Development and Characterization of Corticosteroid loaded Lipid carrier system for Psoriasis

 

Krishna Yadav, Deependra Singh, Manju Rawat Singh*

University Institute of Pharmacy, Pt. Ravishankar Shukla University, Raipur, Chhattisgarh, 492010, India.

*Corresponding Author E-mail: manjursu@gmail.com

 

ABSTRACT:

Psoriasis is a highly complex autoimmune disease of skin with keratinocytes that differentiate abnormally. Because of the inaccessibility of the adequate delivery system, a very favored treatment approaches contemplating topical treatment with current delivery approach are unreliable in targeting disease symptoms. The current work involved the development and characterization of Triamcinolone acetonide loaded solid lipid nanoparticles (TAC-SLNs) and their invitro characterization. The findings revealed that SLNs would substantially increase TA drug permeation across the skin, limiting them to dermis epidermis and helping to achieve the full clinical value of medications. It was also able to sustain the release of drug from lipid carrier. The outcomes of this research concluded that SLNs may efficiently deliver corticosteroids for the efficient treatment of psoriasis.

 

KEYWORDS: Psoriasis, Hyperproliferation, keratinocytes, triamcinolone acetonide, solid lipid Nanoparticles.

 

 


1. INTRODUCTION:

Psoriasis is an immunological skin condition that interferes with T-cells and is predisposed to the creation of an inflamed, thick, hyperproliferative state that is controlled genetically, immunologically and environmentally 1,2. It greatly influences the quality of health of the individual, with an incidence of 2-5 per cent of the world's population. Medication and psoriasis management are highly complex and unpredictable based on the severity and rigor of the condition 1,3,4. Becoming a domain of pathological expression, the topical route is the most favored route for psoriasis-targeted drugs to successfully enter the proximity of the active site below the surface. Psoriasis is a multisystem condition that involves a multi-directional therapy approach that used a specific molecule that works by more than one pathway for optimum clinical benefit 5,6. Of all treatment therapies, topical corticosteroid therapies are ideally used globally for successful psoriasis treatment. Variety of existing methods with the aid of corticosteroids for therapeutic applications are available 7,8.

 

Triamcinolone acetonide (TAC) is by far the most extensively used glucocorticosteroid with a different pharmacological potency as an anti-inflammatory, immunosuppressive, vasoconstrictive and anti-proliferative component in dermal abnormalities that indicates the significance of TA in the treatment of psoriasis. .Pharmacodynamically corticosteroids are capable of inhibiting A2 phospholipase, which is the primary regulating factor for the evocation of arachidonic acid, prostaglandin biosynthesis and leukotriene, which tends to be among the big pathogenic cause 8–12.

 

Technologies have been proposed innovative delivery mechanisms based on lipids for resolving penetration issues, suggesting transdermal delivery for psoriatic skin. It has known that the efficacy of the therapeutic agents can be assured by the delivery approaches. It involves an adequate range of drug carrier schemes. A newer version of this area is the transdermal delivery of drugs by solid lipid nanoparticles (SLNs). SLNs are strengthened with cardinal features, which include effective drug infiltration and relatively high drug capture 13–15. SLNs are nano-sized lipid carriers possessing a profound hydrophobic lipid structure stabilized by a surfactant 6,16–20. Thus, SLNs appear to be an intriguing strategy to the successful delivery of antipsoriatic drugs via the epidermis, which seems to be the main site of psoriasis operation. The key objective of this research was therefore to determine the effectiveness and to explore the capacity of the TA-loaded SLN method for successful transepidermal psoriasis delivery. The drug was loaded into SLNs to facilitate effective penetration of drugs through the skin for maximum medicinal properties. It was further defined by particle size, zeta potential, drug trapping and drug loading, and Fourier-transform infrared spectroscopy (FTIR) and Differential scanning calorimetry (DSC) compatibility. Scanning Electron Microscopy (SEM) also conducted for structure and surface morphology observation. Optimized lipid carriers were also analyzed for in vitro penetration and retention of drugs on pig ears to ensure delivery capacity of SLNs.

 

2. MATERIALS AND METHODS:

2.1. Materials

TA was kindly purchased from Clementia Biotech, New Delhi. Glycerol monostearate (GMS), Phospholipid, Poloxamer 188 and, were obtained from Hi-Media (India). The purified water used throughout the analysis was supplemented with an ultra-pure water device (Synergy UV water purifier system, India). The other materials being used are of analytical standard.

 

2.2. Method of TAC loaded SLNs

SLNs incorporating TAC is developed using a solvent evaporation methodology with marginal modification 10,17. In particular, the lipid phase was processed by the melting of glyceryl monostearate in a water bath at 70°C, when the appropriate TA measure was scattered into the molten lipid phase. An aqueous solution was prepared with 0.1 per cent of Poloxamer 188 and a co-surfactant by warming at 70°C. This mixture was slowly added to this liquid lipid mixture using a mechanical stirrer at 1500 rpm and held at 70°C to build an emulsion. The emulsion was rapidly cooled to around 20°C, immersed in an ice bath and sonicated for 10 minutes to produce sufficient dispersion. Then subsequently, it was lyophilized for 24hrs to acquire TAC-SLNs.

 

2.3. Invitro Characterization of TAC-SLNs:

2.3.1. Compatibility study

The compatibility of TAC with all the components of SLNs were ensured through FTIR and DSC. For FTIR analysis the KBr technique was adopted and scanned for spectra whereas DSC curve was obtained by heating the TAC and TAC-SLNs in differential scanning calorimeter.

 

2.3.2. Particle size and Zeta Potential:

Particle size and zeta potential of TAC-SLNs were investigated by Malvern Zetasizer ZS 90 (Malvern instrument Inc., UK). A partial quantity of dried SLNs was suspended in double-distilled water (10mL) and sonicated for 5 seconds. This suspension was then allowed for particle size and zeta potential analysis. The process was repeated thrice for each prepared batch.

 

2.3.3. Surface morphology:

The shape and surface morphology of the TAC-SLNs were determined by transmission electron microscopy (TEM). A sample of TAC-SLNs was mounted on the standard sample mounting stubs and processed. Photomicrographs were seen for morphological observation.

 

2.3.4. Drug entrapment efficiency and loading efficiency

The TAC entrapment in SLNs was determined by centrifugation of TAC-SLNs dispersion at 8000 rpm, 4°C for 25 min. The supernatants were then isolated and afterward diluted suitably in methanol to inspect under UV spectrophotometer for assurance of unentrapped TAC. Estimations were accomplished at 238 nm for TAC. Drug entrapment efficiency (EE) and drug loading (%DL) were dictated by the equation:

 

% EE = Total drug – Free drug

–––––––––––––––––––––––––––––––––––––X100

Total drug

 

% DL = Total drug – Free drug

–––––––––––––––––––––––––––––––––––––X100

Total lipid

 

 

2.3.5. In-vitro drug release studies

In-vitro drug release from TAC-SLNs and drug aqueous dispersion were studied utilizing the dialysis membrane (dialysis tubing of cellulose film, D9652-100FT, Sigma-Aldrich) technique. Dialysis bag was loaded up with TAC-SLNs dispersion for the assertion of medication release from SLNs. TAC-SLNs and Pure TAC corresponding to 10 mg of medication was submerged in a vial containing 10 ml of release media. A Blend of phosphate buffer saline (PBS) pH 7.4 and methanol (70:30 v/v) was used as release media. Vials were situated on a shaker at 80 rpm and 37oC. At, a fixed time interim, 1 ml of aliquot was withdrawn from the receptor compartment and replenish with an equivalent measure of new discharge media. The release of medication was assessed by UV-spectrophotometry at 238 nm for TAC. The results were mean of three estimations. The release profile of TAC from TAC-SLNs and TA drug dispersion were evaluated by a curve fitting technique.

 

2.5.4 In-vitro drug penetration and distribution studies

In-vitro drug distribution into the skin from TAC-SLNs gel was performed through rat skin. The pig ear were acquired from the local slaughter house (Shastri market, Raipur). To assess the in-vitro drug penetration and skin distribution of medicating by means of TAC-SLNs loaded into carbopol gel 1%. The penetration studies were executed by the Franz diffusion cells with a slight change 21,22. The rat skin (surface region 3.46 cm2) was cut and mounted on Franz cells between donor and receptor compartment with SC fronting to the donor. The formulation containing a drug proportional to 100 mg was packed into the donor compartment. PBS of pH 7.4 with methanol (70:30 v/v) was used as discharge media to keep up the sink condition. The temperature of the medium was settled at 38.7±01oC and stirred at 250 rpm. At foreordained time interim (0, 1, 2, 3, 4, 5, 6, 8, 12 and 24 hours) samples were withdrawn from the receptor compartment and recharged with an equivalent volume of new media. The samples were investigated for medicate penetration through the skin by UV spectroscopy at 238 nm for TAC. After 24 hours, the skin was secluded from Franz cell and the remainder of the gel at the outside of the extracted skin was cleaned out totally with 15 ml water, from there on medicate in the residual gel was separated by adding of 10 ml methanol to the washed sample. The skin was homogenized with 5 ml discharge medium and medication was separated by the addition of 5 ml methanol to all samples. These samples were centrifuged (10000 rpm for 30 min) and the supernatants were measured by UV spectroscopy. The removed medication from dermis-epidermis was estimated for distribution. These measures were rehashed in triplicate.

 

3. RESULTS AND DISCUSSION:

TAC-SLNs were formulated using the solvent evaporation technique and optimized. Further, the optimized formulation was acquainted with different in-vitro investigations.

 

3.1. Compatibility study:

The FTIR spectra and DSC curve of pure TAC and TAC-SLNs showed that drug was compatible with all the component of lipid carriers. The FTIR spectra showed that TAC was capable of maintaining it chemical integrity and characteristic functional groups in the physical mixture of SLNs with fewer shift in peaks (Fig. 1). The DSC curve of TA showed a sharp melting at 294.73°C which was absent in their SLNs form ensuring the proper dispersion of drug inside lipid carrier (Fig. 2).

 

 

 

Fig. 1. FT-IR spectra of A. pure TA and B. TA in physical mixture (TAC-SLNs).

 

Fig. 2. DSC thermogram of A. pure TA and B. TAC-SLNs.

 

3.2. Particle size, Zeta Potential and Surface morphology

The particle and PDI study on optimized TAC-SLNs were estimated as 262.54±1.95 (Fig. 3A) and PDI of 0.293±0.14. The zeta potential was found as -12.6 ± 4.35 (Fig. 3B). The zeta potential estimation of chosen batch showed the stable formulation as the higher estimation of zeta potential (negative or positive) assigns stable SLNs scattering as profoundly charged particles are responsible for the avoidance of particle aggregation because of electric repulsion. The TEM photomicrograph of TAC-SLNs (Fig. 3C) affirmed the oval and round state of SLNs with a narrow size range. The diameter of the SLNs showed in the micrographs were in acceptable concurrence with the information acquired from the DLS study.

 

Fig.3. A. Particle size and PDI of TA-SLNs; B. Zeta Potential of TA-SLNs; C. TEM photomicrograph of TAC-SLNS.

 

3.3. Drug entrapment efficiency and loading efficiency

Percent drug entrapment and drug loading of the TAC-SLNs were assessed at 81.12±1.87% and 12.6±1.32%respectively. High entrapment could be attributed to the lipophilic nature of medication guaranteeing higher proclivity for the picked lipid matrix.

 

3.4 In-vitro drug release studies

In vitro release testing is a significant expository tool that is utilized to examine and build up drug discharge behavior during different phases of product development. The drug release of TAC from drug dispersion and TAC-SLNs dispersion was inspected over a time span of 24 h (Fig. 4). It showed a relative discharge profile of TAC from the drug dispersion and SLNs dispersion. TAC TAC-SLNs dispersion allowed medication release of about 29.135±5.11% and 16.17±4.21% respectively over a time span of 24 h demonstrating the release of medication in transporter grid contrast with drug release from the suspension of plain medication. The consolidation of GMS brought about expanded consistency of medium and more rigid solidified SLNs impeding medication diffusion to the release medium.

 

Fig. 4. Invitro drug release profile of TAC-SLNs.

 

3.5. In-vitro drug penetration and distribution studies

In-vitro skin distribution investigations of TAC-SLNs gel was done utilizing pig ear skin. The outcome observed after 12 h indicated that TAC-SLNs could enter across pig skin, demonstrated that SLNs could be capable to provide TAC and possibly will exert its therapeutic effect. Extraction of medication from dermis-epidermis homogenate tests exhibited that SLNs, containing lipid, could upgrade the deposition of medication in dermis-epidermis compare to free TAC, (Fig. 5) and it was very much coordinated with past research. These results proposed that SLNs are appropriate conveyance systems for TAC to target the dermis-epidermis layer with the ultimate objective of antiinflammatory activity in skin 23.

 

Fig. 5. The deposition of medication in various compartment of skin.

 

4. CONCLUSION:

The promising results of current in vitro examinations acclaimed the potential usage of SLNs for the topical conveyance of drugs intending to a skin issue. In summary, a SLNs containing TAC was formulated for the treatment of psoriasis. The nanosized TAC-SLNs were optimized for size and medication entrapment. This SLNs was further evaluated for drug entrance and skin deposition. Results demonstrated that SLNs outfitted simplicity of utilization, progressively significant passageway, and moderate release of drugs over the epidermal hindrance and limited appropriation to the dermis layer. From these invitro investigations, it was evident that TAC-SLNs may improve the symptoms of inflammatory skin by efficient delivery of TAC. The results of this examination can be a way to advance for the conveyance of TAC using SLNs for effective management of psoriatic skin.

 

5. ACKNOWLEDGMENTS:

The authors are thankful to the Director, University Institute of Pharmacy, Pt. Ravishankar Shukla University, Raipur, Chhattisgarh for providing necessary infrastructure facilities and would like to acknowledge DHR-ICMR project no. V.25011/286-HRD/2016-HR for financial assistance related to this work. I also acknowledge the contributions made by the library of Pt. Ravishankar Shukla University for giving me an opportunity to use their e-resources available through UGC-INFLIBNET.

 

6. DECLARATION OF INTEREST:

The authors declare no conflict of interest.

 

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Received on 19.08.2020           Modified on 14.10.2020

Accepted on 21.11.2020         © RJPT All right reserved

Research J. Pharm. and Tech. 2021; 14(2):966-970.

DOI: 10.5958/0974-360X.2021.00172.4