Santos Thaís Aguiar, Ribeiro Jaqueline Lemes, Battistelli Luisa Souza, Anbinder Ana Lia
Institute of Science and Technology, São Paulo State University (Unesp), Av Engenheiro Francisco José Longo, 777, Jardim São Dimas, São José Dos Campos, SP, CEP: 12245-000, Brazil.
J Bone Miner Metab. 2025 Apr 18. doi: 10.1007/s00774-025-01600-3.
This study aimed to evaluate the effects of Limosilactobacillus reuteri (LR) and its combination with menaquinone-7 (MK-7; K) on ovariectomy-induced bone loss in mice and on bacterial growth in vitro.
In the in vivo study, animals were divided into five groups: sham-operated (SHAM); ovariectomy (OVX); OVX-LR; OVX-K; OVX-LR-K. After 4 weeks of treatment, femur cortical biomechanical properties, vertebral microarchitecture, osteocalcin levels, Occludin and Jam3 expression, and intestinal histomorphometry were evaluated. In vitro, microbial growth was assessed by incubating L. reuteri with MK-7. After incubation, optical densities were measured, and bacteria were cultured on MRS agar for the colony-forming unit (CFU/ml) counting.
L. reuteri, MK-7, and their combination significantly improved femur intrinsic biomechanical properties and cortical vertebral thickness. The combined treatment exhibited a synergistic effect on the modulus of elasticity, and increased cortical vertebral volume and the villus/crypt ratio in comparison to OVX. L. reuteri and its combination with MK-7 restored vertebral trabecular microarchitecture values to SHAM levels. However, no significant differences were observed in serum levels of osteocalcin, Occludin or Jam3 expression among groups. In vitro, a significant increase in optical density and viable cell count was observed after 4 h of incubation.
L. reuteri and its combination with MK-7 improved bone biomechanical and microarchitecture properties. We propose a synergistic preventive action of L. reuteri and MK-7 in estrogen-deficient mice. Additionally, the enhanced survival of L. reuteri in the presence of MK-7 may partially explain the observed benefits of the combined treatment in vivo.
本研究旨在评估罗伊氏乳杆菌（LR）及其与维生素K2（MK-7；K）联合使用对去卵巢诱导的小鼠骨质流失以及体外细菌生长的影响。
在体内研究中，将动物分为五组：假手术组（SHAM）；去卵巢组（OVX）；OVX-LR组；OVX-K组；OVX-LR-K组。治疗4周后，评估股骨皮质生物力学特性、椎体微结构、骨钙素水平、闭合蛋白和连接粘附分子3（Jam3）表达以及肠道组织形态学。在体外，通过将罗伊氏乳杆菌与MK-7共同孵育来评估微生物生长。孵育后，测量光密度，并将细菌接种于MRS琼脂上进行菌落形成单位（CFU/ml）计数。
罗伊氏乳杆菌、MK-7及其组合显著改善了股骨固有生物力学特性和皮质椎体厚度。联合治疗对弹性模量具有协同作用，与OVX组相比，增加了皮质椎体体积和绒毛/隐窝比率。罗伊氏乳杆菌及其与MK-7的组合将椎体小梁微结构值恢复到SHAM组水平。然而，各组间骨钙素、闭合蛋白或Jam3表达的血清水平未观察到显著差异。在体外，孵育4小时后观察到光密度和活菌数显著增加。
罗伊氏乳杆菌及其与MK-7的组合改善了骨生物力学和微结构特性。我们提出罗伊氏乳杆菌和MK-7在雌激素缺乏小鼠中具有协同预防作用。此外，在MK-7存在下罗伊氏乳杆菌存活率的提高可能部分解释了联合治疗在体内观察到的益处。
