Quantification of ocular surface microcirculation by computer assisted video microscopy and diffuse reflectance spectroscopy.

In piglets we tested the applicability of digital video microscopy and diffuse reflectance spectroscopy for non-invasive assessments of limbal and bulbar conjunctival microcirculation. A priori we postulated that the metabolic rate is higher in limbal as compared to bulbar conjunctiva, and that this difference is reflected in microvascular structure or function between the two locations. Two study sites, Oslo University Hospital (OUH), Norway and Cleveland Clinic (CC), USA, used the same video microscopy and spectroscopy techniques to record limbal and bulbar microcirculation in sleeping piglets. Recordings were analyzed with custom-made software to quantify functional capillary density, capillary flow velocity and microvascular oxygen saturation in measuring volumes of approximately 0.1 mm3. The functional capillary density was higher in limbus than in bulbar conjunctiva at both study sites (OUH: 18.1 ± 2.9 versus 12.2 ± 2.9 crossings per mm line, p < 0.01; CC: 11.3 ± 3.0 versus 7.1 ± 2.8 crossings per mm line, p < 0.01). Median categorial capillary blood flow velocity was higher in bulbar as compared with limbal recordings (CC: 3 (1-3) versus 1 (0-3), p < 0.01). Conjunctival microvascular oxygen saturation was 88 ± 5.9% in OUH versus 94 ± 7.5% in CC piglets. Non-invasive digital video microscopy and diffuse reflectance spectroscopy can be used to obtain data from conjunctival microcirculation in piglets. Limbal conjunctival microcirculation has a larger capacity for oxygen delivery as compared with bulbar conjunctiva.

Acute skin trauma induces hyperemia, but superficial papillary nutritive perfusion remains unchanged.

OBJECTIVES: Superficial skin papillary capillaries with blood supply from a superficial vascular plexus and regulated by local metabolic needs supply oxygen and nutrients for epithelial cell proliferation. A deep vascular plexus regulated by autonomous nerves serves body thermoregulation. In healthy volunteers, we assessed circulatory effects of a standardized skin trauma by CAVM, DRS, and LDPM to assess the measuring depth of the three techniques and to describe the acute trauma effects on nutritive and thermoregulatory perfusion. METHODS: Volunteers (n=12) were examined at baseline and after induction of a 5.0 mm×1.0 mm incision on the forearm; 30 minutes after the trauma induction, data were collected at 0-1, 2-3 and 30 mm distances. RESULTS: LDPM showed hyperemia at 2-3 mm distance (35.8±15.2 a.u.), but not at 30 mm distance (7.4±2.5 a.u.) compared to baseline (8.8±1.8 a.u.). The DRS saturation increased at 2-3 mm (71.2±4.8%), but not at 30 mm (49.8±7.9%) compared to baseline (45.8±7.4%). Capillary density and flow velocities were unaffected at all distances. CONCLUSIONS: The results indicate that skin nutritive papillary capillary function can be assessed by CAVM and DRS, but not with LDPM because of its dependence of the deep plexus perfusion.

Effect of Surgical Technique on Corneal Implant Performance.

PURPOSE: Our aim was to determine the effect of a surgical technique on biomaterial implant performance, specifically graft retention. METHODS: Twelve mini pigs were implanted with cell-free, 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC)/N-hydroxysuccinimide (NHS) cross-linked recombinant human collagen type III (RHCIII) hydrogels as substitutes for donor corneal allografts using overlying sutures with or without human amniotic membrane (HAM) versus interrupted sutures with HAM. The effects of the retention method were compared as well as the effects of collagen concentration (13.7% to 15% RHCIII). RESULTS: All implanted corneas showed initial haze that cleared with time, resulting in corneas with optical clarity matching those of untreated controls. Biochemical analysis showed that by 12 months post operation, the initial RHCIII implants had been completely remodeled, as type I collagen, was the major collagenous protein detected, whereas no RHCIII could be detected. Histological analysis showed all implanted corneas exhibited regeneration of epithelial and stromal layers as well as nerves, along with touch sensitivity and tear production. Most neovascularization was seen in corneas stabilized by interrupted sutures. CONCLUSIONS: This showed that the surgical technique used does have a significant effect on the overall performance of corneal implants, overlying sutures caused less vascularization than interrupted sutures. TRANSLATIONAL RELEVANCE: Understanding the significance of the suturing technique can aid the selection of the most appropriate procedure when implanting artificial corneal substitutes. The same degree of regeneration, despite a higher collagen content indicates that future material development can progress toward stronger, more resistant implants.

Skin microvascular morphology and hemodynamics during treatment with veno-arterial extra-corporeal membrane oxygenation.

OBJECTIVES: To explore the potentials of microcirculatory assessments for predicting outcome of patients treated with extra corporeal membrane oxygenation for cardiogenic shock. METHODS: Eight patients with acute cardiogenic shock treated with ECMO and eight healthy controls were examined with skin vital microscopy and laser Doppler perfusion measurements. RESULTS: Three patients died on ECMO (group 1). Five patients were successfully weaned off ECMO (group 2). Four patients were discharged from hospital and one died after successful weaning from bleeding complications. Patients surviving ECMO (group 2) had microcirculatory findings comparable with healthy controls. Patients in group 1 showed major skin microvascular pathology: pericapillary bleedings (n = 1), pericapillary dark haloes (n = 2) and capillary micro thrombi (n = 1). As compared with survivors they had lower functional capillary density (FCD) (n = 3), higher heterogeneity of functional capillary density (n = 3) and significantly reduced capillary mean flow-categorical velocity (n = 2). Laser Doppler measurements in group 1 had non-significant lower laser Doppler flux values as compared with survivors and controls. CONCLUSION: Skin microvascular pathology as detected with video microscopy (pericapillary bleedings or haloes, micro-thrombi/capillaries with "no flow", low FCD with high spatial distribution heterogeneity or low mean flow-categorial velocity) seems to be associated with poor prognosis.