In Vitro Investigation of Bacterial Cellulose/Turmeric Extract (BC-

TE) Nanocomposite for Burn Wound Dressing

Rio Cahyono Siahaan

, Melati Sitorus

, Rayhan Mulia

, Saharman Gea

Departement of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Sumatera Utara, Medan, Indonesia

Faculty of Pharmacy, Universitas Sumatera Utara, Jl. Tri Dharma No. 1, Medan, 20155

Keywords: Bacterial cellulose, turmeric extract, antimicrobial, nanocomposite.

Abstract: Bacterial cellulose is an interesting polymer material for using as a wound dressing. Bacterial cellulose has

an excellent physical and chemical properties such as high biocompatibility, microporosity, transparant,

non-toxic, and also it provides moist environment that made this polymer ideal for wound dressing.

However, bacterial cellulose itself has no antimicrobial activity to prevent wound infection. To achieve

antimicrobial activity, turmeric extract (TE) were impregnated into bacterial cellulose by immersing

bacterial cellulose (BC) in turmeric extract solution to produce BC-TE nanocomposite. A scanning electron

microscope (SEM) was used to examine the surface morphology of BC and BC–TE nanocomposite.

Antimicrobial tests in vitro indicated that BC–TE nanocomposite showed excellent antibacterial activity

against Staphylococcus aureus, and Escherichia coli with the inhibition zone of 12.45 mm and 10 mm,

respectively

1 INTRODUCTION

Burns wound are the most painful wound and can

cause trauma. More than 300,000 people die every

year around the world and 90% of these deaths are

caused by complications due to burns that occur in

many middle and lower income countries (Peck,

2011). First-degree burns usually heal without

complications while partial burns and full burns are

more complex so a clinical challenge must be faced

(Pham et al, 2007). Common complications in the

treatment of burns arise because the area of the

wound that is too large causes a long treatment time.

Contamination of wounds from the external

environment, air, water and hands of health workers

(Mehta et al, 2014). The unavailability of modern

medicine and minimal handling makes this become a

serious health problem. Ointment is less effective in

treating burns. Good treatment of burns must have

an effective wound covering material that can create

an optimal environment for regenerating the outer

skin and prevent infection of chronic wounds and

water loss (McLoughlin, 1995).

Bacterial Cellulose (BC) is one of the promising

polymer compounds produced by several types of

bacteria such as Acetobacter, Rhizobium,

Agrobacterium, Aerobacter, Achromobacter,

Azotobacter, Salmonella, Escherichia, and Sarcina

species (Ullah, Santos, & Khan, 2016). BC has

excellent physical and chemical properties such as

high biocompatibility, hydrophilicity, microporosity,

transparent and non-toxic which makes it ideal in the

treatment of wounds and skin substitutes (Gelin et

al, 2007). However, bacterial cellulose does not have

good antimicrobial activity to prevent infection. The

versatile biomedical characteristics of bacterial

cellulose can be used to make the latest wound

dressing by combining active molecules such as

antimicrobials to improve wound healing properties.

(Maneerung, Tokura, & Rujiravanit 2008).

Turmeric is one type of medicinal plant that has

many benefits and is found in many parts of

Indonesia. Turmeric contains curcumin which can

accelerate wound healing. Curcumin can increase

reepithelialization, suppress inflammation, increase

tissue collagen density and increase proliferation of

fibroblasts (Simanjuntak, 2012). The nature of

turmeric that can heal wounds has been reported

since 1953. The results showed, with turmeric the

rate of wound healing increased 23.3% in rabbits

and 24.4% in mice (Van Schraelen, 2011). Turmeric

Siahaan, R., Sitorus, M., Mulia, R. and Gea, S.

In Vitro Investigation of Bacterial Cellulose/Turmeric Extract (BC-TE) Nanocomposite for Burn Wound Dressing.

DOI: 10.5220/0008927902970300

In Proceedings of the 1st International Conference on Chemical Science and Technology Innovation (ICOCSTI 2019), pages 297-300

ISBN: 978-989-758-415-2

297

has pharmacological effects, such as accelerate

blood circulation and vital energy, eliminates

menstrual blockage, anti-inflammation, facilitates

labor, high antibacterial activity, facilitates the

release of bile (cholagogum), unleash fart and

mousturizier (astringen) (Melin and Soleha 2016).

Composite is a very interesting material because

it combines materials with different properties to

produce new materials with better properties.

Polymer nanocomposites are defined as polymers

containing materials smaller than 100 nm.

Nanocomposites are categorized in nanotechnology

if the resulting composite reflects the superiority of

nanomaterials, which is a significantly improved

performance. (Rudin and Choi, 2013).

2 MATERIALS AND METHODS

2.1 Materials

Acetobacter xylinum, Escherichia coli and

Staphylococcus aureus were purchased from

Microbiological Resources Centre, Thailand

Institute of Scientific and Technological Research

Nutrient broth (Approximate formula*per liter: Beef

extract 3.0 g and Peptone 5.0 g) was purchased from

Difco. Analytical grade D-glucose anhydrous was

purchased from Ajax Fine-chem. Yeast extract

powder and agar powder were bacteriological grade

and purchased from HiMedia. Laboratory grade

calcium carbonate and analytical grade silver nitrate

were purchased from Fisher Scientific. Laboratory

grade sodium borohydride was purchased from

CARLO ERBA. Analytical grade sodium hydroxide

anhydrate pellet and sodium chloride were

purchased from Aldrich Chemical. Analytical grade

glacial acetic acid was purchased from CSL

Chemical. Ethanol was commercial grade and used

without further purification.

2.2 Synthesize of Turmeric Extract

Turmeric used waa a fresh turmeric. The process of

making turmeric extract methanol is carried out

based on Yacob et al. (2010) using maceration and

evaporation methods. First, fresh turmeric washed,

drained, and dried for 3 days until the turmeric is

completely dry. After turmeric has completely dried,

dried turmeric was mashed until it become powder.

Turmeric powder soaked with ethanol until

homogeneous and then turmeric macerated for 3 x

24 hours. Maseration of turmeric powder was

filtered using whatman filter paper No.42. Fractions

containing volatile solvents, were concentrated with

the help of rotary evaporator. The concentrated

extract was unloaded to sterilized collecting tube

2.3 Production of Bacterial Cellulose

2.3.1 Culture-Medium

Culture medium used for the fermentation of A.

xylinum to produce bacterial cellulose consisted of

5 % glucose, 0.5 % bacto-peptone, 0.2 % disodium

phosphate, 0.1 % monocalium phosphate and the

addition of glacial acetic acid until the pH of the

culture medium reached 4. The solution was stirred

for 1 hour at 90⁰ C and followed by autoclave at

121⁰ C for 30 minutes.

2.3.2 Culture Condition

Pre-inoculum for all experiments was prepared by

transferring a single A. xylinum colony grown on

liquid culture medium into a 100 mL beaker glass

filled with liquid culture medium, then inoculated in

an incubator at 28 for 7 days with a rotational speed

of 100 rpm.

2.3.3 Purification of Bacterial Cellulose

After incubation, bacterial cellulose pellicles

produced on the surface of each liquid culture

medium were harvested and purified by soaking

them in 2.5 % NaOH for 24 h, then soaking them in

2.5% NaOCl for 24 h and finally thoroughly washed

in tap water until bacterial cellulose pellicles became

neutral and then immersed in the distilled water

prior to use.

2.3.4 Impregnation of Turmeric Extract

Into Bacterial Cellulose

The impregnation of turmeric extract into bacterial

cellulose using ex-situ method. Turmeric extract

were impregnated into bacterial cellulose fiber by

immersing bacterial cellulose pellicles in turmeric

extract until all the surface of bacterial cellulose

pellicles covered with turmeric extract. After then

bacterial cellulose pellicles impregnated for 24 h,

and then the nanocomposite rinsed with water to

remove turmeric sludge, the obtained nanocomposite

were frozen at and dried in a vacuum at -52⁰ C.

2.4 Characterization

The morphology of bacterial cellulose was observed

by using JEOL JSM-5200 scanning electron

microscope (SEM) operating at 20 kV at a

magnification of 10000.

ICOCSTI 2019 - International Conference on Chemical Science and Technology Innovation

298

2.5 Antimicrobial Activity Studies

Antimicrobial activities of freeze-dried bacterial

cellulose/turmeric extract (BC-TE) nanocomposite

have been investigated against E. coli as the model

Gram-negative bacteria and S. aureus as the model

Gram-positive bacteria. The antimicrobial activities

of freeze-dried bacterial cellulose/turmeric extract

(BC-TE) nanocomposite were carried out by disc

diﬀusion method. This method was performed in

disc diffusion method on the media Muller Hitton

Agar (MHA). Silver sulfadiazine is used as a

standard drug (positive control). Bacterial cellulose

nanocomposite / turmeric extract is placed on sterile

filter paper and also bacterial cellulose. Then placed

on gelatin media containing the selected bacteria

then incubated at 37⁰ C for 24 hours. The inhibition

zone was measured to determine the antimicrobial

ability of bacterial cellulose / turmeric extract

nanocomposite.

3 RESULTS AND DISCUSSION

3.1 Morphology of Bacterial Cellulose

Figure 1 shows the SEM micrographs of bacterial

cellulose and bacterial cellulose/turmeric extract

nanocomposite. As shown in Figure 1a BC had a

fibrous network with highly porous structure. As

shown in Figure 1b, turmeric extract particles

attached on the BC composite membrane, it is

showed that the network of BC was well retained

after impregnation of turmeric extract into BC.

Figure 1: SEM image of (a) surface morphology of

bacterial cellulose (b) bacterial cellulose/

turmeric extract nanocomposite

3.2 Antimicrobial Activity Studies

The antibacterial activity of freeze-dried bacterial

cellulose/turmeric extract nanocomposite for E. coli

and S. aureus was measured by the disc diﬀusion

method. It was found that the freeze-dried bacterial

cellulose/turmeric extract nanocomposite exhibit an

inhibition zone. The growth inhibition ring of E. coli

and S. aureus was 12,45 and 10 mm, respectively.

The growth inhibition zone with the pure bacterial

cellulose as control of E. coli and S. aureus was 6,5

mm and 0 mm, respectively. (see Fig. 2 a and b).

This clearly demonstrates that the antimicrobial

activity is only due to turmeric extract impregnated

inside bacterial cellulose and not due to individual

bacterial cellulose.

Figure 2: Antimicrobial activity against (a) Escherichia

coli and (b) Staphylococcus aureus

Table 1: Antimicrobial activity

E.coli

S. aureus

Impregnated

Pure

Impregnated

Pure

12.45 mm

6.5 mm

10 mm

0 mm

4 CONCLUSIONS

To summarize, we succeeded in the ex situ synthesis

of bacterial cellulose/ turmeric extract

nanocomposite. The preparative procedure is

surprisingly simple. It can provide a facile approach

toward manufacturing of nanocomposites,

antimicrobial materials and other useful materials.

The freeze-dried bacterial cellulose/turmeric extract

nanocomposite exhibited a strong antimicrobial

activity against both S. aureus (Gram-positive

bacteria) and E. coli (Gram-negative bacteria),

which are general bacteria that found on the

contaminated wound. A recent study showed that

impregnation, instead of coating the wound dressing

with turmeric extract improved the antimicrobial

activity of the wound dressing and lowered

possibility of the normal human tissue damage. This

is probably due to the slow and continual release of

turmeric extract and then was slowly changed our

physiological system and interact with bacterial

cells, thus turmeric extract will not be so high

enough to cause the normal human cells damage and

can prolonged the antimicrobial eﬀect.

In Vitro Investigation of Bacterial Cellulose/Turmeric Extract (BC-TE) Nanocomposite for Burn Wound Dressing

299

ACKNOWLEDGEMENTS

Financial support from the Ministry of Research,

Technology and Higher Education of the Republic

of Indonesia is greatly acknowledged.

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