Liron S. Duraku

Re-innervation patterns by peptidergic Substance-P, non-peptidergic P2X3, and myelinated NF-200 nerve fibers in epidermis and dermis of rats with neuropathic pain.

Nerve endings in the epidermis, termed nociceptors, conduct information on noxious stimuli to the central nervous system. The precise role of epidermal nerve fibers in neuropathic pain is however still controversial. Here, we have investigated the re-innervation patterns of epidermal and dermal nerve fibers in a rat neuropathic pain model. After applying the spared nerve injury (SNI) model, we determined the mechanical and thermal withdrawal thresholds in the uninjured lateral (sural) and medial (saphenous) areas of the affected hind paw and investigated the innervations patterns of Substance P (SubP), Neurofilament-200 (NF-200) and P2X3-immunoreactive (IR) nerve fibers in the epidermis and dermis. We found a significant loss in the density of peptidergic (Sub P and NF-200) and non-peptidergic (P2X3) nerve fibers in the center area of the foot sole at 2 weeks postoperatively (PO). The densities of Sub P-IR fibers in the epidermis and upper dermis, and the density of P2X3-IR fibers in the upper dermis were significantly increased at 10 weeks PO as compared to 2 weeks PO, but were still significantly lower than the densities in controls. However, the density of NF-200-IR fibers in the center area reached control levels at 10 weeks PO. No changes were found in the densities of any of the fibers in the medial and lateral parts of the foot sole. The present results suggest that after peripheral nerve injury, specific nerve fibers have different re-innervation patterns in the epidermis and dermis and that they might be involved in the development of neuropathic pain.

A systematic review on the sensory reinnervation of free flaps for tongue reconstruction: Does improved sensibility imply functional benefits?

BACKGROUND Tongue reconstruction after (hemi)glossectomy including sensory recovery is challenging. Although sensory recovery could improve functional outcome, no consensus on the need for reinnervation of the neo-tongue exists. Therefore, a systematic review was performed to determine if sensory reinnervation of free flaps in tongue reconstruction is better than no sensory reinnervation. The secondary study aim was to assess the effect of sensory reinnervation on overall functional outcome, such as speech and deglutition. METHODS Seven databases (Embase, Medline, Web of Science, Scopus, PubMed publisher, Cochrane, and Google Scholar) were searched. Studies that reported the effect of sensory reinnervation on overall functional outcome were identified. RESULTS Fourteen articles were included in the systematic review, concerning a total of 271 tongue reconstructions. Free flaps that were used were the radial forearm (RF) flap (n = 137), the anterolateral thigh (ALT) flap (n = 65), the rectus abdominis (RA) flap (n = 20), and the tensor fascia latae (TFL) flap (n = 5). Seven out of seven articles directly comparing sensory reinnervation with no sensory reinnervation revealed superior sensibility in the reinnervated group. Moreover, the innervated RF and ALT flaps showed superior recovery of sensibility compared to other flaps used for the reconstruction of hemiglossectomy as well as total glossectomy defects. There are indications that sensory reinnervation may have a beneficial effect on overall tongue function. Age, smoking, and sex did not affect sensory recovery. Four out of five articles showed that postoperative radiotherapy does not have a long-term adverse effect on sensory recovery. CONCLUSIONS Sensory reinnervation of free flaps in the reconstruction of (hemi)glossectomy defects improves sensory recovery; however, evidence for beneficial effects on function is poor.

Thermo-sensitive TRP channels in peripheral nerve injury: A review of their role in cold intolerance

One of the sensory complications of traumatic peripheral nerve injury is thermal intolerance, which manifests in humans mainly as cold intolerance. It has a major effect on the quality of life, and adequate therapy is not yet available. In order to better understand the pathophysiological background of thermal intolerance, we focus first on the various transient receptor potential (TRP) channels that are involved in temperature sensation, including their presence in peripheral nerves and in keratinocytes. Second, the role of thermo-sensitive TRP channels in cold and heat intolerance is described showing three different mechanisms that contribute to thermal intolerance in the skin: (a) an increased expression of TRP channels on nerve fibres and on keratinocytes, (b) a lower activation threshold of TRP channels and (c) the sprouting of non-injured nerve fibres. Finally, the data that are available on the effects of TRP channel agonists and antagonists and their clinical use are discussed. In conclusion, TRP channels play a major role in temperature sensation and in cold and heat intolerance. Unfortunately, the available pharmaceutical agents that successfully target TRP channels and counteract thermal intolerance are still very limited. Yet, our focus should remain on TRP channels since it is difficult to imagine a reliable treatment for thermal intolerance that will not involve TRP channels.

Thermoregulation in peripheral nerve injury-induced cold-intolerant rats

PURPOSE Cold intolerance is defined as pain after exposure to non-painful cold. It is suggested that cold intolerance may be related to dysfunctional thermoregulation in upper extremity nerve injury patients. The purpose of this study was to examine if the re-warming of a rat hind paw is altered in different peripheral nerve injury models and if these patterns are related to severity of cold intolerance. METHODS In the spared nerve injury (SNI) and complete sciatic lesion (CSL) model, the re-warming patterns after cold stress exposure were investigated preoperatively and at 3, 6 and 9 weeks postoperatively with a device to induce cooling of the hind paws. Thermocouples were attached on the dorsal side of the hind paw to monitor re-warming patterns. RESULTS The Von Frey test and cold plate test indicated a significantly lower paw-withdrawal threshold and latency in the SNI compared to the Sham model. The CSL group, however, had only significantly lower paw-withdrawal latency on the cold plate test compared to the Sham group. While we found no significantly different re-warming patterns in the SNI and CSL group compared to Sham group, we did find a tendency in temperature increase in the CSL group 3 weeks postoperatively. CONCLUSION Overall, our findings indicate that re-warming patterns are not altered after peripheral nerve injury in these rat models despite the fact that these animals did develop cold intolerance. This suggests that disturbed thermoregulation may not be the prime mechanism for cold intolerance and that, other, most likely, neurological mechanisms may play a more important role. CLINICAL RELEVANCE There is no direct correlation between cold intolerance and re-warming patterns in different peripheral nerve injury rat models. This is an important finding for future developing treatments for this common problem, since treatment focussing on vaso-regulation may not help diminish symptoms of cold-intolerant patients.

Differential Changes in the Peptidergic and the Non-Peptidergic Skin Innervation in Rat Models for Inflammation, Dry Skin Itch, and Dermatitis

Skin innervation is a dynamic process that may lead to changes in nerve fiber density during pathological conditions. We have investigated changes in epidermal nerve fiber density in three different rat models that selectively produce chronic itch (the dry skin model), or itch and inflammation (the dermatitis model), or chronic inflammation without itch (the CFA model). In the epidermis, we identified peptidergic fibers-that is, immunoreactive (IR) for calcitonin gene-related peptide or substance P—and non-peptidergic fibers—that is, IR for P2X3. The overall density of nerve fibers was determined using IR for the protein gene product 9.5. In all three models, the density of epidermal peptidergic nerve fibers increased up to five times when compared with a sham-treated control group. In contrast, the density of epidermal non-peptidergic fibers was not increased, except for a small but significant increase in the dry skin model. Chronic inflammation showed an increased density of peptidergic fibers without itch, indicating that increased nerve fiber density is not invariably associated with itch. The finding that different types of skin pathology induced differential changes in nerve fiber density may be used as a diagnostic tool in humans, through skin biopsies, to identify different types of pathology and to monitor the effect of therapies.

Rotterdam Advanced Multiple Plate: A novel method to measure cold hyperalgesia and allodynia in freely behaving rodents

BACKGROUND To investigate the pathophysiology of temperature hypersensitivity in neuropathic pain rodent models, it is essential to be able to quantify the phenotype as objective as possible. Current temperature sensitivity measuring paradigms are performed during exposure to external factors, i.e. light, sound and smell, which modulate behavior significantly. In addition the present outcome measure for temperature hypersensitivity in rodents is the examination of the hind paw lift upon exposure to a certain temperature, which reflects more a reflex-flexion than an experience of pain. NEW METHOD Therefore the Rotterdam Advanced Multiple Plate (RAMP) was developed to assess cold hyperalgesia and allodynia objectively in freely behaving neuropathic pain rats, which measures the avoidance for certain temperatures and monitoring the location of the rat with an infrared camera while excluding external environmental influences such as light and sound. RESULTS Compared to sham rats, the spared nerve injury (SNI) rats demonstrated a higher preference for the comfortable plate (27 °C) when the other three plates were set at 5 °C, 14 °C, 17 °C and 19 °C. We were unable to detect heat hyperalgesia and allodynia with the RAMP. COMPARISON WITH EXISTING METHOD The paw withdrawal method displays similar results during cold hypersensitivity measurements as observed with the RAMP. The SNI group did display heat hypersensitivity during the paw withdrawal test. CONCLUSIONS The results indicate that the RAMP is able to quantify cold hyperalgesia and allodynia in neuropathic pain rats while resolves some of the problems of conventional temperature sensitivity measuring paradigms in rodents.

Mirror-image pain after nerve reconstruction in rats is related to enhanced density of epidermal peptidergic nerve fibers.

Mirror-image pain is a phenomenon in which unprovoked pain is detected on the uninjured contralateral side after unilateral nerve injury. Although it has been implicated that enhanced production of nerve growth factor (NGF) in the contralateral dorsal root ganglion is important in the development of mirror-image pain, it is not known if this is related to enhanced expression of nociceptive fibers in the contralateral skin. Mechanical and thermal sensitivity in the contralateral hind paw was measured at four different time points (5, 10, 20 and 30weeks) after transection and immediate end-to-end reconstruction of the sciatic nerve in rats. These findings were compared to the density of epidermal (peptidergic and non-peptidergic) nerve fibers on the contralateral hind paw. Mechanical hypersensitivity of the contralateral hind paw was observed at 10weeks PO, a time point in which both subgroups of epidermal nerve fibers reached control values. Thermal hypersensitivity was observed with simultaneous increase in the density of epidermal peptidergic nerve fibers of the contralateral hind paw at 20weeks PO. Both thermal sensitivity and the density of epidermal nerve fibers returned to control values 30weeks PO. We conclude that changes in skin innervation and sensitivity are present on the uninjured corresponding side in a transient pain model. Therefore, the contralateral side cannot serve as control. Moreover, the current study confirms the involvement of the peripheral nervous system in the development of mirror-image pain.

Comments to the term "cold-induced vasodilatation" in "laser doppler perfusion imaging of skin territory to reflect autonomic functional recovery following sciatic nerve autografting repair in rats".

con-tributed to the debate on the influence of nerve injuryand nerve repair on the CIVD reaction. The studyreported a significant decrease in the CIVD reaction ofthe hind paw following a nerve transection of the sciaticnerve as compared to a sham-treated group (same opera-tion procedure, without nerve manipulation). In addition,they measured the CIVD reaction after an autograft repairof the sciatic nerve and found no significant differenceswith the sham-treated group. Therefore, the authors con-cluded that the CIVD reaction is controlled by the nerv-ous system, more specifically, by the autonomic nervoussystem. In this reply, we question if Hu et al. use thedefinition of a CIVD in an accurate manner. It is possiblethat the CIVD reaction defined and measured by Huet al. was not a CIVD reaction but an active rewarmingpattern.A CIVD is currently defined as a cyclic oscillation inblood flow that occurs in extremities on cold exposure.