Design, Evaluation And Antimicrobial Activity Of Egyptian Propolis-Loaded Nanoparticles: Intrinsic Role As A Novel And Naturally Based Root Canal Nanosealer

Figures & data

Table 1 Ingredients And Physicochemical Characteristics Of The Prepared NPs

Ingredients (% w/v)	Formula Code
NP1	NP2		NP5
PLGA	1	0.6		0.6
PVA	2	2		5
ProE	0.5
Parameter	Evaluation (mean ± SD)
	NP1	NP2	Lyo NP2	NP5	Lyo NP5
PZ (nm)	368.4 ± 86.67	200.4 ± 33.4	379.7 ± 3.5	205.7 ± 20.7	369.58 ± 16.8
PDI	0.25 ± 0.093	0.10 ± 0.08	0.269 ± 0.08	0.084 ± 0.05	0.342 ± 0.07
ZP(mV)	−25.8 ± 2.5	−25.23 ± 4.38	−11.77 ± 1.5	−31.8 ± 5.2	−12.73 ± 1.7
% EE	85.48 ± 3.838	86.43 ± 2.771		81.83 ± 6.055
% Yield		52.09 ± 9.315		65.49 ± 21.90
	% DL	41.86 ± 0.8		40.5 ± 1.8

Table

Zero-order model: F = Kot	Korsmeyer-Peppas model: F = Kpt^n
First-order model: ln(1 -F) = -K1t	Hixon-Crowell model: (1-F)^1/3 = 1-Kβt
Higuchi model: F = KHt^1/2

Figure 1 Direct contact test of PE sealer, PE nanosealer and AH plus on (A) Streptococcus mutans and (B) Candida albicans with positive microbial growth and negative control; media only.

Figure 2 GC-MS of ProE.

Table 2 Chemical Composition Assessed By GC/MS Analysis Of Pure ProE

No.	Compounds	*RT	**TIC %
Aliphatic acids
1	2- hydroxyl-propanoic acid	5.77	0.76
2	Propanedioic acid	6.15	0.27
3	Diglycolic acid	13.16	0.21
4	Pentadecenoic acid	22.42	0.21
5	Hexadecanoic acid	39.25	0.32
6	Octadecenoic acid	44.40	0.81
7	Octadecanoic acid	45.18	0.29
Aliphatic acid esters
8	Hexadecanoic acid methyl ester	35.67	0.49
9	10-Octadecenoic acid, methyl ester	41.19	0.85
10	Octadecanoic acid-2,3-hydroxy-propyl ester	57.46	0.14
Phenolic compounds
11	Vanilethanediol	22.42	0.20
12	2H-Furo[2,3h]-1-benzopyran-2-one (Angecin)	32.03	2.56
13	Anthraquinone derivative	53.99	0.12
14	1H-Indole-2-carboxylic acid, 3-methyl-4-oxo-6(3,4,5-trimethoxyphenyl)4,5,6,7-tetrahydro, ethyl ester	58.64	0.67
15	1,3-dihydroxy-5-octenylbenzene	25.03	0.89
Phenolic acids
16	3-hydroxy- Benzoic acid	24.60	0.91
17	4-hydroxy- Benzoic acid	25.03	0.78
18	m-Hydroxymandelic acid	29.15	0.34
19	3,4-Dimethoxy-hydrocinnamic acid	30.01	0.37
20	Ferulic acid	38.07	0.19
21	cis- Caffeic acid	40.95	0.46
22	trans-Caffeic acid	42.07	4.35
Phenolic acids Eeters
23	Prenyl-cis-p-coumarate,	42.93	0.18
24	3-Methyl-3-butenyl-cis-caffeate	43.52	0.24
25	2-Methyl-2-butenyl-cis-caffeate,	44.67	0.23
26	Dimethylallyl coumarate	44.58	0.56
27	Pentanylcaffeate	48.57	7.36
28	3-Methyl-3-butenyl-trans-caffeate	49.63	0.63
29	2-Methyl-2-butenyl-trans-caffeate	50.11	7.88
30	Tetradecatetraenylcaffeate	52.93	0.47
Flavonoids
31	Pinostrobin	51.73	0.38
32	Dihydroxy flavone	53.99	0.34
33	5,7-Dihydroxy-3-butanoyloxyflavone	62.74	0.14
34	Galangin	66.55	0.14
35	3,5,7-Trihydroxy-4ʹ-butanylflavone	68.74	0.16
36	3,5,7-Trihydroxy-4ʹ-butanylflavone (isomer)	70.77	0.39
37	5,7,4ʹ-Trihydroxy-3-butanoyloxyflavone	76.20	0.48
Sugars
38	Mannitol	27.51	0.18
39	Sorbose	30.83	18.30
40	Hexose	30.94	18.61
41	D-Fructose,	32.23	0.31
42	Hexopyranose	33.56	1.60
43	Glucopyranoside,	36.63	1.16
Others
44	Acetamide-N, N-diethyl	5.24	2.11
45	N, N-Dimethylacetoacetamide	5.55	2.27
46	Glycerol	11.48	3.97
47	2,5-Dihydro-3,6-dimethoxy-2-isopropylpyrazine-5-spiro(4-cycloheptene)	22.99	0.50

Notes: *RT=retention time., **TIC=The ion current generated depends on the characteristics of the compound concerned and it is not a true quantitation.

Figure 3 SEM micrographs comparing different batches of NPs; (A) NP₂, (B) Cryoprotected NP₂, (C) NP₅ and (D) Cryoprotected NP_5.

Figure 4 TEM micrographs comparing different batches of NPs; (A) NP₂, (B) Cryoprotected NP₂, (C) NP₅ and (D) Cryoprotected NP_5.

Figure 5 FT-IR spectra of pure ProE (A), PLGA (B), PVA (C), NP₂ (D), NP₅ (E) and trehalose (F).

Figure 6 DSC thermograms of pure ProE (A), PLGA (B), PVA (C), NP₂ (D), NP₅ (E) and trehalose (F).

Figure 7 The in vitro release profiles of ProE from the prepared NPs and free ProE.

Table 3 Mathematical Modeling And Release Kinetics Of ProE From The Prepared NPs

Formulation Model		NP2	NP5	Free PE
Zero	r^2	0.9724	0.9742	0.9959
K0	0.96	0.607	0.33
First	r^2	0.9751	0.9760	0.9978
K1	0.01	0.006	0.004
H-C	r^2	0.9742	0.9754	0.9973
Kβ	0.003	0.002	0.001
Higuchi	r^2	0.9510	0.9582	0.9587
KH	2.89	1.84	2.93
Release mechanism		Dissolution
K-P	r^2	0.9792	0.9848	0.9906
	KP	1.14	0.81	0.53
	n	0.96	0.90	0.90
Drug transport		Super case transport II

Figure 8 The in vitro cytotoxicity estimated as percent cell viability after 48 hr.

Figure 9 Turbidimetric reading of the direct contact test (DCT) of PE sealer, PE nanosealer and AH Plus^® (A and B) Microbial growth of Streptococcus mutans in presence and absence of tested sealers, respectively (C and D) Microbial growth of Candida albicans in presence and absence of tested sealers, respectively (E and F) Microbial growth of Enterococcus faecalis in presence and absence in presence and absence of tested sealers, respectively.