. Peeters L, Sanders NN, Braeckmans K, Boussery K, de Voorde JV, Smedt SCD, et al. Vitreous: a barrier to nonviral ocular gene Therapy. Invest Ophthalmol Vis Sci 2005;46:3553–3561.
51. Lee J, Goh U, Lee HJ, Kim J, Jeong M. Effective retinal penetration of lipophilic and lipid-conjugated hydrophilic agents delivered by engineered liposomes. Mol Pharm 2017;14:423–430.
52. Ravar F, Saadat E, Gholami M, Dehghankelishadi P, Mahdavi M, Azami S. Hyaluronic acid-coated liposomes for targeted delivery of paclitaxel, in-vitro characterization and in-vivo evaluation. J Control Release 2016;10:10–22.
53. Kompella UB, Amrite AC, Pacha Ravi R, Durazo SA. Nanomedicines for back of the eye drug delivery, gene delivery, and imaging. Prog Retin Eye Res 2013;36:172– 198.