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Application of a low-level laser therapy and the purified protein from natural latex (Hevea brasiliensis) in the controlled crush injury of the sciatic nerve of rats: a morphological, quantitative, and ultrastructural study.

Dias FJ, Issa JP, Iyomasa MM, Coutinho-Netto J, Calzzani RA, Iyomasa DM, Sousa LG, de Almeida SR, Cury DP, Watanabe IS - Biomed Res Int (2013)

Bottom Line: After 4 weeks, the myelin density and morphological characteristics improved in groups LG, PG, and LPG compared to IG.After 8 weeks, PG, and LPG were similar to CG and the capillary density was higher in the LG, PG, and LPG.In the ultrastructural analysis the PG and LPG had characteristics that were similar to the CG.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, Institute of Biomedical Sciences, University of Sao Paulo, Sao Paulo, SP, Brazil.

ABSTRACT
This study analyzed the effects of a low-level laser therapy (LLLT, 15 J/cm(2), 780 nm wavelength) and the natural latex protein (P1, 0.1%) in sciatic nerve after crush injury (15 Kgf, axonotmesis) in rats. Sixty rats (male, 250 g) were allocated into the 6 groups (n = 10): CG-control group; EG-nerve exposed; IG-injured nerve without treatment; LG-crushed nerve treated with LLLT; PG-injured nerve treated with P1; and LPG-injured nerve treated with LLLT and P1. After 4 or 8 weeks, the nerve samples were processed for morphological, histological quantification and ultrastructural analysis. After 4 weeks, the myelin density and morphological characteristics improved in groups LG, PG, and LPG compared to IG. After 8 weeks, PG, and LPG were similar to CG and the capillary density was higher in the LG, PG, and LPG. In the ultrastructural analysis the PG and LPG had characteristics that were similar to the CG. The application of LLLT and/or P1 improved the recovery from the nerve crush injury, and in the long term, the P1 protein was the better treatment used, since only the application of LLLT has not reached the same results, and these treatments applied together did not potentiate the recovery.

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Transmission electron microscopy after 8 weeks of nerve injury: (a) CG—general view of the sciatic nerve fibers with myelin (M), groups of unmyelinated fibers (arrows), and Schwann cells with evident nucleus (N) (mag: ×3.000). (b) CG—detailed image of mitochondria in the axoplasm of a myelinated fiber (arrow) and in the cytoplasm of Schwann cells (arrowhead) (mag: ×20.000). (c) EG—myelinated fibers (M) and a grouping of unmyelinated fibers (arrows) (mag: ×3.000). (d) EG—detailed image of the neurofilaments of the axoplasm (*) and the lamellae of myelin (arrows) (mag: ×30.000). (e) IG—degenerated areas (**) among the intact myelin fibers (M) (mag: ×3.000). (f) IG—Details of an area of degeneration of nerve fibers in the presence of myelin debris (thin arrows) (mag: ×5.000). (g) LG—myelinated (arrows) and unmyelinated fibers (arrowheads) arranged in bundles (mag: ×2.500). (h) LG—degenerated area of nerve fibers (**) containing myelin debris (thin arrows)—(mag: ×6.000). (i) PG—grouping of myelinated (arrows) and unmyelinated (arrowheads) nerve fibers, and an area of degeneration (thin arrow) (mag: ×2.500). (j) PG—detailed image of groupings of unmyelinated fibers with mitochondria (arrowheads) in axoplasm (*) and collagen fibers of the endoneurium (C) (mag: ×20.000). (k) LPG—general view of bundles of myelinated fibers (arrows) and an area of degeneration (arrowheads) (mag: ×1.800). (l) LPG—detailed image of a bundle of nerve fibers reveals myelinated and (M) unmyelinated fibers (arrowheads) and Schwann cells (S) (mag: ×5.600).
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fig4: Transmission electron microscopy after 8 weeks of nerve injury: (a) CG—general view of the sciatic nerve fibers with myelin (M), groups of unmyelinated fibers (arrows), and Schwann cells with evident nucleus (N) (mag: ×3.000). (b) CG—detailed image of mitochondria in the axoplasm of a myelinated fiber (arrow) and in the cytoplasm of Schwann cells (arrowhead) (mag: ×20.000). (c) EG—myelinated fibers (M) and a grouping of unmyelinated fibers (arrows) (mag: ×3.000). (d) EG—detailed image of the neurofilaments of the axoplasm (*) and the lamellae of myelin (arrows) (mag: ×30.000). (e) IG—degenerated areas (**) among the intact myelin fibers (M) (mag: ×3.000). (f) IG—Details of an area of degeneration of nerve fibers in the presence of myelin debris (thin arrows) (mag: ×5.000). (g) LG—myelinated (arrows) and unmyelinated fibers (arrowheads) arranged in bundles (mag: ×2.500). (h) LG—degenerated area of nerve fibers (**) containing myelin debris (thin arrows)—(mag: ×6.000). (i) PG—grouping of myelinated (arrows) and unmyelinated (arrowheads) nerve fibers, and an area of degeneration (thin arrow) (mag: ×2.500). (j) PG—detailed image of groupings of unmyelinated fibers with mitochondria (arrowheads) in axoplasm (*) and collagen fibers of the endoneurium (C) (mag: ×20.000). (k) LPG—general view of bundles of myelinated fibers (arrows) and an area of degeneration (arrowheads) (mag: ×1.800). (l) LPG—detailed image of a bundle of nerve fibers reveals myelinated and (M) unmyelinated fibers (arrowheads) and Schwann cells (S) (mag: ×5.600).

Mentions: Eight weeks after the injury, the CG had characteristics similar to those observed in the control group that was analyzed after four weeks, as the groups of myelinated and unmyelinated fibers were present in various sizes and shapes and Schwann cells were present surrounding the fibers (Figure 4(a)). The myelinated fibers adjacent to one group of unmyelinated fibers are shown in detail in the endoneurial area that contains collagen fibers and the cytoplasm of Schwann cells with mitochondria (Figure 4(b)). In the EG, myelin fibers and large amounts of unmyelinated fibers of varying diameters were present (Figure 4(c)). In the axoplasm of unmyelinated and myelinated fibers, neurofilaments and mitochondria were revealed, and the lamellar organization of the myelin sheath was also observed (Figure 4(d)). Characteristics of degeneration were not observed in these first two groups during this analysis period. The IG presented the presence of nerve fibers that were closer to each other compared to the IG 4 weeks after the injury; however, the presence of large areas of nerve fibers degeneration located between intact fibers was visible (Figure 4(e)). Myelin debris could be observed in the areas of nerve fiber degeneration (Figure 4(f)). In the LG, one could observe that the myelinated nerve fibers and unmyelinated diameters and shapes were arranged in bundles, with large spaces between nerve fibers and structures of the linear remnants of Schwann cells (Figure 4(g)). Additionally, the myelin debris in an area of degeneration was surrounded by intact myelinated and unmyelinated fibers (Figure 4(h)). The PG had numerous unmyelinated and myelinated fibers of varying diameters, with few areas of degeneration between the intact fibers (Figure 4(i)). A more detailed view of the myelinated fiber the sheath revealed normal characteristics around the axoplasm, enveloped by a Schwann cell, and collagen fibers were observed in the endoneurial region (Figure 4(j)). The LPG revealed the presence of myelinated and unmyelinated fibers that were arranged in bundles and regions of nerve degeneration with the Schwann cell cytoplasm having a dense appearance; additionally, some areas of fiber degeneration were still observable (Figure 4(k)). The grouping of myelinated and unmyelinated nerve fibers of various diameters could be observed in detail, and the axoplasm contained neurofilaments and mitochondria along with the presence of a Schwann cell with an evident nucleus (Figure 4(l)).


Application of a low-level laser therapy and the purified protein from natural latex (Hevea brasiliensis) in the controlled crush injury of the sciatic nerve of rats: a morphological, quantitative, and ultrastructural study.

Dias FJ, Issa JP, Iyomasa MM, Coutinho-Netto J, Calzzani RA, Iyomasa DM, Sousa LG, de Almeida SR, Cury DP, Watanabe IS - Biomed Res Int (2013)

Transmission electron microscopy after 8 weeks of nerve injury: (a) CG—general view of the sciatic nerve fibers with myelin (M), groups of unmyelinated fibers (arrows), and Schwann cells with evident nucleus (N) (mag: ×3.000). (b) CG—detailed image of mitochondria in the axoplasm of a myelinated fiber (arrow) and in the cytoplasm of Schwann cells (arrowhead) (mag: ×20.000). (c) EG—myelinated fibers (M) and a grouping of unmyelinated fibers (arrows) (mag: ×3.000). (d) EG—detailed image of the neurofilaments of the axoplasm (*) and the lamellae of myelin (arrows) (mag: ×30.000). (e) IG—degenerated areas (**) among the intact myelin fibers (M) (mag: ×3.000). (f) IG—Details of an area of degeneration of nerve fibers in the presence of myelin debris (thin arrows) (mag: ×5.000). (g) LG—myelinated (arrows) and unmyelinated fibers (arrowheads) arranged in bundles (mag: ×2.500). (h) LG—degenerated area of nerve fibers (**) containing myelin debris (thin arrows)—(mag: ×6.000). (i) PG—grouping of myelinated (arrows) and unmyelinated (arrowheads) nerve fibers, and an area of degeneration (thin arrow) (mag: ×2.500). (j) PG—detailed image of groupings of unmyelinated fibers with mitochondria (arrowheads) in axoplasm (*) and collagen fibers of the endoneurium (C) (mag: ×20.000). (k) LPG—general view of bundles of myelinated fibers (arrows) and an area of degeneration (arrowheads) (mag: ×1.800). (l) LPG—detailed image of a bundle of nerve fibers reveals myelinated and (M) unmyelinated fibers (arrowheads) and Schwann cells (S) (mag: ×5.600).
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fig4: Transmission electron microscopy after 8 weeks of nerve injury: (a) CG—general view of the sciatic nerve fibers with myelin (M), groups of unmyelinated fibers (arrows), and Schwann cells with evident nucleus (N) (mag: ×3.000). (b) CG—detailed image of mitochondria in the axoplasm of a myelinated fiber (arrow) and in the cytoplasm of Schwann cells (arrowhead) (mag: ×20.000). (c) EG—myelinated fibers (M) and a grouping of unmyelinated fibers (arrows) (mag: ×3.000). (d) EG—detailed image of the neurofilaments of the axoplasm (*) and the lamellae of myelin (arrows) (mag: ×30.000). (e) IG—degenerated areas (**) among the intact myelin fibers (M) (mag: ×3.000). (f) IG—Details of an area of degeneration of nerve fibers in the presence of myelin debris (thin arrows) (mag: ×5.000). (g) LG—myelinated (arrows) and unmyelinated fibers (arrowheads) arranged in bundles (mag: ×2.500). (h) LG—degenerated area of nerve fibers (**) containing myelin debris (thin arrows)—(mag: ×6.000). (i) PG—grouping of myelinated (arrows) and unmyelinated (arrowheads) nerve fibers, and an area of degeneration (thin arrow) (mag: ×2.500). (j) PG—detailed image of groupings of unmyelinated fibers with mitochondria (arrowheads) in axoplasm (*) and collagen fibers of the endoneurium (C) (mag: ×20.000). (k) LPG—general view of bundles of myelinated fibers (arrows) and an area of degeneration (arrowheads) (mag: ×1.800). (l) LPG—detailed image of a bundle of nerve fibers reveals myelinated and (M) unmyelinated fibers (arrowheads) and Schwann cells (S) (mag: ×5.600).
Mentions: Eight weeks after the injury, the CG had characteristics similar to those observed in the control group that was analyzed after four weeks, as the groups of myelinated and unmyelinated fibers were present in various sizes and shapes and Schwann cells were present surrounding the fibers (Figure 4(a)). The myelinated fibers adjacent to one group of unmyelinated fibers are shown in detail in the endoneurial area that contains collagen fibers and the cytoplasm of Schwann cells with mitochondria (Figure 4(b)). In the EG, myelin fibers and large amounts of unmyelinated fibers of varying diameters were present (Figure 4(c)). In the axoplasm of unmyelinated and myelinated fibers, neurofilaments and mitochondria were revealed, and the lamellar organization of the myelin sheath was also observed (Figure 4(d)). Characteristics of degeneration were not observed in these first two groups during this analysis period. The IG presented the presence of nerve fibers that were closer to each other compared to the IG 4 weeks after the injury; however, the presence of large areas of nerve fibers degeneration located between intact fibers was visible (Figure 4(e)). Myelin debris could be observed in the areas of nerve fiber degeneration (Figure 4(f)). In the LG, one could observe that the myelinated nerve fibers and unmyelinated diameters and shapes were arranged in bundles, with large spaces between nerve fibers and structures of the linear remnants of Schwann cells (Figure 4(g)). Additionally, the myelin debris in an area of degeneration was surrounded by intact myelinated and unmyelinated fibers (Figure 4(h)). The PG had numerous unmyelinated and myelinated fibers of varying diameters, with few areas of degeneration between the intact fibers (Figure 4(i)). A more detailed view of the myelinated fiber the sheath revealed normal characteristics around the axoplasm, enveloped by a Schwann cell, and collagen fibers were observed in the endoneurial region (Figure 4(j)). The LPG revealed the presence of myelinated and unmyelinated fibers that were arranged in bundles and regions of nerve degeneration with the Schwann cell cytoplasm having a dense appearance; additionally, some areas of fiber degeneration were still observable (Figure 4(k)). The grouping of myelinated and unmyelinated nerve fibers of various diameters could be observed in detail, and the axoplasm contained neurofilaments and mitochondria along with the presence of a Schwann cell with an evident nucleus (Figure 4(l)).

Bottom Line: After 4 weeks, the myelin density and morphological characteristics improved in groups LG, PG, and LPG compared to IG.After 8 weeks, PG, and LPG were similar to CG and the capillary density was higher in the LG, PG, and LPG.In the ultrastructural analysis the PG and LPG had characteristics that were similar to the CG.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, Institute of Biomedical Sciences, University of Sao Paulo, Sao Paulo, SP, Brazil.

ABSTRACT
This study analyzed the effects of a low-level laser therapy (LLLT, 15 J/cm(2), 780 nm wavelength) and the natural latex protein (P1, 0.1%) in sciatic nerve after crush injury (15 Kgf, axonotmesis) in rats. Sixty rats (male, 250 g) were allocated into the 6 groups (n = 10): CG-control group; EG-nerve exposed; IG-injured nerve without treatment; LG-crushed nerve treated with LLLT; PG-injured nerve treated with P1; and LPG-injured nerve treated with LLLT and P1. After 4 or 8 weeks, the nerve samples were processed for morphological, histological quantification and ultrastructural analysis. After 4 weeks, the myelin density and morphological characteristics improved in groups LG, PG, and LPG compared to IG. After 8 weeks, PG, and LPG were similar to CG and the capillary density was higher in the LG, PG, and LPG. In the ultrastructural analysis the PG and LPG had characteristics that were similar to the CG. The application of LLLT and/or P1 improved the recovery from the nerve crush injury, and in the long term, the P1 protein was the better treatment used, since only the application of LLLT has not reached the same results, and these treatments applied together did not potentiate the recovery.

Show MeSH
Related in: MedlinePlus