World Health Organization (WHO) in 2005 reported that cases of tuberculosis (TB) in the world occur more than 8 million annually and 5-10% was attacked in spine. The most effective treatment of spinal TB is evacuation of infected bone segments and fills with bone graft. It has been synthesized and characterized of Injectable Bone Substitute (IBS) paste based on hydroxyapatite, gelatin and streptomycin. IBS paste synthesized by mixing hydroxyapatite and gelatin 20% w/v with 75:25, 70:30, 65:35 and 60:40 ratio and streptomycin 10 wt%. The mixture was then added with hydroxypropyl methylcellulose (HPMC) 4% w/v as suspending agent. In vitro characterization performed includes acidity (pH), injectability test, setting time, cytotoxicity (MTT assay) and microbacterium test. Acidity test results indicate a fourth variation of the samples had pH values approaching normal body pH (7.3 to 7.6) and is able to maintain stability when measured in 7 days. Injectability test results indicate IBS paste is injectable with the highest percentage of the injectability value at 97.74% ± 0.19%. IBS paste has been setting within 30 minutes to 1 hour when injected on hydroxyapatite scaffold that resembles the bone cavity and is able to cover the pore scaffold seen from the Scanning Electron Microscope (SEM). Scaffold pore size is smaller from range of 780.8 to 835.4 μm into 225.2 μm. MTT assay results showed that IBS paste is not toxic and experiencing proliferation (viability >100%) that are expected to trigger osteoblast cell growth when applied. Microbacterium test results showed that IBS paste is an antibacterial seen from inhibition zone diameter of Staphylococcus aureus and has a high strength-sensitive antibacterial. Thus, hydroxyapatite, gelatin and streptomycin composites had qualified as injectable bone substitute which applied in cases of spinal tuberculosis.
The most effective treatment for spinal tuberculosis was by eliminating the tuberculosis bacteria and replacing the infected bone with the bone graft to induce the healing process. This study aims to synthesize and characterize nanohydroxyapatite-gelatin-based injectable bone substitute (IBS) with addition of streptomycin. The IBS was synthesized by mixing nanohydroxyapatite and 20 w/v% gelatin with ratio of 40:60, 45:55, 50:50, 55:45, 60:40, 65:35, 70:30, and 75:25 ratio and streptomycin addition as antibiotic agent. The mixture was added by hydroxypropyl methylcellulose as suspending agent. FTIR test showed that there was a chemical reaction occurring in the mixture, between the gelatin and streptomycin. The result of injectability test showed that the highest injectability of the IBS sample was 98.64% with the setting time between 30 minutes and four hours after injection on the HA scaffold that represents the bone cavity and coat the pore scaffold. The cytotoxicity test result showed that the IBS samples were nontoxic towards BHK-21 fibroblast cells and human hepatocyte cells since the viability cell was more than 50% with significant difference (p-value<0.05). The acidity of the IBS was stable and it was sensitive towards Staphylococcus aureus with significantly difference (p-value<0.05). The streptomycin release test showed that the streptomycin could be released from the IBS-injected bone scaffold with release of 2.5% after 4 hours. All the results mentioned showed that IBS was suitable as a candidate to be used in spinal tuberculosis case.
Peripheral nerve injury with gaps between 5 and 30 mm can result in permanent paralysis because axons are cut. The distance between axons, which is more than 1-2 cm, needs graft in the form of nerve connecting pipe in order to repair the defects. A synthesis of hollowfiber polyurethane-collagen coated by chitosan was carried out to identify its potential as treatment accelerator for peripheral nerve injury. The result of Fourier Transform Infrared (FTIR) analysis showed multiple links between chitosan and glutaraldehyde, which can be seen in wavenumber shift from 1080-1100 cm-1 to 1002 cm-1. The degradation test result revealed that the sample displayed mass loss after it was soaked in simulated body fluid(SBF) for seven days. Polyurethanecan be degraded in the body after 30 days. This converges with the nerve mechanism that regenerates at the rate of 1 mm/day or 1 inch/month. The result of tensile test indicated that modulus values of chitosan coating variation of 1%, 1.5%, and 2% were 4.75 MPa, 4.74 MPa, and 7.67 Mpa respectively. The outcome of scanning electron microscope(SEM)showed that hollow fiberhas a diameter of 2.021-2.032 mm, which matches the diameter of peripheral nerves ranging from 1.5 to 3 mm and the membrane pore size of 31.33-39.65 μm. The result of MTT assay demonstrated that the percentage of viable fibroblastBHK-21cells was exceeding 50%, which means that the sample does not have toxic properties. The result of this study is expected to provide theoretical basis for the utilization of polyurethane-collagen coating chitosan as nervegraft for theraphy of peripheral nerve injury. The utilization is possible due to the fact that the composite exhibits biocompatible, regenerative, and easily degradable characteristics. Moreover, it could become an alternative solution to answer the need of a more affordable and easier-to-produce nerve graft, so it can be mass-produced in Indonesia.
ABSTRAK 23.24, 46.55, 53.48, and 55.06
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