In the orthopaedic field, the repair of ariticular cartilage is still a difficult problem, because of the physiological characters of cartilaginous tissues and chondrocytes. To find an effective method of stimulating their regeneration, this in vitro study focuses on the biostimulation of rabbit articular chondrocytes by low-power He-Ne laser. The articular chondrocytes isolated from the cartilage of the medial condyle of the femur of the rabbit were incubated in HamF12 medium. The second passage culture were spread on 24 petri dishes and were irradiated with laser at power density of 2 - 12 mW/cm2 for 6.5 minutes, corresponding to the energy density of 1-6 J/cm2. Laser treatment was performed three times at a 24-hour interval. After lasering, incubation was continued for 24 hours. Non-irradiated cells were kept under the same conditions as the irradiated ones. The cell proliferation activity was evaluated with a XTT colorimetric method. Irradiation of 4 - 6 J/cm2 revealed a considerably higher cell proliferation activity comparing to control cultures. Thereinto, the energy density of 4 and 5 J/cm2 remarkably increased cell growth (P<0.01). The present study showed that a particular laser irradiation stimulates articular chondrocytes proliferation. These findings might be clinically relevant, indicating that low-power laser irradiation treatment is likely to achieve the repair of articular cartilage in clinic.
The lanthanide trivalence ion and its chelates are used for marking substance in time-resolved fluorescence immunoassay (TRFIA), marking the protein, hormone, antibody, nucleic acid probe or biologica alive cell, to measure the concentration of the analysis substance inside the reaction system with time-resolved fluorometry after the reaction system occurred, and attain the quantitative analysis's purpose. TRFIA has been become a kind of new and more sensitive measure method after radioisotope marking, enzymatic marking, chemiluminescence, electrochemiluminescence, it primarily is decided by the special physics and chemistry characteristic of lanthanide trivalence ion and its chelates. In this paper, the result of spectroscopic evaluation of europium trivalence ion and its chelate, and the principle of spectra-resolved technology and a sensitive time-resolved fluorescence immunoassay instrument made by ourselves are reported. In the set, a high frequency Xenon pulsed-light was adopted as exciting light, and two special filters was utilized according to spectra-resolved technique. Thus the influence of scattering light and short-lifetime fluorescence was removed. And the sensitivity is 10-12mol/L (when Eu3+ was used for marking substance), examination repeat is CV ≤ 5%, examination linearity is from 10-8mol/L to 10-12mol/L, correlation coefficient r ≥ 95% (p < 0.01).
The lanthanide trivalence ion and its chelates are used for marking substance in time-resolved fluorescence immunoassay(TRFIA), marking the protein, hormone, antibody, nucleic acid probe or biologic alive cell, to measure the concentration of the analysis substance inside the reaction system with time-resolved fluorometry after the reaction system occurred, and attain the quantitative analysis's purpose. TRFIA has been therefore become a kind of new and more sensitive measure method after radioisotope marking, enzymatic marking, chemiluminescence, electrochemiluminescence, it primarily is decided by the special physics and chemistry characteristic of lanthanide trivalence ion and its chelates. In this paper, the result of spectroscopic evaluation of europium trivalence ion and its chelate, and the principle of time-resolved technology and fluorescence-enhanced technology is reported. At the same time, the experiment shows that excitation wavelength chosen between 336 and 337nm benefits the excitation and the energy transfer of chelate diketone of europium trivalence ion.
An in vitro study was performed to evaluate the laser biostimulatory effect on cartilage using the low-power He-Ne laser. The chondrocytes, isolated from the cartilage of the medial condyle of the femur of the rabbit, were cultivated, incubated and subject to biostimulatory treatment with the laser. The chondrocytes were placed in 96 Petri dishes at the concentration of 0.25×105 cells/ml and divided into 12 groups (3 dishes every group): the first group was used as the control group, other 11 groups (1-11)were treated with the laser. Control cultures received the same treatment with the laser device off. All the groups were evaluated with a MTT test at the end of the laser treatment and then after 5 days. The cultured chondrocytes showed no damage under a fluorescence microscope and immunostaining was operated to confirm the chondrocytic phenotype. The MTT assay results indicated a positive biostimulatory effect of low-power He-Ne laser on cell proliferation. Thereinto, group 5 and 10 provided the best results in terms of cell viability when compared to the other treated groups and the control group. The increase was maintained for 5 days following the end of the laser treatment. The distinct results obtained with the use of the Low-power He-Ne Laser in the biostimulation of the chondrocytes, no damage and no side-effect, permits us to consider the use of the device clinically.
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