Tilakaratne Aruni, Soory Mena
Department of Oral Medicine and Periodontology, University of Peradeniya Sri-Lanka.
Recent Pat Endocr Metab Immune Drug Discov. 2012 Jan;6(1):73-84. doi: 10.2174/187221412799015281.
To study redox responses of cultured osteoblasts, mediated by bacterial lipopolysaccharide (LPS), glucose (G), glucose-oxidised low density lipoprotein (GLDL) and minocycline (M) using radiolabelled steroid markers of redox status and wound healing. The clinical relevance of this concept in periodontitis patients with cardiometabolic risk markers is addressed.
A well differentiated osteoblastic cell-line was cultured in Eagle's MEM in confluent monolayer, in 24 well multiwell plates. Radiolabelled testosterone was used as the steroid substrate. Experiments were set up with controls in the absence of agents, optimal concentrations (previously determined) of G, GLDL, LPS, M, GLDL+LPS and the latter combined with M (n = 8). At the end of a 24h incubation period, the reaction was terminated and the medium analysed for yields of the steroid metabolite 5α-dihydrotestosterone (DHT), the redox marker relevant to wound healing, the weaker androgen 4-androstenedione (4-A) and the diols. Analysis entailed thin layer chromatography and radioisotope scanning.
The yields of DHT showed 1.4-fold and 2.3-fold decreases in response to GLDL and LPS respectively and a 1.3-fold reduction in response to the combination, when compared with controls in the absence of agents. Minocycline stimulated the yield of DHT by 1.4-fold, and when combined with GLDL+LPS, the decreased yield was overcome and raised to 2-fold above the combination in response to the addition of minocycline (n = 8; p < 0.001), when compared with controls. The trends in the yields of 4-A and diols were inversely related to each other with increases and decreases over controls respectively, in keeping with enzymic pathways.
Decreased yields of the oxidative stress marker DHT in response to LPS, G and GLDL were overcome in the presence of minocycline, which demonstrates its potential role as an adjunctive therapeutic agent in an environment of oxidative stress. These applications could be extrapolated to periodontal disease and co-existing cardiometabolic risk markers, in the context of its antiinflammatory and antioxidant actions relevant to healing. In this paper, recent patents relevant to adjunctive therapeutic management of periodontal disease co-existing with cardiometabolic risk markers are addressed. There have been significant advances in therapeutic interventions for overcoming oxidative stress-inducing mechanisms that are common to these disease entities.
使用氧化还原状态和伤口愈合的放射性标记类固醇标志物，研究由细菌脂多糖（LPS）、葡萄糖（G）、葡萄糖氧化的低密度脂蛋白（GLDL）和米诺环素（M）介导的培养成骨细胞的氧化还原反应。探讨这一概念在伴有心脏代谢风险标志物的牙周炎患者中的临床相关性。
将一种分化良好的成骨细胞系在Eagle's MEM中培养至汇合单层，置于24孔多孔板中。使用放射性标记的睾酮作为类固醇底物。实验设置了无试剂对照、G、GLDL、LPS、M、GLDL+LPS的最佳浓度（先前已确定）以及后者与M联合使用的情况（n = 8）。在24小时孵育期结束时，终止反应，并分析培养基中类固醇代谢物5α-二氢睾酮（DHT）的产量，DHT是与伤口愈合相关的氧化还原标志物，较弱的雄激素4-雄烯二酮（4-A）以及二醇。分析采用薄层色谱法和放射性同位素扫描。
与无试剂对照相比，DHT的产量在响应GLDL和LPS时分别下降了1.4倍和2.3倍，在响应联合使用时下降了1.3倍。米诺环素使DHT的产量提高了1.4倍，当与GLDL+LPS联合使用时，添加米诺环素克服了产量下降，并使其提高到联合使用时的2倍以上（n = 8；p < 0.001），与对照相比。4-A和二醇产量的趋势彼此相反，分别比对照增加和减少，符合酶促途径。
在米诺环素存在的情况下，氧化应激标志物DHT因LPS、G和GLDL而降低的产量得到了克服，这证明了其在氧化应激环境中作为辅助治疗剂的潜在作用。鉴于其与愈合相关的抗炎和抗氧化作用，这些应用可外推至牙周疾病和并存的心脏代谢风险标志物。本文讨论了与伴有心脏代谢风险标志物的牙周疾病辅助治疗管理相关的近期专利。在克服这些疾病实体共有的氧化应激诱导机制的治疗干预方面取得了重大进展。
