Maltodextrins that contain both simple sugars and polymers of saccharides have been widely used as ingredients in food products and pharmaceutical delivery systems. To date, no much work has been reported on the applications of maltodextrin from broken rice (RB) sources. Therefore, the objective of this work was to investigate the
Rice constitutes the world’s principal source of food. For example, it is the major source of dietary energy and protein for 80% of the population in Southeast Asia [
Enzymatic modification of starch involves starch hydrolysis using amylolytic enzymes to break the polymer of starch molecules into a lower molecular-weight called maltodextrin, or dextrin, which is widely used in food and pharmaceutical industries [
Wound healing process consists of a series of recovery steps: (a) injured tissue is repaired; (b) specialized tissue is regenerated, and (c) new tissue is reorganized [
Up to date, there is no information on the use of RB maltodextrin as a wound healing agent, reported. Thus, RB maltodextrin with different DE groups was produced and subjected to an
Mature but unripened RB (blends of local varieties, MR 219 and MRR 220) were purchased from the local market (Serdang, Selangor). All starches were prepared at the laboratory scale [
NIH 3T3 cell line was obtained from ATCC, USA, and cultured in RPMI 1640 medium supplemented with 1% penicillin-streptomycin and 10% fetal bovine serum (FBS) in a humidified incubator with 5% CO2. Cell line was detached from the culture flasks using a trypsin-EDTA solution (0.25–0.025%) and resuspended as a single cell suspension in RPMI 1640 culture medium.
NIH 3T3 cells were seeded in a tissue culture 6-well plate at an initial density of 2.4 × 105 cells/cm2 overnight. A micropipette tip was used to create a wound in the monolayer by scraping. A total of 10 (%, w/v) RB maltodextrin were added in each treatment well with or without an addition of 100 ppm of various additives including
Trypan blue cell count was carried out to identify the amount of viable cells present in each sample. After 24 h incubation period, harvested cell suspension (10
Maltodextrin treated and untreated NIH 3T3 cell proliferation was measured using the Bromodeoxyuridine (BrdU) Cell Proliferation Kit (Merck, USA). The cells were seeded in a 96-well plate at a concentration of 0.8 × 105 cells/mL overnight. A total of 10% RB maltodextrin were added separately with or without the addition of 100 ppm of various additives, including
NIH 3T3 cells were seeded on 96-well microtiter plates overnight. A respective amount of 5% and 10% of RB maltodextrin was added in each treatment well separately. An addition of 100 ppm of various additives including
Approximately 2 mL of RBC obtained from a volunteer (Pusat Kesihatan, UPM) and was washed in PBS in a ratio of 1 : 10, followed by centrifuged for 10 min at 1500 rpm with a controlled temperature of 10°C, and this step was repeated triplicate. The RBC was then diluted with PBS to a 1% concentration. Each 100
Data was statistically analyzed by one-way analysis of variance (SPSS statistics version 16). Significant differences (
(a) The percentage of wound closure of NIH 3T3 cells treated with RB maltodextrin DE 10–14 with or without the addition of various additives. (b) The percentage of wound closure of NIH 3T3 cells treated with 5%, 10%, or 20% RB maltodextrin DE 10–14 with or without the addition of 100 ppm various additives. (c) The percentage of wound closure of NIH 3T3 cells treated with RB maltodextrin DE 10–14, 15–19, or DE 20–24 with or without the addition of 100 ppm various additives.
Time, h | RB | RBCUR | RBHYD | RBAA | RBARG | RBLA | RBKA | MUL | CTRL |
---|---|---|---|---|---|---|---|---|---|
(i) The percentage of wound closure of NIH 3T3 cell treated with 10% RB maltodextrin (DE 10–14) with 50 ppm of various additives | |||||||||
0 | 0.00 ± 0.00 |
||||||||
3 |
5.69 ± 3.05 |
9.23 ± 0.00 |
6.43 ± 1.84 |
6.16 ± 3.37 |
8.64 ± 3.92 |
5.81 ± 3.80 |
4.82 ± 4.43 |
0.00 ± 0.00 |
12.04 ± 0.53 |
6 | 15.72 ± 5.95 |
15.38 ± 0.00 |
18.21 ± 8.87 |
13.43 ± 4.89 |
18.06 ± 5.25 |
15.48 ± 3.92 |
10.66 ± 5.63 |
21.17 ± 1.72 |
|
9 | 23.85 ± 3.20 |
20.00 ± 0.00 |
28.21 ± 9.27 |
20.34 ± 0.60 |
24.61 ± 3.51 |
19.35 ± 6.64 |
23.35 ± 9.56 |
29.93 ± 4.63 |
|
12 | 40.38 ± 4.81 |
26.92 ± 0.00 |
35.36 ± 12.31 |
28.92 ± 0.87 |
30.63 ± 2.85 |
32.90 ± 4.06 |
31.98 ± 7.20 |
36.86 ± 1.47 |
|
24 | 100.00 ± 0.00 |
50.77 ± 0.00 |
50.36 ± 2.86 |
47.76 ± 3.33 |
46.86 ± 7.78 |
42.90 ± 0.54 |
41.12 ± 6.36 |
100.00 ± 0.00 | |
|
|||||||||
(ii) The percentage of wound closure of NIH 3T3 cell treated with 10% RB maltodextrin (DE 10–14) with 100 ppm of various additives | |||||||||
0 | 0.00 ± 0.00 |
||||||||
3 | 19.23 ± 1.65 |
15.79 ± 0.00 |
16.40 ± 0.17 |
14.81 ± 1.44 |
16.33 ± 1.79 |
17.74 ± 0.00 |
12.10 ± 0.62 |
0.00 ± 0.00 |
|
6 | 32.69 ± 0.70 |
29.82 ± 2.11 |
28.95 ± 4.96 |
23.33 ± 6.06 |
31.78 ± 0.92 |
28.09 ± 0.75 |
21.66 ± 0.00 |
||
9 | 48.73 ± 2.32 |
42.11 ± 6.88 |
36.36 ± 4.73 |
30.23 ± 1.52 |
38.32 ± 0.78 |
34.83 ± 0.00 |
29.26 ± 0.31 |
||
12 | 61.13 ± 1.39 |
52.97 ± 3.68 |
51.52 ± 2.72 |
47.78 ± 1.90 |
47.66 ± 0.75 |
44.94 ± 4.49 |
32.98 ± 0.56 |
||
24 | 100.00 ± 0.00 |
100.00 ± 0.00 |
100.00 ± 0.00 |
100.00 ± 0.00 |
100.00 ± 0.00 |
100.00 ± 0.00 |
100.00 ± 0.00 |
Time, h | RB | RBCUR | RBHYD | RBAA | RBARG | RBLA | RBKA | MUL | CTRL |
---|---|---|---|---|---|---|---|---|---|
(i) The percentage of wound closure of NIH 3T3 cell treated with 5% RB maltodextrin (DE 10–14) with 100 ppm of various additives | |||||||||
0 | 0.00 ± 0.00 |
||||||||
3 | 3.54 ± 0.76 |
2.51 ± 3.45 |
6.94 ± 3.51 |
7.39 ± 1.21 |
5.01 ± 0.60 |
7.27 ± 1.53 |
3.64 ± 2.49 |
2.04 ± 0.60 |
12.04 ± 0.53 |
6 | 12.96 ± 4.06 |
7.71 ± 1.06 |
13.87 ± 3.85 |
14.43 ± 4.04 |
11.22 ± 4.21 |
20.64 ± 5.11 |
10.75 ± 3.00 |
6.89 ± 3.19 |
21.17 ± 1.72 |
9 | 24.41 ± 5.17 |
21.68 ± 3.48 |
20.65 ± 5.77 |
20.62 ± 2.46 |
23.05 ± 7.83 |
32.27 ± 7.10 |
19.58 ± 2.64 |
8.42 ± 2.17 |
29.93 ± 4.63 |
12 | 33.50 ± 8.81 |
28.14 ± 1.33 |
27.90 ± 8.70 |
26.80 ± 6.94 |
28.86 ± 9.24 |
44.19 ± 4.17 |
27.90 ± 9.47 |
10.20 ± 0.57 |
36.86 ± 1.47 |
24 | 57.91 ± 5.67 |
55.56 ± 10.34 |
53.71 ± 2.41 |
48.97 ± 6.62 |
47.90 ± 3.76 |
46.51 ± 4.05 |
45.58 ± 8.33 |
10.71 ± 1.73 |
100.00 ± 0.00 |
|
|||||||||
(ii) The percentage of wound closure of NIH 3T3 cell treated with 10% RB maltodextrin (DE 10–14) with 100 ppm of various additives | |||||||||
3 | 19.23 ± 1.65 |
15.79 ± 0.00 |
16.40 ± 0.17 |
14.81 ± 1.44 |
16.33 ± 1.79 |
17.74 ± 0.00 |
12.10 ± 0.62 |
0.00 ± 0.00 |
|
6 | 32.69 ± 0.70 |
29.82 ± 2.11 |
28.95 ± 4.96 |
23.33 ± 6.06 |
31.78 ± 0.92 |
28.09 ± 0.75 |
21.66 ± 0.00 |
||
9 | 48.73 ± 2.32 |
42.11 ± 6.88 |
36.36 ± 4.73 |
30.23 ± 1.52 |
38.32 ± 0.78 |
34.83 ± 0.00 |
29.26 ± 0.31 |
||
12 | 61.13 ± 1.39 |
52.97 ± 3.68 |
51.52 ± 2.72 |
47.78 ± 1.90 |
47.66 ± 0.75 |
44.94 ± 4.49 |
32.98 ± 0.56 |
||
24 | 100.00 ± 0.00 |
100.00 ± 0.00 |
100.00 ± 0.00 |
100.00 ± 0.00 |
100.00 ± 0.00 |
100.00 ± 0.00 |
100.00 ± 0.00 |
||
|
|||||||||
(iii) The percentage of wound closure of NIH 3T3 cell treated with 20% RB maltodextrin (DE 10–14) with 100 ppm of various additives | |||||||||
3 | 7.27 ± 0.00 |
11.24 ± 3.08 |
11.11 ± 0.00 |
10.61 ± 4.77 |
12.16 ± 2.66 |
6.93 ± 1.75 |
5.32 ± 4.22 |
0.00 ± 0.00 |
|
6 | 15.64 ± 3.88 |
16.43 ± 0.00 |
16.87 ± 3.18 |
22.07 ± 1.92 |
18.11 ± 0.24 |
20.50 ± 7.62 |
4.98 ± 2.81 |
||
9 | 20.73 ± 1.47 |
20.75 ± 0.00 |
23.46 ± 8.67 |
27.37 ± 2.97 |
22.43 ± 4.92 |
24.65 ± 4.94 |
11.63 ± 3.52 |
||
12 | 22.18 ± 0.84 |
27.38 ± 3.97 |
29.22 ± 9.62 |
33.80 ± 0.00 |
30.54 ± 3.48 |
36.01 ± 1.77 |
25.58 ± 2.42 |
||
24 | 72.73 ± 1.69 |
67.44 ± 0.90 |
66.26 ± 1.78 |
49.16 ± 3.76 |
44.59 ± 4.81 |
43.77 ± 3.35 |
34.55 ± 2.32 |
Time, h | RB | RBCUR | RBHYD | RBAA | RBARG | RBLA | RBKA | MUL | CTRL |
---|---|---|---|---|---|---|---|---|---|
(i) The percentage of wound closure of NIH 3T3 cell treated with 10% RB maltodextrin (DE 10–14) with 100 ppm of various additives | |||||||||
0 | 0.00 ± 0.00 |
||||||||
3 | 19.23 ± 1.65 |
15.79 ± 0.00 |
16.40 ± 0.17 |
14.81 ± 1.44 |
16.33 ± 1.79 |
17.74 ± 0.00 |
12.10 ± 0.62 |
0.00 ± 0.00 |
12.04 ± 0.53 |
6 | 32.69 ± 0.70 |
29.82 ± 2.11 |
28.95 ± 4.96 |
23.33 ± 6.06 |
31.78 ± 0.92 |
28.09 ± 0.75 |
21.66 ± 0.00 |
21.17 ± 1.72 |
|
9 | 48.73 ± 2.32 |
42.11 ± 6.88 |
36.36 ± 4.73 |
30.23 ± 1.52 |
38.32 ± 0.78 |
34.83 ± 0.00 |
29.26 ± 0.31 |
29.93 ± 4.63 |
|
12 | 61.13 ± 1.39 |
52.97 ± 3.68 |
51.52 ± 2.72 |
47.78 ± 1.90 |
47.66 ± 0.75 |
44.94 ± 4.49 |
32.98 ± 0.56 |
36.86 ± 1.47 |
|
24 | 100.00 ± 0.00 |
100.00 ± 0.00 |
100.00 ± 0.00 |
100.00 ± 0.00 |
100.00 ± 0.00 |
100.00 ± 0.00 |
100.00 ± 0.00 |
100.00 ± 0.00 | |
|
|||||||||
(ii) The percentage of wound closure of NIH 3T3 cell treated with 10% RB maltodextrin (DE 15–19) with 100 ppm of various additives | |||||||||
3 | 7.22 ± 1.54 |
10.96 ± 0.00 |
11.41 ± 0.00 |
9.29 ± 2.68 |
16.29 ± 3.23 |
7.02 ± 4.49 |
2.93 ± 0.62 |
0.00 ± 0.00 |
|
6 | 16.58 ± 2.07 |
18.36 ± 0.00 |
21.75 ± 3.75 |
25.70 ± 0.00 |
29.14 ± 4.80 |
18.73 ± 0.75 |
13.20 ± 0.00 |
||
9 | 22.99 ± 6.95 |
26.85 ± 9.73 |
33.42 ± 1.76 |
32.51 ± 1.52 |
39.71 ± 5.30 |
20.40 ± 0.75 |
17.30 ± 0.00 |
||
12 | 56.28 ± 3.40 |
50.41 ± 7.69 |
49.36 ± 5.38 |
45.82 ± 1.80 |
45.14 ± 8.24 |
31.10 ± 0.00 |
20.23 ± 0.00 |
||
24 | 100.00 ± 0.00 |
100.00 ± 0.00 |
100.00 ± 0.00 |
100.00 ± 0.00 |
100.00 ± 0.00 |
46.48 ± 0.00 |
28.15 ± 0.00 |
||
|
|||||||||
(iii) The percentage of wound closure of NIH 3T3 cell treated with 10% RB maltodextrin (DE 20–24) with 100 ppm of various additives | |||||||||
3 | 13.56 ± 3.34 |
9.30 ± 0.00 |
12.33 ± 0.00 |
17.11 ± 0.00 |
8.45 ± 0.00 |
14.10 ± 0.00 |
8.62 ± 0.00 |
0.00 ± 0.00 |
|
6 | 23.73 ± 0.00 |
23.26 ± 6.91 |
24.66 ± 0.00 |
27.63 ± 0.00 |
25.35 ± 0.00 |
26.92 ± 0.00 |
22.41 ± 0.00 |
||
9 | 33.90 ± 0.00 |
35.47 ± 4.24 |
41.10 ± 0.00 |
35.53 ± 0.00 |
25.35 ± 0.00 |
37.18 ± 0.00 |
41.38 ± 0.00 |
||
12 | 47.46 ± 0.00 |
47.67 ± 4.71 |
49.32 ± 0.00 |
44.74 ± 0.00 |
35.21 ± 0.00 |
46.15 ± 0.00 |
53.45 ± 0.00 |
||
24 | 66.67 ± 0.00 |
61.63 ± 3.46 |
65.75 ± 0.00 |
59.21 ± 0.00 |
47.89 ± 0.00 |
57.69 ± 0.00 |
63.79 ± 0.00 |
RB: broken rice maltodextrin; CUR: curcumin; HYD: hydroxyproline; AA: ascorbic acid; ARG: L-arginine; LA: lactic acid; KA: kojic acid; MUL: Multidex; CTRL: control.
Various additives including curcumin, hydroxyproline, ascorbic acid, L-arginine, lactic acid, and kojic acid were selected based on previous publications on their significance to promote wound healing properties [
Additives act as growth factors to promote the tissue repair, migrating cell into the wound site and stimulating cell proliferation [
It is important to identify the concentration at which maltodextrin can perform the best healing capability [
Time, h | Broken rice maltodextrin |
Multidex |
---|---|---|
0 |
|
|
|
||
6 |
|
|
|
||
12 |
|
|
|
||
24 |
|
|
The third parameter of the preliminary study was to investigate the effect of different DE ranges of 10% RB maltodextrin (DE 10–14, DE 15–19, and DE 20–24) on its healing capability of NIH 3T3 cells. Based on the results obtained, RB maltodextrin DE 10–14 (Table
Overall, 10% RB maltodextrin DE 10–14 had shown the best performance in healing NIH 3T3 wounded cells relative to 5%, 20% RB maltodextrin concentrations, and also RB maltodextrin with higher DE value (DE 15–19 and DE 20–24 group). In comparison to various additives effect on the percentage wound closure of NIH 3T3 fibroblast cell, both lactic acid and kojic acid performed significantly the poorest (
To further investigate the wound healing capability of other maltodextrin source on the NIH 3T3 wounded cell, a commercial (COM) maltodextrin from cassava source was studied under the same condition. Cassava maltodextrin was selected as it was known of its high quality maltodextrin and widely used in the food and pharmaceutical applications. A previous comparison study between RB and cassava starch by Koh and Long [
Figure
An occlusive dressing may facilitate a moist wound environment and retains the wound fluid and its various components; however, it also keep oxygen away from the tissues at the same time. Oxygen plays an important role in the collagen synthesis [
A well-recognized essential requirement of most biological investigations using cellular preparations is the assertion of cell viability [
Figure
The percentage of viable cells treated with 10% RB maltodextrin DE 10–14 with 100 ppm of various additives at 24 h incubation period. The values were the means ± SE of ten independent experiments. The differences between the control group and treated group were determined by one-way ANOVA (
Based on the cell counts findings, the cell viability of all maltodextrin treatment groups achieved 100% viability rate, except for RBLA treatment, which only showed the cell viability at the rate of 93%. This phenomenon indicated that the presence of lactic acid had an effect on the cell viability of NIH 3T3 fibroblast cell. Although RB maltodextrin alone was shown to be able to heal the wounds better than the same treatment with addition of other additives, most of the cells treated with additives were able to maintain 100% viability. In all of the results presented, it was found that Multidex, a commercial wound dressing, showed zero percentage of cell viability. This finding indicated that the presence of Multidex in the NIH 3T3 cells most probably had caused substantial cell damage over time after treatment, leading to the increase in the cell death [
Freshney [
BrdU cell proliferation of NIH 3T3 cells treated with 10% RB maltodextrin with 100 ppm of various additives compared to the cell proliferation of the controlled NIH 3T3 cells at 24, 48, and 72 h incubation period. The values were the means ± SE of seven independent experiments. The differences between the control group and treated group were determined by one-way ANOVA (
Overall, all treated NIH 3T3 cells (except COM and Multidex) achieved the highest percentage of cell proliferation after 72 h of incubation. Increase in the percentage of cell proliferation may be attributed to the stimulation by maltodextrin to promote the propagation of fibroblast cells. Cells treated with 10% RB maltodextrin with or without additives showed a higher percentage of cell proliferation than the control group. In general, the highest percentage of cell proliferation at 72 h was RB maltodextrin, which was 47.33% higher in proliferation compared to the control group, followed by RB maltodextrin with 100 ppm curcumin (23.53%), RB maltodextrin with 100 ppm hydroxyproline (13.91%), and RB maltodextrin with 100 ppm
MTT assay was performed to determine the cell viability of NIH 3T3 cell after being treated with maltodextrin. This test involves the conversion of tetrazolium salt, 3-(4,5-dimethylthiazol-2-yl)-2, 5 diphenyl tetrazolium bromide (MTT) to an insoluble formazan product, which is quantitated by spectrophotometric method [
MTT cytotoxic assay of NIH 3T3 cell at 24, 48, and 72 h incubation period treated with (a) 5% RB maltodextrin with 100 ppm of various additives and (b) 10% RB maltodextrin with 100 ppm of various additives. The values were the means ± SE of seven independent experiments. The differences between the control group and treated group were determined by one-way ANOVA (
Briefly, the NIH 3T3 cell treated with 10% RB maltodextrin emerged to have the highest percentage of cell viability after 72 h (121.70%), followed by COM maltodextrin (120.96%), RB maltodextrin with 100 ppm curcumin (116.60%), RB maltodextrin with 100 ppm
The criticism of the classical
Figure
RB maltodextrins with low DE group (DE 10–14) showed better improvement of the wound closure compared to high DE group (DE 15–19 and DE 20–24) as proven in the
The authors declare that there is no conflict of interests regarding the publication of this paper.
the authors gratefully appreciate the financial support of this work by Ministry of Science, Technology and Innovation of Malaysia (06-03-08-SF0320). The authors would like to thank the staffs of the Institute of Bioscience, University Putra Malaysia (UPM), and Malaysian Agricultural Research and Development Institute (MARDI) for their help and the use of all facilities needed to conduct this study.