Absolute quantification (ml blood/sec ∗ mm2 tissue) of normal vs. diabetic foot skin microvascular blood perfusion: Feasibility of FM-PPG measurements under clinical conditions.

Fluorescence-mediated photoplethysmography (FM-PPG) is the first routine clinical methodology by which to quantifiably measure tissue blood perfusion in absolute terms (mL blood/sec ∗ mm2 tissue). The FM-PPG methodology has been described in detail previously in this journal (MVR 114, 2017, 92-100), along with initial proof-of-concept measurements of blood perfusion in both ocular and forearm skin tissues. The motivation for the current study was to investigate whether FM-PPG can be used readily and routinely under realistic clinical conditions. The vehicle for doing this was to measure medial foot capillary blood flow, i.e., tissue perfusion, in 7 normal subjects, mean = 6.76 ±â€¯2.29 E-005 mL/(sec ∙ mm2), and lesion-free areas of 8 type-2 diabetic patients with skin ulceration, mean = 4.67 + 3.15 E-005 mL/(sec ∙ mm2). Thus, perfusion in the diabetics was found to be moderately lower than that in the normal control subjects. Earlier skin perfusion measurements in medial forearms of 4 normal subjects, mean = 2.64 + 0.22 E-005 mL/(sec ∙ mm2), were lower than both the normal and diabetic foot perfusion measurements. Variability in the heartbeat-to-heartbeat blood perfusion pulses in the skin capillaries, defined as the ratio of the standard deviation among beat-to-beat pulses divided by the mean perfusion of those pulses, was determined for each subject. Average variability in foot skin was 21% in the diabetic population, versus 16% for normal subjects; and it was 18% in forearm skin. We conclude that absolute quantitative FM-PPG measurement of skin blood perfusion at the level of nutritive capillaries is feasible routinely under clinical conditions, allowing for quantitative measurement of skin tissue blood perfusion in absolute terms.

Feasibility of extracting velocity distribution in choriocapillaris in human eyes from ICG dye angiograms.

Indocyanine green (ICG) dye angiography has been used by ophthalmologists for routine examination of the choroidal vasculature in human eyes for more than 20 years. In this study, a new approach is developed to extract information from ICG dye angiograms about blood velocity distribution in the choriocapillaris and its feeding blood vessels. ICG dye fluorescence intensity rise and decay curves are constructed for each pixel location in each image of the choriocapillaris in an ICG angiogram. It is shown that at each instant of time the magnitude of the local instantaneous dye velocity in the choriocapillaris is proportional to both the slope of the ICG dye fluorescence intensity curve and the dye concentration. This approach leads to determination of the absolute value of blood velocity in the choriocapillaris, assuming an appropriate scaling, or conversion factor can be determined. It also enables comparison of velocities in different regions of the choriocapillaris, since the conversion factor is independent of the vessel location. The computer algorithm developed in this study can be used in clinical applications for diagnostic purposes and for assessment of the efficacy of laser therapy in human eyes.

Ocular structure and function in an aged monkey with spontaneous diabetes mellitus.

Diabetes mellitus develops spontaneously in middle-aged, obese rhesus monkeys, thus making them a good model for examining the effects of co-morbid factors on the development of end-organ damage. Changes in structure and function in the eyes of one monkey who spontaneously developed type 2 diabetes are reported here. This animal had concomitant hypertension, high levels of triglycerides and serum cholesterol, and a low fraction of high-density lipoprotein. The eyes showed intraretinal hemorrhages and large areas of retinal capillary nonperfusion. Indo-cyanin green (ICG) angiography revealed a large area of non- or poorly perfused choriocapillaris in one eye, and immunohistochemistry showed loss of viable choriocapillaries in this region. Both basal laminar deposits and hard drusen were present on areas of Bruch's membrane adjacent to nonviable choriocapillaris. Blood flow via the nasal posterior ciliary arteries to this section of choroid was not detectable by color duplex Doppler ultrasound, indicating contribution of extraocular vascular disease to ischemia in this eye. There was a severe decline in number of photoreceptor inner and outer segments, and corresponding reductions in the multifocal electroretinogram (ERG), in the areas of choriocapillaris loss. The ganzfeld ERG indicated loss in both inner and outer retinal function. Much of the ganglion cell layer was absent throughout the retina, possibly reflecting the effect of diabetes as well as chronic open angle glaucoma; the latter diagnosis supported by elevated intraocular pressures and excavated optic disks. In summary, high resolution, enzyme histochemical and histopathological analyses of a diabetic hypertensive monkey retina and choroid after serial functional in vivo analyses have demonstrated the relationship between vascular dysfunction and visual function loss. Choroidal vascular dysfunction in both large and small vessels was associated with age-related macular degeneration-like changes in Bruch's membrane and photoreceptor degeneration.

Theoretical investigation of the role of choriocapillaris blood flow in treatment of subfoveal choroidal neovascularization associated with age-related macular degeneration.

PURPOSE: To investigate the relationship between choriocapillaris blood flow and blood flow through an overlying choroidal neovascularization, as it relates to photocoagulation-induced changes in the choriocapillaris circulation. METHODS: A theoretical model that simulates the blood flow in the choriocapillaris and choroidal neovascularization of the human eye was developed, based on histologically determined vascular geometry and experimentally measured blood pressure gradients. The choriocapillaris blood pressure and blood flow were examined before and after simulated photocoagulation of various Sattler layer vessels entering the choriocapillaris in the vicinity of the choroidal neovascularization. (The Sattler layer is the inner layer of medium-sized choroidal vessels that includes both arterioles and venules that supply the choriocapillaris.) RESULTS: The theoretical model showed that both partial and complete occlusion of either Sattler arteriole or venous vessels in the vicinity of the capillary-like vessels connecting a choroidal neovascularization to the underlying choriocapillaris results in significant choroidal neovascularization blood flow reduction. These theoretical results are similar to clinically observed changes induced by laser photocoagulation of feeder vessels. (In this discussion, the term "feeder vessels" refers to those vessels in an indocyanine green angiogram image that appear to supply blood to a choroidal neovascularization; these vessels appear to be Sattler layer vessels, rather than the histologically demonstrated short, capillary-like vessels that form choriocapillaris-choroidal neovascularization communications.) CONCLUSIONS: Reduction of choriocapillaris blood flow underlying a choroidal neovascularization may be sufficient to reduce the blood flow rate in the choroidal neovascularization and thereby reduce the associated retinal edema. The results also suggest that reduction of choriocapillaris blood flow may be the common hemodynamic event associated with the successful application of several currently practiced methods of choroidal neovascularization treatment, including feeder vessel photocoagulation, photodynamic therapy, transpupillary thermotherapy, and prophylactic drusen photocoagulation. Ultimately, this model may be useful in determining optimal placement of laser photocoagulation burns to achieve a desirable perturbation in choroidal blood flow distribution and thereby reduce choroidal neovascularization blood flow to the extent necessary to obliterate associated retinal edema.

An ICG angiogram-based clinical method for characterizing the choroidal circulation used to assess the hemorrheologic effects of pentoxifylline.

PURPOSE: To demonstrate an indocyanine green (ICG) angiography-based clinical method for characterizing choroidal blood flow and for detecting changes in choroidal circulation patterns, and by use of that method, to demonstrate that pentoxifylline affects choroidal blood flow. METHODS: High-speed ICG angiography was performed in rhesus monkeys before and after intravenous administration of pentoxifylline or saline (which served as a control) while monitoring blood pressure and heart rate. From these data, three-dimensional surface maps indicating the instantaneous relative distribution of choroidal blood flow during the peak of intra-ocular pressure pulse systole in a 30 degrees field, centered on the macula, were generated to characterize the state of the choroidal circulation at various times during the experiments. RESULTS: Comparisons of the 3-dimentional surface maps consistently indicated an increase in sub-macular choroidal blood flow occurring within 5 to 10 minutes post-pentoxifylline injection, with a gradual return to baseline level 20-40 minutes later. Injection of equal volumes of saline produced no changes in choroidal blood flow. CONCLUSIONS: Posterior-pole choroidal blood flow can be characterized as by a three-dimensional surface representing the instantaneous relative distribution of choroidal blood flow during the peak of intra-ocular pressure pulse systole. Pentoxifylline does, at least transiently, increase sub-macular choroidal blood flow.

Experimental studies of indocyanine green dye-enhanced photocoagulation of choroidal neovascularization feeder vessels.

PURPOSE: To report a model of choroidal neovascularization feeder vessels that reconciles current histologic, angiographic, and clinical data, and to report experimental studies that investigate the potential of indocyanine-green-dye-enhanced photocoagulation to improve feeder-vessel treatment. METHODS: A model of choroidal neovascularization feeder vessels was conceived to account for current histologic and angiographic data. Based on that model, experimental studies of the efficacy of indocyanine green-dye-enhanced photocoagulation were performed, using pigmented rabbit eyes as a model system. A Zeiss fundus camera was modified to permit visualization of choroidal blood flow by high-speed indocyanine green angiography and to permit simultaneous delivery of 810-nm-wavelength diode laser photocoagulation pulses to specific choroidal vascular targets during indocyanine green-dye bolus transit. RESULTS: Choroidal neovascularization feeder vessels appear to originate in the Sattler layer (that is, that portion of the choroidal vasculature consisting of medium-diameter vessels) and enter the choriocapillaris in close proximity to the small capillary-like vessels that penetrate Bruch membrane and communicate with the choroidal neovascularization. The rabbit eye experiments demonstrated that the presence of high indocyanine green dye concentration in circulating blood enhances uptake of near-infrared laser energy (three eyes); injection of sequential indocyanine green dye boluses results in gradually decreased efficiency of dye-enhanced photocoagulation (two eyes); and by application of laser energy during the initial transit of small-volume, high-concentration indocyanine green dye boluses, dye-enhanced photocoagulation of large diameter choroidal arteries can be accomplished with relatively little concomitant retinal tissue damage (three eyes). CONCLUSIONS: Although future trials will be necessary to substantiate these initial findings in the clinical arena, it appears that the efficiency of choroidal neovascularization feeder-vessel photocoagulation may be enhanced, while minimizing concomitant damage to overlying retinal tissue, by delivery of 810-nm wavelength laser energy immediately upon arrival of a high-concentration indocyanine green dye bolus in a targeted feeder vessel. However, molecules of dye adhering to vessel walls or lying in tissue interstitial spaces appear to divert laser energy from the photocoagulation process, so efficiency of indocyanine green dye-enhanced photocoagulation gradually diminishes as the number of injected dye boluses increases.

Disparity between fundus camera and scanning laser ophthalmoscope indocyanine green imaging of retinal pigment epithelium detachments.

PURPOSE: Indocyanine green (ICG) angiograms of each of five patients with retinal pigment epithelium (RPE) detachments were made using first a Topcon fundus camera and then a Heidelberg scanning laser ophthalmoscope (SLO); for each patient, both types of angiograms were obtained on the same day. In each case, the serous fluid appeared bright throughout the fundus camera studies and dark throughout the SLO studies. This study sought to explain the disparity in the appearance of the lesions in the two kinds of images and to determine whether there was dye in the serous fluid. METHODS: Simple model eyes were constructed to demonstrate the effects of Mie light scatter and integrating sphere behavior of the sclera on ICG image formation by the fundus camera and SLO optics. Analysis was made of both the clinical angiograms and model eye images to structure an explanation for the disparate RPE detachment angiographic images. RESULTS: Indocyanine green fluorescent light from choroidal vessels adjacent to the lesions and scattered by the turbid serous fluid accounted for the lesion brightness seen in the fundus camera images. The models confirmed that SLOs suppress scattered light. CONCLUSIONS: The apparent fluorescence of serous fluid beneath RPE detachments in fundus camera early-phase ICG angiogram images is not attributable to the presence of dye; rather, it appears to be attributable to serous fluid light scatter of fluorescent light arising from adjacent fluorescent structures. This light scatter is a consequence of the fundus camera illumination and recording optics and is not present in SLO-generated images. The necessity of understanding such phenomena as absorption, diffraction, polarization, and scatter of light and routinely applying them to ICG angiogram interpretation is underscored when it is shown that they offer simple explanations for unusual or unexpected angiographic results, as in the case of the patients with RPE detachment discussed here.

Variability in choriocapillaris blood flow distribution.

PURPOSE: To investigate variability of choriocapillaris blood flow patterns. METHODS: After the intravenous injection of indocyanine green, angiograms were recorded at 30 images per second in rhesus monkey eyes using a fundus camera equipped with a pulsed laser diode light source, synchronized with a gated (5 msec), intensified charge-coupled device, or CCD, video camera. Images of choriocapillaris filling alone were extracted. Plastic corrosion casts were made of two of the monkey's choroidal vasculatures for subsequent scanning electron microscopy examination. RESULTS: Pulsed laser indocyanine green fluorescence excitation produced better definition of choriocapillaris filling than had been achieved using continuous illumination. No correlation was found between the choriocapillaris plexus architecture revealed by the plastic corrosion casts and the observed choriocapillaris lobular filling. Overall posterior pole choriocapillaris dye-filling patterns were relatively stable for periods of days, but they changed gradually for periods of weeks. Localized minor pattern changes occurred on a much shorter time scale. Choriocapillaris filling patterns were altered by acutely elevating intraocular pressure, by O2 and CO2 breathing, and by argon laser retinal photocoagulation of adjacent areas. CONCLUSIONS: Choriocapillaris filling patterns appear to be determined by the network of perfusion pressure gradients that exist among the interspersed feeding arterioles and draining venules connected to the choriocapillaris plexus. Changes in intraocular pressure and in blood PO2 and PCO2 levels can produce marked changes in the distribution of choriocapillaris blood flow. Retinal laser photocoagulation of adjacent fundus areas alters choriocapillaris blood flow to the extent that the altered flow might be an important factor in the beneficial results attributed to retinal laser treatment.

Extraction of choriocapillaris hemodynamic data from ICG fluorescence angiograms.

PURPOSE: There are conflicting views about the organization of the posterior pole choriocapillaris, particularly concerning blood flow through it, and there are difficulties associated with attempting to obtain such information using histologic techniques, sodium fluorescein angiography, or both. The present study uses a method of analysis based on high-speed angiograms to investigate posterior pole choroidal blood flow. METHOD: The analysis method employed is based on the premises that dye filling of the choriocapillaris is more rapid (because it is pulsatile) than dye filling of the underlying larger diameter vessels, and that fluorescence from these two overlapping layers is additive. The described analysis algorithm was applied to high-speed ICG fluorescence angiograms to emphasize information about choriocapillaris hemodynamics. RESULTS: The analysis method was demonstrated in rhesus monkeys, and results indicate that the posterior pole choriocapillaris does not behave as a homogeneous structure, consisting of discrete lobular segments. In general, the cycle of dye filling of the choriocapillaris begins in the macular area and progresses radially toward the periphery in a wavelike manner, the filling cycle beginning with and being completed during one cycle of the intraocular pressure pulse. CONCLUSIONS: It is possible to extract information related to choriocapillaris blood flow from high-speed ICG angiograms when a fairly well-defined dye bolus wavefront is present. Interpretation of the results suggests that the choriocapillaris lobules fill in a pulsatile manner, out of phase with each other, and may act thereby to dissipate the blood volume entering the choroid during each cardiac cycle in such a way that the retinal macular is not significantly displaced by expansion of the choroidal vascular volume. Depending upon the distribution of pressure gradients across a group of lobules, blood may flow from one lobule into or even through an adjacent one.

Acute posterior multifocal placoid pigment epitheliopathy. An indocyanine green angiographic study.

Acute posterior multifocal placoid pigment epitheliopathy (APMPPE) is an idiopathic posterior segment inflammatory disorder of young adults. The pathophysiology is poorly understood, and debate persists as to whether it represents a primary pigment epithelial disorder or a choroidal vascular disease. Indocyanine green angiography was used to study choroidal blood flow in two patients with typical APMPPE. The authors demonstrate profound delayed choroidal filling in addition to extensive areas of choroidal vessel nonperfusion in the acute stage of this disease. Recovery of choroidal blood flow was evident during clinical resolution. Choroidal blood flow abnormalities are present in APMPPE and suggest that the clinical findings of this disease reflect a primary choroidal vascular disease.

An image processing approach to characterizing choroidal blood flow.

Indocyanine green (ICG) dye angiography has made possible routine visualization of choroidal blood flow in the human eye; however, to date, its clinical utility has been limited. An overlying layer of densely pigmented tissue and the complex, multilayered vascular structure of the choroid combine to produce angiographic images of low contrast which are difficult to interpret. Conventional image processing can enhance individual images of the blood vessels, but this approach contributes no information about the dynamics of blood flow. Using relatively inexpensive, commercially available personal computer hardware, angiographic image processing algorithms were developed which appear to characterize uniquely a subject choroid in terms of various blood flow parameters. We believe this to be the first successfully demonstrated approach to routinely characterizing the human choroidal circulation in a way that conserves spatial distribution of blood flow dynamics across the entire observed choroidal area. The computer system allows acquisition of digital images from photographic film negatives; alternatively, real-time direct digitization of images from a high-resolution video camera is possible. Once acquired, the digitized data are manipulated according to various algorithms that employ time-sequence analysis to generate two-dimensional curves or three-dimensional surfaces which characterize the choroidal circulation. The unique correspondence of each three-dimensional surface to the subject choroidal circulation from which it was derived is demonstrated. Grouping the characteristic three-dimensional surfaces according to various topographic features in common may provide a basis for discriminating between normal and abnormal choroidal circulations.

Perinatal ocular physiology and ROP in the experimental animal model.

In order to overcome the scarcity of premature human ocular tissues and the enormous obstacles to direct examination of immature human ocular vasculatures, a number of animal models have been employed by investigators in order to study various aspects of ROP. A variety of factors may influence selection of the particular model used, but ultimately it is the faithfulness with which the model mimics human ROP that is most important. The validity of the models has been and remains a controversial subject, but evidence appears strong in favor of the beagle puppy model for studying physiology of the ocular vasculatures during perinatal development. Human ROP pathology usually is defined in terms of static morphological state, physiological dysfunction being considerably more difficult to assess. Most of the animal models fall short of mimicking the pathological lesions found in human eyes, especially those associated with severe, or end-stage ROP, yet they do fairly well in terms of mimicking the retinal vascular physiological changes associated with onset of the disease. Unfortunately, where the physiological aspects of ROP are concerned, focus is primarily on the effects of hyperoxia; other physiological factors as well as the potential role of the choroid are essentially ignored. This paper discusses the potential of physiological changes which occur during the perinatal period to play a role in ROP pathogenesis.

Pulsatile flow in the choroidal circulation: a preliminary investigation.

A preliminary investigation has been made of choroidal blood flow using a computer-aided image analysis approach to interpretation of indocyanine green (ICG) dye choroidal angiograms. The goal of the study was to characterise blood flow through the choroidal arteries vs. choroidal capillaries and veins. The methods of analysis used are briefly reviewed, and preliminary data obtained mainly from monkey eyes are presented. Preliminary conclusions are made regarding the relationship between compliance of choroidal arterial vessels and blood flow through them.