Development of New Localized Drug Delivery System Based on Ceftriaxone-Sulbactam Composite Drug Impregnated Porous Hydroxyapatite: A Systematic Approach for In Vitro and In Vivo Animal Trial
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Present investigation deals with an extensive approach incorporating in vitro and in vivo experimentation to treat chronic osteomyelitis, using hydroxyapatite porous scaffolds.
Materials and Methods
Hydroxyapatite was synthesized in the laboratory by wet chemical method, different porous scaffolds have been fabricated. In vitro studies include variation of porosity with interconnectivity, pore-drug interfacial studies by SEM-EDAX and drug elution studies (by HPLC) both in contact with PBS and SBF at ~37°C. In vivo trials were based on experimental osteomyelitis in rabbit model induced in tibia by Staphylococcus aureus. Characterizations included observation of histopathology, radiology and estimation of drug in both bone and serum for 42 days by HPLC method and subsequent bone-biomaterial interface by SEM.
It was established that lower pore percentage with a distribution of mainly micro-pores were found to be superior over the higher pore percentage both in vitro and in vivo. The criteria was matched with the 50N50H samples which had 50–55% porosity with an average pore size ~110 μm, having higher interconnectivity (10–100 μm), moderately high adsorption efficiency (~50%) when loaded with CFS (drug combinations consisting of irreversible b-lactamase inhibitor and b-lactam antibiotic). CFS release from HAp implants were faster in PBS than SBF. Further, both the results of in vitro and in vivo drug elution after 42 days showed release higher than minimum inhibitory concentration of CFS against Staphylococcus aureus. In vivo studies also proved the superiority of CFS loaded HAp implants than parenteral group based on eradication of infection and new bone formation.
HAp based porous scaffold loaded with CFS and designed porosity (in terms of micro- and macro-porosity, interconnectivity) was found to be an ideal delivery system which could locally, sustainably release the composite antibiotic in reliable manner both in terms of in vitro drug elution behaviour in contact with SBF and in vivo animal trial.
KEY WORDSceftriaxone-sulbactam composite in vivo animal trial osteomyelitis and new bone formation porous hydroxyapatite SBF
American Society for Testing and Materials
Area under the curve
Combination of CFT and SUL drug
Field emission scanning electron microscopy
Fourier-transformed infrared spectroscopy
High performance liquid chromatography
Minimum inhibitory concentration
Phosphate buffered saline
Red blood cell
Simulated body fluid
Scanning electron microscopy-Energy dispersive analysis of X-ray
The authors wish to express their sincere thanks for the financial support by Department of Science and Technology, India [T.1 (7)/TIFA/2006-CGCRI] and the Director, CGCRI, India and Vice Chancellor, West Bengal University of Animal and Fishery Sciences, Kolkata, India for their generous and kind support to this work. All the personnel related to the characterization of the materials are sincerely acknowledged.
- 4.Conterno LO, da Silva Filho CR. Antibiotics for treating chronic osteomyelitis in adults. Cochrane Database Syst Rev:CD004439 (2009).Google Scholar
- 6.Soundrapandian C, Sa B, Datta S. Organic–inorganic composites for bone drug delivery. AAPS PharmSciTech (2009).Google Scholar
- 29.Sinha MK, Sen PS, Basu D, Chattopadhyay S, Basu MK. An improved process for the synthesis of hydroxyapatite powder useful for biomedical applications. In CGCRI (ed.), India, 1998.Google Scholar
- 31.D. Basuand M.K. Sinha. A process for the production of improved porous ocular implants and improved porous ocular implants produced thereby. In CGCRI (ed.), India, 2006.Google Scholar
- 34.Weng W, Baptista JL. Sol-gel derived porous hydroxyapatite coatings. J Mater Sci. 1998;9:159–63.Google Scholar
- 41.Krajewski A, Ravaglioli A, Roncari E, Pinasco P, Montanari L. Porous ceramic bodies for drug delivery. J Mater Sci. 2000;11:763–71.Google Scholar
- 48.Eggli PS, Muller W, Schenk RK. Porous hydroxyapatite and tricalcium phosphate cylinders with two different pore size ranges implanted in the cancellous bone of rabbits. A comparative histomorphometric and histologic study of bony ingrowth and implant substitution. Clin Orthop Relat Res. 1988;232:127–38.PubMedGoogle Scholar
- 60.Joosten U, Joist A, Frebel T, Brandt B, Diederichs S, von Eiff C. Evaluation of an in situ setting injectable calcium phosphate as a new carrier material for gentamicin in the treatment of chronic osteomyelitis: studies in vitro and in vivo. Biomaterials. 2004;25:4287–95.CrossRefPubMedGoogle Scholar
- 62.Girschick HJ, Zimmer C, Klaus G, Darge K, Dick A, Morbach H. Chronic recurrent multifocal osteomyelitis: what is it and how should it be treated? Nat Clin Pract. 2007;3:733–8.Google Scholar