Sustained delivery of thymoquinone and epigallocatechin-3 gallate to potentiate osteogenesis in a femoral rat defect rat model

Abstract

Event Abstract Back to Event Sustained delivery of thymoquinone and epigallocatechin-3 gallate to potentiate osteogenesis in a femoral rat defect rat model Hamed A. Benghuzzi1 and Michelle A. Tucci1 1 Universitry of Mississippi Medical Center, United States Introduction: The goal of this study is to explore, for the first time, the capability of Tricalcium Phosphate Lysine (TCPL) ceramic implants to deliver Thymoquinone (TMQ) and epigallocatechin-3 gallate (EGCG), in a sustained manner using femoral defect animal model. The hypothesis of this project is that the use of alternative anti-oxidant based therapy can provide an ideal means to promote osteogenesis in femoral traumatized model. Black Seed herb contains over 100 components, many of which are yet to be discovered. The common one that is thought to be of medicinal value is TMQ [1]. It is a rich source of unsaturated fatty acids and contains about 35% oil, 21% protein, and 38% carbohydrates. More than 50% of the oil are essential fatty acids. EGCG is a potent anti-inflammatory molecule found in green tea. Previous studies have documented that EGCG intake promoted cardiovascular efficiency along with anti-rheumatic activity. Materials and Methods: Adult male rats were randomly divided into 5 groups of 5 each. Group I were untreated and served as intact control. Group II animals were implanted with the TCPL capsule loaded with antibiotic (30 mg Vancomycin (VC)) alone. Groups III-V animals were implanted with TCPL capsule containing VC+TMQ (20mg), VC+EGCG (20mg) and VC+EGCG (20mg)+TMQ (20mg); respectively. Generally, an incision was made to expose the left hind femur where a hole was drilled using a #6 dental bar (2 mm). Each TCPL capsule was implanted at a close proximity to the femoral defect. Fabrication of Ceramics: The microcrystals of TCP were prepared by following a method developed by Benghuzzi and Bajpai [2][3]. Protein Determination: Total protein content in all experimental and control groups was determined using Pierce BCA method [2]. Morphometric Analysis: All morphological analysis were done utilizing the Image Pro Software. Data Analysis: Sigma Stat Software, Jandel Inc. and Slide Write Software were used to determine and compute statistical significance and produce graphical representations of the results. Analysis of variance was used to compare the groups ad significance differences reported at p<0.05. Results and Discussion: Results obtained from this study demonstrated that TCPL delivery system was capable of releasing EGCG, TMQ alone or in combination at sustained levels for 30 days. The release profiles of both agents ranged between 4.35+/-0.12 and 6.78+/- 0.24 ng/ml. Data obtained revealed that there were no significant changes in body weights of animals treated with either EGCG or TMQ compared to the control group. The levels of proteins in all the groups were observed to be fluctuating as the measurements were done for the four different sampling periods. However, there were no significant differences between the groups for specific sampling dates. The levels of alkaline phosphatase continued to decline with time, however, there were no statistical differences between the groups was noted (p<0.05). All animals exposed to a combination of EGCG+TMQ showed extensive bone ingrowth suggestive of a synergistic effect compared to animals exposed to individual agents (moderate ingrowth). Conclusions: It appears that sustained delivery of TMQ can enhance bone healing with little or no side effects on major vital and reproductive organs. Despite the fact that there is barely any work documented done previously using TMQ+EGCG in an attempt to test its viability in healing fractures, the results obtained in this experiment were promising. Therefore, calls for the need of comprehensive studies to establish mode of action of sustained delivery of TMQ+EGCG.