Spontaneous Myocardial Infarction in a Translational Porcine Model of Coronary Atherosclerosis: A Pilot Study

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Kushwaha, Jessica A. Marathe, John W. Finnie, Joanne T. M. Tan, and 8 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8110478/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 12 You are reading this latest preprint version Abstract This pilot study characterizes the spectrum of atherosclerotic coronary artery pathologies in a porcine model of streptozotocin-induced diabetes, with a particular focus on the natural progression of atherosclerotic plaques. Four pigs (designated P01 - P04) were subjected to an identical streptozotocin regimen to induce diabetes and studied using blood biochemistry, lipid profiling, coronary angiography, optical coherence tomography, and histological analysis of the coronary arteries to assess for the development of coronary atherosclerosis. All animals developed sustained hyperglycemia and dyslipidemia, with total cholesterol levels ranging from 14.9 to 36.7 mmol/L and low-density lipoprotein concentrations reaching up to 32 mmol/L. Notably, the study captured the full pathological continuum: from nearly no coronary abnormality in P01 and early atheromatous plaque formation in P02, to advanced atherosclerosis in P03, and finally, in P04, severe fibrofatty atheroma, and myocardial infarction. Unlike conventional animal myocardial infarction models that rely on interventional triggers, the infarction in P04 occurred spontaneously, demonstrating the natural cascade of plaque development and acute plaque rupture with thrombosis. These findings demonstrate the feasibility of using streptozotocin-treated pigs to model key stages of diabetic coronary artery disease. Health sciences/Cardiology Health sciences/Diseases Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Introduction Coronary artery disease (CAD) is a serious, often life-threatening, condition commonly characterized by atherosclerosis 1 , 2 . Atherosclerosis and its complications, such as myocardial infarction, are major causes of morbidity and mortality in the developed world. Atherosclerosis is defined as a chronic vascular disease and is characterized by fibroinflammatory lipid plaques or atheroma 3 . Despite current treatments, there remains a critical need to develop new tools to better detect, monitor, and manage CAD 4 . To address this need, researchers often use large animal models - particularly porcine models - prior to human studies due to their close anatomical and physiological resemblance to the human coronary system. These models have proven to be highly translational, effectively replicating human-like plaque development, and myocardial infarction 5 . While earlier studies using low-density lipoprotein (LDL) receptor-knockout pigs have successfully modelled plaque formation, no spontaneous myocardial infarction has been demonstrated 6 , 7 . In this pilot study, we describe a large-animal model that replicates the progressive spectrum of CAD under natural conditions. Importantly, this pathology developed without interventional induction, distinguishing it from conventional models that require artificial triggers such as intracoronary coil placement 8 . Through this approach, the model effectively demonstrates the natural history of CAD in a large animal model. By allowing for spontaneous myocardial infarction through biologically driven disease progression - from intimal infiltration by lipid-laden macrophages (foam cells) to mature fibrofatty plaque formation, endothelial disruption with resulting thrombosis, and consequent myocardial infarction - this model offers a powerful platform for studying plaque instability, evaluating interventions, and translating findings into clinical applications. Additionally, the inter-animal variability observed, despite uniform treatment, reflects the heterogeneity characteristic of human CAD, further enhancing the model's translational relevance. Results We report a progressive, temporally mapped spectrum of CAD in a streptozotocin (STZ)-induced swine model of diabetes. Using multimodal assessment, including serum biomarkers, invasive coronary angiography, intravascular optimal coherence tomography (OCT), and histopathology, we documented distinct stages of disease development across four animals (P01 to P04), which were specifically reared with the hope that they developed atherosclerotic disease that could be assessed using multimodality imaging and histological techniques. Blood Biochemistry and Metabolic Monitoring To monitor systemic metabolic status and evaluate the progression of diabetic and atherosclerotic pathology, blood glucose testing was performed on all four STZ-induced diabetic pigs (P01 to P04), and at their final assessment, a full biochemical profile was performed. The biochemical analysis included postprandial glucose levels, lipid profiles (total cholesterol (TC), LDL-C, high-density lipoprotein-cholesterol (HDL-C), triglycerides) (Table 1 ); and liver function enzymes (alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), gamma-glutamyl transferase (GGT), lactate dehydrogenase (LDH), albumin, globulin), renal function markers (creatinine, urea), inflammatory markers [C-reactive protein (CRP)] (Table 2 ). These parameters were selected to assess the physiological impact of chronic hyperglycaemia and an atherogenic diet on vital organ systems and to correlate biochemical changes with coronary imaging and myocardial histological findings. Table 1 Glucose Metabolism and Lipid Profile. Normal porcine ranges taken from 9 . Glucose Metabolism Lipid Profile Pig Glucose (4.7–8.3) TC (4.0-6.5) LDL-C (2.5-4.0) HDL-C (1.2–2.2) TC/HDL-C (2.5–4.5) Triglyceride (1.8-4.0) (mmol/L) mmol/L mmol/L mmol/L - mmol/L P01 17.7 14.9 12.0 2.6 5.7 0.6 P02 28.4 25.7 21.0 4.4 5.8 0.7 P03 21.4 22.1 18.9 2.7 8.2 1.2 P04 13.3 36.7 32.0 3.9 9.4 1.9 All pigs exhibited marked hyperglycaemia following STZ administration, confirming the successful induction of diabetes. Notably, P02 and P03 demonstrated the highest glucose levels (Table 1 ). Persistent hyperglycemia is known to contribute to endothelial dysfunction and accelerate atherosclerotic development 10 . Insulin therapy was administered intermittently to maintain glucose levels within a manageable range and to prevent complications, such as ketoacidosis 11 . Total cholesterol and LDL-C levels were markedly elevated across all animals, with P04 displaying the most extreme lipid profiles, as illustrated in Table 1 . The atherogenic index, calculated as the TC to HDL-C ratio, exceeded 4.5 in all four pigs. These severe lipid abnormalities closely paralleled the worsening atherosclerotic coronary artery pathology observed in imaging and histological assessments. Such dyslipidaemia is known to promote atheroma development through various mechanisms including foam cell formation, endothelial dysfunction, and sustained vascular inflammation 12 . Table 2. Liver Enzymes, Renal Function, Protein and Inflammatory Markers. Normal porcine ranges taken from 9 Liver enzymes, including ALT, AST, ALP, GGT, and LDH, were elevated across several animals, with the most striking abnormalities observed in P04 (Table 2 ). ALT and AST elevations in P04 (134 U/L and 165 U/L, respectively; both above the upper reference limits): whilst this elevation would normally signify hepatocellular injury, in P04 they were thought to be reflective of a recent myocardial infarction, further supported by the profoundly elevated LDH level, reaching 2525 U/L. The serum biochemistry profile also demonstrated consistently low albumin levels (10–13 g/L, below the normal range of 19–39 g/L) across all pigs, while globulin concentrations remained within the reference range (46–59 g/L). This albumin/globulin imbalance, most pronounced in P04, can be a hallmark of chronic systemic inflammation and impaired hepatic protein synthesis, frequently associated with advanced diabetes-related pathology 12 . Creatinine values were within or close to the reference range (99–148 µmol/L, normal 141–239 µmol/L), though P01 was slightly below and P02 - P03 sat toward the lower-normal boundary. Urea levels (4.7–7.2 mmol/L) remained within the expected physiological range. P01 and P03 showed CRP values below the detection limit (< 0.15 mg/L), while P02 and P04 recorded low concentrations of 0.18 mg/L and 0.22 mg/L, respectively. All values were well within the normal physiological range for pigs (5–30 mg/L 9 ). These findings suggest that the animals were in a stable physiological state, and any vascular changes observed are unlikely to have been influenced by acute inflammation. Angiograms, OCT imaging and histopathological analysis Coronary evaluation using invasive coronary angiography, intravascular OCT imaging, and postmortem histology was conducted on all four pigs. In P01, imaging and histology revealed largely normal coronary architecture with almost no atherosclerotic changes. Figure 1 (a) demonstrates a smooth interventricular artery with no evidence of luminal irregularities, findings corroborated by OCT imaging (Fig. 1 (b)), which showed a lumen with clearly delineated vessel wall layers and no evidence of plaque. The optical signal penetrated uniformly, with no signs of fibrotic or calcified tissue, microchannels, or inflammatory infiltration. Histological examination (Fig. 1 (c)) further confirmed preserved arterial architecture, including an intact intima, tunica media and adventitia, and normal adjacent myocardial and epicardial tissues. Biochemically, P01 exhibited relatively mild metabolic disturbance: fasting glucose levels confirmed hyperglycaemia, while lipid abnormalities remained moderate. Collectively, these findings indicate no evidence of cardiovascular or systemic organ damage, positioning P01 as a representative of the baseline or pre-atherosclerotic stage in the diabetic model. P02 demonstrated early signs of coronary atherosclerosis. Coronary angiography revealed subtle luminal irregularities in the left circumflex (LCX) artery (Fig. 2 (a)). The corresponding OCT imaging (Fig. 2 (b)) showed a mildly irregular lumen contour, with eccentric neointimal hyperplasia. The intima appeared mostly intact but exhibited focal discontinuities, likely reflecting endothelial disruption. The media remained thickened yet structurally organized, and the adventitia showed no evidence of inflammation. These features aligned closely with histological findings (Fig. 2 (c)), which confirmed the presence of early intimal changes in the absence of mature plaque formation. Biochemically, P02 exhibited more pronounced metabolic dysfunction than P01, including elevated glucose levels. Taken together, the imaging, histological, and biochemical data support P02 as a representative of early coronary atherosclerosis. P03 exhibited clear evidence of established CAD as can be seen in Fig. 3 . Angiographic imaging (Fig. 3 (a)) revealed luminal narrowing, particularly involving the LCX artery. The OCT analysis (Fig. 3 (b)) confirmed the presence of atherosclerotic disease, demonstrating a markedly abnormal vascular architecture with luminal stenosis and eccentric intimal thickening consistent with an atherosclerotic plaque. The fibrofatty plaque contained abundant lipid-laden macrophages (foam cells) and fibrous connective tissue, appearing in the OCT images as a homogeneous region of densely packed scatterers with uniform speckles [6–10 o’clock in Fig. 3 (b)]. Within this region, signal attenuation and dropout suggested the presence of a necrotic core. Histological examination shown in Fig. 3 (c) corroborated the imaging, revealing a fibrofatty plaque, with a necrotic core comprised of degenerating foam and medial smooth muscle cells, surrounded by hyaline degeneration and abundant mineralisation. OCT imaging demonstrated that the LCX plaque extended into the vessel lumen, resulting in a visible reduction in luminal area. Biochemically, P03 exhibited elevated lipid levels, including a high TC/HDL-C ratio and increased triglycerides compared with the other animals. Hence, P03 represented a state of active fibrofatty plaque formation and highlighting P03 as a representative model of established atherosclerosis. P04 exhibited the most advanced stage of coronary pathology among the studied cohort, intravascular thrombosis leading to spontaneous myocardial infarction. Angiography shown in Fig. 4 (a) revealed subtotal luminal occlusion of the interventricular (left anterior descending (LAD)-equivalent) artery. The OCT image shown in Fig. 4 (b) illustrates dense, highly backscattering material (3–6 o’clock), consistent with fibrotic plaque. The vessel wall appeared markedly thickened (6–12 o’clock), with a homogenous, strong signal suggestive of a fibrotic plaque. These findings were corroborated by histology shown in Fig. 4 (c), which demonstrated plaques on both sides of the vessel wall. Biochemically, P04 displayed pronounced metabolic distress, with persistently elevated glucose levels, and severely deranged liver function tests (ALT 134 U/L, LDH 2525 U/L) likely reflective of recent myocardial infarction. Collectively, P04 represents a useful animal model of advanced atherosclerotic CAD resulting in myocardial infarction. Further cross-sectional analysis of interventricular artery in P04, accompanied by detailed histopathological evaluation, revealed advanced and multifocal coronary pathology consistent with advanced atherosclerotic disease development and myocardial infarction (Fig. 5 ). Discussion This paper reports on a porcine model devised to demonstrate the progressive development of atherosclerotic CAD associated with diabetes, which can lead to spontaneous myocardial infarction. Using a standardized STZ-induction protocol and an atherogenic diet, we achieved metabolic dysfunction in all animals, reflected by sustained hyperglycemia, progressive dyslipidemia, and liver dysfunction. These systemic markers were correlated with advanced coronary artery pathology, as assessed by angiography, intravascular OCT, and histological evaluation. A pathological continuum was observed in our pigs, mimicking the natural history of human coronary atherosclerosis. P01 exhibited mild metabolic disturbance with no detectable coronary artery plaque. P02 progressed to early vascular disease characterized by intimal infiltration of lipid-laden macrophages or foam cells, termed fatty streak formation. P03 demonstrated more advanced atherosclerosis, the fibrofatty plaque breaching the internal elastic lamina and infiltrating, and damaging, the media, with development of a necrotic core. P04, the most pathologically advanced fibrofatty atheromatous plaque, showed endothelial disruption, leading to sub totally occlusive thrombosis, and myocardial infarction. The spontaneous myocardial infarction which occurred in this model is a particularly significant and novel outcome not demonstrated in previous, similar porcine studies. It emphasizes the ability of this model to reproduce biologically-driven coronary atherosclerotic events, unlike other models that require artificial triggers 8 . The inter-animal variability in disease progression, despite identical protocols, further strengthens the translational relevance of this porcine model, as it reflects the heterogeneity found in human diabetic atherosclerosis. OCT provided detailed visualization of atheronecrotic plaque cores, and thrombosis, and proved instrumental in correlating with the development of progressively more severe arterial pathology. Pigs are a valuable model for studies of atherosclerosis since the human and porcine cardiovascular systems share many structural and functional similarities. Many of the differences between pigs and humans are due to their unguligrade (quadruped) and orthograde (bipedal) stance, which results in a laterally and dorsoventrally compressed thorax, respectively. Porcine hearts are valentine in shape, whereas those of humans are trapezoidal, but the ratio of heart to body weight is similar in both. While the coronary circulation of pigs resembles that of humans, the porcine heart has right coronary dominance. The diameter of great vessels such as aorta and pulmonary arteries is proportionately smaller in pigs than humans. Atria in pigs are more prominent and have thicker muscle, and the left atrium receives only two pulmonary veins, whereas humans have four. In pigs, caval veins enter the right atrium at ninety degrees, but the caval veins in humans are aligned in the same axis. The ratio of wall thickness between left and right ventricles is much higher in pigs than humans and trabeculae carnae and papillary muscles are coarser and broader in the porcine heart. Pigs also have a prominent trabecula septomarginalis (moderator band), which contains Purkinje fibres from the atrioventricular bundle, and connects the septal to the free wall of the right ventricle 13 , 14 . In addition, of the domestic animal species, only the pig develops atherosclerosis with any consistency, with fibrofatty plaques leading to stenosis of coronary arteries in animals 8 to 12 years of age. While the degree of vascular lipid deposition can be accelerated by feeding a high fat diet, lesions rarely reach the severity of those seen in humans. Moreover, unlike in the present study, vascular lesions do not routinely result in occlusive thrombosis and its sequelae, principally myocardial infarction 15 . In previous studies of porcine diabetic coronary atherosclerosis (Table 3 ), there has been considerable variation in the signalment (breed and age), the dose of STZ used to induce diabetes, constituents of the high fat diet, and protracted duration of the experimental period (which incurs a high cost for conduct of these experiments). The most common large pig breeds used in these studies are the Yorkshire 16 17 18 19 , Yorkshire-Landrace crosses 20 21 and Landrace 22 , while a few studies have used mini-pigs 23 24 25 . Unlike these earlier models, the present study achieved the full pathological continuum - from early endothelial changes to spontaneous myocardial infarction, using comparatively less STZ (30 mg/kg on Days 1 and 2, followed by 60 mg/kg on Day 3). Table 3 Summary of previous porcine diabetes/high fat diet studies of coronary atherosclerosis. Study Year Duration Pig Strain Diabetes Induction/ Insulin Management Diet Composition Key Outcomes / Findings 23 2012 20 weeks Sinclair mini pigs -STZ 120 mg/kg via splenic/gastroduodenal arteries; repeated if glucose < 10 mmol/L, -Insulin Management: Not Specified High-fat, high-cholesterol (chol.) (Teklad Diet.96366) Consistent diabetes; carotid stenosis led to localized vulnerable plaques 26 2021 15 months Göttingen & Landrace-Yorkshire -STZ 75–180 mg/kg intravenous (IV), -Insulin Management: Not Specified 25% fat, 1% chol., 10% sucrose, 15% fructose DM* group showed advanced coronary atherosclerosis vs controls 16 2021 60 weeks Yorkshire -STZ 50 mg/kg × 3 days; BG > 350 mg/dL, -Insulin Management: Adjusted by glucose 1.5-4% chol., 15–25% lard; target BG 150–350 mg/dL, TC 500–700 mg/dL 2.5× more plaques in DM group; greater lesion complexity 17 2017 20 weeks Yorkshire -STZ 50 mg/kg × 3 days, -Insulin Management: NPH BID; BG 350–500 mg/dL 1–2% chol., 20% tallow, 0.75% cholic acid Lesions at 12 weeks; most severe in DM + high-fat group 27 2017 15 months Yorkshire × Landrace -STZ 140 mg/kg IV (single), -Insulin Management: Insulin to avoid ketosis 10% sucrose, 15% fructose, 25% lard, 1% chol., 0.7% bile salts Chronic hyperglycemia; moderate atheroma formation 28 2011 30 weeks Yorkshire -STZ 50 mg/kg × 3 days, -Insulin Management: Not Specified 1.5% chol. + 15% lard (diet-induced hyperlipidemia) Thin-cap fibroatheromas localized to low-shear regions 18 2001 4–48 weeks Yorkshire -STZ 50 mg/kg × 3 days, -Insulin Management: Not Specified 1.5% chol. + 15% lard vs normal diet Diabetic + hyperlipidemic pigs had 3–4× BG** and advanced plaques 19 2008 8–30 weeks Yorkshire -STZ 50 mg/kg × 3 days, -Insulin Management: Not Specified 1.5% chol. + 15% lard + sucrose Low shear predicted future high-risk plaque regions 29 2016 24 weeks Yorkshire -STZ 150–200 mg/kg IV, -Insulin Management: Not Specified 15% lard + 1.5% chol. One pig developed coronary plaque visible via IVUS/ICG*** 30 2008 1–20 weeks Large White -STZ 150–200 mg/kg IV; with glucose infusion to avoid hypoglycemia, -Insulin Management: Mixed insulin regimen; BG 150–250 mg/dL Normal diet Stable Type 1Diabetes Mellitus model with controlled glycemia 20 2014 7 weeks Yorkshire × Swedish Landrace -STZ 150 mg/kg IV, -Insulin Management: Gradual SC insulin up to 1 IU/kg/day Normal diet Restored carb/fat/amino acid balance post-treatment 31 2015 15 months Landrace × Yorkshire -STZ 140 mg/kg IV, -Insulin Management: Insulin only for ketosis 25% fat + 1% chol. Early microvascular damage and renal pathology 32 2017 12–36 weeks Miniature pigs -No STZ was used, -Insulin Management: Not Specified 4% chol., 14% tallow, 2% bile salts (1 kg/day) Atheroma formation linked to Cyclooxygenase down regulation * DM: Diabetes Mellitus, **BG: Blood Glucose, ***IVUS: Intravascular Ultrasound, ICG: Indocyanine Green One notable aspect of this study is that multiple techniques, including coronary angiography, OCT, and histological staining, enabled comprehensive visualization of disease progression. OCT imaging, achieved using a custom-designed, 3D-printed imaging catheter 33 , enabled in vivo imaging to identify lipid core and necrotic regions of plaque that are not visible by conventional coronary angiography 21 . The combined application of OCT and fluoroscopic angiography enabled precise lesion mapping across epicardial coronary arteries - the right coronary artery (RCA), interventricular artery (analogous to the human LAD artery), and LCX - which enabled comprehensive assessment of the atherosclerotic process. This pilot study has several limitations. The small sample size (n = 4) restricts the generalizability of the findings and prevents robust statistical analysis, even though it allowed observation of the full disease spectrum from minimal atherosclerotic pathology to spontaneous myocardial infarction. Additionally, the exclusive use of male Yorkshire pigs limits the ability to assess sex- or breed-related differences in metabolic response, plaque development, and susceptibility to myocardial infarction. Finally, the study was not designed to assess therapeutic interventions or validate interventional imaging performance: it was an observational study designed to assess the utility of a new intravascular imaging catheter at recognizing atherosclerosis, and future larger studies are required to establish the model’s reproducibility and suitability for translational research. Overall, this study provides foundational evidence for a preclinical platform capable of capturing the full pathological spectrum of diabetic CAD, from subclinical changes to overt myocardial infarction. The model holds promise for future studies aimed at evaluating plaque instability, testing therapeutic interventions, and investigating mechanisms underlying diabetic cardiovascular disease. Methods All animal procedures in this study were conducted in accordance with the Australian Code for the Care and Use of Animals for Scientific Purposes (8th edition, 2013; updated 2021) 34 and were approved by the South Australian Health and Medical Research Institute (SAHMRI) Animal Ethics Committee (protocol number SAM-21-011). All methods were carried out in accordance with relevant guidelines and regulations. Reporting of animal procedures followed the ARRIVE (Animal Research: Reporting of In Vivo Experiments) guidelines 35 . Animal Model and Induction Protocol Four male (n = 4) Yorkshire swine (designated P01 to P04), age between 9 to 12 weeks, each weighing approximately 25–30 kg at baseline, were used in this study. As this was an observational, exploratory pilot study performed as a part of a larger study assessing the utility of a new intravascular imaging catheter, and hence no formal sample-size calculation was performed, nor was a control group required. Randomisation was not applicable, as all animals underwent the same induction protocol and imaging schedule without allocation to different treatment groups. Histological assessments were performed by an experienced pathologist blinded to the imaging findings, while other aspects of the study were not blinded because all animals received identical procedures. Diabetes mellitus (DM) was chemically induced using a STZ protocol tailored to mimic Type 1-like insulin deficiency. Animals received intravenous STZ at a dose of 30 mg/kg 28 on Days 1 and 2, followed by 60 mg/kg on Day 3. Successful diabetic induction was confirmed by persistent fasting blood glucose levels exceeding 8.3 mmol/L. Seven pigs were initially enrolled. The first three animals were used exclusively to develop and optimise the STZ protocol and did not have the full set of blood results required for the planned analyses. Accordingly, these three animals were not included in the final dataset. The study cohort therefore comprised the remaining four animals (P01-P04), which exhibited the full pathological spectrum relevant to this study and were analysed as reported. No animals or individual data points from P01-P04 were excluded from the analyses. Following induction, all animals were transitioned to a high-fat, high-carbohydrate atherogenic diet designed to promote dyslipidaemia and accelerate atherosclerotic CAD development and progression. Animals were housed under standardized conditions and monitored daily for health status. Serum glucose levels were regularly measured, and short-acting insulin was administered as needed to maintain glucose levels below 30 mmol/L, preventing ketoacidosis. Monthly assessments included body weight, blood glucose concentration, general clinical condition, and appetite. The outcome of interest was the development of atherosclerotic coronary disease as assessed by invasive coronary angiography, OCT, and histological analysis. Analgesia and Anaesthesia Protocol To ensure animal welfare, analgesia and anaesthesia protocols were rigorously followed. A fentanyl transdermal patch (25–50 µg/h) was applied 24 hours prior to any imaging procedure for sustained pain control. On the day of imaging, meloxicam (0.4 mg/kg, intramuscular) was administered as supplemental analgesia. Anaesthesia was induced using intramuscular xylazine (2 mg/kg) and ketamine (20 mg/kg) and maintained via inhaled isoflurane (4–5% for induction, 2–3% for maintenance) in 100% oxygen using mechanical ventilation. Intravascular Imaging and Angiography Coronary imaging was conducted to assess the progression of atherosclerotic pathology in vivo . P01 was imaged at Day 124, P02 at Days 163, 213, and 318, P03 at Days 107 and 282, and P04 at Day 240. These time points were selected to represent early, mid, and late stages of disease development. Vascular access was established via the right femoral artery using a 6 French introducer sheath. Coronary angiography was performed using an AL2 guiding catheter under fluoroscopic guidance. An iodinated contrast agent (Ultravist® (iopromide) 370 mg lodine/mL, Bayer, Germany) was injected at a rate of ~ 4mL/s to delineate coronary anatomy, assess luminal patency, and guide intravascular device positioning. For intravascular OCT imaging, a custom-designed, 3D-printed imaging catheter 33 was advanced over a 0.014" coronary guidewire (Runthrough NS Floppy, Terumo, Japan) into the target artery under fluoroscopic guidance. Automated pullback OCT imaging was performed during contrast flushing at 4 mL/s, enabling detailed visualization of luminal structures and vessel wall composition, including detection and composition of plaque. Systemic anticoagulation was maintained throughout the procedure with intravenous heparin at a dose of 100 IU/kg to prevent intraprocedural coronary thrombosis, with no acute complications. OCT images were acquired using a Telesto II spectral-domain OCT scanner (Thorlabs GmbH, Germany), with a central wavelength of 1300 nm and axial resolution of 4.2 µm (in water). Tissue Collection and Histological Analysis Following final imaging, each animal was humanely euthanized under deep anaesthesia. Coronary arteries (interventricular artery, LCX, and RCA) were excised, perfused with 10% neutral buffered formalin, and submitted for gross and histological evaluation. Coronary blood vessels were transversely cut at serial, standard intervals, then paraffin-embedded, and 6µ sections cut and stained with hematoxylin and eosin (H&E). Hearts were incised and immersion fixed in a large volume of 10% neutral buffered formalin and processed for histopathological examination. Histopathological findings were correlated with imaging and biochemical data to confirm the stage and severity of coronary artery atheroma development. Declarations Acknowledgements We acknowledge Dr Ayla Hoogendoorn, Dr. Chris Christou, Ms. Loren Matthews, Ms. Georgia Williams, Ms. Lisa McKenny, Mr. Robb Muirhead, Mr. Jim Manavis, Ms. Sofie Kogoj, and Ms. Yvonne Ciuk for contributing to the management of the diabetic swine, assistance during imaging procedures, and histological preparation and analysis of histology images. We also acknowledge the support of the Optofab node of the Australian National Fabrication Facility and Adelaide Microscopy. Author contributions statement A.K.K: Conceptualization, Probe Fabrication, Methodology, Investigation, Formal analysis, Visualization, Writing - Original Draft. J.A.M.: Methodology, Data curation, Investigation, Writing - Review & Editing. J.F. : Histological analysis, Interpretation of pathology, Writing - Review & Editing. L.X. : Optical system development, Imaging acquisition, Writing - Review & Editing. P.R. : Lens fabrication, Imaging support, Writing - Review & Editing. J.T.M.T. : Experimental planning, Data interpretation, Writing - Review & Editing. C.A.B. : Supervision, Conceptual guidance, Writing - Review & Editing. J.V. : Clinical interpretation, Imaging validation, Writing - Review & Editing. R.A.M. : Supervision, Optical imaging oversight, Writing - Review & Editing. H.G. : Funding acquisition, OCT probe design consultation, Technical input, Writing - Review & Editing. P.J.P. : Conceptualization, Funding acquisition, Cardiovascular expertise, Project supervision, Writing - Review & Editing. J.L. : Conceptualization, Funding acquisition, Supervision, Project administration, Writing - Review & Editing. Funding This work is supported by the National Health and Medical Research Council Development (Grant No. 2022337), the Investigator Grant (Grant No. 2008462), the Heart Foundation Future Leader Fellowships (Grant Nos. 105608 and 106656), and Catalyst (Grant No. 108710-2024_CPG), the Hospital Research Foundation Project (Grant No. 2022-CP-IDMH-014-83100), Australia-Germany Joint Research Co-operation Scheme (UA-DAAD), Baden-Wuerttemberg-Stiftung (Opterial), European Research Council (Advanced Grant Complexplas, PoC Grant 3DPrintedOptics), Bundesministeriumfür Bildung und Forschung (3DPrintedOptics, Integrated3Dprint, QR.X, QR.N), Deutsche Forschungsgemeinschaft (German Research Foundation) (Grant No. 431314977/GRK2642), HORIZON EUROPE European Innovation Council (Grant No. IV-Lab 101115545), Carl-Zeiss Foundation (EndoPrint3D, QPhoton), and University of Stuttgart (Terra Incognita). Competing interests P. J. Psaltis has received research support from Abbott Vascular; consulting fees from Amgen, Esperion, Eli Lilly, Novartis, Novo Nordisk, and Sanofi; and speaker honoraria from Amgen, AstraZeneca, Bayer, Boehringer Ingelheim, Merck Schering-Plough, Pfizer, Novartis, Novo Nordisk, and Sanofi. R. A. McLaughlin is a co-founder and Director of Miniprobes Pty Ltd, a company that develops optical imaging systems. Miniprobes Pty Ltd did not contribute to or participate in this study. J. Li is the founder and Director of Theia Medical Pty Ltd, a company that develops medical imaging devices. Theia Medical Pty Ltd did not contribute to or participate in this study. The remaining authors declare no conflicts of interests. Additional Information Correspondence and requests for materials should be addressed to Dr Jiawen Li (J.L.). Supplementary information accompanies this paper contains ARRIVE checklist as a compliance. References Aronson, D. & Edelman, E. R. 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Adv Photonics 7 (2025). NHMRC. (National Health and Medical Research Council, Canberra, 2021). Percie du Sert, N., Hurst, V., Ahluwalia, A. The ARRIVE guidelines 2.0: Updated guidelines for reporting animal research. BMC Veterinary Research 16 (2020). https://doi.org/https://doi.org/10.1186/s12917-020-02451-y Additional Declarations No competing interests reported. Supplementary Files AuthorChecklistFull.pdf Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 12 Feb, 2026 Reviews received at journal 27 Jan, 2026 Reviews received at journal 14 Jan, 2026 Reviewers agreed at journal 12 Jan, 2026 Reviewers agreed at journal 06 Jan, 2026 Reviewers agreed at journal 28 Dec, 2025 Reviewers agreed at journal 28 Nov, 2025 Reviewers invited by journal 28 Nov, 2025 Editor assigned by journal 28 Nov, 2025 Editor invited by journal 26 Nov, 2025 Submission checks completed at journal 23 Nov, 2025 First submitted to journal 23 Nov, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. 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08:57:16","extension":"html","order_by":15,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":131903,"visible":true,"origin":"","legend":"","description":"","filename":"earlyproof.html","url":"https://assets-eu.researchsquare.com/files/rs-8110478/v1/ee3b79dea957b8c29992d805.html"},{"id":97132792,"identity":"ed0f010b-8bc9-486e-aed6-d863d4004f17","added_by":"auto","created_at":"2025-12-01 08:57:16","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":275390,"visible":true,"origin":"","legend":"\u003cp\u003eRepresentative fluoroscopic angiographic, OCT and histology images obtained in pig P01. (a) Coronary angiography revealed smooth and unobstructed coronary vessels (scale 2 mm) of the interventricular artery as confirmed by its (b) OCT image (scale 500 µm), with no luminal irregularities, findings that were fully supported by (c) histological sections (scale 500 µm) showing structurally normal arterial walls, preserved tunica media, and no evidence of plaque, thrombus, or inflammation. I, intima; M, media; IEL, internal elastic lamina; A, adventitia; G, guidewire.\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-8110478/v1/27d8667b78e3a67121d52909.png"},{"id":97132793,"identity":"bb4c41ef-a1a6-49ab-bf36-faa5ebcb2d3f","added_by":"auto","created_at":"2025-12-01 08:57:16","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":233491,"visible":true,"origin":"","legend":"\u003cp\u003eRepresentative angiography, OCT and histology images obtained in pig P02. (a) P02 exhibited subtle luminal irregularities on angiography (scale 2 mm), particularly in the LCX artery, suggestive of emerging atherosclerotic pathology. (b) The OCT image of P02 reveals a mildly irregular lumen contour (scale 500 µm), an eccentric lesion of neointimal hyperplasia, supported by (c) histological sections (scale 500 µm) showing early intimal foam cell infiltration.\u003c/p\u003e","description":"","filename":"floatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-8110478/v1/788a9612bcec9b374f9a72b7.png"},{"id":97142247,"identity":"3e50d4d8-91e1-4273-aa72-1c2d2a79d77a","added_by":"auto","created_at":"2025-12-01 10:07:27","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":217562,"visible":true,"origin":"","legend":"\u003cp\u003eRepresentative angiography, OCT and histology images obtained in pig P03. (a) The vascular pathology became more pronounced in P03, where angiographic imaging (scale 2 mm) demonstrated moderate luminal narrowing (b) In OCT image (scale 500 µm) of P03, an atherosclerotic plaque is visible, supported by (c) histological sections (scale 500 µm).\u003c/p\u003e","description":"","filename":"floatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-8110478/v1/1c1b92284574f9b2a2f9b622.png"},{"id":97132797,"identity":"08327815-c412-4efe-8150-9c52164e7289","added_by":"auto","created_at":"2025-12-01 08:57:16","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":279099,"visible":true,"origin":"","legend":"\u003cp\u003eRepresentative angiography, OCT and histology images obtained in pig P04. (a) Advanced pathological findings, where angiography (scale 2 mm) identified subtotal luminal occlusion of the interventricular (LAD-equivalent) artery, (b) Optical coherence tomography (OCT) image (scale 500 µm) of the interventricular artery in P04 showing extensive fibrotic plaque. (c) Histologically (scale 500 µm), well-developed fibrofatty plaques were present bilaterally in the vessel wall, consistent with the OCT image.\u003c/p\u003e","description":"","filename":"floatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-8110478/v1/6dc2080f5d454e0e3a241ea0.png"},{"id":97132798,"identity":"54b62b57-4089-4b72-b249-5a7579ce2ac0","added_by":"auto","created_at":"2025-12-01 08:57:16","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":1553742,"visible":true,"origin":"","legend":"\u003cp\u003eHistology images of P04 (a) Well-developed fibrofatty plaque encroaching upon the lumen with extensive mineralization. The zoomed inset highlights the calcified/mineralization region, (b) Severe atheromatous plaque with luminal stenosis and mineralization, (c) Fibrofatty plaque (arrow) with mineralization. Lumen contains a thrombus, (d) Coronary artery thrombosis with attenuation of the vessel wall, (e) Coronary artery contains a thrombus with marked remodeling of the vessel wall. Inset shows endothelial proliferation with invasion of the thrombus (early recanalization), (f) Cardiac muscles showing multifocal myocardial infarction (arrows). Inset shows the region in the white box at higher magnification.\u003c/p\u003e","description":"","filename":"floatimage5.png","url":"https://assets-eu.researchsquare.com/files/rs-8110478/v1/4ea4fb2a555270479f755bae.png"},{"id":97249503,"identity":"ac77301c-d7d1-4aa5-9997-972f038708ff","added_by":"auto","created_at":"2025-12-02 13:12:46","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3458878,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8110478/v1/977c54dd-5670-4884-8824-a0780cf21387.pdf"},{"id":97132790,"identity":"9d4db3e1-a93b-4ce1-9cb9-ea1ff30570f1","added_by":"auto","created_at":"2025-12-01 08:57:16","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"supplement","size":116021,"visible":true,"origin":"","legend":"","description":"","filename":"AuthorChecklistFull.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8110478/v1/db5d48df00bb90a1b9ea3a5b.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Spontaneous Myocardial Infarction in a Translational Porcine Model of Coronary Atherosclerosis: A Pilot Study","fulltext":[{"header":"Introduction","content":"\u003cp\u003eCoronary artery disease (CAD) is a serious, often life-threatening, condition commonly characterized by atherosclerosis \u003csup\u003e1\u003c/sup\u003e, \u003csup\u003e2\u003c/sup\u003e. Atherosclerosis and its complications, such as myocardial infarction, are major causes of morbidity and mortality in the developed world. Atherosclerosis is defined as a chronic vascular disease and is characterized by fibroinflammatory lipid plaques or atheroma \u003csup\u003e3\u003c/sup\u003e. Despite current treatments, there remains a critical need to develop new tools to better detect, monitor, and manage CAD \u003csup\u003e4\u003c/sup\u003e. To address this need, researchers often use large animal models - particularly porcine models - prior to human studies due to their close anatomical and physiological resemblance to the human coronary system. These models have proven to be highly translational, effectively replicating human-like plaque development, and myocardial infarction \u003csup\u003e5\u003c/sup\u003e.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eWhile earlier studies using low-density lipoprotein (LDL) receptor-knockout pigs have successfully modelled plaque formation, no spontaneous myocardial infarction has been demonstrated \u003csup\u003e6\u003c/sup\u003e, \u003csup\u003e7\u003c/sup\u003e. In this pilot study, we describe a large-animal model that replicates the progressive spectrum of CAD under natural conditions. Importantly, this pathology developed without interventional induction, distinguishing it from conventional models that require artificial triggers such as intracoronary coil placement \u003csup\u003e8\u003c/sup\u003e. Through this approach, the model effectively demonstrates the natural history of CAD in a large animal model. By allowing for spontaneous myocardial infarction through biologically driven disease progression - from intimal infiltration by lipid-laden macrophages (foam cells) to mature fibrofatty plaque formation, endothelial disruption with resulting thrombosis, and consequent myocardial infarction - this model offers a powerful platform for studying plaque instability, evaluating interventions, and translating findings into clinical applications. Additionally, the inter-animal variability observed, despite uniform treatment, reflects the heterogeneity characteristic of human CAD, further enhancing the model\u0026apos;s translational relevance.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003eWe report a progressive, temporally mapped spectrum of CAD in a streptozotocin (STZ)-induced swine model of diabetes. Using multimodal assessment, including serum biomarkers, invasive coronary angiography, intravascular optimal coherence tomography (OCT), and histopathology, we documented distinct stages of disease development across four animals (P01 to P04), which were specifically reared with the hope that they developed atherosclerotic disease that could be assessed using multimodality imaging and histological techniques.\u003c/p\u003e\n\u003ch3\u003eBlood Biochemistry and Metabolic Monitoring\u003c/h3\u003e\n\u003cp\u003eTo monitor systemic metabolic status and evaluate the progression of diabetic and atherosclerotic pathology, blood glucose testing was performed on all four STZ-induced diabetic pigs (P01 to P04), and at their final assessment, a full biochemical profile was performed. The biochemical analysis included postprandial glucose levels, lipid profiles (total cholesterol (TC), LDL-C, high-density lipoprotein-cholesterol (HDL-C), triglycerides) (Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e); and liver function enzymes (alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), gamma-glutamyl transferase (GGT), lactate dehydrogenase (LDH), albumin, globulin), renal function markers (creatinine, urea), inflammatory markers [C-reactive protein (CRP)] (Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e). These parameters were selected to assess the physiological impact of chronic hyperglycaemia and an atherogenic diet on vital organ systems and to correlate biochemical changes with coronary imaging and myocardial histological findings.\u003c/p\u003e\n\u003ctable id=\"Tab1\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eGlucose Metabolism and Lipid Profile. Normal porcine ranges taken from \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e9\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003eGlucose Metabolism\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"5\"\u003e\n \u003cp\u003eLipid Profile\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003ePig\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGlucose (4.7\u0026ndash;8.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTC\u003c/p\u003e\n \u003cp\u003e(4.0-6.5)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eLDL-C\u003c/p\u003e\n \u003cp\u003e(2.5-4.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHDL-C\u003c/p\u003e\n \u003cp\u003e(1.2\u0026ndash;2.2)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTC/HDL-C (2.5\u0026ndash;4.5)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTriglyceride\u003c/p\u003e\n \u003cp\u003e(1.8-4.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e(mmol/L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003emmol/L\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003emmol/L\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003emmol/L\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003emmol/L\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eP01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e17.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e14.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eP02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e28.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e25.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e21.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eP03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e21.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e22.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e18.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eP04\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e13.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e36.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e32.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e9.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.9\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003c/p\u003e\n\u003cp\u003eAll pigs exhibited marked hyperglycaemia following STZ administration, confirming the successful induction of diabetes. Notably, P02 and P03 demonstrated the highest glucose levels (Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). Persistent hyperglycemia is known to contribute to endothelial dysfunction and accelerate atherosclerotic development \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e10\u003c/span\u003e\u003c/sup\u003e. Insulin therapy was administered intermittently to maintain glucose levels within a manageable range and to prevent complications, such as ketoacidosis \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e11\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003eTotal cholesterol and LDL-C levels were markedly elevated across all animals, with P04 displaying the most extreme lipid profiles, as illustrated in Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e. The atherogenic index, calculated as the TC to HDL-C ratio, exceeded 4.5 in all four pigs. These severe lipid abnormalities closely paralleled the worsening atherosclerotic coronary artery pathology observed in imaging and histological assessments. Such dyslipidaemia is known to promote atheroma development through various mechanisms including foam cell formation, endothelial dysfunction, and sustained vascular inflammation \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e12\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n \u003cdiv align=\"left\" class=\"colspec\"\u003e\u003cstrong\u003eTable 2. Liver Enzymes, Renal Function, Protein and Inflammatory Markers. Normal porcine ranges taken from\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u003csup\u003e9\u003c/sup\u003e\u003c/strong\u003e\u003c/div\u003e\n \u003cdiv align=\"left\" class=\"colspec\"\u003e\u003cimg 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\"\u003e\u003c/div\u003e\n \u003cdiv align=\"left\" class=\"colspec\"\u003e\u003cbr\u003e\u003c/div\u003e\n \u003cdiv align=\"left\" class=\"colspec\"\u003eLiver enzymes, including ALT, AST, ALP, GGT, and LDH, were elevated across several animals, with the most striking abnormalities observed in P04 (Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e). ALT and AST elevations in P04 (134 U/L and 165 U/L, respectively; both above the upper reference limits): whilst this elevation would normally signify hepatocellular injury, in P04 they were thought to be reflective of a recent myocardial infarction, further supported by the profoundly elevated LDH level, reaching 2525 U/L.\u003c/div\u003e\n\u003c/div\u003e\n\u003cp\u003eThe serum biochemistry profile also demonstrated consistently low albumin levels (10\u0026ndash;13 g/L, below the normal range of 19\u0026ndash;39 g/L) across all pigs, while globulin concentrations remained within the reference range (46\u0026ndash;59 g/L). This albumin/globulin imbalance, most pronounced in P04, can be a hallmark of chronic systemic inflammation and impaired hepatic protein synthesis, frequently associated with advanced diabetes-related pathology \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e12\u003c/span\u003e\u003c/sup\u003e. Creatinine values were within or close to the reference range (99\u0026ndash;148 \u0026micro;mol/L, normal 141\u0026ndash;239 \u0026micro;mol/L), though P01 was slightly below and P02 - P03 sat toward the lower-normal boundary. Urea levels (4.7\u0026ndash;7.2 mmol/L) remained within the expected physiological range.\u003c/p\u003e\n\u003cp\u003eP01 and P03 showed CRP values below the detection limit (\u0026lt;\u0026thinsp;0.15 mg/L), while P02 and P04 recorded low concentrations of 0.18 mg/L and 0.22 mg/L, respectively. All values were well within the normal physiological range for pigs (5\u0026ndash;30 mg/L \u003csup\u003e9\u003c/sup\u003e). These findings suggest that the animals were in a stable physiological state, and any vascular changes observed are unlikely to have been influenced by acute inflammation.\u003c/p\u003e\n\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\n \u003ch2\u003eAngiograms, OCT imaging and histopathological analysis\u003c/h2\u003e\n \u003cp\u003eCoronary evaluation using invasive coronary angiography, intravascular OCT imaging, and postmortem histology was conducted on all four pigs. In P01, imaging and histology revealed largely normal coronary architecture with almost no atherosclerotic changes. Figure \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e (a) demonstrates a smooth interventricular artery with no evidence of luminal irregularities, findings corroborated by OCT imaging (Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e (b)), which showed a lumen with clearly delineated vessel wall layers and no evidence of plaque. The optical signal penetrated uniformly, with no signs of fibrotic or calcified tissue, microchannels, or inflammatory infiltration. Histological examination (Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e (c)) further confirmed preserved arterial architecture, including an intact intima, tunica media and adventitia, and normal adjacent myocardial and epicardial tissues. Biochemically, P01 exhibited relatively mild metabolic disturbance: fasting glucose levels confirmed hyperglycaemia, while lipid abnormalities remained moderate. Collectively, these findings indicate no evidence of cardiovascular or systemic organ damage, positioning P01 as a representative of the baseline or pre-atherosclerotic stage in the diabetic model.\u003c/p\u003e\n \u003cp\u003eP02 demonstrated early signs of coronary atherosclerosis. Coronary angiography revealed subtle luminal irregularities in the left circumflex (LCX) artery (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e (a)). The corresponding OCT imaging (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e (b)) showed a mildly irregular lumen contour, with eccentric neointimal hyperplasia. The intima appeared mostly intact but exhibited focal discontinuities, likely reflecting endothelial disruption. The media remained thickened yet structurally organized, and the adventitia showed no evidence of inflammation. These features aligned closely with histological findings (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e (c)), which confirmed the presence of early intimal changes in the absence of mature plaque formation. Biochemically, P02 exhibited more pronounced metabolic dysfunction than P01, including elevated glucose levels. Taken together, the imaging, histological, and biochemical data support P02 as a representative of early coronary atherosclerosis.\u003c/p\u003e\n \u003cp\u003eP03 exhibited clear evidence of established CAD as can be seen in Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e. Angiographic imaging (Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e (a)) revealed luminal narrowing, particularly involving the LCX artery. The OCT analysis (Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e (b)) confirmed the presence of atherosclerotic disease, demonstrating a markedly abnormal vascular architecture with luminal stenosis and eccentric intimal thickening consistent with an atherosclerotic plaque. The fibrofatty plaque contained abundant lipid-laden macrophages (foam cells) and fibrous connective tissue, appearing in the OCT images as a homogeneous region of densely packed scatterers with uniform speckles [6\u0026ndash;10 o\u0026rsquo;clock in Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e (b)]. Within this region, signal attenuation and dropout suggested the presence of a necrotic core. Histological examination shown in Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e (c) corroborated the imaging, revealing a fibrofatty plaque, with a necrotic core comprised of degenerating foam and medial smooth muscle cells, surrounded by hyaline degeneration and abundant mineralisation. OCT imaging demonstrated that the LCX plaque extended into the vessel lumen, resulting in a visible reduction in luminal area. Biochemically, P03 exhibited elevated lipid levels, including a high TC/HDL-C ratio and increased triglycerides compared with the other animals. Hence, P03 represented a state of active fibrofatty plaque formation and highlighting P03 as a representative model of established atherosclerosis.\u003c/p\u003e\n \u003cp\u003eP04 exhibited the most advanced stage of coronary pathology among the studied cohort, intravascular thrombosis leading to spontaneous myocardial infarction. Angiography shown in Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e (a) revealed subtotal luminal occlusion of the interventricular (left anterior descending (LAD)-equivalent) artery. The OCT image shown in Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e (b) illustrates dense, highly backscattering material (3\u0026ndash;6 o\u0026rsquo;clock), consistent with fibrotic plaque. The vessel wall appeared markedly thickened (6\u0026ndash;12 o\u0026rsquo;clock), with a homogenous, strong signal suggestive of a fibrotic plaque. These findings were corroborated by histology shown in Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e (c), which demonstrated plaques on both sides of the vessel wall. Biochemically, P04 displayed pronounced metabolic distress, with persistently elevated glucose levels, and severely deranged liver function tests (ALT 134 U/L, LDH 2525 U/L) likely reflective of recent myocardial infarction. Collectively, P04 represents a useful animal model of advanced atherosclerotic CAD resulting in myocardial infarction.\u003c/p\u003e\n \u003cp\u003eFurther cross-sectional analysis of interventricular artery in P04, accompanied by detailed histopathological evaluation, revealed advanced and multifocal coronary pathology consistent with advanced atherosclerotic disease development and myocardial infarction (Fig. \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e).\u003c/p\u003e\n\u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eThis paper reports on a porcine model devised to demonstrate the progressive development of atherosclerotic CAD associated with diabetes, which can lead to spontaneous myocardial infarction. Using a standardized STZ-induction protocol and an atherogenic diet, we achieved metabolic dysfunction in all animals, reflected by sustained hyperglycemia, progressive dyslipidemia, and liver dysfunction. These systemic markers were correlated with advanced coronary artery pathology, as assessed by angiography, intravascular OCT, and histological evaluation.\u003c/p\u003e\u003cp\u003eA pathological continuum was observed in our pigs, mimicking the natural history of human coronary atherosclerosis. P01 exhibited mild metabolic disturbance with no detectable coronary artery plaque. P02 progressed to early vascular disease characterized by intimal infiltration of lipid-laden macrophages or foam cells, termed fatty streak formation. P03 demonstrated more advanced atherosclerosis, the fibrofatty plaque breaching the internal elastic lamina and infiltrating, and damaging, the media, with development of a necrotic core. P04, the most pathologically advanced fibrofatty atheromatous plaque, showed endothelial disruption, leading to sub totally occlusive thrombosis, and myocardial infarction. The spontaneous myocardial infarction which occurred in this model is a particularly significant and novel outcome not demonstrated in previous, similar porcine studies. It emphasizes the ability of this model to reproduce biologically-driven coronary atherosclerotic events, unlike other models that require artificial triggers \u003csup\u003e\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e\u003c/sup\u003e. The inter-animal variability in disease progression, despite identical protocols, further strengthens the translational relevance of this porcine model, as it reflects the heterogeneity found in human diabetic atherosclerosis. OCT provided detailed visualization of atheronecrotic plaque cores, and thrombosis, and proved instrumental in correlating with the development of progressively more severe arterial pathology.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003ePigs are a valuable model for studies of atherosclerosis since the human and porcine cardiovascular systems share many structural and functional similarities. Many of the differences between pigs and humans are due to their unguligrade (quadruped) and orthograde (bipedal) stance, which results in a laterally and dorsoventrally compressed thorax, respectively. Porcine hearts are valentine in shape, whereas those of humans are trapezoidal, but the ratio of heart to body weight is similar in both. While the coronary circulation of pigs resembles that of humans, the porcine heart has right coronary dominance. The diameter of great vessels such as aorta and pulmonary arteries is proportionately smaller in pigs than humans. Atria in pigs are more prominent and have thicker muscle, and the left atrium receives only two pulmonary veins, whereas humans have four. In pigs, caval veins enter the right atrium at ninety degrees, but the caval veins in humans are aligned in the same axis. The ratio of wall thickness between left and right ventricles is much higher in pigs than humans and trabeculae carnae and papillary muscles are coarser and broader in the porcine heart. Pigs also have a prominent trabecula septomarginalis (moderator band), which contains Purkinje fibres from the atrioventricular bundle, and connects the septal to the free wall of the right ventricle \u003csup\u003e\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u003c/sup\u003e, \u003csup\u003e14\u003c/sup\u003e. In addition, of the domestic animal species, only the pig develops atherosclerosis with any consistency, with fibrofatty plaques leading to stenosis of coronary arteries in animals 8 to 12 years of age. While the degree of vascular lipid deposition can be accelerated by feeding a high fat diet, lesions rarely reach the severity of those seen in humans. Moreover, unlike in the present study, vascular lesions do not routinely result in occlusive thrombosis and its sequelae, principally myocardial infarction \u003csup\u003e\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eIn previous studies of porcine diabetic coronary atherosclerosis (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e), there has been considerable variation in the signalment (breed and age), the dose of STZ used to induce diabetes, constituents of the high fat diet, and protracted duration of the experimental period (which incurs a high cost for conduct of these experiments). The most common large pig breeds used in these studies are the Yorkshire \u003csup\u003e16 17 18 19\u003c/sup\u003e, Yorkshire-Landrace crosses \u003csup\u003e20 21\u003c/sup\u003e and Landrace \u003csup\u003e\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e\u003c/sup\u003e, while a few studies have used mini-pigs \u003csup\u003e23 24 25\u003c/sup\u003e. Unlike these earlier models, the present study achieved the full pathological continuum - from early endothelial changes to spontaneous myocardial infarction, using comparatively less STZ (30 mg/kg on Days 1 and 2, followed by 60 mg/kg on Day 3).\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eSummary of previous porcine diabetes/high fat diet studies of coronary atherosclerosis.\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"7\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eStudy\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eYear\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eDuration\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003ePig Strain\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eDiabetes Induction/ Insulin Management\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eDiet Composition\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eKey Outcomes / Findings\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003csup\u003e23\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2012\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e20 weeks\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eSinclair mini pigs\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-STZ 120 mg/kg via splenic/gastroduodenal arteries; repeated if glucose\u0026thinsp;\u0026lt;\u0026thinsp;10 mmol/L, -Insulin Management: Not Specified\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eHigh-fat, high-cholesterol (chol.) (Teklad Diet.96366)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eConsistent diabetes; carotid stenosis led to localized vulnerable plaques\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003csup\u003e26\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2021\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e15 months\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eG\u0026ouml;ttingen \u0026amp; Landrace-Yorkshire\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-STZ 75\u0026ndash;180 mg/kg intravenous (IV),\u003c/p\u003e \u003cp\u003e-Insulin Management: Not Specified\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e25% fat, 1% chol., 10% sucrose, 15% fructose\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eDM* group showed advanced coronary atherosclerosis vs controls\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003csup\u003e16\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2021\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e60 weeks\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eYorkshire\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-STZ 50 mg/kg \u0026times; 3 days; BG\u0026thinsp;\u0026gt;\u0026thinsp;350 mg/dL,\u003c/p\u003e \u003cp\u003e-Insulin Management: Adjusted by glucose\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.5-4% chol., 15\u0026ndash;25% lard; target BG 150\u0026ndash;350 mg/dL, TC 500\u0026ndash;700 mg/dL\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e2.5\u0026times; more plaques in DM group; greater lesion complexity\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003csup\u003e17\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2017\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e20 weeks\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eYorkshire\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-STZ 50 mg/kg \u0026times; 3 days,\u003c/p\u003e \u003cp\u003e-Insulin Management: NPH BID; BG 350\u0026ndash;500 mg/dL\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1\u0026ndash;2% chol., 20% tallow, 0.75% cholic acid\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eLesions at 12 weeks; most severe in DM\u0026thinsp;+\u0026thinsp;high-fat group\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003csup\u003e27\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2017\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e15 months\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eYorkshire \u0026times; Landrace\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-STZ 140 mg/kg IV (single),\u003c/p\u003e\u003cp\u003e-Insulin Management: Insulin to avoid ketosis\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e10% sucrose, 15% fructose, 25% lard, 1% chol., 0.7% bile salts\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eChronic hyperglycemia; moderate atheroma formation\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003csup\u003e28\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2011\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e30 weeks\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eYorkshire\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-STZ 50 mg/kg \u0026times; 3 days,\u003c/p\u003e\u003cp\u003e-Insulin Management: Not Specified\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.5% chol. + 15% lard (diet-induced hyperlipidemia)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eThin-cap fibroatheromas localized to low-shear regions\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003csup\u003e18\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e4\u0026ndash;48 weeks\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eYorkshire\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-STZ 50 mg/kg \u0026times; 3 days,\u003c/p\u003e\u003cp\u003e-Insulin Management: Not Specified\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.5% chol. + 15% lard vs normal diet\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eDiabetic\u0026thinsp;+\u0026thinsp;hyperlipidemic pigs had 3\u0026ndash;4\u0026times; BG** and advanced plaques\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003csup\u003e19\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2008\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e8\u0026ndash;30 weeks\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eYorkshire\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-STZ 50 mg/kg \u0026times; 3 days,\u003c/p\u003e \u003cp\u003e-Insulin Management: Not Specified\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.5% chol. + 15% lard\u0026thinsp;+\u0026thinsp;sucrose\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eLow shear predicted future high-risk plaque regions\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003csup\u003e29\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2016\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e24 weeks\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eYorkshire\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-STZ 150\u0026ndash;200 mg/kg IV,\u003c/p\u003e\u003cp\u003e-Insulin Management: Not Specified\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e15% lard\u0026thinsp;+\u0026thinsp;1.5% chol.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eOne pig developed coronary plaque visible via IVUS/ICG***\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003csup\u003e30\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2008\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1\u0026ndash;20 weeks\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eLarge White\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-STZ 150\u0026ndash;200 mg/kg IV; with glucose infusion to avoid hypoglycemia,\u003c/p\u003e \u003cp\u003e-Insulin Management: Mixed insulin regimen; BG 150\u0026ndash;250 mg/dL\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eNormal diet\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eStable Type 1Diabetes Mellitus model with controlled glycemia\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003csup\u003e20\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2014\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e7 weeks\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eYorkshire \u0026times; Swedish Landrace\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-STZ 150 mg/kg IV,\u003c/p\u003e\u003cp\u003e-Insulin Management: Gradual SC insulin up to 1 IU/kg/day\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eNormal diet\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eRestored carb/fat/amino acid balance post-treatment\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003csup\u003e31\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2015\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e15 months\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eLandrace \u0026times; Yorkshire\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-STZ 140 mg/kg IV,\u003c/p\u003e\u003cp\u003e-Insulin Management: Insulin only for ketosis\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e25% fat\u0026thinsp;+\u0026thinsp;1% chol.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eEarly microvascular damage and renal pathology\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003csup\u003e32\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2017\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e12\u0026ndash;36 weeks\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eMiniature pigs\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-No STZ was used,\u003c/p\u003e\u003cp\u003e-Insulin Management: Not Specified\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e4% chol., 14% tallow, 2% bile salts (1 kg/day)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eAtheroma formation linked to Cyclooxygenase down regulation\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\n\u003ch3\u003e* DM: Diabetes Mellitus, **BG: Blood Glucose, ***IVUS: Intravascular Ultrasound, ICG: Indocyanine Green\u003c/h3\u003e\n\u003cp\u003eOne notable aspect of this study is that multiple techniques, including coronary angiography, OCT, and histological staining, enabled comprehensive visualization of disease progression. OCT imaging, achieved using a custom-designed, 3D-printed imaging catheter \u003csup\u003e\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e\u003c/sup\u003e, enabled \u003cem\u003ein vivo\u003c/em\u003e imaging to identify lipid core and necrotic regions of plaque that are not visible by conventional coronary angiography \u003csup\u003e\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u003c/sup\u003e. The combined application of OCT and fluoroscopic angiography enabled precise lesion mapping across epicardial coronary arteries - the right coronary artery (RCA), interventricular artery (analogous to the human LAD artery), and LCX - which enabled comprehensive assessment of the atherosclerotic process.\u003c/p\u003e\u003cp\u003eThis pilot study has several limitations. The small sample size (n\u0026thinsp;=\u0026thinsp;4) restricts the generalizability of the findings and prevents robust statistical analysis, even though it allowed observation of the full disease spectrum from minimal atherosclerotic pathology to spontaneous myocardial infarction. Additionally, the exclusive use of male Yorkshire pigs limits the ability to assess sex- or breed-related differences in metabolic response, plaque development, and susceptibility to myocardial infarction. Finally, the study was not designed to assess therapeutic interventions or validate interventional imaging performance: it was an observational study designed to assess the utility of a new intravascular imaging catheter at recognizing atherosclerosis, and future larger studies are required to establish the model\u0026rsquo;s reproducibility and suitability for translational research.\u003c/p\u003e\u003cp\u003eOverall, this study provides foundational evidence for a preclinical platform capable of capturing the full pathological spectrum of diabetic CAD, from subclinical changes to overt myocardial infarction. The model holds promise for future studies aimed at evaluating plaque instability, testing therapeutic interventions, and investigating mechanisms underlying diabetic cardiovascular disease.\u003c/p\u003e"},{"header":"Methods","content":"\u003cp\u003eAll animal procedures in this study were conducted in accordance with the \u003cem\u003eAustralian Code for the Care and Use of Animals for Scientific Purposes\u003c/em\u003e (8th edition, 2013; updated 2021) \u003csup\u003e\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e\u003c/sup\u003e and were approved by the South Australian Health and Medical Research Institute (SAHMRI) Animal Ethics Committee (protocol number SAM-21-011). All methods were carried out in accordance with relevant guidelines and regulations. Reporting of animal procedures followed the ARRIVE (Animal Research: Reporting of In Vivo Experiments) guidelines\u003csup\u003e\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\n\u003ch3\u003eAnimal Model and Induction Protocol\u003c/h3\u003e\n\u003cp\u003eFour male (n\u0026thinsp;=\u0026thinsp;4) Yorkshire swine (designated P01 to P04), age between 9 to 12 weeks, each weighing approximately 25\u0026ndash;30 kg at baseline, were used in this study. As this was an observational, exploratory pilot study performed as a part of a larger study assessing the utility of a new intravascular imaging catheter, and hence no formal sample-size calculation was performed, nor was a control group required. Randomisation was not applicable, as all animals underwent the same induction protocol and imaging schedule without allocation to different treatment groups. Histological assessments were performed by an experienced pathologist blinded to the imaging findings, while other aspects of the study were not blinded because all animals received identical procedures. Diabetes mellitus (DM) was chemically induced using a STZ protocol tailored to mimic Type 1-like insulin deficiency. Animals received intravenous STZ at a dose of 30 mg/kg 28 on Days 1 and 2, followed by 60 mg/kg on Day 3. Successful diabetic induction was confirmed by persistent fasting blood glucose levels exceeding 8.3 mmol/L. Seven pigs were initially enrolled. The first three animals were used exclusively to develop and optimise the STZ protocol and did not have the full set of blood results required for the planned analyses. Accordingly, these three animals were not included in the final dataset. The study cohort therefore comprised the remaining four animals (P01-P04), which exhibited the full pathological spectrum relevant to this study and were analysed as reported. No animals or individual data points from P01-P04 were excluded from the analyses.\u003c/p\u003e\u003cp\u003eFollowing induction, all animals were transitioned to a high-fat, high-carbohydrate atherogenic diet designed to promote dyslipidaemia and accelerate atherosclerotic CAD development and progression. Animals were housed under standardized conditions and monitored daily for health status. Serum glucose levels were regularly measured, and short-acting insulin was administered as needed to maintain glucose levels below 30 mmol/L, preventing ketoacidosis. Monthly assessments included body weight, blood glucose concentration, general clinical condition, and appetite. The outcome of interest was the development of atherosclerotic coronary disease as assessed by invasive coronary angiography, OCT, and histological analysis.\u003c/p\u003e\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\u003ch2\u003eAnalgesia and Anaesthesia Protocol\u003c/h2\u003e\u003cp\u003eTo ensure animal welfare, analgesia and anaesthesia protocols were rigorously followed. A fentanyl transdermal patch (25\u0026ndash;50 \u0026micro;g/h) was applied 24 hours prior to any imaging procedure for sustained pain control. On the day of imaging, meloxicam (0.4 mg/kg, intramuscular) was administered as supplemental analgesia. Anaesthesia was induced using intramuscular xylazine (2 mg/kg) and ketamine (20 mg/kg) and maintained via inhaled isoflurane (4\u0026ndash;5% for induction, 2\u0026ndash;3% for maintenance) in 100% oxygen using mechanical ventilation.\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eIntravascular Imaging and Angiography\u003c/h3\u003e\n\u003cp\u003eCoronary imaging was conducted to assess the progression of atherosclerotic pathology \u003cem\u003ein vivo\u003c/em\u003e. P01 was imaged at Day 124, P02 at Days 163, 213, and 318, P03 at Days 107 and 282, and P04 at Day 240. These time points were selected to represent early, mid, and late stages of disease development. Vascular access was established via the right femoral artery using a 6 French introducer sheath. Coronary angiography was performed using an AL2 guiding catheter under fluoroscopic guidance. An iodinated contrast agent (Ultravist\u0026reg; (iopromide) 370 mg lodine/mL, Bayer, Germany) was injected at a rate of ~\u0026thinsp;4mL/s to delineate coronary anatomy, assess luminal patency, and guide intravascular device positioning.\u003c/p\u003e\u003cp\u003eFor intravascular OCT imaging, a custom-designed, 3D-printed imaging catheter \u003csup\u003e\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e\u003c/sup\u003e was advanced over a 0.014\" coronary guidewire (Runthrough NS Floppy, Terumo, Japan) into the target artery under fluoroscopic guidance. Automated pullback OCT imaging was performed during contrast flushing at 4 mL/s, enabling detailed visualization of luminal structures and vessel wall composition, including detection and composition of plaque. Systemic anticoagulation was maintained throughout the procedure with intravenous heparin at a dose of 100 IU/kg to prevent intraprocedural coronary thrombosis, with no acute complications. OCT images were acquired using a Telesto II spectral-domain OCT scanner (Thorlabs GmbH, Germany), with a central wavelength of 1300 nm and axial resolution of 4.2 \u0026micro;m (in water).\u003c/p\u003e\n\u003ch3\u003eTissue Collection and Histological Analysis\u003c/h3\u003e\n\u003cp\u003eFollowing final imaging, each animal was humanely euthanized under deep anaesthesia. Coronary arteries (interventricular artery, LCX, and RCA) were excised, perfused with 10% neutral buffered formalin, and submitted for gross and histological evaluation. Coronary blood vessels were transversely cut at serial, standard intervals, then paraffin-embedded, and 6\u0026micro; sections cut and stained with hematoxylin and eosin (H\u0026amp;E). Hearts were incised and immersion fixed in a large volume of 10% neutral buffered formalin and processed for histopathological examination. Histopathological findings were correlated with imaging and biochemical data to confirm the stage and severity of coronary artery atheroma development.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003eAcknowledgements\u003c/p\u003e\n\u003cp\u003eWe acknowledge Dr Ayla Hoogendoorn, Dr. Chris Christou, Ms. Loren Matthews, Ms. Georgia Williams, Ms. Lisa McKenny, Mr. Robb Muirhead, Mr. Jim Manavis, Ms. Sofie Kogoj, and Ms. Yvonne Ciuk for contributing to the management of the diabetic swine, assistance during imaging procedures, and histological preparation and analysis of histology images. We also acknowledge the support of the Optofab node of the Australian National Fabrication Facility and Adelaide Microscopy.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAuthor\u0026nbsp;contributions\u0026nbsp;statement\u003c/p\u003e\n\u003cp\u003eA.K.K: Conceptualization, Probe Fabrication, Methodology, Investigation, Formal analysis, Visualization, Writing - Original Draft. J.A.M.: Methodology, Data curation, Investigation, Writing - Review \u0026amp; Editing. \u003cstrong\u003eJ.F.\u003c/strong\u003e: Histological analysis, Interpretation of pathology, Writing - Review \u0026amp; Editing. \u003cstrong\u003eL.X.\u003c/strong\u003e: Optical system development, Imaging acquisition, Writing - Review \u0026amp; Editing. \u003cstrong\u003eP.R.\u003c/strong\u003e: Lens fabrication, Imaging support, Writing - Review \u0026amp; Editing. \u003cstrong\u003eJ.T.M.T.\u003c/strong\u003e: Experimental planning, Data interpretation, Writing - Review \u0026amp; Editing. \u003cstrong\u003eC.A.B.\u003c/strong\u003e: Supervision, Conceptual guidance, Writing - Review \u0026amp; Editing. \u003cstrong\u003eJ.V.\u003c/strong\u003e: Clinical interpretation, Imaging validation, Writing - Review \u0026amp; Editing. \u003cstrong\u003eR.A.M.\u003c/strong\u003e: Supervision, Optical imaging oversight, Writing - Review \u0026amp; Editing. \u003cstrong\u003eH.G.\u003c/strong\u003e: Funding acquisition, OCT probe design consultation, Technical input, Writing - Review \u0026amp; Editing. \u003cstrong\u003eP.J.P.\u003c/strong\u003e: Conceptualization, Funding acquisition, Cardiovascular expertise, Project supervision, Writing - Review \u0026amp; Editing. \u003cstrong\u003eJ.L.\u003c/strong\u003e: Conceptualization, Funding acquisition, Supervision, Project administration, Writing - Review \u0026amp; Editing.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eFunding\u003c/p\u003e\n\u003cp\u003eThis work is supported by the National Health and Medical Research Council Development (Grant No. 2022337), the Investigator Grant (Grant No. 2008462), the Heart Foundation Future Leader Fellowships (Grant Nos. 105608 and 106656), and Catalyst (Grant No. 108710-2024_CPG), the Hospital Research Foundation Project (Grant No. 2022-CP-IDMH-014-83100), Australia-Germany Joint Research Co-operation Scheme (UA-DAAD), Baden-Wuerttemberg-Stiftung (Opterial), European Research Council (Advanced Grant Complexplas, PoC Grant 3DPrintedOptics), Bundesministeriumf\u0026uuml;r Bildung und Forschung (3DPrintedOptics, Integrated3Dprint, QR.X, QR.N), Deutsche Forschungsgemeinschaft (German Research Foundation) (Grant No. 431314977/GRK2642), HORIZON EUROPE European Innovation Council (Grant No. IV-Lab 101115545), Carl-Zeiss Foundation (EndoPrint3D, QPhoton), and University of Stuttgart (Terra Incognita).\u003c/p\u003e\n\u003cp\u003eCompeting interests\u003c/p\u003e\n\u003cp\u003eP. J. Psaltis has received research support from Abbott Vascular; consulting fees from Amgen, Esperion, Eli Lilly, Novartis, Novo Nordisk, and Sanofi; and speaker honoraria from Amgen, AstraZeneca, Bayer, Boehringer Ingelheim, Merck Schering-Plough, Pfizer, Novartis, Novo Nordisk, and Sanofi.\u0026nbsp;R. A. McLaughlin is a co-founder and Director of Miniprobes Pty Ltd, a company that develops optical imaging systems. Miniprobes Pty Ltd did not contribute to or participate in this study. J. Li is the founder and Director of Theia Medical Pty Ltd, a company that develops medical imaging devices. Theia Medical Pty Ltd did not contribute to or participate in this study. The remaining authors declare no conflicts of interests.\u003c/p\u003e\n\u003cp\u003eAdditional Information\u003c/p\u003e\n\u003cp\u003eCorrespondence and requests for materials should be addressed to Dr Jiawen Li (J.L.).\u003c/p\u003e\n\u003cp\u003eSupplementary information accompanies this paper contains ARRIVE checklist as a compliance.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAronson, D. \u0026amp; Edelman, E. R. Coronary artery disease and diabetes mellitus. \u003cem\u003eCardiol Clin\u003c/em\u003e \u003cstrong\u003e32\u003c/strong\u003e, 439-455 (2014). https://doi.org/10.1016/j.ccl.2014.04.001\u003c/li\u003e\n\u003cli\u003eRajbhandari, J., Fernandez, C. J., Agarwal, M., Yeap, B. X. Y. \u0026amp; Pappachan, J. M. 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(National Health and Medical Research Council, Canberra, 2021).\u003c/li\u003e\n\u003cli\u003ePercie du Sert, N., Hurst, V., Ahluwalia, A. The ARRIVE guidelines 2.0: Updated guidelines for reporting animal research. \u003cem\u003eBMC Veterinary Research\u003c/em\u003e \u003cstrong\u003e16\u003c/strong\u003e (2020). https://doi.org/https://doi.org/10.1186/s12917-020-02451-y\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-8110478/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8110478/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThis pilot study characterizes the spectrum of atherosclerotic coronary artery pathologies in a porcine model of streptozotocin-induced diabetes, with a particular focus on the natural progression of atherosclerotic plaques. Four pigs (designated P01 - P04) were subjected to an identical streptozotocin regimen to induce diabetes and studied using blood biochemistry, lipid profiling, coronary angiography, optical coherence tomography, and histological analysis of the coronary arteries to assess for the development of coronary atherosclerosis. All animals developed sustained hyperglycemia and dyslipidemia, with total cholesterol levels ranging from 14.9 to 36.7 mmol/L and low-density lipoprotein concentrations reaching up to 32 mmol/L. Notably, the study captured the full pathological continuum: from nearly no coronary abnormality in P01 and early atheromatous plaque formation in P02, to advanced atherosclerosis in P03, and finally, in P04, severe fibrofatty atheroma, and myocardial infarction. Unlike conventional animal myocardial infarction models that rely on interventional triggers, the infarction in P04 occurred spontaneously, demonstrating the natural cascade of plaque development and acute plaque rupture with thrombosis. These findings demonstrate the feasibility of using streptozotocin-treated pigs to model key stages of diabetic coronary artery disease.\u003c/p\u003e","manuscriptTitle":"Spontaneous Myocardial Infarction in a Translational Porcine Model of Coronary Atherosclerosis: A Pilot Study","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-12-01 08:57:11","doi":"10.21203/rs.3.rs-8110478/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-02-13T03:12:48+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-01-27T20:24:56+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-01-14T10:06:23+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"42138334838523673202886482284240896142","date":"2026-01-12T19:31:30+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"330935488789149923096371197020639838477","date":"2026-01-06T10:23:00+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"309741219654001281076604654509259006145","date":"2025-12-28T19:26:50+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"337414884734053472134599512712384519312","date":"2025-11-28T12:26:59+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-11-28T11:37:52+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-11-28T05:40:04+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2025-11-26T14:27:18+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-11-23T23:47:21+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2025-11-23T23:44:24+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"e3dc5f12-f445-4e80-81af-11fc0bf22487","owner":[],"postedDate":"December 1st, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[{"id":58844170,"name":"Health sciences/Cardiology"},{"id":58844171,"name":"Health sciences/Diseases"}],"tags":[],"updatedAt":"2026-05-20T07:38:26+00:00","versionOfRecord":[],"versionCreatedAt":"2025-12-01 08:57:11","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8110478","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8110478","identity":"rs-8110478","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}