Rheological properties of the mixture of Lentinan and saliva and its inhibitory effect on Streptococcus mutans

Figures & data

Table 1. Nucleotide sequences of primers used for qPCR and product sizes.

Name	Name Sequence of forward (F) and reverse (R) primers	Size	Gen bank accession No.
16S rRNA	F: GGACGAAAGTCTGACCGAGCR R: AGTTAGCCGTCCCTTTCTGG	138	AY188348.1
ackA	F: ATATTGGTGCTTACGCTGCTGR R: AGCTCCAAGCCAATCAAAGTG	122	AE014133.2
pykf	F: GCTGTTCGTACCATGGCAACR R: CTTTAACAGCTGAAGCGACAAC	124	CP082153.1
pgk	F: ATCGCTATCCTTGGTGGTTCAR R: CCAATTTCGATACCTTGTGCCTT	134	AOCB01000003.1
pfk	F: TAATTGTCAAGGGAGCACGTCAR: R: CTGTGTTCACCACCTTTCCTCT	120	CP082153.1
glk	F: AAGTGGACTTCAAACCGTTGCR R: GATTTCATCCACACGCTTGGT	116	AOCB01000001.1

Figure 1. Effects of the shear rate on the shear stress and apparent viscosity of LNT and AS mixtures at different concentrations. (a, c) LNT; (b, d) LNT + AS (1:1/v:v).

Table 2. Power Law model parameters for the LNT and AS mixture at different concentrations.

Concentration	K/mpa·s^n		n		R^2
	LNT	LNT + AS	LNT	LNT + AS	LNT	LNT + AS
0%	--	0.871	-	0.981	-	0.987
0.125%	1.404	0.974	0.977	0.944	0.989	0.976
0.25%	1.424	0.988	0.967	0.948	0.995	0.967
0.5%	1.545	0.997	0.954	0.967	0.999	0.999
1%	1.586	1.110	0.884	0.968	0.988	0.978
2%	1.623	1.230	0.697	0.977	0.990	0.977
4%	1.652	1.368	0.678	0.978	0.992	0.961
6%	1.828	1.390	0.639	0.989	0.977	0.969
8%	3.991	1.424	0.415	0.989	1.000	0.986

K is the viscosity coefficient, which means that the lower the viscosity, the lower the required stress is. n is the solution flow index, the further away from 1, it means that the fluid is easier to shear and dilute.

Figure 2. Tan δ of the LNT and AS mixture with different concentrations varied with oscillation frequency. (a) LNT; (b) LNT + AS (1:1/v:v).

Figure 3. Particle size distribution of the LNT and AS mixture at different concentrations. (a) LNT; (b) LNT + AS (1:1/v:v).

Table 3. Average particle size of the LNT and AS mixture with different concentration.

concentration	Average particle size (nm)
	LNT	LNT + AS
8%	152.791 ± 0.641^A	156.522 ± 0.431^a*
6%	140.412 ± 0.274^B	142.794 ± 0.202^b*
4%	129.952 ± 0.565^C	137.568 ± 0.434^c*
2%	127.393 ± 0.897^D	132.657 ± 0.582^d*
1%	120.272 ± 0.142^F	129.214 ± 0.173^e*

Mean ± standard deviations are illustrated (n = 3). comparison in the same column is indicated by different uppercase or lowercase letters. Different uppercase or lowercase letters are statistically significant (p < .05); comparison between groups is indicated by *, * indicates p < .05.

Table 4. Surface tension of the LNT and AS mixture at different concentrations.

concentration	surface tension (mN/m)
	LNT	LNT + AS
8%	53.032 ± 3.051^F	71.961 ± 2.531^f**
6%	58.062 ± 1.622^E	71.662 ± 1.872^e**
4%	60.891 ± 0.516^D	70.924 ± 1.395^e**
2%	61.420 ± 1.784^D	70.296 ± 1.713^d**
1%	62.722 ± 0.722^D	63.287 ± 3.261^c*
0.5%	65.391 ± 0.776^C	60.640 ± 0.573^e**
0.25%	71.674 ± 3.901^B	63.172 ± 0.812^d**
0.125%	77.883 ± 0.714^A	66.503 ± 0.495^c**
0%	-	80.646 ± 1.072^e**

Mean ± standard deviations are illustrated (n = 3). Comparisons in the same column are indicated as different uppercase or lowercase letters. Different uppercase or lowercase letters are statistically significant (p < .05); comparisons between groups are * and * *, *p < .05 and * *p < .01.

Table 5. Comparison of solution fracture time at different tensile speeds.

Tensile speed (mm/s)	Break time (s)
	AS	LNT	LNT + AS
0.040	7.261 ± 0.362^a	10.800 ± 0.508^a*	9.532 ± 0.361^a#
0.200	5.413 ± 0.102^b	6.946 ± 0.040^b*	6.141 ± 0.101^b#
1.000	5.382 ± 0.043^b	6.553 ± 0.060^c*	5.883 ± 0.063^c#
5.000	4.260 ± 0.021^c	5.488 ± 0.047^d*	5.294 ± 0.022^d#

Mean ± standard deviations are illustrated (n = 3). Different lowercase letters indicate comparisons in the same column, * indicates p < .05 between LNT and   AS group; # indicates p < .05 between LNT andLNT + AS group.

Table 6. Comparison of solution fracture height at different tensile speeds.

Tensile speed (mm/s)	Fracture height (mm)
	AS	LNT	LNT + AS
0.040	0.448 ± 0.564^d	0.449 ± 0.564^d	0.449 ± 0.564^d
0.200	1.242 ± 0.823^c	1.246 ± 0.821^c	1.245 ± 0.823^c
1.000	5.178 ± 1.062^b	5.187 ± 1.052^b	5.179 ± 0.070^b
5.000	8.687 ± 1.472^a	10.863 ± 1.470^a*	8.787 ± 0.690^a#

Mean ± standard deviations are illustrated (n = 3). Different lowercase letters indicate comparisons in the same column, * indicates p < .05 between LNT and  AS group; # indicates p < .05 between LNT and LNT + AS group.

Figure 4. Solution tensile height and tensile stress at different tensile speeds. (a)5 mm/s; (b) 1 mm/s; (c) 0.2 mm/s; (d) 0.04 mm/s.

Figure 5. Bacteriostasis rate of different concentrations of LNT and AS on oral bacteria. (a) S. mutans; (b) S. salialis; (c) A. viscosus; (d) S. sanguinis; (e) C. albicans. Values represent the mean ± SD (n = 3).

Figure 6. Effects of the mixture of LNT and AS on cell morphology of S. mutans. (a) 0%; (b) 2%; (c) 4%; (d) 8%.

Figure 7. Effects of the mixture of LNT and AS at different concentrations on cell structure of S. mutans. (a) 0%; (b) 2%; (c) 4%; (d) 8%.

Figure 8. Effects of the mixture of LNT and AS on intracellular biomacromolecules in S. mutans. Error bars show standard deviation (n = 3). *, ** and *** indicated respectively p < .05, p < .01 and p < .001 statistically significant difference compared with blank group. a. nucleic acid leakage; b. soluble protein concentration; c. electrical conductivity; d. β -galactosidase activity.

Figure 9. Effects of the mixture of LNT and AS on fluorescence intensity of S. mutans. (a) 0%; (b) 2%; (c) 4%; (d) 8%.

Figure 10. Effects of the LNT and AS mixture on AKP activity and ATP content of S. mutans (Values represent the mean ± SD (n = 3). * and ** indicated respectively p < .05 and p < .01 statistically significant difference compared with blank group.

Figure 11. Effects of the LNT and AS mixture on gene expression involved to the glycolysis pathway in S. mutans (a) glk; (b) pfk; (c) pgk; (d) pykF; (e) ackA. Values represent the mean ± SD (n = 5). Significant differences were expressed in letters (p < .05).

Table 7. Effects of the LNT and AS mixture on acid production of S. mutans.

Time/min	ΔpH
	0%	2%	4%	8%
5	0.240 ± 0.008	0.213 ± 0.001**	0.120 ± 0.012**	0.100 ± 0.012**
10	0.283 ± 0.009	0.377 ± 0.011**	0.130 ± 0.008**	0.110 ± 0.008**
15	0.287 ± 0.004	0.407 ± 0.006**	0.157 ± 0.012**	0.110 ± 0.008**
20	0.367 ± 0.008	0.497 ± 0.013**	0.177 ± 0.005**	0.150 ± 0.004**
25	0.360 ± 0.008	0.557 ± 0.005**	0.180 ± 0.008**	0.167 ± 0.008**
30	0.410 ± 0.016	0.683 ± 0.019**	0.183 ± 0.012**	0.160 ± 0.008**
35	0.465 ± 0.004	0.677 ± 0.024**	0.190 ± 0.008**	0.173 ± 0.012**
40	0.442 ± 0.030	0.753 ± 0.017**	0.193 ± 0.012**	0.183 ± 0.012**
45	0.570 ± 0.008	0.853 ± 0.026**	0.213 ± 0.026**	0.193 ± 0.047**
50	0.593 ± 0.004	0.873 ± 0.025**	0.243 ± 0.012**	0.200 ± 0.008**
55	0.670 ± 0.008	0.980 ± 0.036**	0.283 ± 0.020**	0.227 ± 0.017**
60	0.770 ± 0.012	1.133 ± 0.029**	0.320 ± 0.049**	0.270 ± 0.049**

Mean ± standard deviations are illustrated (n = 3). ** indicates p < .01, different concentrations of LNT + AS were significantly different from 0%.