Available online on 15.01.2024 at http://jddtonline.info

Journal of Drug Delivery and Therapeutics

Open Access to Pharmaceutical and Medical Research

Copyright  © 2024 The  Author(s): This is an open-access article distributed under the terms of the CC BY-NC 4.0 which permits unrestricted use, distribution, and reproduction in any medium for non-commercial use provided the original author and source are credited

Open Access  Full Text Article                                                                                                                                                     Research Article

Preparation and Evaluation of Ketoprofen Loaded Mucoadhesive Microspheres

Suman Pattanayak, Lakshmi Kanta Kanthal, Tamanna Afnan, Shalini Shukla, Pritish Kumar Panda*

Department of Pharmaceutics, Haldia Institute of Pharmacy (An institute of ICARE), ICARE Complex, Hatiberia, Haldia, Purba Medinipur, West Bengal-721657, India

 

 

INTRODUCTION

A mucoadhesive microsphere is a small particle or sphere that is designed to adhere to the mucosal surfaces of the body, such as the gastrointestinal tract, nasal cavity, or ocular surfaces. These microspheres are typically composed of biocompatible polymers that have adhesive properties, allowing them to interact with the mucosal surfaces and remain attached for an extended period of time.

Mucoadhesive microspheres have gained significant attention in the field of drug delivery due to their ability to provide sustained and localized drug release. By adhering to the mucosal surfaces, they can overcome various barriers, such as rapid clearance, enzymatic degradation, and the need for frequent dosing associated with conventional drug formulations. These microspheres can be loaded with a variety of drugs, including small molecules, peptides, proteins, and nucleic acids, and can be formulated to release the drug in a controlled manner over an extended period of time. The mucoadhesive properties of the microspheres ensure prolonged contact with the mucosal surfaces, facilitating enhanced drug absorption and bioavailability ^1,2,3.

Ketoprofen helps relieve pain by inhibiting the synthesis of prostaglandins, which are chemical messengers involved in pain and inflammation. By blocking the action of cyclooxygenase (COX) enzymes, ketoprofen reduces the production of prostaglandins, thereby reducing pain sensation. Ketoprofen exhibits anti-inflammatory effects by reducing the release of inflammatory mediators such as prostaglandins, leukotrienes, and histamines. By suppressing inflammation, it helps alleviate symptoms associated with inflammatory conditions like arthritis, tendinitis, and gout ⁴. Ketoprofen is available as a topical gel or cream for localized application on painful or inflamed areas. When applied to the skin, it penetrates the underlying tissues, providing localized pain relief and reducing inflammation in the affected area ⁵.

Ketoprofen inhibits both cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) enzymes, although it has a higher affinity for COX-1. COX-1 is involved in various physiological processes, including maintaining the integrity of the stomach lining and regulating blood platelets. COX-2, on the other hand, is induced during inflammation and plays a role in pain and swelling ⁶. Ketoprofen can lower fever by inhibiting the production of prostaglandins in the hypothalamus, which is the part of the brain that regulates body temperature. By reducing the production of prostaglandins, ketoprofen helps bring down elevated body temperatures ⁷.

MATERIALS AND METHODS

Ketoprofen was obtained from Infinity Chemicals Pvt. Ltd, Mumbai, India, Carbopol 934P was purchased from Loba chemical Pvt Ltd, Sodium alginate was purchased from Nice chemicals Pvt Ltd. 

 

 

Preparation of Mucoadhesive microsphere ^{8, 9}

1. Ketoprofen, Carbopol 934P, Sodium alginate were pass through 60 no. sieve.
2. Required quantities of Carbopol 934P (mucoadhesive polymer), Sodium alginate were dissolved in purified water and a homogenous polymer solution was made.
3. Ketoprofen was added to the polymeric solution and mixed thoroughly by the help of the magnetic stirrer.
4. The drug-polymeric solution was then added into the calcium chloride solution (2%) drop wise by syringe through needle (size no-18).
5. The resulting droplets were kept into calcium chloride solution up to 15 minutes to obtain its rigidness.
6. Then the prepared microspheres were separated from the solution by filtration.
7. Prepared microspheres were washed and dried at 45⁰C for 12 hours.

Following table represents the composition of mucoadhesive microspheres.

 

 

Table 1: Composition of Ketoprofen loaded mucoadhesive microspheres.

 

 

Characterization of microspheres

Particle size determination

Particle size of prepared microsphere was determined by optical microscopy. Optical microscopy involves observing microspheres under a microscope and measuring their sizes directly ¹⁰. 

Drug encapsulation efficiency

Drug encapsulation efficiency (EE) of each formulationwas determined by following formula. It is a measure of the amount of drug that is successfully encapsulated within microspheres during the formulation process. It is an important parameter in drug delivery systems, as it directly influences the therapeutic efficacy and dosage accuracy of the microsphere formulation. Drug encapsulation efficiency can be calculated using the following formula ¹¹.

EE (%) = (Amount of drug encapsulated / Total amount of drug initially added) × 100

Percentage yield

The percentage yield of microspheres was determined by the following formula. It is referring to the efficiency of the microsphere synthesis process. It indicates the proportion of the desired microspheres obtained compared to the theoretical yield, which is the maximum amount that could be produced under ideal conditions.

Use the following formula to calculate the percentage yield ¹².

Percentage yield = (Actual yield / Theoretical yield) × 100

Morphology

The morphology of prepared microspheres was determined by scanning electron microscopy (SEM) ¹³.

Flow properties

The flow properties of prepared microspheres were determined by porosity, angle of repose, Carr’s index, Hausner’s ratio ^{14, 15}. 

Carr’s index=×100

Hausner’s ratio=

TD= Tapped density, BD=Bulk density

Mucoadhesive properties

The degree of swelling of mucoadhesive microspheres refers to the extent to which these microspheres increase in size when exposed to a surrounding medium, typically biological fluids or mucosal surfaces. To measure the degree of swelling, the microspheres were placed in acid buffer and allowed to swell to equilibrium. The weight of the microspheres is measured at the initial dry state (W_I) and then again after swelling to a certain time point (W_F). Following formula is used to calculate the Degree of swelling ¹⁶.

Degree of swelling=100

W_I= Initial weight of microspheres before swelling, W_F= Final weight of microspheres after swelling

In-vitro drug release studies

In-vitro drug release study of Ketoprofen loaded mucoadhesive microspheres were carried out in USP type II dissolution apparatus. Prepared microsphere was introduced into 900 ml of 0.1 N HCl buffer solutions and allowed to rotate at 50 rpm at 37±0.5⁰C for up to 12 hours. Samples were withdrawn after 1-hour intervals and replace with fresh buffer solution for maintaining the sink condition. Then the samples were analyzed under UV spectrophotometer at- 260 nm ¹⁷.

RESULT AND DISCUSSION

Particle size determination

The particle sizes of ketoconazole loaded mucoadhesive microsphere were observed by optical microscopy and the range of the particle size was between 64-71mm. Among them F4 formulation was having the smaller sizes (Table 2 and Figure 1).

Table 2: Particle size of microspheres

Formulation code	Particle size (mm)
F1	66
F2	68
F3	69
F4	64
F5	71

 

Figure 1: Particle size of microspheres

Drug encapsulation efficiency

By using particular formula drug encapsulation efficiency was calculated and the range was found between 58-85 % (Table 4 and Figure 2).

Table 3: Drug Encapsulation efficiency

Formulation code	Drug encapsulation efficiency (%)
F1	84
F2	74
F3	83
F4	85
F5	58

 

Percentage yield

The percentage yield of prepared microsphere was calculated and among all formulations F4 showed highest percentage yield (Table 4).

Table 4: Percentage yield

Formulation code	Percentage yield (%)
F1	63
F2	59
F3	72
F4	76
F5	68

 

Figure 2: Drug encapsulation efficiency

Morphology

The morphology of prepared microspheres was determined using SEM. All the microspheres were spherical and rough in surface. The graphical representation of the size of the prepared microspheres i.e. the ketoconazole loaded mucoadhesive microspheres were shown in Fig 1.

Flow properties

Angle of repose, Carr’s index, Hausner’s ratio was determined for study the flow property of prepared microspheres. Among all batches, F4 shows good flow property (given below in table 5).

 

 

 

Table 5: Flow properties

Formulation code	Angle of repose  (degrees)	Carr’s index (%)	Hausner’s ratio
F1	26	12	1.16
F2	31	14	1.15
F3	28	13	1.15
F4	26	11	1.12
F5	27	15	1.14

 

 

 

Mucoadhesive properties

The range of swelling index of prepared mucoadhesive microsphere was between 1.22-1.85 % (given in table 6 and figure 3).

Table 6: Swelling index

Formulation code	Weight of microspheres before swelling (mg)	Weight of microspheres after swelling (mg)	Swelling index (%)
F1	70.26	71.12	1.22
F2	71.19	72.25	1.48
F3	68.24	69.46	1.78
F4	69.36	70.65	1.85
F5	67.21	68.17	1.42

 

Figure 3: Swelling Index (%)

 

 

In-vitro drug release studies

Ketoprofen loaded prepared mucoadhesive microspheres undergo the in-vitro dissolution test. In 900 ml of 0.1N HCl, the test was carried out up to 12 hrs and the % CDR were calculated for each formulation. The table 7 and figure 4 show the result and among all formulation batches F4 shows the best % CDR.

 

 

Table 7: Cumulative amount of Drug Release

Time	F1 (%CDR)	F2(%CDR)	F3(%CDR)	F4(%CDR)	F5(%CDR)
1	10	12	19	25	26
2	19	22	33	42	32
3	28	30	41	54	43
4	36	39	47	59	58
5	42	46	50	65	64
6	47	52	60	72	78
7	51	58	67	78	84
8	54	61	74	84	87
9	58	66	82	89	91
10	63	72	84	92	92
11	65	79	90	96	94
12	78	86	94	97	96

 

Figure 4: Cumulative amount of Drug Release

 

 

DISCUSSION 

Ketoprofen loaded microspheres were prepared by ion gelation method by using carbopol934P, sodium alginate, calcium carbonate. After successful preparation they are allowed for different evaluation test like- particle size determination, drug encapsulation efficiency, percentage yield, morphology, flow properties, mucoadhesive properties, in-vitro drug release studies. Prepared mucoadhesive microspheres were found spherical and the range particle size was between 64-71 mm, drug encapsulation efficiency was between 58%-85%, percentage yieldwas between 59%-76%, and swelling index was between1.22-1.85% and the, in-vitro drug release was up to 94% during 12 hrs in acid buffer.

CONCLUSION 

Prepared ketoprofen loaded mucoadhesive microspheres were spherical in shape and showed good flow property as well except that, the microspheres were showed the result of all evaluation test within acceptable range. Among all the formulationsF4 was the best formulation in terms of particle size, drug encapsulation efficiency, percentage yield, flow properties, mucoadhesive properties, in-vitro drug release studies.

ACKNOWLEDGEMENT

The authors of this work were thankful to Infinity Chemicals Pvt. Ltd for providing the Ketoprofen drug and also thankful to Haldia Institute of Pharmacy for all instruments.

REFERENCES:

1. Jain S., et al.. Mucoadhesive microspheres: A review. Journal of Advanced Pharmaceutical Technology & Research, 2014; 5(2):67-81.

2. Nagpal K., et al. Mucoadhesive microspheres: A review. Journal of Drug Delivery Science and Technology, 2019;51:164-180.

3. Rokhade A.P., et al. Mucoadhesive microspheres as a controlled drug delivery system. International Journal of Health Research, 2007;1(2):109-119.

4. https://classic.clinicaltrials.gov/ct2/show/NCT00426985 

5. https://classic.clinicaltrials.gov/ct2/show/NCT00365586 

6. Warner TD, Mitchell JA. Cyclooxygenases: new forms, new inhibitors, and lessons from the clinic. The FASEB journal. 2004 May;18(7):790-804. https://doi.org/10.1096/fj.03-0645rev PMid:15117884

7. Celebi S, Hacimustafaoglu MU, Aygun D, Arisoy ES, Karali Y, Akgoz S, Citak Kurt AN, Seringec M. Antipyretic effect of ketoprofen. The Indian Journal of Pediatrics. 2009 Mar; 76:287-91. https://doi.org/10.1007/s12098-008-0234-z PMid:19129989

8. Rao DS, Reddy SH, Ramu S, Rambabu E. Preparation and evaluation of mucoadhesive microspheres of simvastatin by ionic gelation technique. American Journal of Advanced drug delivery. 2014;2(5):594-608.

9. Patil P, Chavanke D, Wagh M. A review on ionotropic gelation method: novel approach for controlled gastro retentive gelispheres. Int J Pharm Pharm Sci. 2012 Jun 20;4(4):27-32.

10. Garg A, Upadhyay P. Mucoadhesive microspheres - A short review. Asian J Pharm Clin Res 2012;5(3):24-7.

11. Sinha VR, et al. Microspheres: an overview. International Journal of Pharmaceutics, 2004;278(1):1-23. https://doi.org/10.1016/j.ijpharm.2004.01.044 PMid:15158945

12. Ravi S, Peh KK, Darwis Y, Murthy BK, Singh TR, Mallikarjun C. Development and characterization of polymeric microspheres for controlled release protein loaded drug delivery system. Indian journal of pharmaceutical sciences. 2008 May;70(3):303. https://doi.org/10.4103/0250-474X.42978 PMid:20046737 PMCid:PMC2792511

13. Galgatte U and Chaudhari P: Preparation and evaluation of bioadhesive microspheres prepared by ion gelation method and effect of Variables on quality of microspheres. Journal of Pharmaceutical Tech Research 2013; 3(4): 535-546.

14. Carr, R.L. Classifying flow properties of solids. Chem. Eng. 1965; 18:163-168. https://doi.org/10.1001/archopht.1965.00970040165007 PMid:14318489

15. Hausner, H.H. Friction condition in mass of metal powders. Int. J. Powder Metall. 1967; 3:7-13.

16. Mazumder R, Nath LK, Haque A, Maity T, Choudhury PK, Shrestha B. Formulation and in vitro evaluation of natural polymers based microspheres for colonic drug delivery. Int J Appl Pharm 2010;2:211-9.

17. Nayak BS, Nayak UK, Patro KB, Rout PK. Preparation and In vitro Evaluation of Lamivudine Entrapped MOI Microspheres for Oral administration. Research J. Pharm. and Tech. 2008;1(4):437-441