| ObjectiveFractures are one of the most common diseases encountered in clinical setting.They are classified into trabecular fractures and cortical fractures.Fracture not healing or delayed healing is a clinically difficult situation.Fracture healing is a complicated procedure involving knowledge in histology,biology,endocrinology and biodynamics.Factors affecting fracture healing are various,including whole-body and regional factors,iatrogenic factors,bone growth factors,trace elements,biodynamic factors etc.The medical community has been actively focusing on researches and laboratory experiments on how to enhance fracture healing,to understand the fracture healing process and to recover normal bone functions and structure.It is through various researches and laboratory experiments from different perspectives that scholars come to a thorough understanding of fracture healing and its process today.There are various methods to treat bone fractures,some of which are very effective,yet there are existing insufficiencies in this aspect.As such,treatment of bone fractures in traditional Chinese medicine having a long history and experience with cumulative experience can be utilized,with its characteristics being simple and convenient,safe and effective,low adverse effects and low cost.This study aims to investigate and analyze the enhancing effects of Yang-Xue-Gu-Shen-Tang in bone fracture and the processes or interferences and the mechanisms underlying it by observations at different levels.MethodsFirst section on observations in biochemical indicators:80 male S.D.rats weighing 168.5-237.2g with no limb deformities,stable vital signs,normal diets and no infective diseases were reared in SPF-grade laboratory environment throughout the whole investigation.Experiment commenced when S.D.rats reached desirable weight.Experiment model was created by making an open fracture at tibia mid-section,followed by bone marrow cavity fixture using Kirschner Needle and surgical suture without bandaging.72 S.D.rats were selected after successful procedure and were randomly divided into 3 groups(model group,positive group and treatment group)according to weight,with 24 S.D.rats in each group.S.D.rats were given pure water,bone fracture healing agent and Yang-Xue-Gu-Shen-Tang according to weight via stomach gavage in model group,positive group and treatment group respectively.Day 7,Day 14,Day 24 and Day 35 after the fracture-creating procedure,6 S.D.rats were dissected at each time threshold to obtain blood samples.Plasma calcium,plasma phosphorus and ALP were inspected.Second section on histopathological observations:80 male S.D.rats weighing 168.5-237.2g with no limb deformities,stable vital signs,normal diets and no infective diseases were reared in SPF-grade laboratory environment throughout the whole investigation.Experiment commenced when S.D.rats reached desirable weight.Experiment model was created by making an open fracture at tibia mid-section,followed by bone marrow cavity fixture using Kirschner Needle and surgical suture without bandaging.72 S.D.rats were selected after successful procedure and were randomly divided into 3 groups(model group,positive group and treatment group)according to weight,with 24 S.D.rats in each group.S.D.rats were given pure water,bone fracture healing agent and Yang-Xue-Gu-Shen-Tang according to weight via stomach gavage in model group,positive group and treatment group respectively.Day 7,Day 14,Day 24 and Day 35 after the fracture-creating procedure,6 S.D.rats were executed at each time threshold to obtain lower part of left femur to whole tibia to create specimens for histopathological observations and analysis.According to principles in histopathology,specimens were detected by randomly selecting 3 visual points within light microscope.Trabecular area percentage,osteoblast indicator and osteoclast indicator were observed and recorded.Third section on histopathological observations:80 male S.D.rats weighing 168.5-237.2g with no limb deformities,stable vital signs,normal diets and no infective diseases were reared in SPF-grade laboratory environment throughout the whole investigation.Experiment commenced when S.D.rats reached desirable weight.Experiment model was created by making an open fracture at tibia mid-section,followed by bone marrow cavity fixture using Kirschner Needle and surgical suture without bandaging.72 S.D.rats were selected after successful procedure and were randomly divided into 3 groups(model group,positive group and treatment group)according to weight,with 24 S.D.rats in each group.S.D.rats were given pure water,bone fracture healing agent and Yang-Xue-Gu-Shen-Tang according to weight via stomach gavage in model group,positive group and treatment group respectively.Day 7,Day 14,Day 24 and Day 35 after the fracture-creating procedure,6 S.D.rats were executed at each time threshold to obtain lower part of left femur to whole tibia to create specimens for micro computed tomography(micro-CT)detection.Micro-CT was used to detect fracture location by selecting region of interest and collect parameters including BMD,BV/TV,TV,Tb.Sp,Tb.N and SMI using image remodeling software and image detection and analysis.Statistical Analysis:All data were presented using(x±s)and were statistically analyzed using SPSS 21.0 software.Quantitative information with complete variance or data after translation with complete variance was statistically analyzed using ANOVA.Data after translation with incomplete variance was statistically analyzed using t-test.ResultsYang-Xue-Gu-Shen-Tang Group showed increased plasma phosphorus level(P<0.05),increased plasma calcium and phosphorus product(P<0.01),enhanced calcium salt deposit,decreased plasma calcium level(P>0.05)and decreased ALP level(P>0.05).At Day 7,specimens in Yang-Xue-Gu-Shen-Tang Group showed an earlier appearance of cartilage and trabeculae alignment after callus structure formation as compared to Model Group.Proliferation of blood clot was observed in some specimen.Large amount of cartilage cells and sparse trabeculae formation were observed in callus structure.At Day 14,specimens in Yang-Xue-Gu-Shen-Tang Group showed fibrous callus structure and large amount of cartilage and dense trabeculae formation with an increase in amount,as compared to Model Group.At Day 24,most specimens in Yang-Xue-Gu-Shen-Tang Group,as compared to Model Group,showed enhanced transformation of trabeculae and callus manifested by significantly thicker trabeculae,formation of plate bone from external callus,small amount of cartilage cells within callus and individual bone marrow cavity decongestion.At Day 35,both Yang-Xue-Gu-Shen-Tang Group and Model Group showed formation of plate bone from external callus,appearance of bone marrow cells within bone marrow cavity and appearance of small amount of cartilage cells and thickened trabeculae in some specimens.SMI level showed statistical significance in results collected frombone parameters by micro-CT,indicating that Yang-Xue-Gu-Shen-Tang increased bone strength in trabecular structure modeling.ConclusionIn this study,Yang-Xue-Gu-Shen-Tang enhanced fracture healing by elevating plasma calcium and phosphorus,significantly increasing calcium and phosphorus product peak,promoting calcium salt deposition,increasing serum ALP activity and strengthening osteoblast activity.Yang-Xue-Gu-Shen-Tang promoted formation of callus in addition to cartilagous callus and hard callus,and promoted the formation of trabecular structure in terms of increased formation and density.Yan-Xue-Gu-Shen-Tang on an average increased within 0-3,suggesting it can affect bone formation and elevate bone structure strength. |
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