Due to specific structural organization at the molecular level, several biomolecules (e.g., collagen, myosin etc.) which are strong generators of second harmonic generation (SHG) signals, exhibit unique responses depending on the polarization of the excitation light. By using the polarization second harmonic generation (p-SHG) technique, the values of the second order susceptibility components can be used to differentiate the types of molecule, which cannot be done by the use of a standard SHG intensity image. In this report we discuss how to implement p-SHG on a commercial multiphoton microscope and overcome potential artifacts in susceptibility (χ) image. Furthermore we explore the potential of p-SHG microscopy by applying the technique to different types of tissue in order to determine corresponding reference values of the ratio of second-order χ tensor elements. These values may be used as a bio-marker to detect any structural alterations in pathological tissue for diagnostic purposes. The SHG intensity image (red) in (a) shows the distribution of collagen fibers in ovary tissue but cannot determine the type of collagen fiber. However, the histogram distribution (b) for the values of the χ tensor element ratio can be used to quantitatively identify the types of collagen fibers.

Biomolecular changes in the cartilage matrix dur-

Abstract:In a synovial joint, the articular cartilage is directly affected during the progression of Osteoarthritis (OA). The characterization of early stage modification in extra-cellular matrix of cartilage is essential for detection as well as understanding the progression of disease. The objective of this study is to demonstrate the potential and capability of nonlinear optical microscopy for the morphological investigation of early stage osteoarthritic cartilage. ICRS Grade-I cartilage sections were obtained from the femoral condyle of the human knee. The surface of articular cartilage was imaged by second harmonic generation and two-photon excited fluorescence microscopy. Novel morphological features like microsplits and wrinkles were observed, which would otherwise not be visible in other clinical imaging modalities (e.g., CT, MRI, ultrasound and arthroscope. The presence of superficial layer with distinct collagen fibrils parallel to the articular surface in 4 specimens out of 14 specimens, indicates that different phases of OA within ICRS Grade-I can be detected by SHG microscopy. All together, the observed novel morphologies in early stage osteoarthritic cartilage indicates that SHG microscopy might be a significant tool for the assessment of cartilage disorder.

A great deal of effort has been focused on exploring the underlying molecular mechanism of osteoarthritis (OA) especially at the cellular level. We report a confocal Raman spectroscopic investigation on human osteoarthritic chondrocytes. The objective of this investigation is to identify molecular features and the stage of OA based on the spectral signatures corresponding to bio-molecular changes at the cellular level in chondrocytes. In this study, we isolated chondrocytes from human osteoarthritic cartilage and acquired Raman spectra from single cells. Major spectral differences between the cells obtained from different International Cartilage Repair Society (ICRS) grades of osteoarthritic cartilage were identified. During progression of OA, a decrease in protein content and an increase in cell death were observed from the vibrational spectra. Principal component analysis and subsequent cross-validation was able to associate osteoarthritic chondrocytes to ICRS Grade I, II and III with specificity 100.0%, 98.1%, and 90.7% respectively, while, sensitivity was 98.6%, 82.8%, and 97.5% respectively. The overall predictive efficiency was 92.2%. Our pilot study encourages further use of Raman spectroscopy as a noninvasive and label free technique for revealing molecular features associated with osteoarthritic chondrocytes.

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Background Evidence-based recommendations have the potential to improve the quality of health care by promoting interventions of proven benefit and discouraging unnecessary, ineffective, or harmful interventions. Osteoarthritis (OA) treatment recommendations are generally addressing strength training and training to increase aerobic capacity together with referral to, or treatment by, physiotherapists. Previous research has revealed that treatment of patients with OA by medical doctors show low adherence with recommendations (1). Little is not known about the treatment provided by physiotherapists in Norway and whether the treatment provided is in concordance with general recommendations for OA treatment. Objectives To investigate knee OA treatment performed by physiotherapists in Norway and its concordance with EULAR recommendations for non-pharmacological treatment modalities. Methods Patient records in the hospital region of middle Norway were searched to identify patients with knee OA. A letter was sent to the identified patients (N=87) asking for information about which physiotherapist they had seen for treatment. A letter was sent to the physiotherapists asking for the discharge summary, in which they are obliged to send to the referring doctor. A survey of the discharge summaries was conducted independently by 3 health professionals (two physiotherapists, one physician) to categorise and group the treatment given. Results Response rate from patients was 93% (N=81). Of the 81, 13 (15%) had not been referred to physiotherapy treatment. Response rate from physiotherapists (N=68) was 38 (56%). Of the treatment described in the discharge summaries 7 (10%) was partly in line with EULAR recommendations; of these, all 7 used strengthening exercises and 0 used improvement of aerobic capacity as their choice of therapeutic modality. Conclusions According to our findings it may seem that physiotherapy treatments are not always in line with recommendations for knee OA treatment. The incidence of total knee replacement is increasing, raising costs personal and societal. Physiotherapy treatment in line with recommendations may help prevent OA from progression and prevent prosthesis operations to further increase. There seems to be a need for further work to make physiotherapists implement the EULAR guidelines in their day to day work with knee OA patients. References Østerås N, Garratt A, Grotle M, Natvig B, Kjeken I, Kvien TK, Hagen KB. Patient-reported quality of care for osteoarthritis: development and testing of the osteoarthritis quality indicator questionnaire. Arthritis Care Res (Hoboken). 2013 Jul;65(7):1043-51. Acknowledgements This study was given economic support from the central norway regional heath authority. Disclosure of Interest None declared DOI 10.1136/annrheumdis-2014-eular.5148