Poly(γ-Glutamic Acid)/Chitosan Hydrogel Nanoparticles For Effective Preservation And Delivery Of Fermented Herbal Extract For Enlarging Hair Bulb And Enhancing Hair Growth

Figures & data

Figure 1 Schematic illustration of the synthesis of PGA-4HGF and its sustained release under physiological pH condition.

Figure 2 (A) Real image, (B) TEM image, and (C) SEM image of PGA-4HGF, and (D) real image, (E) TEM image, and (F) SEM image of PGA-control without 4HGF.

Table 1 (a) Effect Of Mixing Ratio Of PGA, 4HGF, And CS Or Mg²⁺ On The Synthesis Of Hydrogel Nanoparticles. (b) Effects Of CS Concentrations On The Synthesis Of Hydrogel Nanoparticles

(a)
	Mixing Ratio (PGA: 4HGF: CS or Mg^2+)	Size (nm)	Zeta Potential (mV)	PDI	Encapsulation Efficiency (%)	Loading Capacity (wt%)
PGA/CS with 4HGF	1:1:2	387.8±4.3	16.99±1.23	0.250±0.01	49.5±2.2	6.2±0.5
	1:1:4	381.8±2.4	19.48±1.91	0.255±0.02	51.4±1.9	11.7±0.8
	1:1:8	400.2±6.7	22.18±1.16	0.274±0.01	40.2±2.4	13.1±0.9
PGA/Mg^2+ with 4HGF	1:1:2	643.0±7.7	–38.75±0.25	0.130±0.03	26.7±1.8	<2
	1:1:4	565.4±8.8	–38.75±0.25	0.224±0.03	26.0±1.7	<2
	1:1:8	541.4±10.7	–20.02±3.02	0.140±0.03	24.2±2.0	<2
PGA/CS-control without 4HGF	1:2	250±30.8	12.15±8.91	0.156±0.035	NA	NA
1:4	280±30.4	52.49±1.49	0.005±0.01	NA	NA
(b)
CS (mg/mL) (Mixing Ratio = 1:1:4)		Size (nm)	Zeta Potential (mV)	PDI	Encapsulation Efficiency (%)	Loading Capacity (wt%)
0.1		153.2±3.8	–10.08±1.78	0.250±0.01	2.1±0.2	<2
0.2		176.4±10.8	–2.74±0.66	0.255±0.02	2.5±0.1	<2
0.5		222.4±6.7	3.46±0.64	0.292±0.01	4.7±0.1	<2

Abbreviation: NA, not applicable.

Figure 3 (A) Absorbance spectra of free 4HGF and (B) the corresponding standard curve of the concentration of 4HGF and absorption intensity measured at 400 nm. Release profiles of PGA-4HGF at (C) pH 7.4, (D) pH 4.0, and (E) hair follicles-like conditions (pH 5.5 & 37°C).

Figure 4 Penetration of FITC-labeled PGA-4HGF through the porcine skin after incubation with shaking (150 rpm) for (A) 0.5 hrs, (B) 3 hrs, and (C) 6 hrs, or without shaking for (D) 0.5 hrs, (E) 3 hrs, and (F) 6 hrs. Depth specifications: (A) 1: 15 µm, 2: 35 µm, 3: 85 µm, 4: 135 µm, 5: 185 µm, 6: 235 µm, and 7: 285 µm. (B) 1: 830 µm, 2: 860 µm, 3: 890 µm, 4: 950 µm, 5: 980 µm, 6: 1010 µm, and 7: 1070 µm. (C) 1: 1600 µm, 2: 1650 µm, 3: 1700 µm, 4: 1750 µm, 5: 1800 µm, 6: 1850 µm, 7: 1900 µm, 8: 1950 µm, and 9: 2000 µm. (D) 1: 0 µm, 2: 50 µm, 3: 100 µm, 4: 150 µm, 5: 200 µm, 6: 250 µm, 7: 300 µm, 8: 350 µm, and 9: 400 µm. (E) 1: 720 µm, 2: 750 µm, 3: 780 µm, 4: 840 µm, 5: 870 µm, 6: 900 µm, and 7: 960 µm. (F) 1: 910 µm, 2: 940 µm, 3: 970 µm, 4: 1030 µm, 5: 1060 µm, 6: 1090 µm, and 7: 1150 µm.

Figure 5 (A) Thermal stability by DPPH radical scavenging activity assay. (B) Average size and PDI values of PGA-4HGF during incubation in PBS (pH 7.4).

Figure 6 Effects of PGA-4HGF on: (A) the size of hair bulbs in mice and (B) the morphology of DPCs. In (A), data are represented as mean ± standard deviation (SD) (n > 30 hairs), and values with different alphabets in the same row were significantly different. Scale bar = 100 µm. Data were analyzed by one-way analysis of variance (ANOVA)/Duncan’s t-test. (p < 0.05).