Optical imaging to assess metabolic state of diabetic wound s
Mentor 1
Sandeep Gopalakrishnan
Location
Union 344
Start Date
27-4-2018 1:20 PM
Description
Objective: This study utilizes redox fluorescence imaging to quantitatively assess diabetic wound healing. Introduction: Given the increasing prevalence of diabetes worldwide, lower extremity ulcers and amputations are an increasing problem in individuals living with diabetes. It is critical to understand the underlying mechanisms of these debilitating wounds. In response to this need, we have developed two novel optical imaging techniques (in vivo fluorescence imaging and 3D Cryoimaging) to assess mitochondrial bioenergetics in diabetic wounds. Materials and methods: db/db mice and age matched C57BL/6J mice at the age of 20-22 weeks were used in the experiments. Mice were anesthetized and a 10 mm circular full thickness wound was prepared midline at the shoulder-level. In vivo images of metabolic indices (NADH and FAD) were captured on 1st, 2nd, 3rd, 4th and 5th day post-wounding and the mitochondrial redox-ratio (RR=NADH/FAD) was quantified by calculating redox-ratio images. After 5 days, mice were euthanized, wound biopsies were collected and snap frozen in liquid nitrogen for later 3D cryoimaging of the volumetric mitochondrial redox state. Results: In vivo fluorescence imaging showed that in control wounds, the metabolic marker (RR) increased at day 4th (114% reduced) when compared to day 0 of wound formation; however, in diabetic wounds no difference in RR was detected during wound healing. 3D cryoimages showed that the mean volumetric redox state of diabetic wounds was 61% lower than that measured in non-diabetic control animals. This reduction may be a result from the increased oxidative stress known to be present in diabetic wounds. Conclusion: These findings are consistent with reports of diabetes-induced mitochondrial dysfunction and oxidative stress in the organs and tissues of diabetic animals and humans. They extend the observation of mitochondrial dysfunction and oxidative stress in chronic wounds which are factors contributing to the profound delay in wound healing in diabetes.
Optical imaging to assess metabolic state of diabetic wound s
Union 344
Objective: This study utilizes redox fluorescence imaging to quantitatively assess diabetic wound healing. Introduction: Given the increasing prevalence of diabetes worldwide, lower extremity ulcers and amputations are an increasing problem in individuals living with diabetes. It is critical to understand the underlying mechanisms of these debilitating wounds. In response to this need, we have developed two novel optical imaging techniques (in vivo fluorescence imaging and 3D Cryoimaging) to assess mitochondrial bioenergetics in diabetic wounds. Materials and methods: db/db mice and age matched C57BL/6J mice at the age of 20-22 weeks were used in the experiments. Mice were anesthetized and a 10 mm circular full thickness wound was prepared midline at the shoulder-level. In vivo images of metabolic indices (NADH and FAD) were captured on 1st, 2nd, 3rd, 4th and 5th day post-wounding and the mitochondrial redox-ratio (RR=NADH/FAD) was quantified by calculating redox-ratio images. After 5 days, mice were euthanized, wound biopsies were collected and snap frozen in liquid nitrogen for later 3D cryoimaging of the volumetric mitochondrial redox state. Results: In vivo fluorescence imaging showed that in control wounds, the metabolic marker (RR) increased at day 4th (114% reduced) when compared to day 0 of wound formation; however, in diabetic wounds no difference in RR was detected during wound healing. 3D cryoimages showed that the mean volumetric redox state of diabetic wounds was 61% lower than that measured in non-diabetic control animals. This reduction may be a result from the increased oxidative stress known to be present in diabetic wounds. Conclusion: These findings are consistent with reports of diabetes-induced mitochondrial dysfunction and oxidative stress in the organs and tissues of diabetic animals and humans. They extend the observation of mitochondrial dysfunction and oxidative stress in chronic wounds which are factors contributing to the profound delay in wound healing in diabetes.
