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Evaluation of Stryphnodendron sp. Release Using Natural Rubber Latex Membrane as Carrier



Karoline M. Romeira, Bruno C. Drago, Heitor D. Murbach, Patricia B. Aielo, Regildo M.G. da Silva, Carlos A. Brunello and Rondinelli D. Herculano
 
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ABSTRACT

Natural rubber latex from Hevea brasiliensis has interesting characteristics related to this work such as: it is easy to manipulate, low cost, can stimulate the natural angiogenesis, is a biocompatible material and presents high mechanical resistance. The aim of this study was to develop a novel sustained delivery system for Stryphnodendron sp. based on Natural Rubber Latex (NRL) membranes and to study the Stryphnodendron sp. delivery system behavior. Stryphnodendron sp., commonly known as barbatimão is extensively used in folk medicine for the treatment of diarrhoea, gynaecological problems and for healing wounds. The stem bark of this species is mentioned in the Brazilian Pharmacopeia with a content of at least 20% of tannins. Previous studies showed significant cicatrizant properties, anti-inflammatory activity and gastric anti-ulcerogenic effects for the stem bark crude extract. One possible way to accelerate the tissue repair process, it was incorporated the Stryphnodendron sp. extract in NRL membranes. Stryphnodendron sp extract was incorporated into the NRL, by mixing it in solution for in vitro protein delivery experiments. Results show that the NRL membrane can release Stryphnodendron sp. for up to 49.89% of its Stryphnodendron sp. content for up 400 h. The kinetics of the extract release could be fitted with double exponential function, with two characteristic times of 0.78 and 133.22 h. In this study, we demonstrated that the induced angiogenesis provided by NRL membranes combined with a controlled release of extract is relevant for biomedical applications.

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Karoline M. Romeira, Bruno C. Drago, Heitor D. Murbach, Patricia B. Aielo, Regildo M.G. da Silva, Carlos A. Brunello and Rondinelli D. Herculano, 2012. Evaluation of Stryphnodendron sp. Release Using Natural Rubber Latex Membrane as Carrier. Journal of Applied Sciences, 12: 693-697.

DOI: 10.3923/jas.2012.693.697

URL: https://scialert.net/abstract/?doi=jas.2012.693.697
 
Received: January 13, 2012; Accepted: March 31, 2012; Published: June 19, 2012



INTRODUCTION

The controlled release of drugs/extracts can be efficient if a suitable encapsulation procedure is developed, which requires biocompatible materials to hold and release the drug/extracts. In this study, NRL membrane is used to deliver Stryphnodendron sp extract. Latex membrane from Hevea brasiliensis is an important inductor of wound healing, regeneration of organism. (Ereno et al., 2010; Sampaio et al., 2010; Herculano et al., 2011a; Ciapetti et al., 1994; Mendonca and Coutinho-Netto, 2009). In addition, the treatment of diabetic and phlebopathic ulcers with the biomembrane prepared from the natural latex showed the presence of a vascular growth factor in the latex (Balabanian et al., 2006; Mendonca and Coutinho-Netto, 2009; Ferreira et al., 2009; Frade et al., 2001; Mrue et al., 2004). Stryphnodendron sp., commonly known as barbatimão, is extensively used in folk medicine for the treatment of diarrhoea, gynaecological problems and for healing wounds. The stem bark of this plant contains a considerable amount of tannin (10-37%) and prominent presence of several flavan-3-ols, proanthocyanidins and prorobinetinidins (Lopes et al., 2005; Lima et al., 1998; Holetz et al., 2005; Santos et al., 2002). The crude extract, in the form of decoction or infusion, is traditionally used by the local population for the treatment of leucorrhoea and diarrhea, as well as an anti-inflammatory and healing agent (Di Mambro et al., 2005; Castro et al., 2009; Eurides et al., 1995). Some studies of the crude extract from the stem bark of Stryphnodendron adstringens have revealed significant anti-inflammatory activity and gastric anti-ulcerogenic effects. It has also been observed that a decoction of the stem bark of Stryphnodendron sp. accelerates the healing of wounds and decreases inflammation (Minatel et al., 2010). One possible way to accelerate the tissue repair process, it was incorporated the Stryphnodendron sp. extract in NRL. Stryphnodendron sp. extract was incorporated into the NRL, by mixing it in solution for in vitro protein delivery experiments. NRL membranes have already been used to deliver drugs (Herculano et al., 2006, 2007, 2009, 2010).

In this study, we propose a novel release system based on the encapsulation of Stryphnodendron sp. in NRL membrane for the sustained and controlled delivery of Stryphnodendron sp. for future applications in medicine as wound healing. Results show that the NRL membrane can release Stryphnodendron sp. for up to 400 h, which is relevant for biomedical applications.

MATERIALS AND METHODS

The NRL used in the present study was commercial high-ammonia natural rubber latex (ESALQ-USP, Piracicaba, Brazil) of about 60% dry rubber content (DRC), 4-5% weight of non-rubber constituents such as protein, lipids, carbohydrates and sugar and 35% of water (Hashim and Subramaniam, 1986; Yip et al., 1994; Yip and Sussman, 2000). After extraction, ammonia was used to keep the latex liquid. The deproteinization of natural rubber latex was performed by centrifugation at 8,000 g. The cream fraction after centrifugation was re-dispersed to make 60 wt.% of dry rubber content latex and was washed twice by centrifugation to prepare the deproteinized natural rubber latex and then reduce the cytotoxic protein content on NRL. Stryphnodendron sp., commonly known as barbatimão were collected in March 2011 at Paranapanema Valley region, (Sao Paulo State, Brazil). The barks extract (20 g) was extracted by stirring in an Erlenmeyer at room temperature with acetone/water 7:3 (4x100 mL). These extract was evaporated under vacuum to 30% volume and filtered to remove fats and then freeze-dried. Extract of Stryphnodendron sp. was incorporated by mixing 2 mL of natural rubber latex with 2 mL of extract solution (10 mg mL-1). These membranes were prepared by pouring the latex+extract solution in a stainless steel plate with 5.00±0.05 cm diameter and 200±5.00 μm thick. Typically the membranes were left for 2 days to fully polymerize before use. For the study of the release of extract, latex membrane was placed in 200 mL of an aqueous solution, from which aliquots were collected during an interval ranging from 10 to 24,000 min. The extract released into the solution was monitored by measuring the UV-VIS spectra with a BEL ENGINEERING SF 200 ADV spectrophotometer, as Stryphnodendron sp. extract has a maximum absorption at 297 nm.

RESULTS AND DISCUSSION

The goal of controlled-release delivery systems is to provide desirable delivery patterns so that predictable plasma drug/extract levels can be achieved, which requires the characterization of the basic pharmacodynamic and pharmacokinetic properties of a drug/extract.

Fig. 1(a-b): Absorbance intensity as a function of Stryphnodendron sp. concentration in solution

For Stryphnodendron sp. studied here, it has been found that it absorbs at 297 nm. In Fig. 1 shows the absorbance intensity as a function of Stryphnodendron sp. concentration in solution.

The sustained delivery system for Stryphnodendron sp. has been successfully developed based on NRL. The release profile for Stryphnodendron sp. in a NRL matrix in Fig. 2 shows that the first, fast step of burst release corresponded to the Stryphnodendron sp. near or on the surface of the NRL membrane (Herculano et al., 2011b). The slower release process would be associated with Stryphnodendron sp. diffusing slowly through the matrix. Results demonstrated that the NRL membrane can release Stryphnodendron sp. for 15 days.

The experimental data in Fig. 2 were fitted using a bi-exponential function y(t) = y0+A1e–t/τ1+A2e-t/τ2, where y(t) was the amount of Stryphnodendron sp. in the NRL at a given time, t, y0 is the initial content of Stryphnodendron sp., A1 and A2 are constants equal to -0.030 and -0.035, respectively, the characteristic times are τ1 = 0.78 h and τ2 = 133.22 h.

Fig. 2: Stryphnodendron sp. release as a function of time for the NRL membrane

Upon integrating the curve until 400 h, the total amount of Stryphnodendron sp. released by the membrane in the 200 mL aqueous solution was 17.95 mg (44,89%).

As mentioned earlier the controlled release of drugs/extracts are of interest for medical applications, since the dose can be adjusted according to the necessity of the patient (Shilpa et al., 2003; Kim et al., 2008; Sirianni et al., 2010; Malcolm et al., 2004; Rahimi et al., 2010) . In this work, it was used the method proposed by Langer and Folkman (1976), that is, to mix the drug/extract (Stryphnodendron sp.) with the polymer (latex) in a colloidal state, in order to create a membrane that works as a delivery system.

Several procedures are used to control the release of substances by polymers. For example Woo et al. (2001) used a combination of 3 different biodegradable microspheres of poly (D,L-lactide-co-glycolide (PLGA)), using different molecular weight and terminal endings of the polymeric chain (hydrophilic or hydrophobic) to determine the best delivery system of BMP. They conclude that the best bone healing results were achieved using high dose and slow delivery rate systems.

Already Lobler et al. (2000) developed a device based in polyhydroxyalkanoates (PHA) for implantation of a glaucoma drainage system. In this study, polyhydroxyalkanoates (PHA) based on hydroxybutyric acid were tested in terms of their potential suitability to manufacture mechanically stable tube components of drug delivery drainage systems and in terms of biocompatibility.

Wang et al. (2005) prepared uniform-sized chitosan microspheres by membrane emulsification technique. Uniform chitosan microspheres were further used as a carrier of protein drug (BSA). They observed that BSA loading efficiency was highest when pH value was 8.09 and it decreased with an increase of the crosslinking degree.

Herculano et al. (2011b) proposed a drug release system based on NRL for the sustained and controlled delivery of metronidazole (MET). They concluded that the release time of MET in in vitro tests was very promising for the kinetics of release.

Our results indicate that with very simple changes in preparation of NRL membrane, we could control Stryphnodendron sp. release up to 15 days, according to Woo et al. (2001), Wang et al. (2005), Herculano et al. (2011b), Malcolm et al. (2004) and Langer and Folkman (1976) results, or in other words with a slow release rate.

CONCLUSION

We have prepared natural rubber latex membranes containing Stryphnodendron sp. as a model system for tissue regeneration. The method of preparation is reproducible and the natural rubber latex membrane is very stable. Results demonstrated that the NRL membrane can release Stryphnodendron sp. for 15 days, thus making them promising materials for protein/extract release in in vivo applications. We have observed that 44.89% of the initial Stryphnodendron sp. content inside NRL was released in 400 h. Our results indicate that NRL could be used in the future as an active membrane that could accelerate tissue healing.

ACKNOWLEDGMENTS

This work was supported by PROPe/UNESP and FAPESP (Processes: 2011/17411-8, 2011/06856-9).

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