Effect of nerve repair after unilateral partial facial paralysis on craniofacial growth and development

The effect of partial transection, coaptation, and freezing of the left facial nerve on craniofacial growth and development was investigated. Twelve-day-old New Zealand White rabbits were randomly assigned to four groups: control group (n = 15), nerve ablation group (n = 15) with a surgically induced partial facial paralysis, nerve coaptation group (n = 15) with a surgically induced partial facial paralysis treated by immediate microsurgical nerve coaptation, and nerve freezing group (n = 13) with a freezing-induced partial facial paralysis. All animals were operated on at the age of 12 days; follow-up evaluations were performed at the ages of 2 months (end-point prepubertal growth) and 6 months (full grown). Computerized dorsoventral roentgencephalometric analysis and computer tomographic three-dimensional volumetric measurements were performed at both ages. Roentgencephalometric analysis revealed that the growth disturbances of the nerve ablation group and the nerve coaptation group were analogous. In contrast, in the nerve freezing group, hardly any growth disturbances as compared with the control group were seen. The CT volume measurements revealed a significant left-right difference in maxillary volume at the ages of 2 and 6 months in the nerve coaptation group as compared with the control group. Muscle histomorphometric analysis revealed a shift in muscle fiber composition in the nerve coaptation group and the nerve freezing group, with an increase of type I fibers at the cost of type IIA fibers. This shift was significantly less pronounced in the latter than in the former. Nerve histomorphometric analysis revealed a significant increase in the number of nerve fibers in the nerve coaptation group as compared with the control group. In the nerve freezing group, the increase in the number of nerve fibers was not significantly different as compared with the control group and the nerve coaptation group. Both the equivalent diameter and the myelin area were equally reduced in the nerve coaptation and nerve freezing groups. Thus, the nerve coaptation group and the nerve freezing group did not differ significantly in the extent of nerve recovery, although they differed in the extent of muscle recovery. The extent of muscle recovery, in turn, was related to the extent of abnormal craniofacial growth and development. Indeed, the growth and development were hardly abnormal in the nerve freezing group and as abnormal as after untreated paralysis in the nerve coaptation group. Therefore, factors related to nerve regeneration, other than those assessed by nerve histomorphology, were considered to be responsible for these differences between both groups. The duration of the denervation time was regarded to be considerably shorter in the nerve freezing group than in the nerve coaptation group, resulting in the observed improved muscle recovery. The difference in the degree of axonal malalignment between both groups was considered to be negligible, because of the tolerance toward axonal malalignment at neonatal age.     

Diaphragm muscle fiber function and structure in humans with hemidiaphragm paralysis

Recent studies proposed that mechanical inactivity of the human diaphragm during mechanical ventilation rapidly causes diaphragm atrophy and weakness. However, conclusive evidence for the notion that diaphragm weakness is a direct consequence of mechanical inactivity is lacking. To study the effect of hemidiaphragm paralysis on diaphragm muscle fiber function and structure in humans, biopsies were obtained from the paralyzed hemidiaphragm in eight patients with hemidiaphragm paralysis. All patients had unilateral paralysis of known duration, caused by en bloc resection of the phrenic nerve with a tumor. Furthermore, diaphragm biopsies were obtained from three control subjects. The contractile performance of demembranated muscle fibers was determined, as well as fiber ultrastructure and morphology. Finally, expression of E3 ligases and proteasome activity was determined to evaluate activation of the ubiquitin-proteasome pathway. The force-generating capacity, as well as myofibrillar ultrastructure, of diaphragm muscle fibers was preserved up to 8 wk of paralysis. The cross-sectional area of slow fibers was reduced after 2 wk of paralysis; that of fast fibers was preserved up to 8 wk. The expression of the E3 ligases MAFbx and MuRF-1 and proteasome activity was not significantly upregulated in diaphragm fibers following paralysis, not even after 72 and 88 wk of paralysis, at which time marked atrophy of slow and fast diaphragm fibers had occurred. Diaphragm muscle fiber atrophy and weakness following hemidiaphragm paralysis develops slowly and takes months to occur.     

Nerve and muscle development in paralysé mutant mice

Nerve and muscle development was studied in paralysé mutant mice. The mutant phenotype is first recognizable 6-7 days after birth (PN 6-PN 7) as cessation of muscle growth and weakness and incoordination of movement. Mutant animals die between 2 and 3 weeks of age. Muscle fibers from paralysé mutants had a unimodal distribution of diameters and normal numbers and distributions of acetylcholine receptors. The only structural abnormality seen was a reduced extracellular space within muscle fascicles. Total muscle choline acetyltransferase activity was reduced compared with that of control muscles, indicating that synaptic terminal development was impaired. Light and electron microscopy showed that polyneuronal innervation was retained in mutant endplates, and the normal process of withdrawal of redundant innervation did not occur. The paralysé muscles reacted to experimental denervation with an increase in extrajunctional acetylcholine receptor numbers. Intramuscular axons failed to become myelinated in mutant animals, although sciatic nerve axons were myelinated with a normal myelin thickness/axon diameter ratio. Nodes of Ranvier were elongated and myelin lamellae in the paranodal regions were poorly fused. Sciatic nerves in mutant animals retained the neonatal unimodal distribution of axon diameters, whereas in control animals it became bimodal by 2 weeks of age. Our results are not consistent with a previous suggestion that paralysé mutant muscle endplates are progressively denervated. We conclude that the major expression of the paralysé mutant phenotype is an arrest in development of both nerve and muscle during the first week after birth. The paralysé mutant gene most likely is involved in the general support of development of many or all body tissues from 1 week of age. We found no regression of any aspect of differentiation, once achieved.     

The microscopic morphology of orthotopic free muscle grafts in rabbits

The flexor digitorum superficialis muscle was free grafted (without neurovascular anastomoses) in 122 rabbit forelimbs. Histologic nd histochemical examinations through 6 months after grafting were performed. An early ischemic necrosis of the entire graft, except for a few percent of fibers at the very surface, was consistently seen. Subsequently, there occurred a regeneration of muscle with reconstitution of up to 100 percent of normal numbers of fibers. There was a wide variation in the numbers of fibers regenerated; however, the fiber-free areas were then being replaced by connective tissue. Muscle grafts in 1-month-old rabbits regenerated faster and yielded muscle with evidence of more extensive reinnervation and less connective tissue than 3-month-old animals. In the early postgraft period, minced grafts appeared to be as good as whole ones, but after 1 month, they developed far more connective tissue. Differentiation into fast-twitch and slow-twitch muscle fibers and into high- and low-oxidative fibers began at 2 to 3 weeks after grafting but was not extensive until 3 months. At 6 months, grafts showed areas of normal-appearing muscle interspersed with areas that lacked signs of reinnervation. The earliest sign of regeneration is the appearance of several very elongated nuclei encircling each previously anucleate necrotic muscle fiber. A small amount of basophilic cytoplasm then appears around each new nucleus. As blood vessels grow into the graft, a centripetal wave of phagocytosis is seen, taking 2 to 3 weeks and leaving a bed of immature muscle fibers. We believe this to be the first documentation of regeneration's commencing prior to and thus independently of phagocytosis.     

Ipsilateral and cross-over elongation of the motor nerve by nerve grafting: an experimental study in sheep

In difficult reconstructions, ipsilateral or cross-over nerve grafting is sometimes necessary to achieve reinnervation and motor function. This experimental study in sheep was to answer the question of limitation of elongation of a motor nerve by grafting, the question of the optimal time for suturing the nerve graft to the muscle nerve, and the question of the successful application of this surgical technique in extremities. In 18 sheep, the vastus nerve was elongated by a saphenous nerve graft as long as possible up to 30 cm (step 1). In 10 animals the nerve graft was applied ipsilaterally, and in 8 animals it was used as a cross-over nerve graft to the contralateral limb. The time between nerve grafting and connection of the distal end of the nerve graft to the freshly cut rectus nerve supplying the rectus femoris muscle (step 2) was variable: 0, 3, 6, 9, and 12 months. In all animals, the final experiments (step 3) were performed 6 months after the last operation (step 2). Muscle force measurements in the rectus femoris muscle and quantitative analysis of the number and diameter of myelinated nerve fibers in cross sections of the nerve biopsies at different levels showed that elongation of a motor nerve by nerve grafting is principally not limited. The functional results were rather inhomogeneous and therefore unpredictable (ipsilateral group: maximum tetanic tension = 27 to 172 N; cross-over group: 0 to 227.5 N). Nevertheless, crossover nerve grafting is recommended for selected cases even in extremities. There was no correlation between the time interval between the two operations and the functional or morphologic results, although better functional results were obtained when the distal nerve suture (step 2) was performed some months after nerve grafting (step 1). A clear correlation was found only between the number of regenerated axons in the rectus nerve behind the second suture line and the muscle function.     

Origin of galanin in nerves of cat airways and colocalization with vasoactive intestinal peptide

Galanin is a 29 amino acid residue neuropeptide. In mammalian airways, galanin is found in nerve fibers associated with airway smooth muscle, bronchial glands, and blood vessels, and in nerve cell bodies of airway ganglia. The present study was conducted to determine if galanin-containing fibers in the walls of feline airways originate from the nerve cell bodies of airway ganglia. The colocalization of galanin with vasoactive intestinal peptide was also investigated. Organotypic cultures of cat airways were held in culture for 0 (nonculture control), 3, 5, and 7 days. After each culture period, the distribution of galanin and the colocalization of galanin with vasoactive intestinal peptide were determined by immunocytochemistry. Galanin-containing fibers were found in bronchial smooth muscle, around bronchial glands and in the walls of bronchial arteries and arterioles throughout the culture period. Nerve fibers and cell bodies containing both galanin and vasoactive intestinal peptide were observed after all culture periods. Nerve fibers and cells bodies that contained galanin frequently contained vasoactive intestinal peptide as well, but nerve fibers with only galanin or vasoactive intestinal were also observed. Galanin- and vasoactive intestinal peptide-containing nerve fibers and cell bodies were both well maintained throughout the culture period. The findings show that galanin-containing nerve fibers associated with bronchial smooth muscle, bronchial glands, and bronchial arteries, originate from nerve cell bodies of intrinsic airway ganglia, and that galanin and vasoactive intestinal peptide are frequently colocalized in these neurons.     

Dynamic reconstruction of eye closure by muscle transposition or functional muscle transplantation in facial palsy

For patients with facial palsy, lagophthalmus is often a more serious problem than the inability to smile. Dynamic reconstruction of eye closure by muscle transposition or by free functional muscle transplantation offers a good solution for regaining near-normal eye protection without the need for implants. This is the first quantitative study of three-dimensional preoperative and postoperative lid movements in patients treated for facial paralysis. Between February of 1998 and April of 2002, 44 patients were treated for facial palsy, including reconstruction of eye closure. Temporalis muscle transposition to the eye was used in 34 cases, and a regionally differentiated part of a free gracilis muscle transplant after double cross-face nerve grafting was used in 10 cases. Patients' facial movements were documented by a three-dimensional video analysis system preoperatively and 6, 12, 18, and 24 months postoperatively. For this comparative study, only the data of patients with preoperative and 12-month postoperative measurements were included. In the 27 patients with a final result after temporalis muscle transposition for eye closure, the distance between the upper and lower eyelid points during eye closing (as for sleep) was reduced from 10.33 +/- 2.43 mm (mean +/- SD) preoperatively to 5.84 +/- 4.34 mm postoperatively on the paralyzed side, compared with 0.0 +/- 0.0 mm preoperatively and postoperatively on the contralateral healthy side. In the resting position, preoperative values for the paralyzed side changed from 15.11 +/- 1.92 mm preoperatively to 13.46 +/- 1.94 mm postoperatively, compared with 12.17 +/- 2.02 mm preoperatively and 12.05 +/- 1.95 mm postoperatively on the healthy side. In the nine patients with a final result after surgery using a part of the free gracilis muscle transplant reinnervated by a zygomatic branch of the contralateral healthy side through a cross-face nerve graft, eyelid closure changed from 10.21 +/- 2.72 mm to 1.68 +/- 1.35 mm, compared with 13.70 +/- 1.56 mm to 6.63 +/- 1.51 mm preoperatively. The average closure for the healthy side was from 11.20 +/- 3.11 mm to 0.0 +/- 0.0 mm preoperatively and from 12.70 +/- 1.95 mm to 0.0 +/- 0.0 mm postoperatively. In three cases, the resting tonus of the part of the gracilis muscle transplant around the eye had increased to an extent that muscle weakening became necessary. Temporalis muscle transposition and free functional muscle transplantation for reanimation of the eye and mouth at the same time are reliable methods for reconstructing eye closure, with clinically adequate results. Detailed analysis of the resulting facial movements led to an important improvement of the authors' operative techniques within the last few years. Thus, the number of secondary operative corrections could be significantly reduced. These qualitative and quantitative studies of the reconstructed lid movements by three-dimensional video analysis support the authors' clinical concept of temporalis muscle transposition being the first-choice method in adult patients with facial palsy. In children, free muscle transplantation is preferred for eye closure, so as not to interfere with the growth of the face by transposition of a masticatory muscle. In addition, a higher degree of central plasticity in children might be expected.     

Innervation of the anococcygeus muscle of the rat

Summary The innervation of the anococcygeus muscle of the rat was investigated with regard to the histochemical features of nerve fibers within the muscle and to the location of the postganglionic autonomic neurons which are the source of these fibers. Acetylcholinesterase-positive fibers and catecholaminergic fibers are abundant in the anococcygeus as well as the related retractor penis muscle. Neuronal somata, either between muscle bundles of the anococcygeus or in the connective tissue sheath, are also acetylcholinesterase-positive. Nerve fibers and a minority of the ganglion cells in the anococcygeus and retractor penis muscles are immunoreactive for vasoactive intestinal polypeptide. Injection of the retrogradely transported dye Fluorogold into the anococcygeus muscle filled neurons in the abdominopelvic sympathetic chain, pelvic plexus and a small number of neurons in the inferior mesenteric ganglion. In the pelvic plexus, some neurons were located in the major pelvic ganglion but most were found along the main penile nerve and its branches to the anococcygeus muscle. Immunocytochemistry of these identified neurons indicates that about one half of them are positive for vasoactive intestinal polypeptice. These results raise the possibility that both acetylcholine and vasoactive intestinal polypeptide are important neurotransmitters in autonomic nerves to the anococcygeus muscle.     

Long-term remodeling of vascularized and nonvascularized onlay bone grafts: a macroscopic and microscopic analysis

The present study was performed to compare vascularized and nonvascularized onlay bone grafts to investigate the potential effect of graft-to-recipient bed orientation on long-term bone remodeling and changes in thickness and microarchitectural patterns of remodeling within the bone grafts. In two groups of 10 rabbits each, bone grafts were raised bilaterally from the supraorbital processes and placed subperiosteally on the zygomatic arch. The bone grafts were oriented parallel to the zygomatic arch on one side and perpendicular to the arch on the contralateral side. In the first group, vascularized bone grafts were transferred based on the auricularis anterior muscle, and in the second group nonvascularized bone grafts were transferred. Fluorochrome markers were injected during the last 3 months of animal survival, and animals were killed either 6 or 12 months postoperatively. The nonvascularized augmented zygoma showed no significant change in thickness 6 months after bone graft placement and a significant decrease in thickness 1 year after graft placement (p < 0.01). The vascularized augmented zygoma showed a slight but statistically significant decrease in thickness 6 months after graft placement (p < 0.003), with no significant difference relative to its initial thickness 1 year after graft placement. In animals killed 6 months after bone graft placement, both the rate of remodeling and the bone deposition rate measured during the last 3 months of survival were significantly higher in the vascularized bone grafts compared with their nonvascularized counterparts (p < 0.02). By 1 year postoperatively, there were no significant differences in thickness, mineral apposition rate, or osteon density between bone grafts oriented perpendicular and parallel to the zygomatic arch. These findings indicate that the vascularity of a bone graft has a significant effect on long-term thickness and histomorphometric parameters of bone remodeling, whereas the direction of placement of a subperiosteal graft relative to the recipient bed has minimal effect on these parameters. In vascularized bone grafts, both bone remodeling and deposition are accelerated during the initial period following graft placement. Continued bone deposition renders vascularized grafts better suited for the long-term maintenance of thickness and contour relative to nonvascularized grafts.     

Transplantation and transposition of skeletal muscles into the faces of monkeys

Restoration of normal facial movement after long-term facial paralysis with muscle atrophy has not yet been achieved reliably by either free grafts, in which fibers degenerate and regenerate, or by grafts made with microneurovascular repair, in which most fibers survive. Our purpose was to compare the structural and functional properties of free muscle grafts and continuously perfused muscle flaps transplanted into the faces of monkeys. In adult monkeys, the facial muscles were replaced by either a free graft of a donor muscle from the lower limb or a denervated flap of ipsilateral temporalis muscle. Each graft or flap was reinnervated with the preserved buccal branch of the facial nerve. The control muscles, grafts, and flaps were examined 90 days later for gross appearance, contractile properties, and fiber areas. Compared with muscle flaps, free grafts showed greater adaptability to the new location and innervation and a closer approximation to the structural and functional properties of the original facial musculature.     

Prior running reduces hypertrophic growth of skeletal muscle grafts

Run training can increase the mass of soleus muscle grafts, yet values remain lower than nongrafted muscle even with continued training. Thus we tested the hypothesis that nerve-implant soleus grafts of rats previously run trained would be refractory to the hypertrophic stimulus of ablation of synergistic muscle. We also compared the magnitude of growth of the nerve-implant soleus graft after ablation with that reported by others for the nerve-intact soleus graft. We studied eight groups that differed relative to the combination and order of treatments (running and ablation of synergistic muscle) and the graft age at the time of the ablation operation and study. Graft mass, protein concentration, and histochemical fiber composition were measured. Compared with grafts from cage-sedentary rats, the mass and protein content of the nerve-implant soleus grafts were higher (16-63%) at all times after ablation. When the ablation operation was performed at 56 days postgrafting, there was a 33% increase in protein content of the soleus graft by 84 days for cage-sedentary animals. This increase was twofold greater (P less than or equal to 0.02) than the 15% increase that followed ablation for the grafts from the animals that had been run trained before the ablation operation. Four weeks of run training before the ablation operation impaired the adaptive response of muscle grafts to the ablation of synergistic muscles, which may reflect alterations in motor unit recruitment and/or satellite cell activity. Ablation of synergistic muscles resulted in an absolute growth of the nerve-implant soleus grafts that was comparable with that reported for nerve-intact soleus grafts.     

Muscle spindle formation and differentiation in regenerating rat muscle grafts

Muscle spindle development and function are dependent upon sensory innervation. During muscle regeneration, both neural and muscular components of spindles degenerate and it is not known whether reinnervation of a regenerating muscle results in reestablishment of proper neuromuscular relationships within spindles or whether sensory neurons may exert an influence upon differentiation of these spindles. Muscle spindle regeneration was studied in bupivacaine-treated grafts of rat extensor digitorum longus (EDL) muscles. Three types of EDL graft were performed in order to manipulate the extent to which regenerating spindles might be reinnervated: (1) grafts reinnervated following severance of their nerve supply (standard grafts); (2) grafts in which intact nerve sheaths appear to facilitate reinnervation (nerveintact grafts); and (3) grafts in which reinnervation was prevented (nonreinnervated grafts). Complete degeneration of muscle fibers occurred in all grafts prior to regeneration. Initial formation of spindles in regenerating EDL grafts is independent of innervation; intrafusal muscle fibers degenerate and regenerate within spindle capsules that remain intact and viable. The extent of spindle differentiation was evaluated in each type of graft using criteria that included nucleation and ATPase activity, both of which have been shown to be regulated by sensory innervation, as well as the number of muscle fibers/spindle and morphology of spindle capsules.While most spindles contained normal numbers of muscle fibers, most of these fibers were morphologically and histochemically abnormal. Alterations of ATPase activity occurred in all spindles, but were least severe in nerve-intact grafts. While fully differentiated nuclear bag and chain fibers were not observed in regenerated spindles, large, vesicular nuclei, similar to those of normal intrafusal fibers, were present in a small number of spindles in nerve-intact grafts. Sensory nerve terminations were observed only in those spindles that also contained the distinctive nuclei. This study suggests that a specific neurotrophic influence is necessary for regeneration of normal intrafusal muscle fibers and that this influence corresponds to the properly timed sensory neuron-muscle interaction which directs muscle spindle embryogenesis. However, the infrequent occurrence of characteristics unique to intrafusal muscle fibers indicates that reinnervation of regenerating muscle grafts by sensory neurons is inadequate and/or faulty.     

"Donor" muscle structure and function after end-to-side neurorrhaphy

End-to-end nerve coaptation is the preferred surgical technique for peripheral nerve reconstruction after injury or tumor extirpation. However, if the proximal nerve stump is not available for primary repair, then end-to-side neurorrhaphy may be a reasonable alternative. Numerous studies have demonstrated the effectiveness of this technique for muscle reinnervation. However, very little information is available regarding the potential adverse sequelae of end-to-side neurorrhaphy on the innervation and function of muscles innervated by the "donor" nerve. End-to-side neurorrhaphy is hypothesized to (1) acutely produce partial donor muscle denervation and (2) chronically produce no structural or functional deficits in muscles innervated by the donor nerve. Adult Lewis rats were allocated to one of two studies to determine the acute (2 weeks) and chronic (6 months) effects of end-to-side neurorrhaphy on donor muscle structure and function. In the acute study, animals underwent either sham exposure of the peroneal nerve (n = 13) or end-to-side neurorrhaphy between the end of the tibial nerve and the side of the peroneal nerve (n = 7). After a 2-week recovery period, isometric force (F(0) was measured, and specific force (sF(0) was calculated for the extensor digitorum longus muscle ("donor" muscle) for each animal. Immunohistochemical staining for neural cell adhesion molecule (NCAM) was performed to identify populations of denervated muscle fibers. In the chronic study, animals underwent either end-to-side neurorrhaphy between the end of the peroneal nerve and the side of the tibial nerve (n = 6) or sham exposure of the tibial nerve with performance of a peroneal nerve end-to-end nerve coaptation approximately 6), to match the period of anterior compartment muscle denervation in the end-to-side neurorrhaphy group. After a 6-month recovery period, contractile properties of the medial gastrocnemius muscle ("donor" muscle) were measured. Acutely, a fivefold increase in the percentage of denervated muscle fibers (1 +/0 0.7 percent to 5.4 +/-2.7 percent) was identified in the donor muscles of the animals with end-to-side neurorrhaphy (p < 0.001). However, no skeletal muscle force deficits were identified in these donor muscles. Chronically, the contractile properties of the medial gastrocnemius muscles were identical in the sham and end-to-side neurorrhaphy groups. These data support our two hypotheses that end-to-side neurorrhaphy causes acute donor muscle denervation, suggesting that there is physical disruption of axons at the time of nerve coaptation. However, end-to-side neurorrhaphy does not affect the long-term structure or function of muscles innervated by the donor nerve.     

Immunohistochemistry and nerve lesion experiments on the methionine-enkephalin immunopositive neurons in the small intestine of the bullfrog (Rana catesbeiana)

Summary Nerve elements in the small intestine of the bullfrog. Rana catesbeiana, were studied by immunohistochemistry with anti-methionine enkephalin antisera and by nerve lesion experiments, using laser irradiation. Methionine-enkephalin immunopositive nerve fibers occur in the myenteric plexus, circular muscle layer, submucosa, and mucosa. Immunopositive nerve cell bodies in the myenteric plexus have dendrite-like and a long axon-like processes. In the froglet (3 months after metamorphosis), these axon-like processes lead posteriorly in the nerve strand of the myenteric plexus. Some bifurcate, one branch continuing posteriorly, the other doubling back to lead anteriorly; both form terminal varicose fibers in the circular muscle layer. Nerve lesion experiments, in the adult bullfrog, resulted in accumulations of methionine-enkephalin immunoreactivity at the oral and hinder edges of the laser-irradiated necrotic area; there were sprouting and nonsprouting immunopositive stumps. It is suggested that bidirectional flow of methionine-enkephalin in the myenteric plexus is mediated via the anterior and posterior branches of the axon-like process. The difference in sprouting behavior of immunopositive nerve fiber stumps, after nerve lesion, is discussed with reference to regional differences of the axon-like process.     

Tissue engineering of peripheral nerves: A comparison of venous and acellular muscle grafts with cultured Schwann cells

Bioengineering is considered to be the laboratory-based alternative to human autografts and allografts. It ought to provide "custom-made organs" cultured from patient's material. Venous grafts and acellular muscle grafts support axonal regeneration only to a certain extent because of the lack of viable Schwann cells in the graft. We created a biologic nerve graft in the rat sciatic nerve model by implanting cultured Schwann cells into veins and acellular gracilis muscles, respectively. Autologous nerve grafts and veins and acellular muscle grafts without Schwann cells served as controls. After 6 and 12 weeks, regeneration was assessed clinically, histologically, and morphometrically. The polymerase chain reaction analvsis showed that the implanted Schwann cells remained within all the grafts. The best regeneration was seen in the control; after 12 weeks the number of axons was increased significantly compared with the other grafts. A good regeneration was noted in the muscle-Schwann cell group, whereas regeneration in both of the venous grafts and the muscle grafts without Schwann cells was impaired. The muscle-Schwann cell graft showed a systematic and organized regeneration including a proper orientation of regenerated fibers. The venous grafts with Schwann cells showed less fibrous tissue and disorganization than the veins without Schwann cells, but failed to show an excellent regeneration. This might be attributed to the lack of endoneural-tube-like components serving as scaffold for the sprouting axon. Although the conventional nerve graft remains the gold standard, the implantation of Schwann cells into an acellular muscle provides a biologic graft with basal lamina tubes as pathways for regenerating axons and the positive effects of Schwann cells producing neurotrophic and neurotropic factors, and thus, supporting axonal regeneration.     

Sensory restoration of the skin graft on a free muscle flap: experimental rabbit study.

Transplantation of a muscle flap with free skin graft for wound coverage is a common procedure in reconstructive microsurgery. However, the grafted skin has little or no sensation. Restoration of the sensibility of the grafted skin on the transferred muscle is critically important, especially in palmar hand, plantar foot, heel, and oral cavity reconstruction. The purpose of this study was to investigate the possibility of sensory restoration of the grafted skin on a trimmed muscle surface that has been sensory neurotized after sensory nerve-to-motor nerve transfer, using the rabbit gracilis muscle as an animal model. The ipsilateral saphenous nerve (sensory) was transferred to the motor nerve of the gracilis muscle for sensory neurotization. A 4 x 4-cm2 area of skin island over the midportion of the gracilis muscle was harvested as a full-thickness skin graft. The upper half of the gracilis muscle was then excised, becoming a rough surface. The harvested skin was reapplied on the trimmed rough surface of the muscle. After 6 months, retrograde and antegrade horseradish peroxidase labeling studies were performed through skin and muscle injection. The group with a free skin graft was compared with the group with an intact surface of the gracilis muscle. This study clearly shows that sensory nerves can regenerate and penetrate into the trimmed muscle surface and grow into the overlying grafted skin. However, if the muscle surface is intact as with the compared group, sensory reinnervation of the grafted skin is not possible.     

Effects of denervation and colchicine treatment on the chloride conductance of rat skeletal muscle fibers.

Membrane potentials, cable parameters, and component resting conductances were measured in extensor digitorum longus (EDL) muscle fibers from adult rats in vitro at 24 degrees C, after 15 to 18 days of denervation by nerve section, and at seven to ten days following epineural injection of 100 to 450 mug of colchicine in the peroneal nerve. The denervated muscles were paralyzed throughout the experimental period, whereas the colchicine-treated preparations showed no clinical paralysis except for the first day or two. The EDL from the untreated side served as a control. Both the denervated and colchicine-treated fibers were depolarized, showed signs of fibrillation, had tetrodotoxin-resistant potentials, and membrane resistance was increased two- to sevenfold. In the denervated fibers, mean chloride conductance GC1 dropped from a control value of 3196 to 596 mumhos/cm2 while mean potassium conductance GK showed a tendency to rise from 260 to 332 muhos/cm2. Colchicine-treated fibers while showing a similar fall in mean GC1 from 2993 to 1066 mumhos/cm2, also showed a significant fall in mean GK from 213 to 116 mumhos/cm2. It was concluded that factors transported by the microtubular system are important for the maintenance of the high resting GC1 of mammalian skeletal muscle fibers.     

An analysis of the interactions between sympathetic nerve fibers and smooth muscle cells in tissue culture

The interactions between sympathetic nerve fibers and smooth muscle cells and fibroblasts from the newborn guinea pig vas deferens were studied in tissue culture with phase contrast microscopy, time-lapse microcinematography, catecholamine fluorescence histochemistry and scanning and transmission electron microscopy. The amount of sympathetic nerve fiber growth, its catecholamine fluorescence reaction and the size of the nerve cell bodies and their nuclei all increased in the presence of vas deferens tissue. Specific growth of nerve fibers to large clumps of vas deferens tissue was seen from distances of up to 2 mm. In contrast, no specific growth from a distance occurred to single cells or small groups of cells. However, random contact with a muscle cell often led to close, extensive, and long-lasting associations. Contact with fibroblasts was always transitory.The rate of sympathetic nerve fiber growth over individual muscle cells was faster than over fibroblasts, which, in turn, was faster than over the collagen-coated surface of the coverslip. Palpation of a muscle cell by a nerve fiber growth cone increased the rate of spontaneous contraction of the muscle cell, the extent of the increase being dependent on the number of nerve fibers involved. Multiple innervation of a smooth muscle cell occurred if nerve fibers reached the cell at about the same time, but not if there was a close association already established. These results are discussed in relation to possible interactions of sympathetic nerve fibers with smooth muscle cells in vivo.     

Clinical application of peripheral nerve transplantation.

Surgical reconstruction of extensive peripheral nerve injuries frequently exhausts the patient's own source of expendable autogenous nerve grafts. Nerve allografts would offer a limitless supply of graft material. A 23-cm, 10-cable sciatic nerve allograft was performed in an 8-year-old boy in September of 1988. The patient was managed with Cyclosporin A for 2 years. Forty-four months after the transplant surgery and 19 months after the cessation of Cyclosporin A therapy, the patient has evidence of nerve regeneration across the allograft with recovery of functional sensibility in his foot. In the selected patient with an otherwise irreparable nerve injury, consideration can be given to the use of a nerve allograft.     

Mandibular condylar growth alterations after unilateral partial facial paralysis: an experimental study in the rabbit.

In a previous study in the rabbit, the authors defined the macroscopic growth alterations after unilateral partial facial paralysis. Dry skull measurements revealed a reduced premaxillary, maxillary, mandibular, and anterior corpus length with a simultaneous increase in mandibular ramal height on the paralyzed side. The authors hypothesize that these mandibular growth alterations are, among others, caused by alterations in condylar growth activity and that an altered occlusal relationship may be involved in the adaptive condylar growth response after facial paralysis.A total of 84 New Zealand White rabbits were used for this study. The animals were randomly assigned to either a control group that was not operated on (n = 28), a group that underwent a sham-operation (n = 28), or an experimental group (n = 28). In the sham-operation group, the facial nerve was dissected as in the experimental group but was left intact. In the experimental group, a left-side partial facial paralysis involving the midfacial muscles was induced by an operation at the age of 12 days. After different follow-up time intervals of 3.5, 7, 14, 21, 28, 42, and 56 days, four control, four sham-operation, and four experimental animals (all randomly selected) were killed for histomorphometric measurements of the left control and sham condyles and the left-side and right-side experimental condyles.No significant differences between the control and sham-operation groups were found. The other results revealed that shortly after the paralysis in the experimental group, as compared with the controls, a decrease in condylar growth activity was seen before a catch-up increase in activity, as expressed by the time-sequenced decrease and increase in the height of the functional and hypertrophic chondroblast layer. The response on the right side was analogous, though less intense.It is suggested that the mandibular ramal growth alterations might be the result of a chain of adaptations involving the lateral pterygoid muscle and the condylar growth activity. The unilaterally restricted length increment of the maxillary snout, as a result of the loss of tensile forces caused by paralysis of the midfacial musculature, necessitated an adaptation in the position of the mandible to maintain a normal occlusal relationship. Subsequently, the function of muscles involved or influenced by an altered mandibular position, such as the lateral pterygoid muscle, were changed. These altered muscle activities induced condylar growth adaptations, which in turn explained the alterations in mandibular ramal growth.