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Peripheral Nerve Transplantation Combined with Acidic Fibroblast Growth Factor and Chondroitinase Induces Regeneration and Improves Urinary Function in Complete Spinal Cord Transected Adult Mice.

DePaul MA, Lin CY, Silver J, Lee YS - PLoS ONE (2015)

Bottom Line: Cystometry analysis and external urethral sphincter electromyograms reveal that treatment with PNG+aFGF+ChABC reduced bladder weight, improved bladder and external urethral sphincter histology, and significantly enhanced LUT function, resulting in more efficient voiding.Regeneration of serotonin axons correlated with LUT recovery.These results suggest that our mouse model of LUT dysfunction recapitulates the results found in the rat model and may be used to further investigate genetic contributions to regeneration failure.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurosciences, Case Western Reserve University, Cleveland, Ohio, United States of America.

ABSTRACT
The loss of lower urinary tract (LUT) control is a ubiquitous consequence of a complete spinal cord injury, attributed to a lack of regeneration of supraspinal pathways controlling the bladder. Previous work in our lab has utilized a combinatorial therapy of peripheral nerve autografts (PNG), acidic fibroblast growth factor (aFGF), and chondroitinase ABC (ChABC) to treat a complete T8 spinal cord transection in the adult rat, resulting in supraspinal control of bladder function. In the present study we extended these findings by examining the use of the combinatorial PNG+aFGF+ChABC treatment in a T8 transected mouse model, which more closely models human urinary deficits following spinal cord injury. Cystometry analysis and external urethral sphincter electromyograms reveal that treatment with PNG+aFGF+ChABC reduced bladder weight, improved bladder and external urethral sphincter histology, and significantly enhanced LUT function, resulting in more efficient voiding. Treated mice's injured spinal cord also showed a reduction in collagen scaring, and regeneration of serotonergic and tyrosine hydroxylase-positive axons across the lesion and into the distal spinal cord. Regeneration of serotonin axons correlated with LUT recovery. These results suggest that our mouse model of LUT dysfunction recapitulates the results found in the rat model and may be used to further investigate genetic contributions to regeneration failure.

No MeSH data available.


Related in: MedlinePlus

PNG+aFGF+ChABC decreases collagen scaring in the lesion.(A) Gross observation of the spinal cord at 18 weeks after SCI in TX-only and PNG+aFGF+ChABC animals. White arrow marks the lesion. (B) Representative photomicrographs of the spinal cord lesion stained with Masson’s trichrome from TX-only and PNG+aFGF+ChABC mice 18 weeks post injury. The morphology of the PNG can be visualized in the PNG+aFGF+ChABC treated animals. Arrow marks the PNG and area of expanded view. Scale bar, 200μm. Rostral is to the left, and caudal to the right. Collagen is stained blue. (C) Collagen scar area (μm2). The PNG+aFGF+ChABC group shows a significant reduction of collagen scarring compared to the TX-only group. Five serial sections per animal were quantified, n = 5 animals per group. ****p<0.0001.
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pone.0139335.g003: PNG+aFGF+ChABC decreases collagen scaring in the lesion.(A) Gross observation of the spinal cord at 18 weeks after SCI in TX-only and PNG+aFGF+ChABC animals. White arrow marks the lesion. (B) Representative photomicrographs of the spinal cord lesion stained with Masson’s trichrome from TX-only and PNG+aFGF+ChABC mice 18 weeks post injury. The morphology of the PNG can be visualized in the PNG+aFGF+ChABC treated animals. Arrow marks the PNG and area of expanded view. Scale bar, 200μm. Rostral is to the left, and caudal to the right. Collagen is stained blue. (C) Collagen scar area (μm2). The PNG+aFGF+ChABC group shows a significant reduction of collagen scarring compared to the TX-only group. Five serial sections per animal were quantified, n = 5 animals per group. ****p<0.0001.

Mentions: We next examined the lesion of the TX-only and PNG+aFGF+ChABC for differences in gross anatomy and scarring. In the TX-only group, the rostral and caudal stumps of the lesioned cord were connected by thin, translucent tissue containing a dense collagen matrix (Fig 3A and 3B). The distribution of collagen dense deposits can be observed in the lesion gap, in the periphery around the cord, and inside the spinal cord tissue from both rostral and caudal end. In contrast, the lesioned cord of animals receiving PNG+aFGF+ChABC treatment were thick and opaque with dense peripheral nerve grafts bridging the rostral and caudal stumps completely filling the lesion, giving an appearing of a continuous interface across the lesion (Fig 3A). The collagen matrix of PNG+aFGF+ChABC treated animals was less dense and covered significantly less area than that of the TX-only treatment (Fig 3B and 3C). To compare the mouse response to injury to that of the rat, we collected available complete spinal cord transection tissue in our laboratory and stained for collagen. The rat lesion had very little collagen scarring compared to the mouse, illustrating the difference in scar composition between the two species (S1 Fig). These results suggest that PNG+aFGF+ChABC treatment can significantly reduce collagen scarring in the mouse, which can facilitate axonal regeneration by decreasing the physical scar barrier.


Peripheral Nerve Transplantation Combined with Acidic Fibroblast Growth Factor and Chondroitinase Induces Regeneration and Improves Urinary Function in Complete Spinal Cord Transected Adult Mice.

DePaul MA, Lin CY, Silver J, Lee YS - PLoS ONE (2015)

PNG+aFGF+ChABC decreases collagen scaring in the lesion.(A) Gross observation of the spinal cord at 18 weeks after SCI in TX-only and PNG+aFGF+ChABC animals. White arrow marks the lesion. (B) Representative photomicrographs of the spinal cord lesion stained with Masson’s trichrome from TX-only and PNG+aFGF+ChABC mice 18 weeks post injury. The morphology of the PNG can be visualized in the PNG+aFGF+ChABC treated animals. Arrow marks the PNG and area of expanded view. Scale bar, 200μm. Rostral is to the left, and caudal to the right. Collagen is stained blue. (C) Collagen scar area (μm2). The PNG+aFGF+ChABC group shows a significant reduction of collagen scarring compared to the TX-only group. Five serial sections per animal were quantified, n = 5 animals per group. ****p<0.0001.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4591338&req=5

pone.0139335.g003: PNG+aFGF+ChABC decreases collagen scaring in the lesion.(A) Gross observation of the spinal cord at 18 weeks after SCI in TX-only and PNG+aFGF+ChABC animals. White arrow marks the lesion. (B) Representative photomicrographs of the spinal cord lesion stained with Masson’s trichrome from TX-only and PNG+aFGF+ChABC mice 18 weeks post injury. The morphology of the PNG can be visualized in the PNG+aFGF+ChABC treated animals. Arrow marks the PNG and area of expanded view. Scale bar, 200μm. Rostral is to the left, and caudal to the right. Collagen is stained blue. (C) Collagen scar area (μm2). The PNG+aFGF+ChABC group shows a significant reduction of collagen scarring compared to the TX-only group. Five serial sections per animal were quantified, n = 5 animals per group. ****p<0.0001.
Mentions: We next examined the lesion of the TX-only and PNG+aFGF+ChABC for differences in gross anatomy and scarring. In the TX-only group, the rostral and caudal stumps of the lesioned cord were connected by thin, translucent tissue containing a dense collagen matrix (Fig 3A and 3B). The distribution of collagen dense deposits can be observed in the lesion gap, in the periphery around the cord, and inside the spinal cord tissue from both rostral and caudal end. In contrast, the lesioned cord of animals receiving PNG+aFGF+ChABC treatment were thick and opaque with dense peripheral nerve grafts bridging the rostral and caudal stumps completely filling the lesion, giving an appearing of a continuous interface across the lesion (Fig 3A). The collagen matrix of PNG+aFGF+ChABC treated animals was less dense and covered significantly less area than that of the TX-only treatment (Fig 3B and 3C). To compare the mouse response to injury to that of the rat, we collected available complete spinal cord transection tissue in our laboratory and stained for collagen. The rat lesion had very little collagen scarring compared to the mouse, illustrating the difference in scar composition between the two species (S1 Fig). These results suggest that PNG+aFGF+ChABC treatment can significantly reduce collagen scarring in the mouse, which can facilitate axonal regeneration by decreasing the physical scar barrier.

Bottom Line: Cystometry analysis and external urethral sphincter electromyograms reveal that treatment with PNG+aFGF+ChABC reduced bladder weight, improved bladder and external urethral sphincter histology, and significantly enhanced LUT function, resulting in more efficient voiding.Regeneration of serotonin axons correlated with LUT recovery.These results suggest that our mouse model of LUT dysfunction recapitulates the results found in the rat model and may be used to further investigate genetic contributions to regeneration failure.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurosciences, Case Western Reserve University, Cleveland, Ohio, United States of America.

ABSTRACT
The loss of lower urinary tract (LUT) control is a ubiquitous consequence of a complete spinal cord injury, attributed to a lack of regeneration of supraspinal pathways controlling the bladder. Previous work in our lab has utilized a combinatorial therapy of peripheral nerve autografts (PNG), acidic fibroblast growth factor (aFGF), and chondroitinase ABC (ChABC) to treat a complete T8 spinal cord transection in the adult rat, resulting in supraspinal control of bladder function. In the present study we extended these findings by examining the use of the combinatorial PNG+aFGF+ChABC treatment in a T8 transected mouse model, which more closely models human urinary deficits following spinal cord injury. Cystometry analysis and external urethral sphincter electromyograms reveal that treatment with PNG+aFGF+ChABC reduced bladder weight, improved bladder and external urethral sphincter histology, and significantly enhanced LUT function, resulting in more efficient voiding. Treated mice's injured spinal cord also showed a reduction in collagen scaring, and regeneration of serotonergic and tyrosine hydroxylase-positive axons across the lesion and into the distal spinal cord. Regeneration of serotonin axons correlated with LUT recovery. These results suggest that our mouse model of LUT dysfunction recapitulates the results found in the rat model and may be used to further investigate genetic contributions to regeneration failure.

No MeSH data available.


Related in: MedlinePlus