Correlation of synovial caspase-3 concentration and the photodynamic effectiveness in osteoarthritis treatment.

BACKGROUND: The present study focuses on investigation of Intra-articular PDT mechanisms for OA treatment. Also, a search for determination of the most effective dose of chlorin e6 (Ce6) for anti-inflammatory PDT of OA was carried out. METHODS: The study was carried out on laboratory animals (11 Chinchilla rabbits, 1 year, 2.5 kg) with a gonarthritis model of post-traumatic OA. According to the instructions for using Photoditazin (Ce6 based PS) for PDT of human oncological and non-oncological diseases, the recommended dose is 0.7-1.2 mg/kg. For studies on rabbits, taking into account the conversion coefficient (3.2), the PS doses of 2.4, 3.2 and 6.4 mg/kg were selected. Fluorescence spectra were measured intra-articular before and after PDT using spectrometer with fiber-optic probe. The intrajoint PDT was carried out using a laser (662 ± 10 nm) and a fiber-optic catheter with a cylindrical diffuser inside a sapphire needle for a uniform distribution of the laser radiation. The immunohistochemical study was carried out by staining the samples with caspase-3. RESULTS: Histological and immunohistochemical analysis showed that the best PS dose for intravenous administration for PDT of rabbit gonarthritis is 3.2 mg/kg. The PS concentration directly in the synovial tissue was 0.5 mg/kg, and this was enough to achieve the most positive results to reduce the caspase-3 level. CONCLUSION: The caspase-3 level correlates well with other signs of inflammation in the synovial membrane (edema, etc.). Therefore, to assess the PDT effectiveness in the treatment of gonarthritis accompanied by synovitis, it is sufficient to analyze only for caspase-3. The efficacy of PDT with Ce6 showed that 3.2 mg/kg PS dose (1 mg/kg for a human) is the most effective.

Gonarthritis photodynamic therapy with chlorin e6 derivatives.

BACKGROUND: The new methods of osteoarthritis treatment are in constant demand due to the complexity of the early diagnosis and therapy. Specific features of Сhlorin e6 derivative (Ce6) accumulation in knee joint tissues and the efficiency of photodynamic therapy (PDT) of gonarthritis were studied. METHODS: The experimental research was conducted on the model of posttraumatic gonarthritis on rabbits. The analysis of dynamics of change of Ce6 concentration in tissues of a knee joint was carried out by the method of fluorescent diagnostics. The intra-joint PDT was carried out using 662nm laser with energy density of 120-150J/cm(2) and a sapphire diffuser. An analysis of slices was conducted to confirm the anti-inflammatory effect through apoptosis. RESULTS: The method of fluorescent spectroscopy revealed that the highest amount of Ce6 was accumulated in the synovial membrane of a damaged knee joint 2.5h after its intravenous introduction. On 14th day after gonarthritis modeling but before PDT the synovial membrane showed signs of synovitis. On 21st day after PDT the synovial membrane possessed noticeable villous structure, and no cells of inflammatory nature were observed. CONCLUSION: Fluorescent diagnostics in knee joint tissues can be used in clinical practice of gonarthritis before, during and after PDT for monitoring the Ce6 accumulation and for treatment control. Optimal radiation energy density was determined to be 150J/cm(2). In the studied time intervals (5-25min) no dependency of PDT effect on irradiation time at the same energy density was observed. The analysis of results of clinical and morphological research shows that PDT is a low-invasive method of gonarthritis treatment with a high degree of efficiency and selectivity.