Formulation and in vitro/in vivo evaluation of chitosan-based film forming gel containing ketoprofen

Figures & data

Table 1. Formulation of CbFG containing permeation enhancer.

weight ratio %	CbFG-OA 5%	CbFG-NMP 5%	CbFG-Tw80 5%	CbFG-Cr40 5%	CbFG-OA 3%	CbFG-OA 2%	CbFG-OA 1%	CbFG-OA 0.5%
Ketoprofen	3
Chitosan	3
Lactic acid	4
Propylene glycol	2
Oleic acid	5	–	–	–	3	2	1	0.5
NMP	–	5	–	–	–	–	–	–
Tween 80	–	–	5	–	–	–	–	–
Cremophor RH 40	–	–	–	5	–	–	–	–
95% Ethanol	40	40	40	40	40	40	40	40
D.W.	43	43	43	43	45	46	47	47.5
Total	100

Table 2. Inter- and Intra-data on ketoprofen analysis using HPLC.

Concentration of Ketoprofen (μg/ml) (n = 3)	Inter day		Intra day
	Mean ± SD	% RSD	Mean ± SD	% RSD
	15.60	15.72 ± 0.20	1.25	15.34 ± 0.08	0.53
62.50	62.26 ± 0.35	0.57	62.76 ± 0.04	0.07
250.00	249.71 ± 0.94	0.38	249.62 ± 0.03	0.01

Figure 1. Transparency changes of CbFG-film during film formation; (A) initial state after CbFG application, (B) 5 min after CbFG application; (C) 10 min after CbFG application.

Figure 2. Mechanical properties of CbFG-films; (A) film tensile strength, (B) maximum elongation at break. One-way ANOVA analysis, *p < 0.05 (n = 4) compared with film containing OA 0%.

Figure 3. SEM micrographs of CbFG-films without or with OA.

Figure 4. FT-IR spectra of CbFG-films without or with OA.

Figure 5. XRD diffractograms of CbFG-films without or with OA, raw chitosan and raw ketoprofen.

Figure 6. DSC thermograms of CbFG-films without or with OA and raw ketoprofen.

Figure 7. In vitro skin permeation profiles of CbFG-film containing ketoprofen; (A) with different kinds of enhancers through excised SD-rat skin, (B) with different amount oleic acid as an enhancer through excised hairless mouse skin. One-way ANOVA analysis,* p < 0.05 (n = 3) compared with Commercial product.

Table 3. In vitro skin permeation rate and lag time of CbFG-film through excised SD-rat skin and hairless mouse skin.

	Commercial product	CbFG Non enhancer	CbFG OA 5% *	CbFG NMP 5%	CbFG Tw80 5%	CbFG Cr40 5%
In vitro skin permeation rate through excised SD-rat skin
Flux (μg/cm^2/h)	39.9 ± 6.1	12.6 ± 5.1	75.4 ± 5.0	13.6 ± 1.0	24.3 ± 1.5	15.2 ± 3.3
Lag time (h)	3.3 ± 1.7	3.2 ± 1.3	3.7 ± 0.8	2.8 ± 0.3	3.4 ± 0.8	4.1 ± 0.7
	Commercial product	CbFG OA 3%*	CbFG OA 2%*	CbFG OA 1%*	CbFG OA 0.5%*
In vitro skin permeation rate through excised hairless mouse skin
Flux (μg/cm^2/h)	95.5 ± 28.0	209.9 ± 17.6	281.7 ± 77.0	264.9 ± 7.3	308.6 ± 11.5
Lag time (h)	1.7 ± 0.1	1.7 ± 0.9	1.8 ± 0.4	1.5 ± 0.5	1.0 ± 0.6

One-way ANOVA analysis.

* p < 0.05 (n = 3) compared with Commercial product.

Figure 8. In vivo efficacy study of CbFG-film-OA 0.5% and commercial product; (A) difference in swelling of each hind paw, (B) difference in weight of each hind paw, (C) PGEM levels in joint tissue for 2 days after administration. One-way ANOVA analysis: *p < 0.05 (n = 6), compared with positive control; **p < 0.05 (n = 6), compared with positive control; #p < 0.05 (n = 6), compared with negative control; ##p < 0.05 (n = 6), compared with negative control; +p < 0.05 (n = 6), compared with Commercial product; ++p < 0.01 (n = 6), compared with Commercial product.