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N Danielsen

Publications and source records attributed to N Danielsen.

At least 37 records · Page 2Linked to original sources
PubMed1997

Nerve regeneration in a 'pseudo-nerve' graft created in a silicone tube.

The pseudo-nerve, which contains longitudinal Schwann cell columns without axons and surrounded by perineurium-like tissue but no axons (Q. Zhao, L.B. Dahlin, M. Kanje, G. Lundborg, Brain Res. 592 (1992) 106-114), was applied as a graft to repair nerve defect in rats. Creation of the pseudo-nerve was accomplished by inserting the proximal and distal stumps of a cut sciatic nerve into a silicone tube. The proximal insert was cut far proximally to prevent axons from entering the tube. After 4 weeks, the pseudo-nerve was harvested, trimmed into a 10-mm long graft and transplanted into a corresponding defect of the contralateral sciatic nerve. Nerve regeneration through the pseudo-nerve was examined by pinch reflex test and neurofilament staining after 6 days or by morphology after 4, 6 or 8 weeks. The results showed that the pseudo-nerve could induce nerve regeneration to a similar extend as a real nerve graft. The neurobiological composition of the pseudo-nerve and the factors influencing its formation were also studied. By double staining of S-100 and laminin we found that the longitudinally organized Schwann cell columns in the pseudo-nerve were surrounded by basal laminae and ensheathed by a layer of vascularized perineurium-like tissue. Macrophages (ED1 and ED2) and their products interleukin-1beta (IL-1beta) and transforming growth factor-beta1 (TGF-beta1) were constantly present in the pseudo-nerve. Besides, the size of tube was a crucial factor in influencing pseudo-nerve formation, e.g. a thicker pseudo-nerve was formed in tubes with larger diameters or shorter gap lengths. No pseudo-nerve was formed when the gap was 15 mm long. When both proximal and distal inserts were isolated nerve segments the pseudo-nerve was still formed but thin, probably because of compromised vascular supply. Taken together, the results suggested that the pseudo-nerve contains the essential neurobiological elements to induce successful axonal elongation.

Animals↗
PubMed1997

S-100beta stimulates neurite outgrowth in the rat sciatic nerve grafted with acellular muscle transplants.

S-100beta promotes neurite extension in vitro and motoneuron survival in the chicken embryo. We demonstrate here that local administration of S-100beta stimulates the sciatic nerve regeneration into acellular muscle grafts. Normally there is a 8-10 day delay in the regeneration of axons into such grafts. Local administration of S-100beta (0.5-1.0 microg/h) significantly stimulated regeneration into the grafts. In S-100beta treated grafts, the regeneration distance was increased with a factor of about 2.3 times as compared to vehicle treated grafts. The distance of regeneration was monitored with pinch test which detects sensory axons. Regenerating axons were growing outside the necrotic muscle cells as revealed with immunohistochemistry for the neurofilament light weight polypeptide. S-100beta was demonstrated immunocytochemically in motor neurons of the rat lumbar spinal cord and in large and medium sized neurons of the dorsal root ganglia. The results suggest that S-100beta is a physiological growth factor for peripheral nerve axons.

Animals↗
PubMed1997

Rat sciatic nerve regeneration through a micromachined silicon chip.

The capacity of regenerating nerve fibres to grow through a perforated silicon chip was tested using the silicone chamber model for nerve regeneration. The chips were fabricated as circular membranes, 4 mm in diameter, thickness 60 microns, with a perforated area, 2 mm in diameter, in the centre. Three types of chips were fabricated utilizing anisotropic etching. The chips were glued with silicone adhesive between two halves of silicone rubber tubing (total length 8 mm, inner diameter 1.8 mm, outer diameter 3.0 mm) which was used to bridge a 4 mm gap between the proximal and distal nerve stumps of a transected rat sciatic nerve. The capacity of regenerating nerve fibres to grow through the holes of the chip was analysed by light and scanning electron microscopy after 4 or 16 weeks of regeneration. Furthermore, the muscle contractility force of the gastrocnemius muscle was measured after 16 weeks of regeneration and compared as a percentage of the contralateral uninjured side. Nerves generated through chips with hole diameters of 10 or 50 microns were morphological and functional failures. The nerve structures distal to chips with hole diameters of 100 microns contained many myelinated nerve fibres in a minifascicular pattern after both 4 and 16 weeks of regeneration. The muscle contractility force was 56% of that of contralateral control muscles.

Animals↗
PubMed1997

Inflammatory reaction dependence on implant localization in rat soft tissue models.

This study compares two different implantation models in soft tissue in rat abdominal wall with regard to inflammatory reactions. Titanium rods and discs, penetrating or not penetrating the peritoneal wall respectively, were implanted. After 3, 10 or 30 days the distribution of monocytes/macrophages and cytokines (interleukin-1 and transforming growth factor-beta) in the tissue adjacent to the implants was investigated under immunohistochemistry. The macrophage-specific antibody, ED1, was used for the identification of newly recruited macrophages and the ED2 antibody was used for the mature tissue macrophages. After 10 days the non-penetrating implants had a larger number of cells close to the implant than the penetrating implants. The opposite was seen after 30 days implantation, with a larger number of cells around the penetrating implants. At all time intervals the penetrating implants had a thicker reactive capsule. The cytokines interleukin-1beta and transforming growth factor-beta could be detected in the reactive tissue adjacent to both types of implants, without obvious differences for the two implant situations. The biocompatibility of a material appears to be influenced by the localization of the implant. In addition, it seems to be of importance to extend the follow-up periods further, as we cannot assume that steady state is reached at 30 days implantation.

Abdominal Muscles↗
PubMed1997

Islet amyloid polypeptide and calcitonin gene-related peptide expression are down-regulated in dorsal root ganglia upon sciatic nerve transection.

Islet amyloid polypeptide (IAPP) is structurally related to calcitonin gene-related peptide (CGRP) and has been implicated in glucose homeostasis and diabetes pathogenesis because it is expressed in insulin cells and forms amyloid in pancreatic islets from type II diabetic patients. IAPP is also constitutively co-expressed with CGRP in rat sensory neurons. Whether expression of IAPP is altered by nerve injury with or without regeneration was investigated in adult rats subjected to unilateral sciatic axotomy; IAPP and CGRP expression were determined by quantitative in situ hybridization and immunocytochemistry at days 3, 10 and 30 after axotomy. In ipsilateral L4-L5 dorsal root ganglia (DRG), the percentages of nerve cell profiles labelled for IAPP and CGRP mRNA were reduced at all time points studied. IAPP and CGRP mRNA expression were lower in nerve cell profiles in ipsilateral DRGs compared to the contralateral side after axotomy alone whereas epineurial nerve suture maintained or restored IAPP and CGRP expression. The numbers of IAPP- and CGRP-immunoreactive DRG nerve cell profiles and dorsal horn fibers were reduced on the ipsilateral side at all time points. Thus, IAPP and CGRP expression are down-regulated upon axotomy. Nerve repair maintains or restores IAPP and CGRP expression in individual neurons but does not prevent the loss of CGRP/IAPP phenotype of some of these neurons in response to axotomy.

Amyloid↗
PubMed1997

Tubular versus conventional repair of median and ulnar nerves in the human forearm: early results from a prospective, randomized, clinical study.

Injury to a peripheral nerve is followed by local synthesis and release of neurotrophic factors of importance for the regeneration process. This concept was adopted for repair of transected human median and ulnar nerves in the forearm. As an alternative to conventional microsurgical repair of the nerve trunk, silicone tubes of appropriate size were used to enclose the injury zone, intentionally leaving a gap measuring 3-4 mm between the nerve ends inside the tube. The early results from a prospective, randomized, clinical study comparing this principle with conventional microsurgical technique for repair of human median and ulnar nerves, is presented. Eighteen patients (14 men and 4 women), aged 12-72 (mean, 29.5) years, were randomized to either tubulization (11 cases) or conventional microsurgical repair (7 cases). A battery of tests for sensory and motor functions of the hand were carried out at regular intervals for up to 1 year after surgery. The results show no difference between the both techniques, with the exception of perception of touch, which showed a significant difference (p < .05) at the 3-month checkup in favor of the tubulization technique. At re-exploration 11 months after the initial procedure (1 case), the former gap was replaced by regenerated nerve tissue in direct continuity with the proximal and distal parts of the nerve trunk, the exact level of the former injury being impossible to identify. Study data demonstrate an intrinsic capacity of human major nerve trunks to reconstruct themselves in a preformed space when an optimal environment is offered and the surgical trauma is minimized.

Adolescent↗
PubMed1997

Supersensitivity in rat micro-arteries after short-term denervation.

Contractile responses to phenylephrine and high-K+ were investigated in vitro in microvascular preparations from the rat medial plantar artery, a branch from the saphenous artery, obtained after short-term denervation in vivo. Two groups of animals were studied: (1) animals undergoing surgical resection of the saphenous nerve, and (2) animals undergoing surgical resection of both the sciatic and saphenous nerves. The animals were operated on one side only. Microvascular preparations (diameter about 325 microns) were obtained 10 days after surgery. Vessels from the non-operated side served as controls. Immunocytochemistry showed a decreased number of both neuropeptide Y (NPY) and calcitonin gene-related peptide (CGRP) immunoreactive nerve fibres in vessels after resection of the saphenous nerve only. Resection of both the saphenous and the sciatic nerve caused a complete loss of immunoreactive nerve fibres. Mechanical measurements were performed using a wire myograph. In vessels subjected to resection of the saphenous nerve the sensitivity to phenylephrine was similar to controls. Vessels denervated by resection of both the saphenous and sciatic nerves showed significant increases in phenylephrine and potassium sensitivity. When depolarized in high-K+ solution the denervated vessels showed an increased sensitivity to extracellular Ca2+. The results show that complete short-term denervation of the rat medial plantar artery in vivo causes a pronounced supersensitivity in the vascular smooth muscle. The supersensitivity appears not to be restricted to the sympathetic alpha-receptors but also associated with changes in the cellular excitation-contraction coupling. Such altered reactivity of the vascular smooth muscle may contribute to vascular disturbances observed in vivo after nerve damage or surgical denervation.

Adrenergic alpha-Agonists↗
PubMed1997

Nerve injury induced by vibration: prevention of the effect of a conditioning lesion by D600, a Ca2+ channel blocker.

OBJECTIVES: Exposing a hind leg of a rat to vibration induces an injury to the sciatic nerve--a so called conditioning lesion. After such injury induced by vibration the regenerative capacity of the nerve is improved and can be detected as an increased axonal outgrowth from a test crush lesion to the same nerve. The purpose was to study whether the effect of a conditioning lesion induced by vibration can be prevented by local treatment with a Ca2+ channel blocker D600. METHODS: D600 (methoxyverapamil) or Ringer's solution was locally applied to the sciatic nerve on one side through a silicone tube connected to a miniosmotic pump, which was implanted subcutaneously. During the same period the hind leg was exposed to vibration (80 Hz; 32 m/s2 root mean squared) for five hours daily for five consecutive days. The other hind leg was not vibrated. After the end of exposure to vibration the sciatic nerves were crushed bilaterally (test crush lesions) and three or six days later the regeneration distances of sensory axons were measured by the pinch reflex test. RESULTS: Nerves in the control animals (without implanted miniosmotic pumps and nerves on to which Ringer's solution was locally applied) that were exposed to vibration showed a significantly increased outgrowth length of sensory axons from the test crush lesion compared with the non-vibrated side. Such an effect of a conditioning lesion from the exposure to vibration was suppressed by local application of D600. CONCLUSIONS: Local administration of a Ca2+ channel blocker D600 can prevent the effect of a conditioning lesion-that is, the nerve injury induced by vibration can be inhibited by D600. This may have implications for the treatment of patients with neuropathy of the hand induced by vibration.

Animals↗
PubMed1997

Role of macrophages in the stimulation and regeneration of sensory nerves by transposed granulation tissue and temporal aspects of the response.

Application of granulation tissue, which is rich in macrophages, to a peripheral nerve induces a conditioning effect, in that it enhances the regeneration capability of peripheral nerves after a test crush lesion. The temporal aspects of this response and the role of macrophages and interleukin-1 beta (IL-1 beta) were studied in the sciatic nerves of 71 rats. Granulation tissue was implanted close to the sciatic nerve and test crush lesions were applied after various periods of time (0-21 days). Regeneration was evaluated after an additional two, three, four, or six days. Regeneration distances were longer in granulation-treated nerves than in nerves treated with subcutaneous tissue. Furthermore, in animals in which the test crush lesion was made at the same time as the granulation tissue was implanted (n = 6), regeneration distances were longer, 8.1 (0.8) mm compared with 7.2 (0.6), than those in which the crush was made after conditioning intervals of 3 (n = 6, 7.6 (0.4) compared with 6.9 (0.4), p = 0.03); 7 (n = 6, 7.4 (0.4) compared with 6.6 (0.1), p = 0.03); and 21 days [(n = 8, 7.2 (0.6) compared with 6.4 (0.5)]. Inactivation of the granulation tissue by freezing suppressed the conditioning effect. There were numerous ED1 and ED2 positive macrophages as well as positive staining for IL-1 beta in the granulation tissue on day 0. Positive staining for IL-1 beta was also seen in nerve fibres as well as in non-neuronal cells after a conditioning interval. The results suggest that regeneration is stimulated by factors released from the cells of the granulation tissue, and that the amount of factors released or the responsiveness of the regenerating nerve change during the conditioning interval.

Animals↗
PubMed1996

Nerve regeneration and repair.

A cut peripheral nerve can be repaired by either suturing the two nerve stumps together directly or by interposing a nerve graft between the two stumps. Several factors such as the patient's age, type of injury, and proximity of the lesion will influence the functional results. In our own experiments we have encased the two nerve stumps in various chambers intentionally leaving a short gap between the nerve stumps. In these chambers neurotrophic factors will accumulate and in a prospective randomized clinical trial it has been demonstrated that the chamber principle has some advantages as compared to established technique. Other experiments utilizing predegenerated experimental nerve grafts have demonstrated that regenerating axons enter these grafts more rapidly than in ordinary nerve grafts.

Age Factors↗
PubMed1996

Immunohistochemical studies on the distribution of albumin, fibrinogen, fibronectin, IgG and collagen around PTFE and titanium implants.

Time-dependent distribution of extracellular proteins (albumin, fibrinogen, fibronectin, collagen-I and IgG) in the interface zone between implant and soft tissue has been investigated utilizing a recently developed method. Commercially pure (c.p.) titanium and polytetrafluoroethylene (PTFE) implants were inserted in the abdominal wall of rats for 1, 6 and 12 weeks followed by a mild fixation, cryoprotection, rapid freezing in LN2-cooled propane, cryosubstitution and low-temperature infiltration with UV curing of the methacrylate LR-Gold. Before sectioning, the bulk part of the titanium was removed by an electrolytical dissolution technique (electropolishing), while the PTFE implants were removed by a fracture technique. Employing a cryosubstitution method combined with postembedding immunohistochemistry, a light microscopic analysis was allowed. The selected proteins had an apparently varying distribution in the implant-close tissue and their distribution changed during the follow-up period. There was also a difference in the distribution pattern for each protein around titanium and PTFE implants. Insertion of the c.p. titanium implants elicited an inflammatory reaction in many respects similar to a normal wound healing response, while the PTFE implants caused a more pronounced, persistent inflammation.

Abdominal Muscles↗
PubMed1996

Pituitary adenylate cyclase activating peptide expression in the rat dorsal root ganglia: up-regulation after peripheral nerve injury.

Pituitary adenylate cyclase activating peptide (PACAP) is expressed in a population of capsaicin-sensitive primary sensory neurons of small to medium size in the rat. In the present report we have examined the effect of sciatic nerve injury (unilateral transection) on PACAP expression (immunocytochemistry, radioimmunoassay, in situ hybridization and northern blot analysis) in dorsal root ganglia at the lumbar level and on immunoreactive PACAP in the spinal cord and in the sciatic nerve stump. For comparison, calcitonin gene-related peptide was examined. In dorsal root ganglia of the intact side immunoreactive PACAP and PACAP messenger RNA were localised to a population of nerve cell bodies of small to medium size. In dorsal root ganglia on the injured side, PACAP-immunoreactive nerve cell bodies were more numerous and PACAP messenger RNA was considerably more abundant as studied 14 days after sciatic nerve transection. By contrast, calcitonin gene-related peptide-containing nerve cell bodies were numerous and rich in calcitonin gene-related peptide messenger RNA in dorsal root ganglia on the intact side, while after transection both the number of immunoreactive nerve cell bodies and their content of messenger RNA were markedly reduced. There were indications of axotomy-induced expression of PACAP messenger RNA in larger neurons. In the dorsal horn of the spinal cord on the intact side PACAP and calcitonin gene-related peptide-immunoreactive fibres were densely accumulated in the superficial layers. On the transected side the densities of both PACAP and calcitonin gene-related peptide-immunoreactive nerve fibres were reduced in the medial part. The data obtained indicate a marked up-regulation of PACAP in sensory neurons following peripheral nerve injury. Since PACAP depresses a C-fibre evoked flexion reflex, this may have implications for sensory transmission. Further, in view of the known promoting effects of PACAP on neuronal survival and differentiation and non-neuronal cell growth as well as its proinflammatory effects a role of PACAP in the neuronal and periaxonal tissue restoration after injury is not inconceivable.

Animals↗
PubMed1996

Regeneration across a partial defect in rat sciatic nerve encased in a silicone chamber.

Forty-four rat sciatic nerves with partial defects were repaired with a silicone chamber. Each partial defect was created by resecting a 10 mm segment from the tibial fascicle leaving the peroneal fascicle intact. The proximal and the distal stumps of the tibial fascicle together with the intact peroneal fascicle were encased in a single silicone chamber. After seven days a fibrin matrix had surrounded the peroneal fascicle and spanned the defect between the tibial stumps. This matrix was later invaded by non-neuronal cells and regenerating axons. Non-myelinated nerve fibres had almost regenerated across the defect by 16 days. The tetanic force of the gastrocnemius muscle 120 days after repair showed 80% recovery, which was no different from that of partial defects repaired with conventional nerve grafts. The results suggest that the silicone chamber technique could be applicable to the treatment of partially transected nerve trunks.

Animals↗
PubMed1995

Predegeneration enhances regeneration into acellular nerve grafts.

In the present study, we determined the regeneration rate and the initial delay in rat sciatic nerve grafts first made hypercellular by predegeneration then acellular by freeze-thawing. 7-day predegenerated nerve pieces from the distal nerve stump on the right side were made acellular by repeated freeze-thawing and inserted as grafts into a 10-mm long freshly created defect on the left contralateral side. Freshly made (no predegeneration period) acellular nerve grafts were used as controls. Both types of grafts supported outgrowth of regenerating axons as demonstrated by the sensory pinch test. However, the predegenerated acellular nerve grafts had a significantly shorter initial delay period (2.7 days) as compared with freshly made acellular nerve grafts (9.5 days). The initial delay period for predegenerated acellular nerve grafts was similar to that for fresh cellular nerve grafts but significantly longer than that for predegenerated cellular nerve grafts [24]. The rate of regeneration appeared independent of the type of grafts used. We suggest that modifications of the basal lamina and/or factors produced during the predegeneration period by non-neuronal cells survive the freeze-thawing cycle and account for the decrease in the initial delay period.

Animals↗
PubMed1995

Characterization of neurotrophic activity in the silicone-chamber model for nerve regeneration.

In the present study, the authors reevaluated the temporal course and properties of neurotrophic activities present in the fluid accumulating in the silicone-chamber model for nerve regeneration. The fluid collected from silicone chambers was tested in four different dissociated neuronal cell cultures. Furthermore, the activity of the chamber fluid was examined, using a cell blot technique. There was one major peak of neurotrophic activity and this activity peaked early, about 3 to 6 hr after nerve injury. Results also indicate that the chamber fluid contains at least two types of neurotrophic activities, namely nerve growth factor and ciliary neurotrophic factor.

Animals↗
PubMed1995

Sympatho-adrenergic inhibition of basal and acid-induced changes in duodenal motility, mucosal net fluid and alkaline secretion in the anaesthetized cat.

Experiments were performed on chloralose anaesthetized cats. A 2-cm segment of the proximal duodenum was isolated between two luminally situated balloons and perfused with isotonic saline containing [14C]-PEG 4000 as a non-absorbable marker. The perfusate was analysed with regard to alkalinity (back titration) and concentration of marker (liquid scintillation). Net alkalinization and net fluid transport were calculated with conventional equations. Motor activity in the duodenal wall was recorded as changes in volume of the proximal balloon. In presence of sympathetic neural activity (spontaneous or electrically stimulated) basal motor activity and mucosal alkaline secretion was low and increased minimally in response to luminal HCl (30 mM). Net fluid transport was in an absorptive state and shifted to a small secretion upon the acid-exposure. Subsequent to bilateral acute splanchnicotomy, or the administration of the adrenolytic guanethidine (3-4 mg kg-1, i.v.), spontaneous duodenal contractions occurred and the alkaline secretion was increased. Furthermore, both parameters were then markedly stimulated by luminal perfusion with 30 mM HCl. Basal net fluid transport was zero and turned into secretion upon the acid-exposure. No morphological changes of the duodenal surface epithelium could be detected. The study demonstrates the existence of splanchnic nerve-mediated, adrenergic inhibition of basal, as well as of acid-induced duodenal motility, fluid and alkaline secretion.

Animals↗
PubMed1994

Pre-degenerated nerve grafts enhance regeneration by shortening the initial delay period.

In the present study we tested how nerve grafts with different pre-degeneration periods (1-28 days) influenced the early regenerative response in the rat sciatic nerve. The sciatic nerve on the right side was crushed and after 1-28 days of pre-degeneration, a 10 mm segment was used as an autologous nerve graft and transposed to a freshly made 10 mm long nerve defect on the left side. The regeneration distance was measured by the sensory pinch test 2-10 days after nerve repair. A newly developed mathematical model was used to calculate regeneration rates and initial delay periods from the measured regeneration distances. Pre-degenerated nerve grafts improved nerve regeneration by decreasing the initial delay period as compared to fresh nerve grafts without affecting the regeneration rate. Only one day of pre-degeneration was sufficient to reduce the initial delay period from 3.6 days to 1.7 days. The maximal effect on the initial delay period was achieved after 3 days of pre-degeneration. The initial delay period at later pre-degeneration intervals (7-14 days) was about 1 day. The effect persisted for at least 28 days of pre-degeneration. The regeneration rate was 1.5 mm/day for fresh nerve grafts and between 1.8-2.1 mm/day for pre-degenerated grafts. The results suggest that the effects of pre-degeneration are not only due to the increased cell proliferation in the graft, but that also trophic and/or inflammatory mechanisms may be of importance. Grafts pre-degenerated by crush may have clinical implications since they are easy to perform if an elective nerve grafting procedure is planned.

Animals↗