Punica granatum L. Hydrogel for Wound Care Treatment: From Case Study to Phytomedicine Standardization - PubMed (original) (raw)

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Punica granatum L. Hydrogel for Wound Care Treatment: From Case Study to Phytomedicine Standardization

Aline Fleck et al. Molecules. 2016.

Abstract

The pharmacological activities of many Punica granatum L. components suggest a wide range of clinical applications for the prevention and treatment of diseases where chronic inflammation is believed to play an essential etiologic role. The current work reports a case study analyzing the effect produced by a magistral formulation of ethanolic extracts of Punica granatum peels on a non-healing chronic ulcer. The complete closure of the chronic ulcer that was initially not responsive to standard medical care was observed. A 2% (w/w) P. granatum peels ethanolic extract hydrogel-based formulation (PGHF) was standardized and subjected to physicochemical studies to establish the quality control parameters using, among others, assessment criteria such as optimum appearance, pH range, viscosity and hydrogel disintegration. The stability and quantitative chromatographic data was assessed in storage for six months under two temperature regimes. An efficient HPLC-DAD method was established distinguishing the biomarkers punicalin and punicalagin simultaneously in a single 8 min run. PGHF presented suitable sensorial and physicochemical performance, showing that punicalagin was not significantly affected by storage (p > 0.05). Formulations containing extracts with not less than 0.49% (w/w) total punicalagin might find good use in wound healing therapy.

Keywords: Punica granatum; hydrogel; punicalagin; quality control; wound healing.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1

Figure 1

Chemical structures of the ellagitannins punicalin (a) and punicalagin (b).

Figure 2

Figure 2

Photographs of the patient’s wound. PGMF: Punica granatum magistral formulation; (a): 1 day before PGMF treatment; (b): week 1 of PGMF treatment; (c): week 6 of PGMF treatment; (d): week 10 of PGMF treatment.

Figure 3

Figure 3

Analytical curves of the biomarkers (a) punicalin and (b) punicalagin.

Figure 4

Figure 4

HPLC chromatographs of punicalin (a) and punicalagin (b) standards. Chromatograph was run on a Shimadzu LC-20AT HPLC equipped with a SPD-M20A PDA detection system. Separations were carried out on a Dr. Maisch GmbH C18 (Ammerbuch-Entringen, Germany) column (5 μm, 250 mm × 4.6 mm), 25 °C, with a mobile phase of 0.1% (v/v) aqueous solution of phosphoric acid and acetonitrile (80:20). The mobile phase flow rate was 1 mL/min with 10 min run time. The standards were detected using a wavelength of 254 nm. Sample injection volume was 10 μL. Retention times for punicalin and punicalagin were 4.203 and 5.987 min, respectively.

Figure 5

Figure 5

Spreadability test of hydrogel-based formulation of ethanolic extract of P. granatum peels (2% w/w) in different storage conditions. (a) Climatic chamber; (b) Temperature test chamber; T0: after production; T3: 3 months; T6: 6 months. Results are represented by means (n = 3).

Figure 6

Figure 6

Share stress—share rate curves of hydrogel-based formulation of ethanolic extract of P. granatum (2%, w/w); T0: after production, T3: 3 months and T6: 6 months of storage under different storage conditions. (a) Temperature test chamber; (b) Climatic chamber. Results are represented by means (n = 3).

Figure 7

Figure 7

Viscosity—share rate curves of hydrogel-based formulation of ethanolic extract of P. granatum (2%, w/w); T0: after production, T3: 3 months and T6: 6 months of storage at different storage conditions. (a) Climatic chamber; (b) Temperature test chamber. Results are represented by means (n = 3).

Figure 8

Figure 8

Disintegration time of hydrogel-base formulations. Blank; after production (T0); temperature test chamber-polyethylene-time: 3 months (TTCPolyT3); temperature test chamber-polyethylene-time: 6 months (TTCPolyT6); temperature test chamber-aluminum-time: 3 months (TTCAlT3); temperature test chamber-aluminum-time: 6 months (TTCAlT6); climatic chamber-polyethylene-time: 3 months (CCPolyT3); climatic chamber-polyethylene-time 3months (CCPolyT6); climatic chamber-aluminum-time: 3 months (CCAlT3); climatic chamber-aluminum-time: 6 months (CCAlT6). Results are represented by means ± S.E.M. (n = 3).

Figure 9

Figure 9

HPLC chromatographs of Punica granatum peel ethanolic extract hydrogel-based formulation (PGHF), showing retention times for punicalin (4.203 min) and punicalagin (5.987 min). Chromatography was run on a Shimadzu LC-20AT HPLC with SPD-M20A PDA detection. Separations were carried out on a Dr. Maisch GmbH C18 column (5 μm, 250 mm × 4.6 mm), 25 °C, with a mobile phase of 0.1% (v/v) aqueous solution of phosphoric acid and acetonitrile (80:20). The mobile phase flow rate was 1 mL/min with 10 min run time. The standards were detected using a wavelength of 254 nm. Sample injection volume was 10 μL.

Figure 10

Figure 10

Punicalagin (a) and punicalin (b) contents in PGHF; HGT3TTC-hydrogel, time: 3 months, temperature test chamber; HGT6TTC-hydrogel, time: 6 months, temperature test chamber; HGT3CC -hydrogel, time: 3 months, climate chamber; HGT6CC-hydrogel, time: 6 months, climate chamber. Results are represented by means ± S.E.M. (n = 3). No statistical significant difference was detected. Statistical analysis was performed by two-way ANOVA followed by Bonferroni’s test of multiple comparisons (p < 0.05).

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