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Thermoreversible poly(ethylene glycol)-g-chitosan hydrogel as a therapeutic T lymphocyte depot for localized glioblastoma immunotherapy.

Tsao CT, Kievit FM, Ravanpay A, Erickson AE, Jensen MC, Ellenbogen RG, Zhang M - Biomacromolecules (2014)

Bottom Line: Local immunotherapy has the potential to eradicate the residual infiltrative component of these tumors.Nuclear magnetic resonance spectroscopy confirmed the chemical structure of poly(ethylene glycol)-g-chitosan, and rheological studies revealed that the sol-to-gel transition of the PCgel occurred around ≥32 °C.Our study suggests that this unique PCgel depot may offer a viable approach for localized immunotherapy for glioblastoma.

View Article: PubMed Central - PubMed

Affiliation: Department of Materials Science and Engineering, University of Washington , Seattle, Washington 98195, United States.

ABSTRACT
The outcome for glioblastoma patients remains dismal for its invariably recrudesces within 2 cm of the resection cavity. Local immunotherapy has the potential to eradicate the residual infiltrative component of these tumors. Here, we report the development of a biodegradable hydrogel containing therapeutic T lymphocytes for localized delivery to glioblastoma cells for brain tumor immunotherapy. Thermoreversible poly(ethylene glycol)-g-chitosan hydrogels (PCgels) were optimized for steady T lymphocyte release. Nuclear magnetic resonance spectroscopy confirmed the chemical structure of poly(ethylene glycol)-g-chitosan, and rheological studies revealed that the sol-to-gel transition of the PCgel occurred around ≥32 °C. T lymphocyte invasion through the PCgel and subsequent cytotoxicity to glioblastoma were assessed in vitro. The PCgel was shown to be cellular compatible with T lymphocytes, and the T lymphocytes retain their anti-glioblastoma activity after being encapsulated in the PCgel. T lymphocytes in the PCgel were shown to be more effective in killing glioblastoma than those in the Matrigel control. This may be attributed to the optimal pore size of the PCgel allowing better invasion of T lymphocytes. Our study suggests that this unique PCgel depot may offer a viable approach for localized immunotherapy for glioblastoma.

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Related in: MedlinePlus

Biocompatibility of the PCgel. The numbersof T lymphocytes culturedon a TCP, PCgel, and Matrigel quantified using AlamarBlue after a24 h culture.
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fig3: Biocompatibility of the PCgel. The numbersof T lymphocytes culturedon a TCP, PCgel, and Matrigel quantified using AlamarBlue after a24 h culture.

Mentions: Cell viabilityis the most fundamental feature of cellular compatibility for biomedicalmaterials. The viability of genetically modified T lymphocytes inthe PCgel was assessed with the AlamarBlue assay. Specifically, 3× 105 T lymphocytes were loaded evenly in 400 μLof the liquid PCgel onto 24-well cell culture plates. The T lymphocyte-loadedPCgel was then placed in an incubator for solidification. An additional400 μL of culture medium was added on top of the solidifiedPCgel 2 h after T lymphocyte cell seeding. A blank standard TCP andMatrigel were used as two-dimensional and three-dimensional controls,respectively. Twenty-four hours after T lymphocyte seeding, the culturemedium was replaced with AlamarBlue for 8 h. The AlamarBlue solutionwas then collected, and the centrifuged supernatant was transferredto a 96-well black-walled plate for quantifying the number of livecells. Figure 3 shows the number of T cellson a TCP, in the PCgel, and in Matrigel. Though the number of T lymphocytesin the PCgel was observed to be slightly lower than that of a TCP(89%; p = 0.353) and slightly higherthan that of Matrigel (113%; p = 0.184), there wasno statistical difference among the three tested materials (p = 0.156). This confirmed that the PCgel had good cellularcompatibility with T lymphocytes.


Thermoreversible poly(ethylene glycol)-g-chitosan hydrogel as a therapeutic T lymphocyte depot for localized glioblastoma immunotherapy.

Tsao CT, Kievit FM, Ravanpay A, Erickson AE, Jensen MC, Ellenbogen RG, Zhang M - Biomacromolecules (2014)

Biocompatibility of the PCgel. The numbersof T lymphocytes culturedon a TCP, PCgel, and Matrigel quantified using AlamarBlue after a24 h culture.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4215871&req=5

fig3: Biocompatibility of the PCgel. The numbersof T lymphocytes culturedon a TCP, PCgel, and Matrigel quantified using AlamarBlue after a24 h culture.
Mentions: Cell viabilityis the most fundamental feature of cellular compatibility for biomedicalmaterials. The viability of genetically modified T lymphocytes inthe PCgel was assessed with the AlamarBlue assay. Specifically, 3× 105 T lymphocytes were loaded evenly in 400 μLof the liquid PCgel onto 24-well cell culture plates. The T lymphocyte-loadedPCgel was then placed in an incubator for solidification. An additional400 μL of culture medium was added on top of the solidifiedPCgel 2 h after T lymphocyte cell seeding. A blank standard TCP andMatrigel were used as two-dimensional and three-dimensional controls,respectively. Twenty-four hours after T lymphocyte seeding, the culturemedium was replaced with AlamarBlue for 8 h. The AlamarBlue solutionwas then collected, and the centrifuged supernatant was transferredto a 96-well black-walled plate for quantifying the number of livecells. Figure 3 shows the number of T cellson a TCP, in the PCgel, and in Matrigel. Though the number of T lymphocytesin the PCgel was observed to be slightly lower than that of a TCP(89%; p = 0.353) and slightly higherthan that of Matrigel (113%; p = 0.184), there wasno statistical difference among the three tested materials (p = 0.156). This confirmed that the PCgel had good cellularcompatibility with T lymphocytes.

Bottom Line: Local immunotherapy has the potential to eradicate the residual infiltrative component of these tumors.Nuclear magnetic resonance spectroscopy confirmed the chemical structure of poly(ethylene glycol)-g-chitosan, and rheological studies revealed that the sol-to-gel transition of the PCgel occurred around ≥32 °C.Our study suggests that this unique PCgel depot may offer a viable approach for localized immunotherapy for glioblastoma.

View Article: PubMed Central - PubMed

Affiliation: Department of Materials Science and Engineering, University of Washington , Seattle, Washington 98195, United States.

ABSTRACT
The outcome for glioblastoma patients remains dismal for its invariably recrudesces within 2 cm of the resection cavity. Local immunotherapy has the potential to eradicate the residual infiltrative component of these tumors. Here, we report the development of a biodegradable hydrogel containing therapeutic T lymphocytes for localized delivery to glioblastoma cells for brain tumor immunotherapy. Thermoreversible poly(ethylene glycol)-g-chitosan hydrogels (PCgels) were optimized for steady T lymphocyte release. Nuclear magnetic resonance spectroscopy confirmed the chemical structure of poly(ethylene glycol)-g-chitosan, and rheological studies revealed that the sol-to-gel transition of the PCgel occurred around ≥32 °C. T lymphocyte invasion through the PCgel and subsequent cytotoxicity to glioblastoma were assessed in vitro. The PCgel was shown to be cellular compatible with T lymphocytes, and the T lymphocytes retain their anti-glioblastoma activity after being encapsulated in the PCgel. T lymphocytes in the PCgel were shown to be more effective in killing glioblastoma than those in the Matrigel control. This may be attributed to the optimal pore size of the PCgel allowing better invasion of T lymphocytes. Our study suggests that this unique PCgel depot may offer a viable approach for localized immunotherapy for glioblastoma.

Show MeSH
Related in: MedlinePlus