Abstract：
OBJECTIVE: The present study aimed to investigate the histomorphometric and immunohistochemical impacts of vitamin K2 on guided bone regeneration (GBR) in calvarial critical-size defects (CSDs) in diabetic rats.
METHODS: A total of 30 rats were used in this study, comprising 12 non-diabetic (control) rats and 18 with streptozotocin-nicotinamide-induced experimental Diabetes mellitus (DM). In all rats, two calvarial CSDs were created: one defect was left empty (E), the other was treated with bovine-derived bone graft and collagen-based resorbable membrane (GM). Study groups were as follows: control rats administered saline (n = 6, C-E and C-GM groups) or vitamin K2 (n = 6, CK-E and CK-GM groups) and diabetic rats administered saline (n = 6, DM-E and DM-GM groups) or vitamin K2 (n = 6, DMK-E and DMK-GM groups). After 4 weeks of saline or vitamin K2 administration, the rats were euthanized. Bone defect healing and new bone formation were assessed histomorphometrically, and osteocalcin and osteopontin levels were examined immunohistochemically.
RESULTS: Percentage of new bone formation was greater in CK-GM vs. CK-E and in DMK-GM vs. DMK-E [d = 3.86 (95% CI = 16.38-28.61), d = 1.86, (95% CI = 10.74-38.58), respectively, p < .05]. Bone defect healing scores were higher in CK-GM vs. CK-E and in DMK-GM vs. DMK-E [d = 2.69 (95% CI = -2.12 to -0.87), d = 3.28 (95% CI = 0.98-1.91), respectively, p < .05]. Osteocalcin expression levels were elevated in CK-GM vs. CK-E, in DMK-GM vs. DMK-E [d = 1.19 (95% CI = 0.08-1.41), d = 1.10 (95% CI = 0.02-1.22), respectively p < .05]. Vitamin K2 enhanced osteocalcin expression levels in DMK-E vs. DM-E [d = 2.78, (95% CI = 0.56-1.53), p < .05] and in DMK-GM vs. DM-GM [d = 2.43, (95% CI = 0.65-2.10), p < .05]. Osteopontin expression was enhanced in defects treated with GM vs. E defects [C-GM vs. C-E, d = 1.56 (95% CI = 0.38-2.01); CK-GM vs. CK-E, d = 1.91 (95% CI = 0.49-1.72); DM-GM vs. DM-E, d = 2.34 (95% CI = -1.12 to -0.50); DMK-GM vs. DMK-E, d = 2.00 (95% CI = 0.58-1.91), p < .05].
CONCLUSIONS: The research findings suggest that administering vitamin K2 in GBR for rats with DM favorably impacts bone healing in CSDs, presenting an adjunctive strategy for bone regeneration.
METHODS: A total of 30 rats were used in this study, comprising 12 non-diabetic (control) rats and 18 with streptozotocin-nicotinamide-induced experimental Diabetes mellitus (DM). In all rats, two calvarial CSDs were created: one defect was left empty (E), the other was treated with bovine-derived bone graft and collagen-based resorbable membrane (GM). Study groups were as follows: control rats administered saline (n = 6, C-E and C-GM groups) or vitamin K2 (n = 6, CK-E and CK-GM groups) and diabetic rats administered saline (n = 6, DM-E and DM-GM groups) or vitamin K2 (n = 6, DMK-E and DMK-GM groups). After 4 weeks of saline or vitamin K2 administration, the rats were euthanized. Bone defect healing and new bone formation were assessed histomorphometrically, and osteocalcin and osteopontin levels were examined immunohistochemically.
RESULTS: Percentage of new bone formation was greater in CK-GM vs. CK-E and in DMK-GM vs. DMK-E [d = 3.86 (95% CI = 16.38-28.61), d = 1.86, (95% CI = 10.74-38.58), respectively, p < .05]. Bone defect healing scores were higher in CK-GM vs. CK-E and in DMK-GM vs. DMK-E [d = 2.69 (95% CI = -2.12 to -0.87), d = 3.28 (95% CI = 0.98-1.91), respectively, p < .05]. Osteocalcin expression levels were elevated in CK-GM vs. CK-E, in DMK-GM vs. DMK-E [d = 1.19 (95% CI = 0.08-1.41), d = 1.10 (95% CI = 0.02-1.22), respectively p < .05]. Vitamin K2 enhanced osteocalcin expression levels in DMK-E vs. DM-E [d = 2.78, (95% CI = 0.56-1.53), p < .05] and in DMK-GM vs. DM-GM [d = 2.43, (95% CI = 0.65-2.10), p < .05]. Osteopontin expression was enhanced in defects treated with GM vs. E defects [C-GM vs. C-E, d = 1.56 (95% CI = 0.38-2.01); CK-GM vs. CK-E, d = 1.91 (95% CI = 0.49-1.72); DM-GM vs. DM-E, d = 2.34 (95% CI = -1.12 to -0.50); DMK-GM vs. DMK-E, d = 2.00 (95% CI = 0.58-1.91), p < .05].
CONCLUSIONS: The research findings suggest that administering vitamin K2 in GBR for rats with DM favorably impacts bone healing in CSDs, presenting an adjunctive strategy for bone regeneration.
摘要：
目的：本研究旨在研究维生素K2对糖尿病大鼠颅骨临界尺寸缺损（CSDs）引导骨再生（GBR）的组织形态和免疫组织化学影响。
方法：本研究共使用30只大鼠，包括12只非糖尿病（对照）大鼠和18只链脲佐菌素-烟酰胺诱导的实验性糖尿病（DM）。在所有的老鼠中，创建了两个颅骨CSD：一个缺陷为空（E），另一种是用牛衍生的骨移植物和基于胶原蛋白的可吸收膜（GM）治疗。研究组如下:给予生理盐水(n=6,C-E和C-GM组)或维生素K2(n=6,CK-E和CK-GM组)的对照大鼠和给予生理盐水(n=6,DM-E和DM-GM组)或维生素K2(n=6,DMK-E和DMK-GM组)的糖尿病大鼠。生理盐水或维生素K2给药4周后，对大鼠实施安乐死。骨缺损愈合和新骨形成进行组织形态学评估，免疫组化检测骨钙蛋白和骨桥蛋白水平。
结果：新骨形成的百分比在CK-GM与CK-E和DMK-GM与DMK-E[d=3.86（95%CI=16.38-28.61），d=1.86，（95%CI=10.74-38.58），分别,p<.05]。骨缺损愈合评分较高的CK-GM与CK-E和DMK-GM与DMK-E[d=2.69（95%CI=-2.12至-0.87），d=3.28（95%CI=0.98-1.91），分别,p<.05]。CK-GM与骨钙蛋白表达水平升高CK-E,在DMK-GMvs.DMK-E[d=1.19（95%CI=0.08-1.41），d=1.10（95%CI=0.02-1.22），分别p<.05]。维生素K2增强DMK-E与骨钙蛋白的表达水平DM-E[d=2.78，（95%CI=0.56-1.53），p<.05]和DMK-GM与DM-GM[d=2.43，（95%CI=0.65-2.10），p<.05]。骨桥蛋白表达在用GM治疗的缺损中增强。E缺陷[C-GMvs.C-E,d=1.56（95%CI=0.38-2.01）；CK-GM与CK-E,d=1.91（95%CI=0.49-1.72）；DM-GM与DM-E,d=2.34（95%CI=-1.12至-0.50）；DMK-GM与DMK-E,d=2.00（95%CI=0.58-1.91），p<.05]。
结论：研究结果表明，在糖尿病大鼠的GBR中施用维生素K2有利于CSD的骨愈合，提出了骨再生的辅助策略。
方法：本研究共使用30只大鼠，包括12只非糖尿病（对照）大鼠和18只链脲佐菌素-烟酰胺诱导的实验性糖尿病（DM）。在所有的老鼠中，创建了两个颅骨CSD：一个缺陷为空（E），另一种是用牛衍生的骨移植物和基于胶原蛋白的可吸收膜（GM）治疗。研究组如下:给予生理盐水(n=6,C-E和C-GM组)或维生素K2(n=6,CK-E和CK-GM组)的对照大鼠和给予生理盐水(n=6,DM-E和DM-GM组)或维生素K2(n=6,DMK-E和DMK-GM组)的糖尿病大鼠。生理盐水或维生素K2给药4周后，对大鼠实施安乐死。骨缺损愈合和新骨形成进行组织形态学评估，免疫组化检测骨钙蛋白和骨桥蛋白水平。
结果：新骨形成的百分比在CK-GM与CK-E和DMK-GM与DMK-E[d=3.86（95%CI=16.38-28.61），d=1.86，（95%CI=10.74-38.58），分别,p<.05]。骨缺损愈合评分较高的CK-GM与CK-E和DMK-GM与DMK-E[d=2.69（95%CI=-2.12至-0.87），d=3.28（95%CI=0.98-1.91），分别,p<.05]。CK-GM与骨钙蛋白表达水平升高CK-E,在DMK-GMvs.DMK-E[d=1.19（95%CI=0.08-1.41），d=1.10（95%CI=0.02-1.22），分别p<.05]。维生素K2增强DMK-E与骨钙蛋白的表达水平DM-E[d=2.78，（95%CI=0.56-1.53），p<.05]和DMK-GM与DM-GM[d=2.43，（95%CI=0.65-2.10），p<.05]。骨桥蛋白表达在用GM治疗的缺损中增强。E缺陷[C-GMvs.C-E,d=1.56（95%CI=0.38-2.01）；CK-GM与CK-E,d=1.91（95%CI=0.49-1.72）；DM-GM与DM-E,d=2.34（95%CI=-1.12至-0.50）；DMK-GM与DMK-E,d=2.00（95%CI=0.58-1.91），p<.05]。
结论：研究结果表明，在糖尿病大鼠的GBR中施用维生素K2有利于CSD的骨愈合，提出了骨再生的辅助策略。
