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Research Papers

Development of a Nanoparticle-Embedded Chitosan Sponge for Topical and Local Administration of Chemotherapeutic Agents

[+] Author and Article Information
Manijeh Goldberg

Massachusetts Institute of Technology,
Cambridge, MA 02139
David H. Koch Institute for Integrative Cancer Research,
Department of Chemical Engineering,
Cambridge, MA 02142
University of Massachusetts Lowell,
Department of Mechanical Engineering,
Lowell, MA 01854
Privo Technologies, LLC,
Cambridge, MA 02141

Aaron Manzi

Massachusetts Institute of Technology,
Cambridge, MA 02139
David H. Koch Institute for Integrative Cancer Research,
Department of Chemical Engineering,
Langer Lab,
Cambridge, MA 02142
Privo Technologies, LLC,
Cambridge, MA 02141

Erkin Aydin

Massachusetts Institute of Technology,
Cambridge, MA 02139
David H. Koch Institute for Integrative Cancer Research,
Department of Chemical Engineering,
Langer Lab,
Cambridge, MA 02142
Abdullah Gul University,
Department of Mechanical Engineering,
Kocasinan, Turkey

Gurtej Singh, Robert Langer

Massachusetts Institute of Technology,
Cambridge, MA 02139
David H. Koch Institute for Integrative Cancer Research,
Department of Chemical Engineering,
Langer Lab,
Cambridge, MA 02142

Payam Khoshkenar, Amritpreet Birdi

Massachusetts Institute of Technology,
Cambridge, MA 02139
David H. Koch Institute for Integrative Cancer Research,
Department of Chemical Engineering,
Langer Lab,
Cambridge, MA 02142
Privo Technologies, LLC,
Cambridge, MA 02141

Brandon LaPorte

Massachusetts Institute of Technology,
Cambridge, MA 02139
David H. Koch Institute for Integrative Cancer Research,
Department of Chemical Engineering,
Langer Lab,
Cambridge, MA 02142
Privo Technologies, LLC,
Cambridge, MA 02141

Alejandro Krauskopf

Massachusetts Institute of Technology,
Cambridge, MA 02139

Geralle Powell

Wellesley College,
Department of Biology,
Wellesley, MA 02481

Julie Chen

University of Massachusetts Lowell,
Department of Mechanical Engineering,
Lowell, MA 01854

Manuscript received April 12, 2015; final manuscript received June 17, 2015; published online July 15, 2015. Assoc. Editor: Jianping Fu.

J. Nanotechnol. Eng. Med 5(4), 040905 (Nov 01, 2014) (11 pages) Paper No: NANO-15-1033; doi: 10.1115/1.4030899 History: Received April 12, 2015; Revised June 17, 2015; Online July 15, 2015

The following work describes the development of a novel noninvasive transmucosal drug delivery system, the chitosan sponge matrix (CSM). It is composed of cationic chitosan (CS) nanoparticles (NPs) that encapsulate cisplatin (CDDP) embedded within a polymeric mucoadhesive CS matrix. CSM is designed to swell up when exposed to moisture, facilitating release of the NPs via diffusion across the matrix. CSM is intended to be administered topically and locally to mucosal tissues, with its initial indication being oral cancer (OC). Currently, intravenous (IV) administered CDDP is the gold standard chemotherapeutic agent used in the treatment of OC. However, its clinical use has been limited by its renal and hemotoxicity profile. We aim to locally administer CDDP via encapsulation in CS NPs and deliver them directly to the oral cavity with CSM. It is hypothesized that such a delivery device will greatly reduce any systemic toxicity and increase antitumor efficacy. This paper describes the methods for developing CSM and maintaining the integrity of CDDP NPs embedded in the CSM.

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Figures

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Fig. 1

Molecular structures and formation conditions for chitin and CS. The acetyl groups (C2H3O) are removed from chitin to form CS. Note how the CS primary amine groups (NH2) become protonated in acidic conditions.

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Fig. 2

Schematic diagrams of the CSM. CSM can be molded to have several sizes, however the current size of 1.5 × 1.5 cm2 provides adequate coverage of a tumor that is 1 cm in diameter. The sponge is 2 mm in height. Note that the backing ethyl cellulose can be brushed on the back of CSM to create a unidirectional application. The CSM is designed to remain on the tumor for 1 hr and the remaining CSM with its non degradable ethylcellulose backing will be discarded (a). CS (CDDP NPs) (b). Ex vivo study: CSM is placed on the freshly harvested lamb's tongue for permeation studies. Transition of the CSM into a hydrogel after 5 mins of exposure to a wetting agent (c).

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Fig. 3

Molecular structures of FITC, CS, and FITC-CS. FITC-CS is synthesis is catalyzed by the addition of EDC. The primary amine of CS acts as a nucleophile to the carboxylic acid of FITC and forms a covalent bond between the two molecules. The secondary amine on FITC-CS is unable to become protonated and will remain neutral.

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Fig. 4

(a) Effect of increasing the TPP concentration on NP zeta potential. CS concentration was kept constant at 1.0 mg/mL (b) impact of increasing TPP concentration on NP size. CS concentration was kept constant at 1.0 mg/mL (c) results of increasing CS concentration on NP zeta potential. TPP concentration remained at 1.0 mg/mL (d) effect of increasing CS concentration on NP size. TPP concentration was fixed at 1.0 mg/mL (e) impact of increasing the CS solution pH used for NP synthesis with 0.1 M NaOH (f) results of increasing the CS solution pH used for NP synthesis with 0.1 M NaOH. When the TPP concentration was increased, there was a clear trend in the reduction of NP zeta potential, but no predictable impact was made on NP size. The zeta potential and size both increased when the CS concentration was heightened. The NPs produced with higher pH CS solutions showed both larger size and lower zeta potential.

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Fig. 5

(a) pH of the CDDP-TPP solution measured at 15 mins intervals for the initial 2 hrs and at 30 mins intervals over the course of an additional 5 hrs and (b) rate of the hypothesized CDDP-TPP reaction. The rate was measured by plotting out the % decrease in pH over the first hour and analyzing the slopes of the lines for each condition. We believe that the greater pH depression rates can be correlated with a more rapid production of phosphoric acid.

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Fig. 6

The percentage cell viability observed when free CDDP, BLK NPs, and CDDP NPs were incubated with HCPC-1 cells (a) and FaDu cells (b) over the course of 48 hrs at 37 °C and 5% CO2. Both cells lines showed the lowest viability when exposed to the CDDP NPs, followed by free CDDP and BLK NPs, respectively.

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Fig. 7

(a) NP cell uptake microscopy image after 1 hr incubation of OC TR-146 cells with ALX-CS NPs and DAPI stained cell nuclei shown and (b) evaluation of cell uptake by flow cytometry, percentage of positive cells after 30 and 60 mins incubation with NPs at a concentration of 0.03 g/L. NPs were taken up by more than 95% of the cells in both cases.

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Fig. 8

Release profile of CDDP NPs (2.88:1.00) at pH 7.2 in PBS. Percentage release was determined using Eqs. (1)–(4) using Pt concentrations quantified with ICP-AES.

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Fig. 9

Photograph of CSM containing CDDP NPs. Note the smooth surface texture and pure white color. 1.5 cm × 1.5 cm × 2 mm.

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Fig. 10

SEM images of lyophilized free CDDP NPs (a), CDDP NPs embedded within CSM (b), and an image of CS sponge synthesized without any NPs (c). The morphology of the free CDDP NPs and CDDP NPs in CSM are nearly identical, with both exhibiting a granulated texture. In contrast, the sponge composed of only CS showed a flat, smooth surface texture with no abnormal structures. This suggests that the NPs remain intact within CSM.

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Fig. 11

The concentration and % size increase in FITC NPs released from CSM in artificial human saliva (pH 7.0) taken using NanoSight (Malvern). This data confirm the presence of intact NPs within CSM. The increased size can be attributed to the higher pH of the saliva compared to the NP synthesis conditions.

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