In Vitro and In Vivo Uptake Study of Escherichia coli Nissle 1917 Bacterial Ghosts: Cell-Based Delivery System to Target Ocular Surface Diseases
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2013
Authors
Stein, Elisabeth
Inić-Kanada, Aleksandra

Belij, Sandra

Montanaro, Jacqueline
Bintner, Nora
Schlacher, Simone
Mayr, Ulrike Beate
Lubitz, Werner
Stojanović, Marijana

Najdenski, Hristo
Barisani-Asenbauer, Talin

Article (Published version)

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PURPOSE. For the successful topical administration of drugs or vaccines to treat ocular surface diseases, efficient and well-tolerated delivery systems/carriers for conjunctival delivery are crucial in the development of new treatment strategies. The present study investigated the efficiency of internalization of bacterial ghosts (BGs) produced from probiotic Escherichia coli Nissle 1917 (EcN) by human conjunctival epithelial (HCjE) cell line, the EcN BGs cytotoxicity for HCjE cells, and in vivo uptake of EcN BGs by conjunctival guinea pig epithelial cells. METHODS. The uptake of EcN BGs by HCjE cells was analyzed by laser scanning microscopy and flow cytometry. Immunohistochemistry was used to localize the EcN BGs in the guinea pig conjunctival tissue. Cytotoxicity of EcN BGs on HCjE cells was evaluated by measurement of LDH. RESULTS. Laser scanning microscopy and flow cytometry revealed that EcN BGs internalization by HCjE cells was time-and dose dependent. No cytotoxic effect on HCj...E cells was observed after EcN BGs inoculation for 30 and 120 minutes, as well as 24 hours. In addition, the uptake of EcN BGs was detected in the conjunctival cells after in vivo administration of EcN BGs into the eye of the guinea pig. CONCLUSIONS. The findings that EcN BGs are nontoxic and effectively internalized in vitro by human and in vivo by guinea pig conjunctival cells comprise an important contribution to the future use of BGs as a system for conjunctival delivery of drugs and vaccines, either to treat or prevent ocular surface diseases.
Keywords:
bacterial ghosts / ocular surface / human conjunctival cell line / drug delivery / guinea pigSource:
Investigative Ophthalmology & Visual Science, 2013, 54, 9, 6326-6333Publisher:
- Assoc Research Vision Ophthalmology Inc, Rockville
Funding / projects:
- Laura Bassi Centers of Expertise (FFG) [822768] - Austrian Research Promotion Agency
DOI: 10.1167/iovs.13-12044
ISSN: 0146-0404
PubMed: 23920373
WoS: 000325169500049
Scopus: 2-s2.0-84884540406
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TorlakTY - JOUR AU - Stein, Elisabeth AU - Inić-Kanada, Aleksandra AU - Belij, Sandra AU - Montanaro, Jacqueline AU - Bintner, Nora AU - Schlacher, Simone AU - Mayr, Ulrike Beate AU - Lubitz, Werner AU - Stojanović, Marijana AU - Najdenski, Hristo AU - Barisani-Asenbauer, Talin PY - 2013 UR - http://intor.torlakinstitut.com/handle/123456789/381 AB - PURPOSE. For the successful topical administration of drugs or vaccines to treat ocular surface diseases, efficient and well-tolerated delivery systems/carriers for conjunctival delivery are crucial in the development of new treatment strategies. The present study investigated the efficiency of internalization of bacterial ghosts (BGs) produced from probiotic Escherichia coli Nissle 1917 (EcN) by human conjunctival epithelial (HCjE) cell line, the EcN BGs cytotoxicity for HCjE cells, and in vivo uptake of EcN BGs by conjunctival guinea pig epithelial cells. METHODS. The uptake of EcN BGs by HCjE cells was analyzed by laser scanning microscopy and flow cytometry. Immunohistochemistry was used to localize the EcN BGs in the guinea pig conjunctival tissue. Cytotoxicity of EcN BGs on HCjE cells was evaluated by measurement of LDH. RESULTS. Laser scanning microscopy and flow cytometry revealed that EcN BGs internalization by HCjE cells was time-and dose dependent. No cytotoxic effect on HCjE cells was observed after EcN BGs inoculation for 30 and 120 minutes, as well as 24 hours. In addition, the uptake of EcN BGs was detected in the conjunctival cells after in vivo administration of EcN BGs into the eye of the guinea pig. CONCLUSIONS. The findings that EcN BGs are nontoxic and effectively internalized in vitro by human and in vivo by guinea pig conjunctival cells comprise an important contribution to the future use of BGs as a system for conjunctival delivery of drugs and vaccines, either to treat or prevent ocular surface diseases. PB - Assoc Research Vision Ophthalmology Inc, Rockville T2 - Investigative Ophthalmology & Visual Science T1 - In Vitro and In Vivo Uptake Study of Escherichia coli Nissle 1917 Bacterial Ghosts: Cell-Based Delivery System to Target Ocular Surface Diseases EP - 6333 IS - 9 SP - 6326 VL - 54 DO - 10.1167/iovs.13-12044 ER -
@article{ author = "Stein, Elisabeth and Inić-Kanada, Aleksandra and Belij, Sandra and Montanaro, Jacqueline and Bintner, Nora and Schlacher, Simone and Mayr, Ulrike Beate and Lubitz, Werner and Stojanović, Marijana and Najdenski, Hristo and Barisani-Asenbauer, Talin", year = "2013", abstract = "PURPOSE. For the successful topical administration of drugs or vaccines to treat ocular surface diseases, efficient and well-tolerated delivery systems/carriers for conjunctival delivery are crucial in the development of new treatment strategies. The present study investigated the efficiency of internalization of bacterial ghosts (BGs) produced from probiotic Escherichia coli Nissle 1917 (EcN) by human conjunctival epithelial (HCjE) cell line, the EcN BGs cytotoxicity for HCjE cells, and in vivo uptake of EcN BGs by conjunctival guinea pig epithelial cells. METHODS. The uptake of EcN BGs by HCjE cells was analyzed by laser scanning microscopy and flow cytometry. Immunohistochemistry was used to localize the EcN BGs in the guinea pig conjunctival tissue. Cytotoxicity of EcN BGs on HCjE cells was evaluated by measurement of LDH. RESULTS. Laser scanning microscopy and flow cytometry revealed that EcN BGs internalization by HCjE cells was time-and dose dependent. No cytotoxic effect on HCjE cells was observed after EcN BGs inoculation for 30 and 120 minutes, as well as 24 hours. In addition, the uptake of EcN BGs was detected in the conjunctival cells after in vivo administration of EcN BGs into the eye of the guinea pig. CONCLUSIONS. The findings that EcN BGs are nontoxic and effectively internalized in vitro by human and in vivo by guinea pig conjunctival cells comprise an important contribution to the future use of BGs as a system for conjunctival delivery of drugs and vaccines, either to treat or prevent ocular surface diseases.", publisher = "Assoc Research Vision Ophthalmology Inc, Rockville", journal = "Investigative Ophthalmology & Visual Science", title = "In Vitro and In Vivo Uptake Study of Escherichia coli Nissle 1917 Bacterial Ghosts: Cell-Based Delivery System to Target Ocular Surface Diseases", pages = "6333-6326", number = "9", volume = "54", doi = "10.1167/iovs.13-12044" }
Stein, E., Inić-Kanada, A., Belij, S., Montanaro, J., Bintner, N., Schlacher, S., Mayr, U. B., Lubitz, W., Stojanović, M., Najdenski, H.,& Barisani-Asenbauer, T.. (2013). In Vitro and In Vivo Uptake Study of Escherichia coli Nissle 1917 Bacterial Ghosts: Cell-Based Delivery System to Target Ocular Surface Diseases. in Investigative Ophthalmology & Visual Science Assoc Research Vision Ophthalmology Inc, Rockville., 54(9), 6326-6333. https://doi.org/10.1167/iovs.13-12044
Stein E, Inić-Kanada A, Belij S, Montanaro J, Bintner N, Schlacher S, Mayr UB, Lubitz W, Stojanović M, Najdenski H, Barisani-Asenbauer T. In Vitro and In Vivo Uptake Study of Escherichia coli Nissle 1917 Bacterial Ghosts: Cell-Based Delivery System to Target Ocular Surface Diseases. in Investigative Ophthalmology & Visual Science. 2013;54(9):6326-6333. doi:10.1167/iovs.13-12044 .
Stein, Elisabeth, Inić-Kanada, Aleksandra, Belij, Sandra, Montanaro, Jacqueline, Bintner, Nora, Schlacher, Simone, Mayr, Ulrike Beate, Lubitz, Werner, Stojanović, Marijana, Najdenski, Hristo, Barisani-Asenbauer, Talin, "In Vitro and In Vivo Uptake Study of Escherichia coli Nissle 1917 Bacterial Ghosts: Cell-Based Delivery System to Target Ocular Surface Diseases" in Investigative Ophthalmology & Visual Science, 54, no. 9 (2013):6326-6333, https://doi.org/10.1167/iovs.13-12044 . .