Accuracy of 3D fluoro-navigated anterior transpedicular screws in the subaxial cervical spine: an experimental study on human specimens.

INTRODUCTION: Pedicle screw stabilization, the standard technique in the thoracic and lumbar spine, is increasingly used in the cervical spine. Initial studies on the use of anterior pedicle screws (ATPS) in the cervical spine have been recently published. ATPS use has theoretical advantages over posterior stabilization. We have already established a 3D-fluoroscopy navigation setup in a study of artificial bones. The aim of the current study was to evaluate the positioning quality/accuracy of ATPS introduced to human specimens. METHODS: 36 cannulated screws (3.5 mm) were implanted anteriorly into the C3-C7 segments of four spines (unfixed, frozen, cadaveric specimens) using a 3D-fluoroscopy navigation system. Placement accuracy was evaluated using a recently published classification on postoperative CT scans. Grade 1 is perfect position with pedicle wall perforation <1 mm, grade 2 is perforation <2 mm, etc., and finally grade 5 is cortical perforation of >4 mm and/or transverse foramen entry. RESULTS: 36 anterior pedicle screws were inserted into four human cervical spine specimens. Of these, seven screws were introduced to C3, five to C4 and eight each to C5, C6, and C7. Classified with the modified G&R, 21 of 36 (58.3%) were grade 1. Ten screws (27.8%) were grade 2. Grade 4 was assessed for two screws and grade 5 for three. Customary "good" positioning, combining grades 1 and 2, was thus found in 86.1%. Five screws (13.9%) did not meet this criterion (grade ≥3). CONCLUSIONS: With 86.1% of good positioning (grade 2 or better), a 3D-fluoroscopy navigation of ATPS screws into human c-spine specimens achieved a satisfying results. These are at least comparable to results presented in the literature for posteriorly introduced subaxial pedicle screws.

[Lateral ulnar collateral ligament reconstruction : A biomechanical analysis of posterolateral rotatory instability of the elbow]. / Laterale ulnare Kollateralbandplastik : Biomechanische Untersuchung zur posterolateralen Rotationsinstabilität des Ellenbogens.

BACKGROUND: Chronic posterolateral rotatory instability (PLRI) of the elbow is the result of an insufficiency of the lateral collateral ligament (LCL). Lateral ulnar collateral ligament (LUCL) reconstruction represents a well-established treatment method for PLRI. However, recurrent instability remains a problem. OBJECTIVES: The goal of this in-vitro study was to evaluate the posterolateral rotatory stability of the intact elbow, after sectioning of the LCL and after LUCL reconstruction with a triceps tendon autograft and double BicepsButton(TM) fixation. MATERIALS AND METHODS: Posterolateral rotatory stability of 6 fresh-frozen elbow specimens at a torque of 3 Nm was analyzed at 0, 45, 90 and 120° of flexion for the intact LCL, after sectioning of the LCL and after LUCL reconstruction. Moreover, cyclic loading (1000 cycles) of the intact specimens and after LUCL reconstruction was performed. RESULTS: The intact LCL and the LUCL reconstruction provided equal primary stability (0.250 ≤ p ≤ 0.888). Sectioning of the LCL significantly increased PLRI (p < 0.001). The stability of the intact specimens and after LUCL reconstruction did not differ after cyclic loading (p = 0.218). During cyclic loading, posterolateral rotation increased significantly more after LUCL reconstruction (3.2 ± 0.8°) when compared to the native LCL (2.0 ± 0.7°, p = 0.020). CONCLUSIONS: LUCL reconstruction with BicepsButton(TM) fixation provides comparable stability to the native LCL. Further clinical results are necessary to evaluate whether this technique can decrease the complication rate.

[The bare area of the proximal ulna : An anatomical study on optimizing olecranon osteotomy]. / Die Bare Area der proximalen Ulna : Eine anatomische Studie zur Optimierung der Olekranonosteotomie.

BACKGROUND: Olecranon osteotomy is an established approach for the treatment of distal humerus fractures. It should be performed through the bare area of the proximal ulna to avoid iatrogenic cartilage lesions. OBJECTIVES: The goal of this study was to analyze the anatomy of the proximal ulna with regard to the bare area and, thereby, to optimize the hitting area of the bare area when performing olecranon osteotomy. MATERIALS AND METHODS: The bare areas of 30 embalmed forearm specimens were marked with a radiopaque wire and visualized three-dimensionally with a mobile C­arm. By means of 3D reconstructions of the data sets, the following measurements were obtained: height of the bare area; span of the bare area-hitting area in transverse osteotomy; ideal angle for olecranon osteotomy to maximize the hitting area of the bare area; distance of the posterior olecranon tip to the entry point of the transverse osteotomy and the ideal osteotomy. RESULTS: The height of the bare area was 4.92 ± 0.81 mm. The hitting area of the transverse osteotomy averaged 3.73 ± 0.89 mm. The "ideal" angle for olecranon osteotomy was 30.7° ± 4.19°. The distance of the posterior olecranon tip to the entry point was 14.08 ± 2.75 mm for the transverse osteotomy and 24.21 ± 3.15 mm for the ideal osteotomy. The hitting area of the bare area in the ideal osteotomy was enhanced significantly when compared to the transverse osteotomy (p < 0.0001). CONCLUSIONS: This study provides guide values for correct osteotomy of the olecranon. Moreover, a 30° angulation of the osteotomy can significantly increase the hitting area of the bare area.

Identification of overlengthening after replacement of the radial head with a bipolar prosthesis.

INTRODUCTION: Overlengthening of the radial column leads to insufficient functionality and increased capitellar wear. Methods to detect or prevent overlengthening have been described for monopolar prostheses. The aim of this study was to evaluate whether one such method described by Athwal et al. is also applicable for a bipolar prosthesis. MATERIALS AND METHODS: The radial heads of six fresh frozen upper extremities were resected. A bipolar radial head prosthesis was implanted in each, and the effects of sequential overlengthening on the alignment of the radiocapitellar and ulnohumeral joint line were recorded by fluoroscopic images. Digital image analysis and estimation of overlengthening followed according to the method described by Athwal et al. RESULTS: Statistical analysis of the estimated and actual differences between the native state and bipolar replacement of the radial head with stepwise overlengthening of 1.5, 3, 4.5, and 6 mm showed a specificity of 86 % but consistently underestimated the amount of overlengthening with a sensitivity of only 61 %. DISCUSSION: The method described by Athwal et al. for the identification of overlengthening by a monopolar prosthesis was not found to be reliable for ruling out or quantifying overlengthening of the tested bipolar prosthesis. However, the use of the method to detect (rule in) overlengthening may be acceptable in certain circumstances. A reliable method for postoperative quantification of overlengthening by bipolar prostheses has still to be found.

Lumbar lordosis and sacral slope in lumbar spinal stenosis: standard values and measurement accuracy.

STUDY DESIGN: Radiological study. PURPOSE: To asses standard values, intra- and interobserver reliability and reproducibility of sacral slope (SS) and lumbar lordosis (LL) and the correlation of these parameters in patients with lumbar spinal stenosis (LSS). METHODS: Anteroposterior and lateral X-rays of the lumbar spine of 102 patients with LSS were included in this retrospective, radiologic study. Measurements of SS and LL were carried out by five examiners. Intraobserver correlation and correlation between LL and SS were calculated with Pearson's r linear correlation coefficient and intraclass correlation coefficients (ICC) were calculated for inter- and intraobserver reliability. In addition, patients were examined in subgroups with respect to previous surgery and the current therapy. RESULTS: Lumbar lordosis averaged 45.6° (range 2.5°-74.9°; SD 14.2°), intraobserver correlation was between Pearson r = 0.93 and 0.98. The measurement of SS averaged 35.3° (range 13.8°-66.9°; SD 9.6°), intraobserver correlation was between Pearson r = 0.89 and 0.96. Intraobserver reliability ranged from 0.966 to 0.992 ICC in LL measurements and 0.944-0.983 ICC in SS measurements. There was an interobserver reliability ICC of 0.944 in LL and 0.990 in SS. Correlation between LL and SS averaged r = 0.79. No statistically significant differences were observed between the analyzed subgroups. CONCLUSION: Manual measurement of LL and SS in patients with LSS on lateral radiographs is easily performed with excellent intra- and interobserver reliability. Correlation between LL and SS is very high. Differences between patients with and without previous decompression were not statistically significant.

Arterial supply of the distal humerus.

PURPOSE: Distal humeral fractures are rare, but severe injuries, the treatment of which is often accompanied by serious complications and its outcome strongly depends on the quality of surgical therapy. Non-union is a common entity, compromising clinical results and requiring revision surgery. Osteonecrosis is an underestimated etiologic factor in the development of non-union. The present study aims to display the distribution patterns of the arterial vessels at the distal humerus, to correlate the displayed vessels with local nutrient foramina and to disclose an endangerment of these structures by common osteosynthetic implants. METHODS: Eight plastinated fresh frozen upper extremities were digitally analyzed regarding the vascular density of the cancellous bone, by calculating the ratio of area comprised by arterial vessels and the area comprised by cancellous bone on sagittal cuts of the distal humerus. Possible differences in the vascular density of the medial epicondylar region, the lateral epicondylar region and a watershed area between the epicondyles and distal to the supracondylar region were investigated. On the basis of 200 macerated humeri, the distribution pattern of cortical nutrient foramina and their anatomic relation to properly applied common distal humerus plates were documented. RESULTS: The data show a significantly higher density of vessels per cancellous bone in the epicondylar regions than in the watershed region (p < 0.000, median 0.148 vs. 0.103). The analysis of the nutrient foramina showed distinct distribution patterns with a single foramen over the medial epicondyle (55 specimens, 27.5 %) and an area of several foramina at the posterior part of the lateral epicondyle (200 of the specimens, 100 %). In almost every specimen, the application of the osteosynthetic implants led to an overlay over the investigated nutrient foramina. DISCUSSION: Osteonecrosis and non-union are severe complications in the surgical treatment of distal humeral fractures. The biology of the bone, especially the blood supply, has to be respected as much as possible during open procedures, to optimize bony healing. This has to be considered when performing periosteal stripping or applying osteosynthetic plates over the postero-lateral and medial epicondyle. The watershed area of the distal humerus has to be considered as being prone to minor arterial blood supply and thereby non-union is possible, if the arterial vessels coming from the epicondyles are destroyed.

Biomechanical evaluation of the primary stability of pedicle screws after augmentation with an innovative bone stabilizing system.

INTRODUCTION: In today's aging population, diminished bone quality often affects the outcome of surgical treatment. This occurs especially when surgical implants must be fixed to bone, as it occurs when lumbar fusion is performed with pedicle screws. Besides Polymethylmethacrylate (PMMA) injection, several techniques have been developed to augment pedicle screws. The aim of the current study was to evaluate the primary stability of an innovative system (IlluminOss™) for the augmentation of pedicle screws in an experimental cadaveric setup. IlluminOss™ is an innovative technology featuring cement with similar biochemical characteristics to aluminum-free glass-polyalkenoate cement (GPC). MATERIALS AND METHODS: IlluminOss™ was inserted transpedicularly via a balloon/catheter system in 40 human cadaveric lumbar vertebrae. For comparability, each vertebra was treated bilaterally with pedicle screws, augmented and non-augmented. The maximum failure load during pull out test was documented by a universal material testing machine. RESULTS: The results showed significantly higher failure loads for the augmented pedicle screws (Median 555.0 ± 261.0 N, Min. 220.0 N, Max. 1,500.0 N), compared to the native screws (Median 325.0 ± 312.1 N, Min. 29.0 N, Max. 1,400.0 N). CONCLUSIONS: Based on these data, we conclude the IlluminOss™ system can be used to augment primary screw stability regarding axial traction, compared to native screws. The IlluminOss™ monomer offers ease of control for use in biological tissues. In contrast to PMMA, no relevant heat is generated during the hardening process and there is no risk of embolism. Further studies are necessary to evaluate the usefulness of the IlluminOss™ system in the in vivo augmentation of pedicle screws in the future.

[Proximal carpal row carpectomy]. / Die Entfernung der proximalen Handwurzelreihe.

OBJECTIVE: Resection of the proximal carpal row, termed proximal row carpectomy (PRC), is performed in order to treat pathologies of the proximal carpal row or radiocarpal joint between the scaphoid and scaphoid facet. It entails the articulation of the capitate and the lunate facet. INDICATIONS: Lunate necrosis, carpal collapse, joint infection with concomitant intercarpal ligament lesions. CONTRAINDICATIONS: Severe cartilage lesions of the lunate facet and the capitate, wrist capsule laxity, rheumatoid arthritis, neuromuscular dysbalance of the wrist-covering soft tissue structures. SURGICAL TECHNIQUE: Dorsal approach to the wrist, incision of the third and fourth extensor compartments, resection and coagulation of the dorsal interosseous nerve, usage of a ligament-sparing capsule incision, identification of the proximal carpal row and inspection of cartilage of the lunate facet and capitate, mobilization and excision of the lunate, scaphoid and triquetrum, articulation of lunate facet and capitate is controlled clinically and fluoroscopically, wound closure, application of plaster slabs. POSTOPERATIVE MANAGEMENT: Immobilization of the wrist on plaster slabs for 2 weeks, removal of sutures after 14 days. RESULTS: PRC is a surgical procedure with few complications. Satisfactory range of motion and grip strength could be preserved without limiting function of the upper extremity. Postoperative osteoarthritis of capitate and lunate facet did not correlate with the good clinical outcome.

[Surgical decompression of the superficial radial nerve: Wartenberg syndrome]. / Die operative Dekompression des Ramus superficialis des Nervus radialis: Das Wartenberg-Syndrom.

OBJECTIVE: Open decompression of the superficial radial nerve is performed at the distal forearm in cases of circumscript entrapment. Broad-based entrapments can be treated by endoscopically assisted decompression. INDICATIONS: Entrapment of the nerve along its course between the brachioradialis and extensor carpi radialis longus muscles or tendons. Persistent neuropathic pain with Tinel's sign. Numbness distal to the entrapment in accordance to nerval innervation. Futile conservative treatment. Pathological electrophysiological findings. CONTRAINDICATIONS: Endogeneous neuropathy, bleeding disorders, anticoagulation medication. SURGICAL TECHNIQUE: Longitudinal skin incision at the Tinel's sign at the forearm. The subcutaneous tissue is dissected until forearm fascia is detected. The fascia is opened cautiously under direct visualization and the superficial radial nerve is identified. In cases of broad-based entrapments, endoscopically assisted decompression can be performed. The dissector with attached optical device is introduced proximally and distally. Blunt mobilization using the dissector and preparation with the Metzenbaum scissors release the superficial radial nerve sufficiently. POSTOPERATIVE MANAGEMENT: Bandaging allowing immediate motion, removal of sutures after 14 days, avoidance of excessive use for 2 weeks. RESULTS: Surgical decompression of the superficial radial nerve yields good to excellent results regarding pain reduction and sensory function.

Axonal branching and recovery of coordinated muscle activity after transection of the facial nerve in adult rats.

Facial nerve surgery inevitablyleads to pareses, abnormally associated movements, and pathologically altered reflexes. The reason for this "post-paralytic syndrome" is the misdirected reinnervation of targets, which consists of two major components. First, due to malfunctioning axonal guidance, a muscle gets reinnervated by a "foreign" axon, that has been misrouted along a "wrong" fascicle. Second, the supernumerary collateral branches emerging from all transected axons simultaneously innervate antagonistic muscles and cause severe impairment of coordinated activity. Since it is hardly possible to influence the first major component and improve the guidance of several thousands of axons, we concentrated on the second major component and tried to reduce the collateral axonal branching. The efficiency of various treatments was evaluated in rats by determining: (1) the degree of post-operative axonal branching as estimated by the number of double-or triple-labeled perikarya after application of crystalline DiI, Fluoro-Gold (FG), and Fast Blue (FB) to the zygomatic, buccal, and marginal mandibular branch of the facial nerve respectively; (2) the accuracy of reinnervation as estimated by the number of double-labeled perikarya innervating the whisker pad muscles before and after surgery as shown by intramuscular injections of FG and FB respectively; (3) the recovery of vibrissal motor performance, estimated by a video based motion analysis. So far, we have tried to reduce branching by alteration of the afferent trigeminal input to the axotomized facial motoneurons and by focal application of: (1) neurite outgrowth fostering ECM proteins; (2) neutralizing antibodies to NGF, BDNF, CNTF, GDNF, IGF-I, and FGF-II; (3) suspensions of olfactory ensheathing cells, Schwann cells, and bone marrow stroma cells; and (4) pieces of autologous olfactory mucosa to the transection site. Although most of these manipulations do influence peripheral nerve regeneration to some extent, only the application of autologous olfactory mucosa yielded a major improvement, i.e., better function.

Axotomy induces transient calbindin D28K immunoreactivity in hypoglossal motoneurons in vivo.

Calbindin D28K, an intracellular calcium-binding protein, acts as Ca2+ buffering system in the cytoplasm. By means of this property, calbindin may protect neurons against large fluctuations in free intracellular Ca2+ and, hence, may prevent cell death. Although axotomy causes a massive influx of calcium into the lesioned neurons, resection of the hypoglossal nerve does not induce extensive neuronal cell death in rats. Even several weeks after axotomy, about 70% of the motoneurons survive despite permanent target deprivation. The mechanisms responsible for this remarkable survival rate are unknown. In this study, we have looked at the modification of calbindin immunoreactivity in axotomized hypoglossal motoneurons. In non-axotomized motoneurons, no calbindin is detectable by immunocytochemistry. Axotomy induced an increase of calbindin immunoreactivity in lesioned motoneurons. This increase, visualised by the number of calbindin-immunoreactive neurons extended from 1 day to 28 days. At this time most, but not all, motoneurons located on the side of the lesion were calbindin-positive as shown by retrograde labeling and immunoquenching. From 14 days post operation, calbindin immunoreactivity decreased and reached its basal value after 35 days post operation. At that time, only fibres were still calbindin immunoreactive. Interestingly, calbindin-immunoreactivity was also increased in almost all cell nuclei, compatible with a nuclear regulation. These data are consistent with the hypothesis that, as a reaction to axotomy, motoneurons trigger an increase in calbindin expression which acts as a compensatory Ca(2+)-buffering system, enabling neurons to maintain Ca2+ homeostasis and the survival of many motoneurons after axotomy.

Expression of different isoforms of nitric oxide synthase in experimentally denervated and reinnervated skeletal muscle.

Denervated muscle fibers express enhanced levels of stress and apoptosis-associated proteins and undergo apoptosis. In experimentally denervated and reinnervated rat facial muscle, we now evaluate changes in the expression patterns of different isoforms of nitric oxide synthase (NOS)-generating nitric oxide (NO), which mediates oxidative stress and apoptosis. Physiological expression of NOS corresponds to a constant sarcolemmal staining pattern for neuronal NOS (nNOS) and a patchy sarcolemmal and weak sarcoplasmic labeling for the endothelial NOS-isoform, with no expression for inducible NOS (iNOS). Denervated muscle displayed distinct downregulation of nNOS with preserved expression of dystrophin. Also, denervated and immediately reinnervated muscle fibers showed decreased expression of nNOS. However, muscle fibers reinnervated for 10 weeks revealed a restored physiological expression of nNOS. There were no changes in the expression of endothelial and inducible NOS. As NO is known to induce growth arrest and collapse of neuronal growth cones, downregulation of NOS may contribute to promotion of axonal regeneration by aiding formation of new endplates. NO is upregulated in reinnervated muscle fibers and thus prevents polyneural hyperinnervation by extrajunctional synapses. Furthermore, downregulation of NOS during denervation is compatible with the finding that low levels of NO contribute to apoptosis instead of necrosis in disease states of oxidative stress.

Slow axonal regrowth but extreme hyperinnervation of target muscle after suture of the facial nerve in aged rats.

Unilateral transection and suture of the facial nerve was performed in 60 old rats (20 months of age). The time course of mimetic reinnervation was studied by counting all retrogradely labeled motoneurons in the facial nucleus after injection of HRP into the whiskerpad muscles for 14-112 days post operation. The comparison between these neuron counts and data for young rats yielded four conclusions. First, the qualitative equivalent of the phenomenon "misdirected reinnervation" in aged rats was the same as in young adults: HRP-labeled motoneurons were scattered throughout the facial nucleus lacking myotopic organization from 18 until 112 days post operation. Second, no age-related loss of motoneurons was detected. Third, the axonal regrowth was delayed in aged rats. Fourth, the postoperative hyperinnervation (the projection of more motoneurons into a muscle than under normal conditions, i.e., the quantitative aspect of misdirected reinnervation) was more than two times higher than in young rats. These data may provide reasonable explanations for the poor functional recovery after reconstructive surgery on the facial nerve in old patients.

Recovery of original nerve supply after hypoglossal-facial anastomosis causes permanent motor hyperinnervation of the whisker-pad muscles in the rat.

Hypoglossal-facial anastomosis (HFA), used in humans for the treatment of facial palsy, was experimentally performed in adult female Wistar rats. The time course of facial reinnervation and the extent of the new motor nerve supply of the vibrissal muscles that develops after HFA were estimated by counting all motoneurons in the brainstem labeled by injection of horseradish peroxidase (HRP) into the whisker pad; muscle innervation by motor endplates was not studied. In untreated animals, HRP injection labels 1,254 +/- 54 (mean +/- S.D.; n = 6) motoneurons, localized exclusively in the lateral subdivision of the facial nucleus. Immediately following HFA, this number drops to zero. The first HRP-labeled motoneurons appear in the hypoglossal nucleus at 28 days postoperation (dpo) and at 56 dpo their number reaches 1,096 +/- 48. Unexpectedly, the facial nerve, whose proximal stump has been left as blind end during surgery, additionally sends axons to the facial periphery. This resprouting is first detected at 42 dpo with HRP-marked neurons throughout the facial nucleus lacking somatotopic organization. The number of these labeled neurons also rises with time, and at 56 dpo, a total of 1,797 +/- 142 facial and hypoglossal motoneurons, that is, 43% more motoneurons than in normal animals, supplies the whisker pad. This hyperinnervation, that is, the projection of more motoneurons into the target muscle than under normal conditions--further increases to 1,978 +/- 92 motoneurons at 224 dpo and may provide a new animal model for studying the competitive relationships between motoneurons in their search for peripheral targets.

Ultrastructural three-dimensional reconstruction of group III and group IV sensory nerve endings ("free nerve endings") in the knee joint capsule of the cat: evidence for multiple receptive sites.

The noncorpuscular endings ("free nerve endings") of thinly myelinated group III and nonmyelinated group IV afferent nerve fibers have been examined in the knee joint capsule of sympathectomized cats by transmission electron microscopy and three-dimensional reconstruction of series of semi- and ultrathin sections. The sensory ending is the most distal part of a group III or IV nerve fiber that consists only of the sensory axon and associated Schwann cells but lacks a myelin sheath and is not surrounded by perineurium. The sensory axon divides into several branches and forms a terminal tree. The branches run either as single fibers or within small Remak bundles in parallel to sensory axons of other endings; they spread along vessel walls and also extend into dense connective tissue. Each sensory axon consists of a series of spindle-shaped thick segments ("beads") connected by waist-like thin segments. Thus all axons of sensory endings have a string-of-beads appearance, which resembles that of efferent sympathetic nerve fibers. The beads of the sensory axon and the end bulb at its tip show the same ultrastructural features which are characteristic of receptive sites: an accumulation of mitochondria and glycogen particles and various vesicles in the axoplasm and "bare" areas of axolemma that are not covered by Schwann cell processes. Group III and group IV sensory endings differ in the length of their branches (up to 200 microM in group III vs. more than 300 microM in group IV), number of beads per 100 microM axon length (about seven vs. nine or ten), mean diameter of axons (0.9-1.5 microM vs. 0.3-0.6 microM), and the presence of a neurofilament core consisting of bundles of parallel microfilaments only in group III. In conclusion, we propose that the sensory part of noncorpuscular "free nerve endings" is formed by the entire terminal tree of group III or group IV nerve fibers and that the beads in the course of the sensory axon represent multiple receptive sites.

Delayed hypoglossal-facial nerve suture after predegeneration of the peripheral facial nerve stump improves the innervation of mimetic musculature by hypoglossal motoneurons.

Surgical reconstruction of the facial nerve is common clinical practice following destruction of the intracranial facial nerve. Delayed hypoglossal-facial anastomosis (HFA) is the procedure of choice, although the effect of delay on outcome remains unclear. To study the effect of delayed anastomosis on reinnervation, we sutured the proximal stump of a freshly transected hypoglossal nerve of Wistar rats to the distal stump of the ipsilateral facial nerve, which had been transected 7-56 days earlier. Animals that had received HFA without delay served as the control group. Forty days after HFA, horseradish peroxidase (HRP) was injected into the whisker pad; 2 days later, the animals were killed. Reinnervation was assessed by determining the proportion of labeled neuronal cell bodies in the brainstem. The control group had 68% reinnervation of these muscles by hypoglossal neurons and had 32% reinnervation by facial neurons. When the distal facial nerve had been allowed to degenerate for 7 days before HFA, reinnervation of the hypoglossal nerve decreased to 54%, and reinnervation by the facial nerve increased to 46%. However, after a delay of 10-56 days, the hypoglossal fraction increased and stabilized at 77%, and the facial motoneuron fraction decreased to 23%. The presence of new neuromuscular junctions was confirmed by HRP labeling of motor end plates in vivo and by electromyography. We conclude that, under the conditions of hypoglossal-facial crossed nerve suture, the predegeneration of the distal stump of a transected facial nerve enhances the reinnervation of facial muscles by hypoglossal axonal sprouts.

Fine sensory innervation of the knee joint capsule by group III and group IV nerve fibers in the cat.

Afferent group III and IV nerve fibers of the knee joint markedly differ in their responsiveness to mechanical stimulation, which may be reflected in the structure and location of their terminals. Therefore, in sympathectomized cats, the fine afferent innervation of the knee joint capsule was studied via ultrastructural three-dimensional reconstructions over distances of up to 300 microns. Small peripheral nerves and "free" (noncorpuscular) sensory nerve endings were found in a superficial layer of the outer fibrous part of the capsule, in the patellar retinaculum, and in the outer and inner surface layers of the medial collateral and patellar ligaments. Group III nerve fibers showed a proximal myelinated portion inside the nerve, an intermediate portion that lacks a myelin sheath and is only surrounded by perineurium, and a distal portion outside of the perineurium that forms the sensory ending proper. Group IV fibers showed only two distinct portions, an intraperineurial (proximal) and an extraperineurial (distal) portion without any further morphological differences. Outside of the perineurium, a network formed by Schwann cells ("Schwann cell reticulum") provides a pathway for the distal portion of the sensory axons. No distinct subgroups of the sensory terminal fibers could be defined according to the configuration of the Schwann cells and the nerve fiber terminals. Sensory terminals were located adjacent to different structures such as venous and lymphatic vessels, fat cells, and collagenous fibers. Distinct parts of the same terminal nerve fiber were found in close contact to a vessel wall; others were surrounded by dense collagenous tissue. Close to sensory endings, mast cells and mast cell-like cells were frequently found, indicating a functional relationship.

Hypoglossal and reticular interneurons involved in oro-facial coordination in the rat.

Chewing, swallowing, breathing, and vocalization in mammals require precise coordination of tongue movements with concomitant activities of the mimetic muscles. The neuroanatomic basis for this oro-facial coordination is not yet fully understood. After the stereotaxic microinjection of retrograde and anterograde neuronal tracers (biotin-dextran, Fluoro-Ruby, Fluoro-Emerald, and Fluoro-Gold) into the facial and hypoglossal nuclei of the rat, we report here a direct bilateral projection of hypoglossal internuclear interneurons onto facial motoneurons. We also confirm the existence of a small pool of neurons in the dorsal part of the brainstem reticular formation that project ipsilaterally to both facial and hypoglossal nuclei. For precise tracer injections, both motor nuclei were located and identified by the electrical antidromic activation of their constituent motoneurons. Injections of retrograde tracers into the facial nucleus consistently labeled neurons in the hypoglossal nucleus. These neurons prevalently lay in the ipsilateral side, were small in size, and, like classic intrinsic hypoglossal local-circuit interneurons, had several thin dendrites. Reverse experiments - injections of anterograde tracers into the hypoglossal nucleus - labeled fine varicose nerve fiber terminals in the facial nucleus. These fiber terminals were concentrated in the intermediate subdivision of the facial nucleus, with a strong ipsilateral prevalence. Double injections of different tracers into the facial and the hypoglossal nuclei revealed a small, but constant, number of double-labeled neurons located predominantly ipsilateral in the caudal brainstem reticular formation. Hypoglossal internuclear interneurons projecting to the facial nucleus, as well as those neurons of the parvocellular reticular formation that project to both facial and hypoglossal nuclei, could be involved in oro-facial coordination.

Exogenous antigen containing perivascular phagocytes induce a non-encephalitogenic extravasation of primed lymphocytes.

Recent evidence suggests that T-lymphocyte extravasation and CNS-parenchymal infiltration during autoimmune disease might be regulated by antigen-presenting (ED2(+)) cerebral/spinal perivascular phagocytes (CPP/SPP). Since the massive erythrocytic and leukocytic infiltrates in the CNS of rats with experimental allergic encephalomyelitis do not allow a precise differentiation between CPP/SPP and the invading cells in the Virchow-Robin space, we developed a new immune-response model whereby the extravasation of T-lymphocytes was not followed by other blood cells. Adult Lewis rats were sensitized to horseradish peroxidase (HRP). Subsequent intracerebroventricular (i.c.v.) injections of HRP and/or Fluoro-Emerald (FE) served to: (1) challenge the primed T-lymphocytes and (2) label the CPP/SPP for additional immunocytochemical analysis. We found that 24 h and 3 days after single, double, or triple antigen boosting T-lymphocytes (R73(+), W3/25(+), OX50(+)) entered the Virchow-Robin space but did not break through the astrocytic glia limitans. Instead they adhered to HRP-containing activated CPP/SPP (mabs OX-6(+), SILK6(+), CD40(+), CD80(+), CD86(+)). This selective contact was mediated neither by cell adhesion molecules (P-selectin, ICAM-1, VCAM-1), nor promoted by chemokine receptors (CCR1, CCR5) or chemokines (monocyte chemoattractant protein (MCP)-1, MIP-1alpha, MIP-1beta, RANTES). This non-inflammatory, but antigen-dependent lymphocyte extravasation provides optimal conditions to further study the CNS immune response.

Changes in eye blink responses following hypoglossal-facial anastomosis in the cat: evidence of adult mammal motoneuron unadaptability to new motor tasks.

Hypoglossal-facial anastomosis is used in humans to restore the activity of the mimic musculature following irrecoverable facial nerve lesions. As eyelid movement kinetics is very well known, we have used this experimental model in cats to follow the evolution of blink responses and the adaptability of hypoglossal motor pools to new motor tasks. Although the electromyographic activity of the orbicularis oculi muscle in response to corneal air puffs, flashes of light or electrical stimulation of the supraorbital nerve was not recovered in the seven months following this crossed anastomosis, reflex blinks were got back by the increased activity of the retractor bulbi and extraocular recti muscles. The lid of the anastomosed side oscillated in perfect synchronization with tongue movements during licking, while it was severely affected in its motor function during optokinetic stimulation because of the spontaneous appearance of tongue-related hypoglossal activity. Present results suggest that adult mammal motoneurons are unable to readapt their motor programs to the kinetic needs of new motor targets and that most of the functional recovery observed in the cat was achieved by the compensatory hyperactivity of motor systems not directly affected by the surgery.