Functionalized poly( l -lactide) single crystals coated with antigens in development of vaccines

Casini, G.; Petrone, Linda; Bakry, Ahmed; Francolini, Iolanda; Bonito, Paola Di; Giorgi, Colomba; Martinelli, Andrea; Piozzi, Antonella; D’Ilario, L.

doi:10.1016/j.jconrel.2010.07.080

Cited by 9 publications

(13 citation statements)

References 6 publications

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“…In fact, beside the good processability, they do not elicit adverse body tissue reaction and are able to degrade into non-toxic products [21]. Depending on polymer and drug features, different strategies, including drug entrapment, absorption or grafting, have been proposed to obtain drug-loaded PHA nano-or micro-particles [22][23][24]. Particularly, thanks to simplicity, good loading efficiency and flexibility, the water-in-oil emulsion and the water-in-oil-in-water double emulsion/evaporation methods are the most applied for hydrophilic and lipophilic drugs, respectively [25].…”

Section: Introductionmentioning

confidence: 99%

Release behavior and antibiofilm activity of usnic acid-loaded carboxylated poly(l-lactide) microparticles

Martinelli

Bakry

D’Ilario

et al. 2014

European Journal of Pharmaceutics and Biopharmaceutics

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Section: Introductionmentioning

confidence: 99%

Release behavior and antibiofilm activity of usnic acid-loaded carboxylated poly(l-lactide) microparticles

Martinelli

Bakry

D’Ilario

et al. 2014

European Journal of Pharmaceutics and Biopharmaceutics

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“…After washing with p ‐xylene, PLLA sc were dispersed in ethanol. PLLA sc were aminolyized under moderate conditions than reported previously to keep their features. A total of 500 mg of PLLA sc was reacted with 3 mL of 3% (wt/vol) distilled TEPA solution in isopropanol for 3 min at 50 °C.…”

Section: Methodsmentioning

confidence: 99%

Synergistic effects of surface grafting with heparin and addition of poly(d,l‐lactide) microparticles on properties of poly(l‐lactide) single crystals scaffolds

Bakry

2019

J of Applied Polymer Sci

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Mixtures of heparin grafted poly(l‐lactide) single crystals (Hep‐PLLAsc) and poly(d,l‐lactide) (PDLLA) microparticles are used for assembling hydrophilic and cytocompatible three‐dimensional tissue engineering scaffolds simply by compression molding/salt leaching technique. PLLA is grafted with heparin to promote its surface cytocompatability, while PDLLA is added to reinforce the scaffolds. PLLAsc are aminolyzed with tetraethylenepentamine (13‐atom spacer) to generate sterically accessible amino groups at the surface allowing the covalent attachment of heparin by aqueous carbodiimide chemistry. Morphological and compressive strength studies manifest the integrity and compactness of scaffolds. Toluidine blue assay assures the consistency of heparin distribution throughout the scaffolds, which is in contrary to the conventional grafting reactions by immersing the scaffold in heparin solution. The scaffolds applicability is examined by fibroblast cells seeding experiments. Contrary to pristine scaffolds, Hep‐PLLAsc scaffolds display a hydrophilic and cytocompatible interface for cell adhesion, spreading, growth, and migration. Therefore, combining presurface grafted PLLAsc with PDLLA microparticles could offer durable scaffolds exhibiting bioactive interface and controllable spatial distribution of the grafted biomacromolecules. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2019, 136, 47797.

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“…The samples were successively centrifuged at low speed (300× g) and washed twice with 1 mL of Tris-NaCl buffer to remove unbound E7. The quantity of E7 adsorbed on the PLLA sc and APLLA sc samples was estimated by SDS-PAGE after Coomassie brilliant blue staining of the gel and using known amounts of bovine serum albumin protein as a standard, as described in Casini et al 26 The E7-containing samples were designated E7-PLLA sc and E7-APLLA sc . These complexes were used to immunize mice, as described in the Immunization and tumor protection experiments section.…”

Section: Expression Purification and Analysis Of E7 Proteinmentioning

confidence: 99%

“…The amount, structural integrity, and stability of the E7 protein absorbed on pristine PLLA sc and APLLA sc were analyzed by SDS-PAGE (data not shown). 26 Despite the different chemical surface composition, the two E7-loaded samples bound similar quantities of protein. In particular, a value of 300±30 ng of E7 per mg of APLLA sc or PLLA sc was determined as described in Casini et al 26 Nevertheless, the protein released from the two substrates by controlled wash steps was different (Figure 3).…”

mentioning

confidence: 98%

“…26 Despite the different chemical surface composition, the two E7-loaded samples bound similar quantities of protein. In particular, a value of 300±30 ng of E7 per mg of APLLA sc or PLLA sc was determined as described in Casini et al 26 Nevertheless, the protein released from the two substrates by controlled wash steps was different (Figure 3). In fact, a suitable quantity of either E7-APLLA sc or E7-PLLA sc complexes was repeatedly rinsed (five times) with 1 mL aliquots of phosphate-buffered saline.…”

mentioning

confidence: 98%

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Amino-functionalized poly(L-lactide) lamellar single crystals as a valuable substrate for delivery of HPV16-E7 tumor antigen in vaccine development

Bonito¹,

Petrone²,

Casini³

et al. 2015

IJN

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Background Poly( l -lactide) (PLLA) is a biodegradable polymer currently used in many biomedical applications, including the production of resorbable surgical devices, porous scaffolds for tissue engineering, nanoparticles and microparticles for the controlled release of drugs or antigens. The surfaces of lamellar PLLA single crystals (PLLA sc ) were provided with amino groups by reaction with a multifunctional amine and used to adsorb an Escherichia coli -produced human papillomavirus (HPV)16-E7 protein to evaluate its possible use in antigen delivery for vaccine development. Methods PLLA single crystals were made to react with tetraethylenepentamine to obtain amino-functionalized PLLA single crystals (APLLA sc ). Pristine and amino-functionalized PLLA sc showed a two-dimensional microsized and one-dimensional nanosized lamellar morphology, with a lateral dimension of about 15–20 μm, a thickness of about 12 nm, and a surface specific area of about 130 m 2 /g. Both particles were characterized and loaded with HPV16-E7 before being administered to C57BL/6 mice for immunogenicity studies. The E7-specific humoral-mediated and cell-mediated immune response as well as tumor protective immunity were analyzed in mice challenged with TC-1 cancer cells. Results Pristine and amino-functionalized PLLA sc adsorbed similar amounts of E7 protein, but in protein-release experiments E7-PLLA sc released a higher amount of protein than E7-APLLA sc . When the complexes were dried for observation by scanning electron microscopy, both samples showed a compact layer, but E7-APLLA sc showed greater roughness than E7-PLLA sc . Immunization experiments in mice showed that E7-APLLA sc induced a stronger E7-specific immune response when compared with E7-PLLA sc . Immunoglobulin G isotyping and interferon gamma analysis suggested a mixed Th1/Th2 immune response in both E7-PLLA sc -immunized and E7-APLLA sc -immunized mice. However, only the mice receiving E7-APLLA sc were fully protected from TC-1 tumor growth after three doses of vaccine. Conclusion Our results show that APLLA single crystals improve the immunogenicity of HPV16-E7 and indicate that E7-APLLA sc could be used for development of an HPV16 therapeutic vaccine against HPV16-related tumors.

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Functionalized poly( l -lactide) single crystals coated with antigens in development of vaccines

Cited by 9 publications

References 6 publications

Release behavior and antibiofilm activity of usnic acid-loaded carboxylated poly(l-lactide) microparticles

Release behavior and antibiofilm activity of usnic acid-loaded carboxylated poly(l-lactide) microparticles

Synergistic effects of surface grafting with heparin and addition of poly(d,l‐lactide) microparticles on properties of poly(l‐lactide) single crystals scaffolds

Amino-functionalized poly(L-lactide) lamellar single crystals as a valuable substrate for delivery of HPV16-E7 tumor antigen in vaccine development

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