M. Carmen Acosta

Ocular surface wetness is regulated by TRPM8-dependent cold thermoreceptors of the cornea

Basal tearing is crucial to maintaining ocular surface wetness. Corneal cold thermoreceptors sense small oscillations in ambient temperature and change their discharge accordingly. Deletion of the cold-transducing ion channel Transient receptor potential cation channel subfamily M member 8 (TRPM8) in mice abrogates cold responsiveness and reduces basal tearing without affecting nociceptor-mediated irritative tearing. Warming of the cornea in humans also decreases tearing rate. These findings indicate that TRPM8-dependent impulse activity in corneal cold receptors contributes to regulating basal tear flow.

Sensory experiences in humans and single‐unit activity in cats evoked by polymodal stimulation of the cornea

1 The cornea of human subjects and of anaesthetised cats was stimulated with a jet of air of controlled flow, temperature and CO2 concentration delivered by a gas aesthesiometer. 2 In humans, the intensity and magnitude of various components of the sensory experience (intensity of the sensation, degree of irritation, magnitude of burning and stinging pain, magnitude of the cold and warm components of the sensation) were measured using separate visual analog scales. In anaesthetised cats, the impulse response to the same stimuli was recorded from single mechanosensory, polymodal and cold‐sensitive corneal fibres in the ciliary nerves. 3 Intensity‐response curves for mechanical stimulation showed that all parameters of the sensation experienced by humans increased with the intensity of the stimulus. Mechanical stimuli recruited mainly phasic mechanosensory and polymodal afferents in the cat. 4 Acidic stimulation with gas mixtures of increasing CO2 concentration evoked irritation, burning and to a lesser extent stinging pain of a magnitude roughly proportional to the intensity of the stimulus in humans. CO2 primarily recruited polymodal afferents and weakly excited cold‐sensitive fibres in the cats cornea. 5 Heat stimuli evoked in humans a sensation profile similar to CO2 but accompanied by a warmth component. In the cats cornea, heat excited only polymodal fibres and silenced cold‐sensitive corneal units. 6 Cold stimuli applied to the human cornea elicited a sensation of cooling that became irritant at the lowest temperatures. Corneal cold‐sensitive fibres of the cat were activated in a manner proportional to the temperature drop, while polymodal nociceptor fibres were recruited only by the lowest temperatures. Topical menthol (0.2 mm) applied to humans evoked and later eliminated cold sensations produced by cold stimuli while the irritation sensation caused by low temperature stimuli still persisted. 7 Human subjects were able to identify masked mechanical, thermal and chemical stimuli applied to the cornea. 8 Irritation and cold sensations can therefore be evoked separately from the cornea by selective activation of mechanosensory, polymodal and cold corneal sensory afferents. Stimulation with different forms of energy usually leads to combined activation and/or inhibition of the different populations of sensory afferent fibres, evoking blended sensations that include irritation and thermal components in a variable degree.

Nerves and Sensations from the Eye Surface

Because vision plays a critical role in obtaining information from the external world, evolutionary development has provided the structures that sustain this function with special protection against injury. Thus, the cornea possesses the richest sensory innervation of the body to detect noxious stimuli. The trigeminal sensory neurons that innervate the eye vary in their chemical composition and electrophysiological properties, and can be classified according to the stimuli that activate them preferentially: mechanical forces, temperature, or irritant chemicals. Different classes of noxious stimuli (mechanical injuries, heat, extreme cold) activate to a different degree the various populations of sensory fibers of the ocular surface and evoke unpleasant sensations of distinct quality. When injured either accidentally or following ocular surgery, sensory nerve fibers of the ocular surface may form neuromas that develop abnormal activity and become the source of unpleasant sensations, such as pain, dryness, grittiness, etc. In parallel, their response to natural stimuli is diminished. The possibility of hypesthesia and dysaesthesias must be considered in the assessment of the risks of therapeutic procedures that involve damage to ocular sensory nerves.

Recovery of Corneal Sensitivity to Mechanical and Chemical Stimulation After Laser in situ Keratomileusis

PURPOSE To evaluate the time course of changes in corneal sensitivity to mechanical and chemical stimuli produced by laser in situ keratomileusis (LASIK) in humans. METHODS We performed a cross-sectional study of 17 LASIK-operated eyes (VisX S2, equipped with version 2.50-3.10 software) and 15 control eyes of 17 individuals to evaluate regeneration of corneal sensitivity after LASIK. Gas pulses of variable flow and compositions were applied to the cornea by a non-contact gas esthesiometer. Mechanical stimuli consisted of air puffs at flows from 0 to 200 ml/min. Chemical stimulation was made with gas pulses containing 0% to 80% CO2 in air at subthreshold flow. Mechanical and chemical thresholds and intensity-response curves for the evoked sensations were determined prior to surgery, and 7 to 9 days, 3 to 5 months, and 1.5 to 2.5 years after surgery. RESULTS Corneal sensitivity to mechanical stimulation was enhanced 7 to 9 days after surgery but subsequently dropped markedly and remained significantly below control levels 3 to 5 months after LASIK. Sensitivity to both mechanical and chemical types of stimuli was close to normal 2 years postoperatively. CONCLUSIONS Corneal sensitivity decreased immediately after LASIK but mechanical sensitivity showed a transient hyperesthesia 7 to 9 days afterward. Subsequently, a long-lasting and deep hypoesthesia to mechanical and chemical stimuli was observed. Gas esthesiometry revealed that disturbances of corneal sensation still exist at times when coarse mechanical sensitivity appeared to be normal.

What Causes Eye Pain

Eye pain is an unpleasant sensory and emotional experience including sensory-discriminative, emotional, cognitive, and behavioral components and supported by distinct, interconnected peripheral and central nervous system elements. Normal or physiological pain results of the stimulation by noxious stimuli of sensory axons of trigeminal ganglion (TG) neurons innervating the eye. These are functionally heterogeneous. Mechano-nociceptors are only excited by noxious mechanical forces. Polymodal nociceptors also respond to heat, exogenous irritants, and endogenous inflammatory mediators, whereas cold thermoreceptors detect moderate temperature changes. Their distinct sensitivity to stimulating forces is determined by the expression of specific classes of ion channels: Piezo2 for mechanical forces, TRPV1 and TRPA1 for heat and chemical agents, and TRPM8 for cold. Pricking pain is evoked by mechano-nociceptors, while polymodal nociceptors are responsible of burning and stinging eye pain; sensations of dryness appear to be mainly evoked by cold thermoreceptors. Mediators released by local inflammation, increase the excitability of eye polymodal nociceptors causing their sensitization and the augmented pain sensations. During chronic inflammation, additional, long-lasting changes in the expression and function of stimulus-transducing and voltage-sensitive ion channels develop, thereby altering polymodal terminal’s excitability and evoking chronic inflammatory pain. When trauma, infections, or metabolic processes directly damage eye nerve terminals, these display aberrant impulse firing due to an abnormal expression of transducing and excitability-modulating ion channels. This malfunction evokes ‘neuropathic pain’ which may also result from abnormal function of higher brain structures where ocular TG neurons project. Eye diseases or ocular surface surgery cause different levels of inflammation and/or nerve injury, which in turn activate sensory fibers of the eye in a variable degree. When inflammation dominates (allergic or actinic kerato-conjunctivitis), polymodal nociceptors are primarily stimulated and sensitized, causing pain. In uncomplicated photorefractive surgery and moderate dry eye, cold thermoreceptors appear to be mainly affected, evoking predominant sensations of unpleasant dryness.

Regeneration of functional nerves within full thickness collagen–phosphorylcholine corneal substitute implants in guinea pigs

Our objective was to evaluate promotion of tissue and nerve regeneration by extracellular matrix (ECM) mimics, using corneal implantation as a model system. Porcine type I collagen and 2-methacryloyloxyethyl phosphorylcholine (MPC) were crosslinked using 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS) and moulded into appropriate corneal dimensions to serve as substitutes for natural corneal ECM. These were implanted as full thickness grafts by penetrating keratoplasty into the corneas of guinea pigs after removal of the host tissue, and tracked over eight months, by clinical examination, slit-lamp biomicroscopy, and esthesiometry. Histopathology and ex vivo nerve terminal impulse recordings were performed at three months and at eight months. The implants promoted regeneration of corneal cells, nerves and the tear film, while retaining optical clarity. After three months, electrophysiological recordings showed evidence of mechano-nociceptors, and polymodal units inside the implants, while cold-sensitive units were present only on the peripheral host cornea. Following eight months, the incidence of nerve activity and the frequency of spontaneous firing were higher than in control eyes as reported for regenerating fibers. Active cold nerve terminals also innervated the implant area. We show that ECM mimetic materials can promote regeneration of corneal cells and functional nerves. The simplicity in fabrication and demonstrated functionality shows potential for ECM substitutes in future clinical applications.

Decreased corneal sensitivity and tear production in fibromyalgia.

PURPOSE To investigate corneal sensitivity to selective mechanical, chemical, heat, and cold stimulation in patients with fibromyalgia (FM). METHODS Twenty patients with FM (18 women, 2 men; 51.9 +/- 2.3 years old) and 18 control subjects (16 women, 2 men; 51.7 +/- 2.4 years) participated voluntarily in the study. Subjective symptoms of ocular dryness were explored and a Schirmer I test was performed. The response to selective stimulation of the central cornea with the Belmonte gas esthesiometer was measured. RESULTS The majority (18/20) of patients with FM reported dry eye symptoms, with the ocular dryness score significantly higher in affected subjects than in healthy ones (2.3 +/- 0.1 vs. 0.05 +/- 0.02; P < 0.001). The Schirmer test results were significantly lower in patients with FM than in those in the control group (10.5 +/- 2.2 and 30.6 +/- 1.6 mm, respectively; P < 0.001). Mean corneal threshold sensitivity values to chemical stimulation (31.16% +/- 2.04% CO(2) FM; 15.72% +/- 0.67% CO(2) control), heat (1.87 +/- 0.11 degrees C FM; 0.99 +/- 0.05 degrees C control), and cold (-2.53 +/- 0.11 degrees C FM; -0.76 +/- 0.05 degrees C control) were increased in patients with FM, whereas threshold responses to mechanical stimulation did not vary significantly (123.0 +/- 8.0 mL/min FM; 107.8 +/- 4.4 mL/min control). CONCLUSIONS The reduced corneal sensitivity of patients with fibromyalgia is attributable to a moderate decrease in corneal polymodal and cold nociceptor sensitivity, which may be the consequence or the cause of the chronic reduction in tear secretion also observed in these patients.

Changes in sensory activity of ocular surface sensory nerves during allergic keratoconjunctivitis.

Summary Ocular discomfort sensations accompanying experimental allergic keratoconjunctivitis are caused by increased responsiveness of corneal polymodal nociceptors, while cold thermoreceptor activity is reduced. Abstract Peripheral neural mechanisms underlying the sensations of irritation, discomfort, and itch accompanying the eye allergic response have not been hitherto analyzed. We explored this question recording the changes in the electrical activity of corneoconjunctival sensory nerve fibers of the guinea pig after an ocular allergic challenge. Sensitization was produced by i.p. ovalbumin followed by repeated application in the eye of 10% ovalbumin on days 14 to 18. Blinking and tearing rate were measured. Spontaneous and stimulus‐evoked (mechanical, thermal, chemical) impulse activity was recorded from mechanonociceptor, polymodal nociceptor and cold corneoscleral sensory afferent fibers. After a single (day 14) or repeated daily exposures to the allergen during the following 3 to 4 days, tearing and blinking rate increased significantly. Also, sensitization was observed in mechanonociceptors (transient reduction of mechanical threshold only on day 14) and in polymodal nociceptors (sustained enhancement of the impulse response to acidic stimulation). In contrast, cold thermoreceptors showed a significant decrease in basal ongoing activity and in the response to cooling. Treatment with the TRPV1 and TRPA1 blockers capsazepine and HC‐030031 reversed the augmented blinking. Only capsazepine attenuated tearing rate increase and sensitization of the polymodal nociceptors response to CO2. Capsazepine also prevented the decrease in cold thermoreceptor activity caused by the allergic challenge. We conclude that changes in nerve impulse activity accompanying the ocular allergic response, primarily mediated by activation of nociceptor’s TRPV1 and to a lesser degree by activation of TRPA1 channels, explain the eye discomfort sensations accompanying allergic episodes.

Abnormal activity of corneal cold thermoreceptors underlies the unpleasant sensations in dry eye disease

Abstract Dry eye disease (DED) affects >10% of the population worldwide, and it provokes an unpleasant sensation of ocular dryness, whose underlying neural mechanisms remain unknown. Removal of the main lachrymal gland in guinea pigs caused long-term reduction of basal tearing accompanied by changes in the architecture and density of subbasal corneal nerves and epithelial terminals. After 4 weeks, ongoing impulse activity and responses to cooling of corneal cold thermoreceptor endings were enhanced. Menthol (200 &mgr;M) first excited and then inactivated this augmented spontaneous and cold-evoked activity. Comparatively, corneal polymodal nociceptors of tear-deficient eyes remained silent and exhibited only a mild sensitization to acidic stimulation, whereas mechanonociceptors were not affected. Dryness-induced changes in peripheral cold thermoreceptor responsiveness developed in parallel with a progressive excitability enhancement of corneal cold trigeminal ganglion neurons, primarily due to an increase of sodium currents and a decrease of potassium currents. In corneal polymodal nociceptor neurons, sodium currents were enhanced whereas potassium currents remain unaltered. In healthy humans, exposure of the eye surface to menthol vapors or to cold air currents evoked unpleasant sensations accompanied by increased blinking frequency that we attributed to cold thermoreceptor stimulation. Notably, stimulation with menthol reduced the ongoing background discomfort of patients with DED, conceivably due to use-dependent inactivation of cold thermoreceptors. Together, these data indicate that cold thermoreceptors contribute importantly to the detection and signaling of ocular surface wetness, and develop under chronic eye dryness conditions an injury-evoked neuropathic firing that seems to underlie the unpleasant sensations experienced by patients with DED.

Corneal sensitivity in diabetic patients subjected to retinal laser photocoagulation.

PURPOSE To determine the changes in corneal sensitivity to different stimulus modalities in diabetes mellitus (DM)1 and DM2 patients with retinopathy, and to explore whether argon laser photocoagulation exacerbates sensitivity loss in diabetic patients. METHODS Corneal sensitivity to different modalities of stimulus was determined in one randomized eye in 52 patients with DM1 (n = 35) or DM2 (n = 17), and in 27 healthy subjects. Medical history was obtained from all the patients, including age, sex, time from DM diagnosis, type of diabetes, time from onset of retinopathy, type of diabetic retinopathy, and type of argon laser treatment. Corneal sensitivity was determined using a gas esthesiometer. Mechanical, chemical, and thermal (heat and cold) stimuli were applied on the central cornea. RESULTS Sensitivity thresholds to selective mechanical, chemical, and cold stimulation were significantly higher in DM patients compared to controls. Sensitivity threshold to mechanical and chemical stimuli was higher in DM2 than in DM1 patients. In DM1 patients, mechanical threshold increased with time after DM diagnosis. No correlation was found between sensitivity thresholds to chemical or thermal stimulation and the age of the patient, type of retinopathy, or time from its diagnosis. Laser treatment generated a further impairment of corneal sensitivity. CONCLUSIONS Corneal sensitivity to mechanical, chemical, and thermal stimulation is decreased in DM patients, suggesting that diabetes affects homogeneously the different types of sensory neurons innervating the cornea. Corneal sensitivity appears to be more disturbed in DM2 than in DM1. Laser treatment of DM patients generates a further impairment in corneal sensitivity, probably as the result of physical damage to ciliary nerves.