Vilma Campana

Evaluation of inflammatory biomarkers associated with oxidative stress and histological assessment of low-level laser therapy in experimental myopathy

The objective of the present work was to study the effect of helium–neon (He–Ne) and gallium–arsenide (Ga–As) laser upon inflammatory biomarkers associated with oxidative stress: fibrinogen, nitric oxide (NO), L‐citrulline, and superoxide dismutase (SOD). These were evaluated through histological assessment, in rats with experimental myopathy. Materials and Methods: The groups studied were: (A) control, (B) injured, (C) injured and treated with He–Ne laser, (D) injured and treated with Ga–As laser, (E) irradiated with He–Ne; and (F) irradiated with Ga–As laser. Myopathy was induced by injecting 0.05 mg/rat/day of adrenaline in the left posterior limb muscle at the same point on 5 consecutive days, in groups B, C, and D. Low‐level laser therapy (LLLT) was applied with 9.5 J/cm2 daily for 7 consecutive days with each laser. The determination of the biomarkers was made by spectrophotometry. The muscles (5/8, single blinded) were stained with Gomori Trichrome and examined by optic microscopy. The quantitative variables were statistically analyzed by the Fishers test and categorical data by the Axionvision 4.8 program. Pearsons chi‐squared test was applied, setting significant difference at P < 0.05 for all cases. Results: In group B, the biomarkers were significantly increased compared to the other groups (P < 0.001), except for NO which in group B decreased significantly (P < 0.001). In group B, there was a higher inflammatory infiltration level (80.67%) in relation to destroyed fibers. Conclusions: LLLT caused significant changes in inflammatory biomarkers and oxidative stress: decreased levels of fibrinogen, L‐citrulline and SOD as opposed to the increase of NO in rats with experimental myopathies and significant muscle recovery. Lasers Surg. Med. 42:577–583, 2010.

He-Ne laser on microcrystalline arthropathies.

OBJECTIVE The objective of this work is to assess the anti-inflammatory capacity of He-Ne laser therapy as determined by the plasmatic levels of inflammatory markers, fibrinogen, and TNFalpha and by histopathological study in rats with arthropathy induced by calcium pyrophosphate crystals. BACKGROUND DATA Microcrystalline arthropathies are a group of diseases characterized by the deposit of different crystals in joints. MATERIALS AND METHODS Two milligrams of dicalcium pyrophosphate crystals (DCPP) were injected in both joints of the lower limbs of rats during 2 days. A group was treated with laser of He-Ne (6 mW) on the injected joints during 3 consecutive days. After 96 h of the first injection, animals were sacrificed to determine TNFalpha using the ELISA method and fibrinogen was assessed using spectrophotometry. Sections from the lower limbs were used for histopathology. RESULTS A statistically significant increase (p < 0.001) in plasma fibrinogen levels and TNFalpha was noted between the control group and the laser-treated group. The histological transversal section of a posterior limb joint of a rat injected with DCPP showed fibroadipose tissue with diffuse chronic infiltrate. The histopathology of the group of rats injected with DCPP and subsequently treated with He-Ne laser showed no inflammatory response. CONCLUSION He-Ne laser treatment in the microcrystalline arthropathy induced in rats by DCPP injection might have an antiinflammatory effect, evaluated by fibrinogen plasma levels and TNF-alpha (inflammatory markers) and by the histopathology regressive process.

Helium-Neon Laser Reduces the Inflammatory Process of Arthritis

OBJECTIVE A histological study of the anti-inflammatory effect of helium-neon laser in models of arthropathies induced by hydroxyapatite and calcium pyrophosphate in rats. BACKGROUND Crystal deposition diseases are inflammatory pathologies induced by cellular reaction to the deposit of crystals in the joints. METHODS Fifty-six Suquia strain rats were distributed in seven groups. Two mg of each crystal diluted in 0.05 ml physiologic solution were injected six times in each back limb joint, during two weeks on alternate days. Eight J/cm(2) were applied daily to the crystal-injected joints on five consecutive days. The joints were cut and put in 10% formaldehyde, stained with hematoxylin-eosin and observed by light microscopy. The percentage of area with inflammatory infiltrates was determined in five optical microscopy photographs (100X) for each group and analyzed using the Axionvision 4.6 program. A Pearsons Chi Squared test was applied, with significance level set at p < 0.05. RESULTS Both crystals produced an inflammatory process in the osteoarticular structures, consisting of predominantly mononuclear infiltration, fibrosis, and granulomas of foreign body-type giant cells containing phagocytosed remains of crystals. In the arthritic joints treated with laser, a marked decrease (p < 0.0001) was found in the percentage of area with inflammatory infiltrates, although the granulomas remained in a less ostensible form, with adipose tissue cells, fibrosis bands with light residual inflammation, and an absence of or very few crystals. Laser alone or physiologic solution injection did not produce histological changes. CONCLUSIONS Helium-neon laser reduced the intensity of the inflammatory process in the arthritis model induced by hydroxyapatite and calcium pyrophosphate crystals.

Inflammatory and oxidative stress markers in experimental crystalopathy: their modification by photostimulation.

Crystalopathies are inflammatory pathologies caused by cellular reactions to the deposition of crystals in the joints. The anti-inflammatory effect of the helium-neon (He-Ne) laser and that of the nonsteroidal anti-inflammatory drugs (NSAIDs) diclofenac, meloxicam, celecoxib, and rofecoxib was studied in acute and chronic arthritis produced by hydroxyapatite and calcium pyrophosphate in rats. The presence of the markers fibrinogen, L-citrulline, nitric oxide, and nitrotyrosine was determined. Crystals were injected into the posterior limb joints of the rats. A dose of 8 J/cm(2) of energy from an He-Ne laser was applied for 3 d in some groups and for 5 d in other groups. The levels of some of the biomarkers were determined by spectrophotometry, and that of nitrotyrosine was determined by ELISA. For statistical analysis, Fishers exact test was used, and p +/- 0.05 was considered significant. In arthritic rats, the fibrinogen, L-citrulline, nitric oxide, and nitrotyrosine levels increased in comparison to controls and to the laser-treated arthritic groups (p +/- 0.001), (p +/- 0.001), (p +/- 0.02), and (p +/- 0.01), respectively. When comparing fibrinogen from arthritic rats with disease induced by hydroxyapatite with undiseased and arthritic rats treated with NSAIDs, the He-Ne laser decreased levels to values similar to those seen in controls (p +/- 0.01). Inflammatory and oxidative stress markers in experimental crystalopathy are positively modified by photobiostimulation.

Evaluation of inflammatory biomarkers associated with oxidative stress and histological assessment of magnetic therapy on experimental myopathy in rats.

The effect of pulsed electromagnetic field (PEMF) therapy, also called magnetic therapy, upon inflammatory biomarkers associated with oxidative stress plasma fibrinogen, nitric oxide (NO), L-citrulline, carbonyl groups, and superoxide dismutase (SOD) was evaluated through histological assessment, in rats with experimental myopathy. The groups studied were: (A) control (intact rats that received PEMF sham exposures); (B) rats with myopathy and sacrificed 24 h later; (C) rats with myopathy; (D) rats with myopathy and treated with PEMF; and (E) intact rats treated with PEMF. Groups A, C, D, and E were sacrificed 8 days later. Myopathy was induced by injecting 50 μl of 1% carrageenan λ (type IV) once sub-plantar. Treatment was carried out with PEMF emitting equipment with two flat solenoid disks for 8 consecutive days in groups D and E, at 20 mT and 50 Hz for 30 min/day/rat. The biomarkers were determined by spectrophotometry. The muscles (5/8) were stained with Hematoxylin-Eosin and examined by optic microscopy. Quantitative variables were statistically analyzed by the Fisher test, and categorical applying Pearsons Chi Squared test at p < 0.05 for all cases. In Groups B and C, the biomarkers were significantly increased compared to A, D, and E groups: fibrinogen (p < 0.001); NO, L-citrulline and carbonyl groups (p < 0.05); SOD (p < 0.01) as well as the percentage of area with inflammatory infiltration (p < 0.001). PEMF caused decreased levels of fibrinogen, L-citrulline, NO, SOD, and carbonyl groups and significant muscle recovery in rats with experimental myopathies.

Simvastatin: pharmacological response in experimental hyperfibrinogenaemias.

Through a disorder in the endothelial haemostatic balance, hyperfibrinogenaemia could generate endothelial dysfunction. Statins would have antiinflammatory effects on injured endothelium. Objective — Simvastatin pharmacological response in rats with hyperfibrinogenaemias induced by laparotomies was studied. Methods and results — Rats were subjected to multiple injuries (MI) for 30 days (1 laparotomy/week) and for 60 days (1 laparotomy/2 weeks). Simvastatin (0.035 mg/kg) was administered orally to the 30-day multiple injuries group after the third injury for a period of 10 days. A similar dose was administered to the 60-day multiple injuries group after the second injury for a period of 45 days. Blood samples of all the groups were obtained 72 hours after the last injury. In the 30 and 60-day multiple injuries groups, a statistically significant fibrinogen increase was observed (336.6 ± 7.5 and 358.7 ± 9.9, respectively) compared with the control group (207.0 ± 3.0) (p < 0.001).There were no significant differences in the plasmatic fibrinogen (PF) levels between the control and simvastatin treated groups (224.9 ± 1.4 and 216.3 ± 4.3, respectively).There were significant differences between the 30 or 60-day MI untreated groups compared with the 30 or 60-day multiple injuries + simvastatin treated group (p < 0.001). Endothelial denudation and intima widening were observed in the untreated injured groups, whereas in the 60 day multiple injuries group + simvastatin, a regression of histopathological lesions was observed. Conclusions — the decrease of the inflammatory component that would accompany early atherogenesis processes and the regression of the histopathological lesions after treatment could be attributed to the decreased plasmatic fibrinogen.

Helium-neon effects of laser radiation in rats infected with thromboxane B 2

In previous investigations it was found that prostaglanding E1 (PGE1) and Bradiquinine (B), liberated in the inflammatory process, produced a significant increment on the Plasma fibrition Level (PFL), indicative of inflammatory process. The mentioned increment was completely abolished by the irradiation with HeNe laser in the area of infection of the mentioned substances. In the current investigation it was studied the effect of HeNe laser radiation on the P.F.L. of rats injected with another substance related with the inflammatory process and tissular injury: Thromboxane (Tx). It is though that the signal to increase P.F.L. is though an adrenal and an extra adrenal pathways. To study it we injected normal and medullectomized animals and both showed marked increment of P.F.L. Then we repeated the experiment but followed immediately by HeNe laser radiation and we noted a complete blockage of the P.F.L. increment in both groups which suggest that the effect is extra-adrenal. All substances were injected I.M. once daily X 3 days. Immediately after injection the area was irradiated with HeNe laser, 1.5 J total energy. In the normal, non injected non irradiated animals the P.F.L. reached 210.3 + 1.15 mg%. The single injection of Tx did not modify the P.F.L. compared with the previous group. The HeNe irradiation alone did not modify the P.F.L. in the animals injected with Tx only. The animals injected with PGE1+B showed a marked increment of P.F.L. to 337.6 + 14.5 mg%; the HeNe laser radiation completely abolished the increment (231 + 22.3 mg%). But in the animals injected with PGE1+B+Tx, the P.F.L. reached even larger values: 375.2 + 15.3 mg%. The HeNe laser radiation produced a partial blockage in P.F.L. increment (270.3 +/- 13.4 mg%). Showing a significant difference (p < 0.001) compared with normal rats or with rats injected with PGE1+B+Tx without radiation. In conclusion Tx potentiate the effect of PGE+B on the P.F.L. The HeNe laser blocks completely the interaction of PGE1+B on the P.F.L. but only partially the interaction PGE(subscript 1+B+Tx.