Joo Yup Lee

The effect of collagen nerve conduits filled with collagen- glycosaminoglycan matrix on peripheral motor nerve regeneration in a rat model

BACKGROUND Bioabsorbable unfilled synthetic nerve conduits have been used in the reconstruction of small segmental nerve defects with variable results, especially in motor nerves. We hypothesized that providing a synthetic mimic of the Schwann cell basal lamina in the form of a collagen-glycosaminoglycan (GAG) matrix would improve the bridging of the nerve gap and functional motor recovery. METHODS A unilateral 10-mm sciatic nerve defect was created in eighty-eight male Lewis rats. Animals were randomly divided into four experimental groups: repair with reversed autograft, reconstruction with collagen nerve conduit (1.5-mm NeuraGen, Integra LifeSciences), reconstruction with collagen nerve conduit filled with collagen matrix, and reconstruction with collagen nerve conduit filled with collagen-GAG (chondroitin-6-sulfate) matrix. Nerve regeneration was evaluated at twelve weeks on the basis of the compound muscle action potential, maximum isometric tetanic force, and wet muscle weight of the tibialis anterior muscle, the ankle contracture angle, and nerve histomorphometry. RESULTS The use of autograft resulted in significantly better motor recovery compared with the other experimental methods. Conduit filled with collagen-GAG matrix demonstrated superior results compared with empty conduit or conduit filled with collagen matrix with respect to all experimental parameters. Axon counts in the conduit filled with collagen-GAG matrix were not significantly different from those in the reversed autograft at twelve weeks after repair. CONCLUSIONS The addition of the synthetic collagen basal-lamina matrix with chondroitin-6-sulfate into the lumen of an entubulation repair significantly improved bridging of the nerve gap and functional motor recovery in a rat model. CLINICAL RELEVANCE Use of a nerve conduit filled with collagen-GAG matrix to bridge a motor or mixed nerve defect may result in superior functional motor recovery compared with commercially available empty collagen conduit. However, nerve autograft remains the gold standard for reconstruction of a segmental motor nerve defect.

Does the addition of a nerve wrap to a motor nerve repair affect motor outcomes

The purpose of this study was to evaluate the effect of wrapping bioabsorbable nerve conduit around primary suture repair on motor nerve regeneration in a rat model. Forty rats were randomly divided into two experimental groups according to the type of repair of the rat sciatic nerve: group I had primary suture repair; group II had primary suture repair and bioabsorbable collagen nerve conduit (NeuraGen® 1.5 mm, Integra LifeSciences Corp., Plainsboro, NJ) wrapped around the repair. At 12 weeks, no significant differences in the percentage of recovery between the two groups were observed with respect to compound muscle action potentials, isometric muscle force, and muscle weight (P = 0.816, P = 0.698, P = 0.861, respectively). Histomorphometric analysis as compared to the non‐operative sites was also not significantly different between the two groups in terms of number of myelinated axons, myelinated fiber area, and nerve fiber density (P = 0.368, P = 0.968, P = 0.071, respectively). Perineural scar tissue formation was greater in primary suture repair group (0.36 ± 0.15) than in primary repair plus conduit wrapping group (0.17 ± 0.08). This difference was statistically significant (P < 0.001). Wrapping bioabsorbable nerve conduit around primary nerve repair can decrease perineural scar tissue formation. Although the scar‐decreasing effect of bioabsorbable nerve wrap does not translate into better motor nerve recovery in this study, it might have an effect on the functional outcome in humans where scar formation is much more evident than in rats.

Functional evaluation in the rat sciatic nerve defect model: a comparison of the sciatic functional index, ankle angles, and isometric tetanic force.

Background: The sciatic functional index has long been the standard method of assessing motor recovery in the rat sciatic nerve model. The relative subjective nature of the assessment has led to development of newer methods, including video gait analysis and quantitative measurement of isometric tetanic muscle force. Methods: Forty male Lewis rats, each with a 10-mm segmental defect in the sciatic nerve, were divided randomly into two groups: rats in group I underwent repair with reversed autograft, and those in group II received a collagen conduit. Video gait analyses were performed at 0, 4, 8, and 12 weeks, and sciatic functional index and ankle angles in four different walking phases were recorded. Isometric tetanic force of the tibialis anterior muscle was also measured at 12 weeks and correlated with sciatic functional index and video gait analysis data. Results: The sciatic functional index results did not correlate with isometric tetanic force. Significantly, the sciatic functional index could not be measured in 26 percent of the rats at 8 weeks and 59 percent of the rats at 12 weeks secondary to toe contractures. Among various ankle angle measurements, only the ankle angle in toe-off phase correlated well with isometric tetanic force. Conclusions: Toe contractures occurred more frequently in rats with better nerve recovery, and interfered with evaluation of the motor recovery using the sciatic functional index method. Ankle angle in toe-off phase measured from video gait analysis is a useful parameter that reflects functional recovery of the muscle force.

Return of motor function after repair of a 3-cm gap in a rabbit peroneal nerve: a comparison of autograft, collagen conduit, and conduit filled with collagen-GAG matrix.

BACKGROUND The purpose of this study was to evaluate the motor nerve recovery in a rabbit model after repair of a 3-cm gap in the peroneal nerve with a conduit filled with a collagen-GAG (glycosaminoglycan) matrix and compare the results with those after reconstruction with an autograft or an empty collagen conduit. METHODS Forty-two male New Zealand rabbits were divided into three experimental groups. In each group, a unilateral 3-cm peroneal nerve defect was repaired with a nerve autograft, an empty collagen conduit, or a conduit filled with a collagen-GAG matrix. At six months, nerve regeneration was evaluated on the basis of the compound muscle action potentials, maximum isometric tetanic force, and wet muscle weight of the tibialis anterior muscle as well as nerve histomorphometry. RESULTS The autograft group had significantly better motor recovery than the conduit groups. The empty collagen conduits and conduits filled with the collagen-GAG matrix led to results that were similar to each other. CONCLUSIONS On the basis of this rabbit model, autologous nerve grafting remains the gold standard in the reconstruction of 3-cm segmental motor nerve defects. CLINICAL RELEVANCE Segmental motor nerve defects should be reconstructed with autograft nerves. The use of a collagen conduit filled with a collagen-GAG matrix for motor nerve reconstruction should be limited until additional animal studies are performed.

Effect of Vascular Endothelial Growth Factor Administration on Nerve Regeneration after Autologous Nerve Grafting

BACKGROUND The aim of this study was to evaluate the effect of vascular endothelial growth factor (VEGF) administration around the autologous nerve graft on nerve recovery in a rat model. METHODS A total of 69 rats were randomly divided into three experimental groups. A 10-mm sciatic nerve defect was made and reconstructed with the reversed nerve segment. Group I received an osmotic pump with saline, group II received an osmotic pump with VEGF, and group III added a silicone tube around the nerve graft to decrease the surrounding blood supply. Nine animals in each group were sacrificed on day 3 to evaluate improvement in new vessel formation. In each group 14 animals were sacrificed at 16 weeks after the initial procedure to evaluate the functional motor nerve regeneration using compound muscle action potential, isometric tetanic force, wet muscle weight, and nerve histomorphometry. RESULTS The average vascular density on day 3 was 10.7% in group I, 21.4% in group II, and 0.9% in group III. These differences were significant. However, the average maximum isometric tetanic force at 16 weeks was 54.4% in group I, 57.5% in group II, and 47.6% in group III. No difference was found with or without VEGF administration. Histomorphometric analysis was also not significantly different between the groups. CONCLUSIONS New vessel formation on autologous nerve graft was enhanced by VEGF administration. However, the neovascularization effect of VEGF administration did not translate into better motor nerve recovery.

Magnolin targeting of ERK1/2 inhibits cell proliferation and colony growth by induction of cellular senescence in ovarian cancer cells

Ras/Raf/MEKs/ERKs and PI3 K/Akt/mTOR signaling pathways have key roles in cancer development and growth processes, as well as in cancer malignance and chemoresistance. In this study, we screened the therapeutic potential of magnolin using 15 human cancer cell lines and combined magnolin sensitivity with the CCLE mutaome analysis for relevant mutation information. The results showed that magnolin efficacy on cell proliferation inhibition were lower in TOV‐112D ovarian cancer cells than that in SKOV3 cells by G1 and G2/M cell cycle phase accumulation. Notably, magnolin suppressed colony growth of TOV‐112D cells in soft agar, whereas colony growth of SKOV3 cells in soft agar was not affected by magnolin treatment. Interestingly, phospho‐protein profiles in the MAPK and PI3 K signaling pathways indicated that SKOV3 cells showed marked increase of Akt phosphorylation at Thr308 and Ser473 and very weak ERK1/2 phosphorylation levels by EGF stimulation. The phospho‐protein profiles in TOV‐112D cells were the opposite of those of SKOV3 cells. Importantly, magnolin treatment suppressed phosphorylation of RSKs in TOV‐112D, but not in SKOV3 cells. Moreover, magnolin increased SA‐β‐galactosidase‐positive cells in a dose‐dependent manner in TOV‐112D cells, but not in SKOV3 cells. Notably, oral administration of Shin‐Yi fraction 1, which contained magnolin approximately 53%, suppressed TOV‐112D cell growth in athymic nude mice by induction of p16Ink4a and p27Kip1. Taken together, targeting of ERK1 and ERK2 is suitable for the treatment of ovarian cancer cells that do not harbor the constitutive active P13 K mutation and the loss‐of‐function mutations of the p16 and/or p53 tumor suppressor proteins.