| Objective Diabetes is endangering people's health and disease currently following heart disease,cancer seriously.With the advent of an aging society,the incidence of diabetes has increased year by year.Skeletal system is a dynamic organ,diabetes and osteoporosis(DOP)are secondary OP,DOP is one of the most common complications of DM.Along with nutrition,metabolism and endocrine factors such as changes in the series of pathological changes,DM reduced to their BMD and slow process of osteoclast-based.In recent years,with oral and maxillofacial plastic surgery and oral cultivation technologies mature and perfect, there was indication of more extensive operation.How to deal with diabetes mellitus associated with osteoporosis,dentists are facing one of the important tasks.Therefore,the oral disease prevention and treatment must take into account systemic factors,and systemic disease treatment is also an important component of the oral diseases treatment.On the one hand,diabetic osteoporosis should be considered on the impact of oral diseases;on the other hand,the relationship between secondary osteoporosis and dental disease should be analysed from the perspective of the treatment,in order to further improve oral disease prevention and treatment.This study focused on the establishment of the two experimental T1DM and T2DM rats.There were glucose metabolism,bone metabolism and serum biochemical markers and urine calcium determination tested.Right alveolar bone of mandible of rats was to BMD measurement.Left alveolar bone of mandible was to observe bone tissue morphology under fluorescence microscope and measured the dynamic parameters of bone tissue morphology.Understand the bone metabolism,BMD,bone micro-structure of the two types of diabetes which lead to or not to the occurrence of osteoporosis and explore its possible mechanism.Methods:The study is divided into three partsPart 1 Preparation of rats with experimental diabetes66 rats(♀20,♂46),average weight of 250±3g,were purchased from the Experimental Animal Center of Shandong University(Animals Zhilianghegezheng book #:LU RP20021024),keeping in Shandong Province Institute of Endocrinology and Metabolism Center Laboratory(two grade animal rooms).The test divided into two groups:the first large group was made from 20 female rats and 21male rats selected randomly for the establishment of T2DM;the second large group was made from 25 male rats remained randomly for the establishment of T1DM.Since the establishment of experimental DM model,the test didn't terminate until 20 weeks later.Part 2 Collection and production of specimen from experimental animal modelRats were anesthetized by intraperitoneal injection of Ketamine hydrochloride(100 mg/kg),executed by ventricular blood.Serum was extracted.Left and right mandibles were severed completely,peeling soft tissue carefully.The right mandible covered with isotonic saline gauze,-20℃refrigerator preserved,reserved for measuring BMD,see photographs 1-3.The left side of the mandible at 70%alcohol solution, 4℃refrigerator preserved,reserved for observating bone morphogenetic indicators and analysis of the dynamic indicators.Part 3 Test on experimental animal models of specimens in vitroThere were glucose metabolism,bone metabolism and serum biochemical markers and urine calcium determination.Right alveolar bone of mandible of rats was to BMD measurement.Left alveolar bone of mandible was to make demineralized bone.Regional alveola alveolar bone below molar root was cut into 10um mesio-distal slices which could be observed bone tissue morphology under fluorescence microscope and measured the dynamic parameters of of bone tissue morphology.Part 4 Statistical AnalysisAll data are shown in the list,with the average±standard deviation(x±S), were dealed with SPSS11.5 statistical package and the results were determinated with normality and homogeneity of variance test.For further comparison,t test of variance was tested between two groups.P＜0.05 was considered as significant difference.Results1.Obervation of the simulation model of diabetes ratsT1DM rats began to drink more,eat more and polyuria after intraperitoneal injection of STZ in 5-7 days.Three-more and one less symptom in T2DM rats was small lighter than in T1DM,injected STZ in 8-10 days.Model eight weeks,the special symptoms began to come up to two DM group rats.T1DM(N)rats were significantly thinner coat bleak,dull.The spirit of T1DM(N)rats was worse than T2DM rats and T1DM(I)rats.Normal control rats kept to drinking,eating normally,increasing weight,but the body weight in N♀and N♂Group was higher than that of N rats,with the representatives of two types of DM control group by age requirements,color glossy.2 T1DM(N)rats were completely blind because of cataract,5 rats with T2DM suffered from cataract but not completely blind,bright flu.DM rats' tail fester in different degrees,to T1DM(N)in the most serious,only 1/3-1/5 long compared with the original tail.In the course of the experiment,the normal rats' weight was fast-growing-up,rats with diabetes disease extended the growing thinner,showing a growth inhibition with their weight decreased significantly.2.Study of the simulation model of diabetes,Model 20 weeks,kinds of rats' weight changes in comparison among the different group of the rats: T2DM,weight of N♂and T2DM♂rats was higher than that of N♀and T2DM♀rats,there were significant differences(P＜0.05).Weight of T2DM♀and T2DM♂rats was lower than thatof N♀and N♂,a significant difference(P＜0.05).T1DM,weight of T1DM(N)rats was less than that of N and T1DM(I),there were significant differences(P＜0.05);Weight of T1DM(I)rats was also less than N group,there were significant differences(P＜0.05).Weight of T2DM♂and T1DM group in comparison:N♂＞N,T2DM♂＞T1DM(I)＞T1DM(N),a significant difference(P＜0.05).See tablel-3.3.Study of the simulation model of diabetes,Model 20 weeks,to compare of BMD in alveolar bone of mandible among the different group of the rats:T2DM group,there was significant difference that BMD in alveolar bone of mandible of N♂and T2DM♂rats was higher than that of N♀and T2DM♀rats respectively(P＜0.05).BMD in alveolar bone of mandible of T2DM♀and T2DM♂rats also had more significant differences than that of N♀and N♂(P＜0.05).T2DM♂and N♂rats carred out the alveolar bone of mandible and the whole body bone tissue reaches regular rat bone density height comparatively, being in proper order:thigh-bone＞alveolar bone＞lumbar vertebra.T2DM rats bones density height in proper order:alveolar bone≈thigh-bone＞lumbar vertebra.T1DM group,there was significant difference that BMD in alveolar bone of mandible of T1DM(N)rats was lower than that of N and T1DM(I)rats(P＜0.05). There was no significant difference that BMD in alveolar bone of mandible between N and T1DM(I)rats(P＞0.05).BMD in alveolar bone of mandible of T2DM♂and T1DM rats in comparison:N♂≈N,T2DM♂≈T1DM(I)＞T1DM(N).See table 4-6.4.Study of the simulation model of diabetes,Model 20 weeks,to compare of glucose and lipid metabolism,bone metabolism,biochemical markers and calcium in urine among the different group of the rats:Glucose Metabolism:Blood sugar of T1DM and T2DM rats were higher than that of normal control rats,there were significant differences(P＜0.05).Blood sugar of T1DM(N)vs T1DM(I)and T2DM♀vs T2DM♂was no significant difference(P＞0 0.05).Insulin levels of T2DM♀rats was slightly higher than that of normal control rats,there were significant differences(P＜0.05).T2DM♂rats with normal insulin levels had no significant difference(P＞0.05).There was also no significant difference between T2DM♀and T2DM♂(P＞0.05).Model 9 weeks later,there were significant differences that T2DM♀and T2DM♂rats' HOMA-IR compared with the normal control rats'(P＜0.05).HOMA-IR of T1DM(N)vs T1DM(I)rats was lower than normal control rats, there were significant differences(P＜0.05).There was also significant difference between T1DM(N)and T1DM(I)(P＜0.05).HOMA-IR of T1DM rats vs the normal control rats,there was no significant difference(P＞0.05).The results of glucose metabolism showed that T2DM rats after nine weeks of modeling were found insulin resistance,setting up a successful T1DM and T2DM rats respectively.Lipid Metabolism:Serum TG,TC of T2DM♀and T2DM♂rats were higher than those of N♀and N♂ones,there were significant differences(P＜0.05). Serum TC in T1DM(N)was less than that of N and T1DM(I)rats,there was significant difference(P＜0.05);but there was no significant difference in the TG in T1DM rats(P＞0.05).Bone Metabolism:Serum OCN of DM rats decreased,in addition to T1DM(I) rats,were significantly different(P＜0.05).Serum TRACP of DM rats significantly increased,there was significant difference(P＜0.05).Serum IGF-1 of DM rats was lower than the control group rats,with the exception of T1DM(I) and T1DM(N)groups,there were significantly different(P＜0.05).Biochemical indicators:there was no significant difference hyperphosphatemia from two types of diabetic rats and normal control groups(P＞0.05).Calcium in addition to T1DM(N)group fell and the rest have no significant difference(P＞0.05).Calcium:Urine Ca of DM rats was increased,there were significant differences(P＜0.05).See table 7-8.5.Study of the simulation model of diabetes,model 20 weeks,to compare of morphology dynamic parameter measurement results in alveolar bone of mandible among different groups of the rats:T2DM,MS/BS,BFR,MAR in alveolar bone of mandible of DM♀rats were lower than that of N♀rats,MLT was longer,there were significantly different(P＜0.05).MAR in alveolar bone of mandible of DM♂rats was lower than that of N♂rats,showing significant differences(P＜0.05),and the remaining showed no significant difference(P＞0.05).There were significantly different that MS/BS,BFR in alveolar bone of mandible of DM♂rats were higher than that of DM♀rats,and MLT was shorter than DM♀rats(P＜0.05).T1DM,MS/BS,BFR,MAR in alveolar bone of mandible of DM(N)rats were lower than Normal rats;MLT was longer,there were significantly different respectly(P＜0.05).MAR in alveolar bone of mandible of DM(I)rats was higher than that of N rats,showing significant differences(P＜0.05),and the remaining showed no significant difference(P＞0.05).There were significantly different that MS/BS,BFR,MAR in alveolar bone of mandible of DM(I)rats were higher than that of DM(N)rats,and MLT was shorter than DM(N)rats(P＜0.05).See table 9.6.Study of the simulation model of diabetes,to compare of alveolar bone of mandible formation observed under fluorescence microscope among the different groups of the rats:Yellow-green fluorescent marker in the dark green background was visible under fluorescence microscope.Fluorescent display area from T1DM(N),T2DM♀,T2DM♂rats alveolar bone of mandible showed osteogenesis is inactive with absorpted lacunae apparent,followed by reducing the degree one by one.Those in T1DM(I)and the normal control rats showed osteoblast active,no obvious absorpted lacunae.See Figure 1-7.7 The relationships among disorders of blood biochemistry,glucose metabolism,lipid metabolism,bone metabolism,BMD,BHM changes caused by T1DM and T2DM and change in alveolar bone of mandible were as follows: ①T2DM,there was positive correlation among serum OCN,IGF-1 and MS/BS, BFR in alveolar bone of mandible with a negative correlation as MLT in alveolar bone of mandible.There was negative correlation among blood sugar,serum TG,TC,TRACPand MS/BS,BFR in alveolar bone of mandible with a positive correlation as MLT in alveolar bone of mandible.②T1DM,there was positive correlation among serum Ca,OCN,TC,IGF-1 and MS/BS,BFR,MAR in alveolar bone of mandible with a negative correlation as MLT in alveolar bone of mandible.There was negative correlation among TRACPand MS/BS,BFR,MAR in alveolar bone of mandible with a positive correlation as MLT in alveolar bone of mandible.Conclusions1.Rat models of type 1 and 2 diabetes were successfully established.2.DOP in the mandibular alveolar bone of female and male T2DM rats:T2DM♀＜T2DM♂.3.DOP did not happen to the mandibular alveolar bone of T1DM(I)rats, and T1DM(N)rats failed in DOP.4.BMD in the mandibular alveolar of DM rats:T2DM♂≈T1DM(I)＞T1DM(N).5.T2DM:BMD in N♂rats:femur＞mandibular alveolar bone＞lumbar. BMD in DM♂rats:mandibular alveolar bone≈femur＞lumbar spine.6.BMD in alveolar bone of 2 kinds of diabetes is reduced into decompensated status:new bone formation＜resorption.7.Blood biochemistry,sugar,fat,bone metabolism disorder and bone structure of micro-dynamic parameters of the change and osteoporosis caused by two types of diabetes,can be used as early detection of osteoporosis indicators.
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