Araújo Rita, Bernardino Raquel L, Monteiro Mariana P, Gomes Pedro S
BoneLab, Faculdade de Medicina Dentária, Universidade do Porto, Rua Dr. Manuel Pereira da Silva, 4200-393, Porto, Portugal.
LAQV/REQUIMTE, Faculdade de Medicina Dentária, Universidade do Porto, Rua Dr. Manuel Pereira da Silva, 4200-393, Porto, Portugal.
Mol Med. 2025 May 17;31(1):194. doi: 10.1186/s10020-025-01251-0.
Bone fragility due to poor glycemic control is a recognized complication of diabetes, but the mechanisms underlying diabetic bone disease remain poorly understood. Despite the importance of bioenergetics in tissue functionality, the impact of hyperglycemia on bone bioenergetics has not been previously investigated.
To determine the effects of high glucose exposure on energy metabolism and structural integrity in bone tissue using an ex vivo organotypic culture model of embryonic chick femur.
Femora from eleven-day-old Gallus gallus embryos were cultured for eleven days under physiological glucose conditions (5.5 mM, NG), chronic high glucose exposure (25 mM, HG-C), or acute high glucose exposure (25 mM, HG-A). Bioenergetic assessments (Seahorse assays), proteomic analysis (liquid chromatography-mass spectrometry), histomorphometric and microtomographic evaluations, and oxidative stress measurements (carbonyl content assay) were performed. Statistical analyses were conducted using IBM® SPSS® Statistics (v26.0). The Mann-Whitney nonparametric test was used for group comparisons in microtomographic analysis, ALP activity, and carbonyl content assays. For Seahorse assay results, ANOVA with Tukey's post-hoc test was applied after confirming data homoscedasticity with Levene's test.
Chronic high glucose exposure reduced bone mineral deposition, altered histomorphometric indices, and suppressed key osteochondral development regulators. Acute high glucose exposure enhanced glycolysis and oxidative phosphorylation, while chronic exposure caused oxygen consumption uncoupling, increased ROS generation, and downregulated mitochondrial proteins critical for bioenergetics. Elevated oxidative stress was confirmed in the chronic high glucose group.
Chronic high glucose exposure disrupted bone bioenergetics, induced mitochondrial dysfunction, and compromised bone structural integrity, emphasizing the metabolic impact of hyperglycemia in diabetic bone disease.
血糖控制不佳导致的骨脆性是糖尿病公认的并发症，但糖尿病骨病的潜在机制仍知之甚少。尽管生物能量学在组织功能中很重要，但高血糖对骨生物能量学的影响此前尚未得到研究。
使用鸡胚股骨的体外器官型培养模型，确定高糖暴露对骨组织能量代谢和结构完整性的影响。
将11日龄鸡胚的股骨在生理葡萄糖条件（5.5 mM，NG）、慢性高糖暴露（25 mM，HG-C）或急性高糖暴露（25 mM，HG-A）下培养11天。进行生物能量评估（海马实验）、蛋白质组分析（液相色谱-质谱联用）、组织形态计量学和显微断层扫描评估以及氧化应激测量（羰基含量测定）。使用IBM® SPSS® Statistics（v26.0）进行统计分析。在显微断层扫描分析、碱性磷酸酶活性和羰基含量测定中，采用曼-惠特尼非参数检验进行组间比较。对于海马实验结果，在使用莱文检验确认数据同质性后，应用带有图基事后检验的方差分析。
慢性高糖暴露减少了骨矿物质沉积，改变了组织形态计量学指标，并抑制了关键的骨软骨发育调节因子。急性高糖暴露增强了糖酵解和氧化磷酸化，而慢性暴露导致氧消耗解偶联，增加了活性氧生成，并下调了对生物能量学至关重要的线粒体蛋白。慢性高糖组氧化应激升高得到证实。
慢性高糖暴露破坏了骨生物能量学，诱导线粒体功能障碍，并损害了骨结构完整性，强调了高血糖在糖尿病骨病中的代谢影响。
