Frozen vein wrapping for chronic nerve constriction injury reduces sciatic nerve allodynia in a rat model.

BACKGROUND: Autologous vein wrapping (VW) is used in the treatment of recurrent chronic constriction neuropathy and traumatic peripheral nerve injury. However, use of autologous veins is limited by the inability to obtain longer veins of sufficient length for larger sites. Frozen allograft tissue has several advantages, including its availability for large grafts, avoidance of donor-site morbidity, and shorter operation time. Here, we investigated the effect of frozen vein wrapping (FVW) in Wistar rats as a model of sciatic nerve injury. RESULTS: The rats were grouped by treatment as (i) untreated after chronic constriction injury surgery (CCI; control group), (ii) treated with vein wrapping using freshly isolated vein (VW), and (iii) treated with vein wrapping using frozen vein (FVW). Mechanical allodynia was assessed with von Frey filaments on postoperative days (PODs) 1, 3, 5, 7, and 14. Gene expression of HO-1 was evaluated by quantitative polymerase chain reaction (qPCR). The response of heme oxygenase-1 gene, Hmox-1, expression to VW and FVW was assessed by RT-PCR. Both VW and FVW significantly increased withdrawal threshold levels compared to the untreated control group on POD 1, 3, and 5. Both VW and FVW also showed increased HO-1 expression compared to the CCI group. CONCLUSIONS: FVW increased the withdrawal threshold similar to VW in a rat CCI model for short periods. Frozen vein wrapping using vein allograft without donor site morbidity may be an alternative therapeutic option.

Evaluation of behavior and expression of NaV1.7 in dorsal root ganglia after sciatic nerve compression and application of nucleus pulposus in rats.

PURPOSE: The pathomechanisms of pain resulting from lumbar disc herniation have not been fully elucidated. Prostaglandins and cytokines generated at the inflammatory site produce associated pain; however, non-steroidal anti-inflammatory drugs and steroids are sometimes ineffective in patients. Tetrodotoxin-sensitive voltage-gated sodium (NaV) channels are related to sensory transmission in primary sensory nerves. The sodium channel NaV1.7 has emerged as an attractive analgesic target. The purpose of this study was to evaluate pain-related behavior and expression of NaV1.7 in dorsal root ganglia (DRG) after combined sciatic nerve compression and nucleus pulposus (NP) application in rats. METHODS: Rats were divided into three groups and underwent either sciatic nerve compression with NP for 2 s using forceps (n = 20), sham operation with neither compression nor NP (n = 20), or no operation (controls, n = 20). Mechanical hyperalgesia was measured every second day for three weeks using von Frey filaments. NaV1.7 expression in L5 DRG was examined 7 and 14 days after surgery using immunohistochemistry. The number of neurons immunoreactive for NaV1.7 was compared among the three groups. RESULTS: Mechanical hyperalgesia was found over the 14-day observation in the nerve compression plus NP application group, but not in the sham-operated or control groups (P < 0.05). NaV1.7 expression in L5 DRG was up-regulated in the nerve compression plus NP application group, compared with sham-operated and control rats (P < 0.01). CONCLUSIONS: Our results indicate that nerve compression plus NP application produces pain-related behavior. We conclude that NaV1.7 expression in DRG neurons may play an important role in mediating pain from sciatic nerves after compression injury and exposure to NP.

Combined effect of DBM, PRP, and bone marrow fluid on bone union in a rat posterolateral fusion model.

Platelet-rich plasma (PRP) promotes bone union through osteoinduction. We investigated whether adding demineralized bone matrix (DBM), derived naturally from biomaterial and with various growth factors, for osteoconductivity and bone marrow fluid for osteogenesis results in different bone unions. Eight-week-old male Sprague-Dawley rats were divided into four groups of five based on transplantation material: sham control (C group); DBM alone (D group); DBM + PRP (DP group); and DBM + PRP + bone marrow fluid (DPB group). After posterolateral fusion at L3-5, postoperative weekly CT imaging determined average number of bone union in facet joints (4 joints × 5 animals = 20 joints) and bone formation. Pathological evaluation and bone strength were assessed using 3-point bending two weeks postoperatively. Facet joint bone union at four weeks postoperatively was 4/20 (20%, DP group) and 8/20 (40%, DPB group) joints. Six weeks postoperatively, it was 7/20 (35%, D group), 12/20 (60%, DP group), and 16/20 (80%, DPB group). Eight weeks postoperatively, it was 13/20 (65%, D group), 17/20 (85%, DP group), and 20/20 (100%, DPB group), suggesting that DPB > DP > D > C. Bone formation and bone strength showed a similar DPB > DP > D > C group trend. Adding PRP and bone marrow fluid to DBM promotes bone union and strength.

Nerve decompression surgery suppresses TNF-ɑ expression and T cell infiltration in a rat sciatic nerve chronic constriction injury model.

Decompression surgery (DS) is a standard treatment for chronic nerve compression injuries; however, the mechanisms underlying its effects remain unclear. Here, we investigated the effects of DS on messenger RNA (mRNA) expression of tumor necrosis factor-α (TNF-α) and T cell recruitment in a rat sciatic nerve (SN) chronic constriction injury (CCI) model. Male Wistar rats were subjected to CCI to establish a model of SN injury (CCI group). DS, in which all ligatures were removed, was performed 3 days after CCI surgery (CCI + dec group). Mechanical sensitivity was assessed using the von Frey test 3, 7, and 14 days after the CCI surgery. Gene expression of Tnfa, Cd3, Cxcl10, and immunolocalization of TNF-α and the pan T cell marker, CD3, was evaluated using quantitative polymerase chain reaction (qPCR) and immunohistochemistry, respectively. In addition, the effects of TNF-α on Cxcl10 expression and CXCL10 protein production were evaluated using qPCR and enzyme-linked immunosorbent assay in SN cell culture. Rats that received DS had significantly higher withdrawal threshold levels than those in the CCI group. In addition, Tnfa, Cd3, and Cxcl10 mRNA expression increased following CCI. DS suppressed this elevated expression, with the CCI + dec group showing significantly reduced expression levels compared to the CCI group. Furthermore, TNF-α induced Cxcl10 expression and CXCL10 protein production in SN cell culture. Therefore, DS reduced TNF-α expression and T cell recruitment in the rat SN CCI model. These observations may partly explain the mechanism underlying the therapeutic effects of DS.

Biomechanics of callus in the bone healing process, determined by specimen-specific finite element analysis.

As fractures heal, immature callus formed in the hematoma is calcified by osteoblasts and altered to mature bone. Although the bone strength in the fracture-healing process cannot be objectively measured in clinical settings, bone strength can be predicted by specimen-specific finite element modeling (FEM) of quantitative computed tomography (qCT) scans. FEM predictions of callus strength would enable an objective treatment plan. The present study establishes an equation that converts material properties to bone density and proposes a specimen-specific FEM. In 10 male New Zealand white rabbits, a 10-mm long bone defect was created in the center of the femur and fixed by an external fixator. The callus formed in the defect was extracted after 3-6 weeks, and formed into a (5 × 5 × 5 mm3) cube. The bone density measured by qCT was related to the Young's modulus and the yield stress measured with a mechanical tester. For validation, a 10-mm long bone defect was created in the central femurs of another six New Zealand white rabbits, and fixed by an external fixator. At 3, 4, and 5 weeks, the femur was removed and subjected to Computed tomography (CT) scanning and mechanical testing. A specimen-specific finite element model was created from the CT data. Finally, the bone strength was measured and compared with the experimental value. The bone mineral density σ was significantly and nonlinearly correlated with both the Young's modulus E and the yield stress σ. The material-property conversion equations were E = 0.2391e8.00ρ and ρ = 30.49σ2.41. Moreover, the experimental bone strength was significantly linearly correlated with the prospective FEM. We demonstrated the Young's moduli and yield stresses for different bone densities, enabling a FEM of the bone-healing process. An FEM based on these material properties is expected to yield objective clinical judgment criteria.

A1 pulley stretching treats trigger finger: A1 pulley luminal region under digital flexor tendon traction.

BACKGROUND: A1 pulley stretching is recognized as a clinically beneficial treatment for trigger finger. It is thought to lead to an increase in the cross-sectional area of the A1 pulley luminal region, thus improving trigger finger symptoms. The purpose of the present study was thus to evaluate the resultant forces during stretching that increase the CSA of the A1 pulley luminal region using fresh-frozen cadavers. METHODS: Using seven fingers from three fresh-frozen cadavers to replicate A1 pulley stretching, we investigated the resultant forces during stretching that increase the cross-sectional area of the A1 pulley luminal region. The traction forces of the flexor digitorum profundus tendons were increased in steps to 150 N, and the cross-sectional area and height of the A1 pulley luminal region were measured using ultrasonography. FINDINGS: The cross-sectional area of the A1 pulley luminal region increased with step-wise increases in the flexor digitorum profundus traction. On average, the cross-sectional area and height of the A1 pulley luminal region showed increases of 31.4% and 43.6%, respectively, compared to the unloaded condition. INTERPRETATION: These results confirmed that A1 pulley stretching increases the cross-sectional area of the A1 pulley luminal region. A1 pulley stretching has the potential to reduce the severity of trigger finger in patients facing surgery.

Wrapping With Basic Fibroblast Growth Factor-Impregnated Collagen Sheet Reduces Rat Sciatic Nerve Allodynia.

Autologous vein wrapping is used to treat recurrent chronic constriction neuropathy and traumatic peripheral nerve injury. However, its use is restricted due to the inability to obtain sufficiently long veins for larger grafts. We previously reported that vein-derived basic fibroblast growth factor (bFGF) promotes heme oxygenase-1 (HO-1), which reduces allodynia via its anti-inflammatory properties. To mimic vein wrapping, we developed a collagen sheet impregnated with bFGF. Chronic constriction injury (CCI) was induced in male Wistar rats as a model of sciatic nerve injury, and the rats were divided into three groups: (i) untreated after CCI surgery (control group), (ii) treated with a collagen sheet wrap impregnated with phosphate-buffered saline (PBS/CS group), and (iii) treated with a collagen sheet wrap impregnated with bFGF (bFGF/CS group). Pain behavior (von Frey test) was evaluated on postoperative days (PODs) 1, 5, 7, and 14. Quantitative polymerase chain reaction was conducted on sciatic nerve RNA to quantify HO-1 gene, Hmox1, expression. Enzyme-linked immunosorbent assay were used to determine HO-1 protein levels on POD 1. von Frey testing showed significantly greater pain hypersensitivity in the control and PBS/CS groups than the bFGF/CS group. In the bFGF/CS group, Hmox1 messenger RNA and HO-1 protein levels were significantly increased in the sciatic nerve compared with the control and PBS/CS groups on PODs 1 and 5 and POD 1, respectively. The bFGF/CS group showed decreased allodynia and HO-1 induction, as observed with vein wrapping. Therefore, local application of bFGF may be an alternative treatment strategy for compressive neuropathy and peripheral nerve trauma in clinical settings. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 37:2258-2263, 2019.