Drug Release and Cytotoxicity of Hyaluronic Acid and Zinc Oxide Gels, An In-Vitro Study

Figures & data

Figure 1. Study design.

Table 1. Study groups.

Group A – Zinc Oxide Gels	Group B – Hyaluronic acid Gels
I. 87% w/v (POSITIVE CONTROL) II. 36% w/v zinc Oxide gel. III. 18% w/v zinc Oxide gel. IV. 9% w/v zinc oxide gel.	Part 1 – HMW-HA
	0.1% 1% 5% 10%
	Part 2 – LMW-HA
	0.1% 1% 5% 10%

(LMW-HA: Low molecular weight hyaluronic acid; HMW-HA: High molecular weight hyaluronic acid.)

Figure 2. Drug release pattern of different concentrations of LMW-HA by UV spectrophotometry. (LMW-HA: Low molecular weight hyaluronic acid; UV: Ultraviolet).

Table 2. Comparison of viability values at different time intervals among different concentrations within the groups.

		24 hr			48 hr			72 hr
	Conc (%)	Median (Min – Max)	IQR	p value	Median (Min – Max	IQR	p value	Median (Min – Max	IQR	P value
LMW-HA-HPDL	0.10	131.93(89.2 − 239.2)	130.5	0.8	103.41 (71.22–198.68)	99.44	0.4	134.68 (130.67 142.59)	9.68	0.008*
	1	125.47 (68.2–141.9)	57.65		95.04 (73.08–106.18)	25.96		67.83 (40.54–86.17)	38.01
	5	116.74 (78.36–168.49)	78.86		112.74(91.22–119.39) 93.65(78.45–108.39	23.45		95.65 (78.75–107.21)	25.54
	10	107.65 (87.45–117.24)	24.72			28.16		93.5 (80.82–95.19)	11.05
LMW-HA-HGF	0.10	97.76 (56.95–162.17)	93.48	0.6	70.69 (51.3–81.95)	23.61	0.9	95.95 (57.86–115.95)	43.65	0.54
	1	106.08 (60.89–150.93)	72.85		62.32 (57.77–104.8)	36.83		91.1 (57.61–112.93)	42.73
	5	129.56 (52.59–158.23)	91.73		73.68 (52.21–115.14)	49.92		107.38 (72.18–134.43)	46.82
	10	82.34 (55.0–99.2)	36.59		70.92 (49.83–85.09)	26.65		107.91 (87.81–130.76)	35.26
HMW-HA-HPDL	0.10	99.15 (81.06–111.33)	24.82	0.02*	99.15 (81.06–111.33)	24.82	0.02*	119.22 (104.51–141.96)	30.14	0.01*
	1	109.39 (93.84–160.59)	51.49		109.39 (93.84–160.59)	51.49		77.22 (62.32–127.31)	50.77
	5	127.11 (63.73–213.9)	114.38		127.11 (63.73–213.9)	114.38		88.61 (84.05–91.56)	6.05
	10	50.52 (40.32–59.91)	15.41		50.52 (40.32–59.91)	15.41		59.87 (50.68–61.94)	8.82
HMW-HA-HGF	0.10	87.62 (46.8–132.9)	70.89	0.03*	87.62 (46.8–132.9)	70.89	0.03*	105.76 (75–136.73)	59.08	0.25
	1	97.99 (89.39–104.17)	11.41		97.99 (89.39–104.17)	11.41		80.71 (71.69–87.79)	13.3
	5	91.68 (57.78–106.6)	39.65		91.68 (57.78–106.6)	39.65		78.58 (67.51–84.66)	15.64
	10	22.36 (17.9–33.31)	11.83		22.36 (17.92–33.31)	11.83		65.21 (57.23–91.78)	26.87

P value calculated using Kruskal–Wallis test; *Significant

(LMW-HA: Low molecular weight hyaluronic acid; HMW-HA: High molecular weight hyaluronic acid; HPdLF: Human periodontal ligament fibroblasts; hGFs: Human gingival fibroblasts)

Table 3. Comparison of the viability values within the groups at subsequent time intervals.

Variables		0.10%	1.00%	5%	10%
LMW-HA	PDL	0.77	0.17	0.17	0.47
	HGF	0.36	0.36	0.36	0.039*
HMW-HA	PDL	0.039*	0.17	0.17	0.039*
	HGF	0.47	0.36	0.36	0.018*

P value calculated using Friedman test; *Significant (LMW-HA: Low molecular weight hyaluronic acid; HMW-HA: High molecular weight hyaluronic acid; HPdLF: Human periodontal ligament fibroblasts; hGFs: Human gingival fibroblasts)

Table 4. Comparison of cell viability between the LMW-HA and HMW-HA groups.

Time intervals	Concentrations	Chi-Square value	p value
24 hr	0.10%	1.985	0.575
	1%	1.963	0.580
	5%	1.875	0.599
	10%	13.125	0.004*
48 hr	0.10%	7.743	0.052
	1%	1.875	0.599
	5%	6.684	0.083
	10%	10.891	0.012*
72 hr	0.10%	6.296	0.098
	1%	2.713	0.438
	5%	5.382	0.146
	10%	11.272	0.010*

*Significant

(LMW-HA: Low molecular weight Hyaluronic acid; HMW-HA: High molecular weight Hyaluronic acid; HPdLF: Human periodontal ligament fibroblasts; hGFs: Human gingival fibroblasts)

Figure 3. 96-Well microplates used for cytotoxicity with all the experimental groups.