POLYELECTROLYTE COMPLEX OF SILK FIBROIN WITH

ALGINATE AS THERAPEUTIC PROTEIN DELIVERY MODEL

1,2 2 3
Gita Cahya Eka Darma , Sundani Nurono Soewandhi , Chandra Risdian
1
Department of Pharmacy, Bandung Islamic University, Jl. Tamansari No.1 Bandung 40116, Indonesia
2
School of Pharmacy, Bandung Institute of Technology, Jl. Ganesha No.10 Bandung 40132, Indonesia
3
Department of Chemistry, The Indonesian Institute of Sciences, Jl. Cisitu Komplek LIPI Bandung 40135, Indonesia
Corresponding author email: g.c.ekadarma@gmail.com

Silk Fibroin (SF) and Alginate (ALG) are biomaterials that have therapeutic effect in treatment of wounds dressing which are hemostatic and the ability upon absorption of wound exudates. This
research aims to make delivery model for wounds dressing in the form of fiber membrane results from polyelectrolyte complexation (PEC) through coacervation method. SF from silkworm
cocoons (Bombyx mori Linn) used as polycations, and ALG as polyanions agent. Characterization of PEC fiber membranes are conducted with analysis of the functional groups (FTIR), crystallinity
degree (XRD), thermal properties (DSC), microscopic (SEM). Then, the study continued to the measurement of coacervation yield. SDS PAGE was used to verify the protein. This research has
obtained SF with better characteristics that suitable for the treatment of skin wounds, which is responsive to the pH environment. The membrane is a modified form from the hydrogel-forming
of SF/ALG coacervate. Characterization analysis of the membrane shown that the electrostatic bonds occurs between SF with ALG (PEC) and approved that transformation polymorphic forms of
SF from Silk I (metastable) to Silk II (stable) was happened. Polymorphism, thermal properties, and crystallinity degree of SF can be modulated and controlled to obtain the appropiate final
result of the membrane.
Keywords: Silk fibroin, sodium alginate, coacervation, polyelectrolyte complex, fiber membrane

Step 01
Silk degumming Formulation
Step 05 Step 02 F1 = SF/ALG (1:1); F2 = SF/ALG (1:3)
PEC fiber membrane SF solution (*RSF 1) F3 = SF/ALG (3:1)

Fibroin protein
Cocoons weight = 25,051 g
Methods

Degummed silk = 18,132 g

SF characteristic & properties Recovery = 72,528%
biomaterial modulation
Final delivery form modification SF solution (RSF) = 954 mL
*RSF (Regenerated Silk Fibroin) SF concentration = 18,132 g/954 mL
= 1,9%
Step 03
SF paste (*RSF 2) Fibroin identification (SDS-PAGE)
SF is containing of Light chain/L-chain
(~25 kDa) and Heavy chain/H-chain
(~325 kDa) with ratio 1:1 (Inoue, 2000;
Step 04 Zhou, 2000).
Coacervate SF/ALG

SDS PAGE analysis Hydrogel-forming of SF/ALG coacervate SF/ALG PEC fiber membrane
Discussion
Result &

Fiber membrane-forming of SF/ALG coacervate

Fourier Transformed Infrared (FTIR) X-Ray Diffraction (XRD) Differential Scanning Calorimetry (DSC) Coacervation yield

Scanning Electron Microscope (SEM)

(a) Degummed silk; (b) SF paste; (c) ALG SF/ALG PEC fiber membrane (a) F1 (1:1);
(b) F2 (1:3); (c) F3 (3:1)

(References of SF IR-band: Pelton, 2000; Barth, 2002; Garidel, 2006; Hu, 2006; Lu, 2010 | SF difractogram: Shimizu, 1941; Hirabayashi, 1967; Magoshi, 1977; Ayub, 1994 | SF DSC: Sashina et al., 2007)

1. Ayub, Z.H., Arai, M., Hirabayashi, K. (1994), Quantitative structural analysis

Silk fibroin from silkworm cocoons (Bombyx mori
Linn) with sodium alginate can be formulated
as a therapeutic protein delivery model into
PEC fiber membrane through coacervation.
Conclusion
SF have polymorphic form that could be regula-
te and controlled upto Silk I or Silk II. Polyeletro-
lyte complex of SF/ALG obtained from coacervation (hydrogel-forming) could modified
into fiber membrane. Crystallinity degree from electrostatic crosslinking could be setting
by the ratio of SF/ALG that we used. Characterization of PEC fiber membrane had proved
there is crosslinking between SF and ALG, SF polymorpism is in stable form (Silk II), crys-
tallinity degree SF decreased after forming complexation with ALG which is suitable for
wound dressing because it could swelling and dissolve depend on pH system.
and physical-properties of silk fibroin hydrogels, Polymers, 35, 2197-2200.
2. Barth, A., Zscherp, C. (2002), What vibrations tell us about proteins, Quart.
Rev. Biophys, 35, 369–430.
04 05 References 3. Garidel, P., Schott, H. (2006), Fourier-transform midinfrared spectroscopy for
analysis and screening of liquid protein formulations, Bio-Process
International, June, 48−55.
4. Hirabayashi, K., Ishikava, H. (1967), Studies on the fine structure of silk fibroin.
IV Studies on the alfa-form of Bombyx mori silk fibroin, Sen-i Gakkaishi, 23,
538.
5. Hu, X., Kaplan, D.L., Cebe, P. (2006), Determining beta-sheet crystallinity in fibrous proteins by thermal analysis and infrared spectroscopy,
Macromolecules, 39 (18), 6161–6170.
6. Inoue, S.I., Tanaka, T., Magoshi, J., Magoshi, Y., Kobayashi, M., Tsukada, H., Nakamura, S. (2001), Abstr. Pap. —Am. Chem. Soc., 221, 123.
7. Jacquet, M., Janin, J., Duguet, M., Perasso, R., Li, Z.G. (2000), Fine organization of Bombyx mori fibroin heavy chain gene, Nucleic Acids Res, 28,
2413–2419.
8. Lu, Q., Hu, X., Wang, X., Kluge, J., Lu, S., and Cebe, P. (2010), Water-insoluble silk films with silk I structure, Acta Biomaterialia, 6(4), 1380–1387.
9. Magoshi, J., Magoshi, Y. (1977), Physical properties and structure of silk. Physical properties and structure of silk, J Polym Sci Polym Phys, 15, 1675-1683.
10. Pelton, J.T., McLean, L.R. (2000), Spectroscopic methods for analysis of protein secondary structure, Analytical Biochemistry, 277, 167-176.
11. Sashina, E.S., Janowska, G., Zaborski, M., Vnuchkin, A.V. (2007), Compatibility of fibroin/chitosan and fibroin/cellulose blends studied by thermal
analysis, J. Therm. Anal. Calorimet., 89, 887–891.
12. Shimizu, M. (1941), Sanshi Shikenjo Houkoku, 10, 475.
13. Zhou, C.Z., Confalonieri, F., Medina, N., Zivanovic, Y., Esnault, C., Yang, T., Jacquet, M., Janin, J., Duguet, M., Perasso, R., Li, Z.G. (2000), Fine
organization of Bombyx mori fibroin heavy chain gene, Nucleic Acids Res, 28, 2413–2419.

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Publish at International Seminar In Pharmaceutical Technology 27-28 January 2016, Bandung, West Java, Indonesia