Adaptation of cancellous bone to overloading in the adult rat: a single photon absorptiometry and histomorphometry study

Nine-month-old female rats were subjected to right hindlimb immobilization or served as controls for 0, 2, 10, 18, and 26 weeks and were double-labeled with bone markers. The right limb was immobilized against the abdomen and considered unloaded, while the left limb was overloaded during ambulation. Single-photon absorptiometry was performed on intact femur; static and dynamic histomorphometry were performed on 20 microns thick undecalcified frontal sections of the proximal tibial metaphysis. Changes in the continuously overloaded limb was compared to that in both limbs of age-matched control animals. Single-photon absorptiometry detected increases of bone mineral density of +6%, +6%, and +5% in the proximal and +9%, +7%, and +10% in the distal femoral metaphyses after 10, 18, and 26 weeks of continuous overloading. Morphometrically, significant changes occurred in proximal tibial metaphyses compared to age-matched controls: trabecular area increased +41% and +45%, trabecular number increased +31% and +32%, and trabecular separation decreased -30% and -31% after 18 and 26 weeks of overloading. A significant increase in mineral apposition rate (+38%) was found only at 26 weeks of overloading. Insignificant decreases in both eroded and labeled bone surfaces occurred at all time periods. The histomorphometric changes indicated that increased cancellous bone mass was caused by an increase in bone formation activity (i.e., increases in mineral apposition and bone formation rates) and a decrease in remodeling space (i.e., decrease in bone eroded surface). These findings indicate that the adult skeleton can quickly adapt to the increased biomechanical needs by increasing its cancellous bone mass with an adequate structural pattern.(ABSTRACT TRUNCATED AT 250 WORDS)     

Adaptation of cancellous bone to aging and immobilization in growing rats

Two-and-half month-old female rats were subjected to right hindlimb immobilization or served as controls for 0, 1, 2, 8, 14, and 20 weeks. The right hindlimb was immobilized by bandaging it against the abdomen, thus unloading it. Cancellous bone histomorphometry was performed on microradiographs and double-fluorescent labeled 20 μm sections of the distal femoral metaphyses. Primary spongiosa bone loss occurred rapidly by 2 weeks, and secondary spongiosa bone loss occurred rapidly by 8 weeks of immobilization, and then equilibrated at 60% less bone mass than age-related controls. The negative bone balance induced by immobilization was caused by transient increase in bone resorption, decrease in bone formation, and longitudinal bone growth. The dynamic data of secondary spongiosa cancellous bone showed that percent eroded perimeter was transiently elevated by 55 to 82% between 1 and 8 weeks, percent labeled perimeter was transiently depressed by 32% to 50% between 1 and 14 weeks, mineral apposition rate was depressed by 23% and 19% at 1 and 2 weeks, and bone formation rate-bone area referent was transiently depressed by 35% and 59% at 1 and 2 weeks. All the above parameters were at age-related control levels by 20 weeks of immobilization. However, bone formation rate-tissue area refent was depressed (-65%) throughout the study. Immobilization depressed completely longitudinal bone growth by 2 weeks and remained so. Only 0.65 mm of new metaphysis was generated in the immobilized versus 2.1 mm in controls during the study period. The immobilization induced an early cancellous bone loss which equilibrated at a new steady state with less bone and a normal (age-related control) bone turnover rate. When these findings were compared to an earlier study of 9-month-old virgin females subjected to right hindlimb immobilization up to 26 weeks, we found the adaptive responses of the cancellous bone were identical except that they occurred earlier and equilibrated sooner in younger rats.© Willey-Liss, Inc.     

Adaptation of cancellous bone to aging and immobilization in growing rats.

Two-and-half-month-old female rats were subjected to right hindlimb immobilization or served as controls for 0, 1, 2, 8, 14, and 20 weeks. The right hindlimb was immobilized by bandaging it against the abdomen, thus unloading it. Cancellous bone histomorphometry was performed on microradiographs and double-fluorescent labeled 20 microns sections of the distal femoral metaphyses. Primary spongiosa bone loss occurred rapidly by 2 weeks, and secondary spongiosa bone loss occurred rapidly by 8 weeks of immobilization, and then equilibrated at 60% less bone mass than age-related controls. The negative bone balance induced by immobilization was caused by transient increase in bone resorption, decrease in bone formation, and longitudinal bone growth. The dynamic data of secondary spongiosa cancellous bone showed that percent eroded perimeter was transiently elevated by 55 to 82% between 1 and 8 weeks, percent labeled perimeter was transiently depressed by 32% to 50% between 1 and 14 weeks, mineral apposition rate was depressed by 23% and 19% at 1 and 2 weeks, and bone formation rate-bone area referent was transiently depressed by 35% and 59% at 1 and 2 weeks. All the above parameters were at age-related control levels by 20 weeks of immobilization. However, bone formation rate-tissue area referent was depressed (-65%) throughout the study. Immobilization depressed completely longitudinal bone growth by 2 weeks and remained so. Only 0.65 mm of new metaphysis was generated in the immobilized versus 2.1 mm in controls during the study period. The immobilization induced an early cancellous bone loss which equilibrated at a new steady state with less bone and a normal (age-related control) bone turnover rate. When these findings were compared to an earlier study of 9-month-old virgin females subjected to right hindlimb immobilization up to 26 weeks, we found the adaptive responses of the cancellous bone were identical except that they occurred earlier and equilibrated sooner in younger rats.     

去势大鼠骨形态计量学、生物性能变化

目的 通过观察大鼠血清学、骨形态计量学测定、骨生物性能等方法研究去卵巢大鼠骨变化.方法 采用7-12月龄SD雌性大鼠30只,随机分为对照组、模型组和尼尔雌醇组.模型组和尼尔雌醇组大鼠切除双侧卵巢进行造模.尼尔雌醇组大鼠造模后45天开始灌服尼尔雌醇,连续90天;对照组和模型组大鼠手术后第45天开始灌服去离子水,连续90天.所有大鼠给药第80、87天时两次ip四环素20mg/kg进行骨荧光标记.实验结束时,取出大鼠股骨及第四腰椎骨进行股骨形态计量学测定、股骨骨密度测量、股骨三点弯曲实验及椎骨压缩实验,同时测定血清碱性磷酸酶(ALP)、抗酒石酸酸性磷酸酶(StrACP)水平.结果 大鼠去双侧卵巢后,股骨骨密度、骨最大载荷、最大应力、弹性模量、刚度水平和骨小梁体积显著性提高;椎体压缩性能、股骨形态计量学、股骨弯曲性能有明显改变;而雌激素对以上变化有不同程度改善作用.结论 骨组织形态、骨生物性能是评价骨质疏松模型或药物疗效的重要指标;发生骨质疏松时,骨组织形态学变化先于骨生物性能变化.     

In vivo measurement of bone quality in the horse: Estimates of precision for ultrasound velocity measurement and single photon absorptiometry

The in vivo precision of ultrasound velocity measurement and single photon absorptiometry for the assessment of equine bone quality is discussed. In vivo precisions for ultrasound velocity measurements were less than 0.5 per cent, whereas cortical cross-sectional area, compact bone density and modulus of elasticity were around 1 per cent, and bone mineral content and density were just over 2 per cent. Except for ultrasound velocity, substantial improvements could be achieved by taking the mean of five readings for each measurement. The long-term precision of the techniques was also high, with all precision values being less than 3 per cent. The possible sources of variation in ultrasound velocity measurements were also investigated. The method for combining ultranonic and photon absorptiometric data for equine bone quality assessment has been proven to be highly accurate and precise. There appears to the no reason why the same principles cannot be applied with equal success to the noninvasive assessment of bone quality in humans.     

Adaptation of diaphyseal structure to aging and decreased mechanical loading in the adult rat: a densitometric and histomorphometric study

Nine-month-old female rats were subjected to right hindlimb immobilization or served as controls for 0, 2, 10, 18, and 26 weeks. They were double-labeled with bone markers prior to sacrifice. Experimental unloading was produced by immobilizing the right limb against the abdomen with an elastic bandage. Single-photon absorptiometry was performed on the intact femurs; static and dynamic histomorphometry were performed on 20-micron thick toluidine blue-stained, undecalcified cross sections of the tibial shafts. Changes in the continuously immobilized tibiae were compared to those in both tibiae of age-matched controls. Unloading shut off nearly all periosteal bone formation and accelerates bone marrow expansion over that which occurs in age-related controls. The effect of unloading appeared to be mediated by recruiting fewer osteoblasts which showed inhibited activity. Furthermore, unloading increased endocortical percentage eroded surface. These histological changes lowered cortical bone mass by inhibiting diaphyseal cross sectional expansion and enlarging the bone marrow cavity. The results support Frost's suggestion that decrease mechanical usage depresses bone modeling-dependent bone gain by decreasing activation of modeling in the formation mode. It also stimulates bone remodeling-dependent bone loss by increasing activation of remodeling in the resorption mode.     

Dual photon absorptiometry using a gadolinium-153 source applied to measure equine bone mineral content

The application of the dual photon absorptiometry (DPA) technique, using gadolinium-153 as the photon source, to evaluate the bone mineral density (BMD) of the third metacarpal bone of horses is presented. The radiation detector was implemented with a NaI(TI) scintillator coupled to a 14 stage photomultiplier. A modular mechanical system allows the position of the prototype to be adjusted in relation to the animal. A moveable carrier makes it possible to scan the third metacarpal with a velocity adjustable between 1 and 12 mm s(-1), in steps of 1 mm s(-1), for a total distance of 250 mm. The prototype was evaluated with a phantom of the third metacarpal bone made of perspex and aluminium, and in vitro with a transverse slice of the third metacarpal bone of a horse. The tests showed that the prototype has an accuracy and precision of, approximately, 10% and 6%, respectively, for a 6 s acquisition time. Preliminary studies carried out in three foals from birth to one year of age indicated that the prototype is well suited to in vivo and in situ analysis of the BMD of the third metacarpal bones of horses, making it possible to evaluate the changes of BMD levels on a monthly basis. Also, results indicated an exponential behaviour of the BMD curve during the first year of life of the studied horses.     

Dynamic histomorphometry of alveolar bone remodeling in the adult rat

Alveolar bone normally undergoes remodeling on one side of the socket and modeling on the opposite side as the tooth migrates at a rate of 6.7 μm per day. Periodontal ligament width, however, remains constant. Because of this very high turnover rate, this bone is a good model to study bone modeling and remodeling activities. This study was undertaken in order to measure the different cellular events occurring during tooth migration along the alveolar bone of the rat. The histomorphometric measurements performed on this model permitted us to calculate the duration of each phase of the remodeling cycle, i.e., resorption lasts about 1.5 days and reversal about 3.5 days. Since the duration of the forming phase is about 1 day (Guyomard and Baron, ′74), the total duration of each remodeling cycle is about 6 days. This time is very short compared to 60–120 days in adult human trabecular bone. Additionally, in this model each osteoclast resorbs 2–4 times its own volume of bone per day. Based on this knowledge, it will be possible to measure accurately the effects of experimental conditions on bone cells and bone remodeling in this rat alveolar bone model.     

Theoretical and experimental limits of triple photon energy absorptiometry in the measurement of bone mineral

Theoretical and experimental studies have been conducted to explore the possibilities of triple photon energy absorptiometry in the measurement of bone mineral content. The purpose of this technique is to correct the measured bone mineral density for fat and soft tissues. However, theoretical considerations lead us to doubt the precision and accuracy of such measurements. In a first approximation the absorption coefficient can be split into Compton and photoelectric energy-independent factors. A consequence of such a model is the impossibility of finding more than two independent mass attenuation coefficients for different energies. The existence of an energy-dependent third factor may justify the use of triple photon energy absorptiometry but experimental tests and numerical simulations have shown that its value is too low for triple photon energy absorptiometry to be considered as an adequate method for the measurement of bone mineral content.     

Adaptation of diaphyseal structure with aging and increased mechanical usage in the adult rat: a histomorphometrical and biomechanical study

The experimental increase in mechanical usage or overloading of the left hindlimb was produced by immobilization of the contralateral hindlimb. The right hindlimb was placed in a flexed position against the body and was immobilized using an elastic bandage. Some control animals were sacrificed initially at time zero and increased mechanical usage and age-matched control animals were sacrificed after 2, 10, 18, and 26 weeks of treatment. All animals received double bone fluorochrome labeling prior to sacrifice. Cortical bone histomorphometry and cross-sectional moments of inertia were determined. Marrow cavity enlargement and total cross-sectional area expansion represented the age-related cortical bone changes. Increased mechanical usage enhanced periosteal bone modeling in the formation mode and dampened endocortical bone remodeling and bone modeling in the resorption mode (resorption drift) to create a slight positive bone balance. These observations are in general agreement with Frost's postulate for mechanical effects on bone modeling and remodeling (Frost, H.M. 1987b. Bone "mass" and the "mechanostat." A proposal. Anat. Rec. 219: 1-9). The maximum moment of inertia did not change significantly in either control or overloaded tibial shafts. The minimum and polar moment of inertias in overloaded bones increases over those of controls at 18 and 26 weeks of the experiment.     

Comparison of osteopenic changes in cancellous bone induced by ovariectomy and/or immobilization in adult rats

Background: Ovariectomy (OVX) and immobilization (IMM) in rats are useful models of osteopenia, replicating some aspects of osteoporosis in humans. The purpose of this study was to compare changes in cancellous bone after OVX and/or IMM. Methods: Differences in cancellous bone were determined at 6 and 12 weeks after OVX or IMM. Comparisons were also made when rats were ovariectomized or immobilized for 6 weeks and then immobilized (OVX/IMM) and ovariectomized (IMM/OVX), respectively, for 6 more weeks. The femurs were used to determine bone mineral content (BMC) using single photon absorptiometry (SPA) and for scanning electron microscopy (SEM). Tibias were collected for microradiography, image analysis, and histomorphometry of metaphyseal cancellous bone. Results: Six and 12 weeks after OVX, there was less cancellous bone mass, compared with controls, as indicated by SPA, SEM, microradiography, image analyses, and histomorphometry. Bone was lost primarily from the central metaphyseal regions in the OVX animals, whereas the loss occurred throughout the metaphyses in the IMM animals. There were more rodlike bone spicules and fewer platelike trabecule in the OVX and IMM groups compared with controls. Differences in the structural aspects of the cancellous bone, including differences in the types of bone struts and marrow star volumes, indicated less trabecular connectivity and greater trabecular separation in the OVX and IMM animals, compared with controls. Endochondral growth indices in the IMM groups tended to be less, whereas the OVX groups tended to be greater than controls. Cancellous bone formation rates were generally greater in the OVX groups but less in the IMM groups compared with controls. Osteoclastic resorption surfaces were substantially elevated in the IMM and OVX groups, particularly the IMM groups. Changes reflecting OVX and IMM, independently, were apparent in the OVX/IMM and IMM/OVX groups and indices of osteopenia were different from controls, including less bone mass, trabecular connectivity, and greater trabecular separation, bone turnover rates, and osteoclastic surface. Conclusions: These results demonstrate differences in the osteopenic changes that occur in cancellous bone following OVX or IMM. The changes were generally more dramatic in the IMM than in the OVX animals. When OVX and IMM were applied in combination, the osteopenic changes are particularly severe, emphasizing the importance of mechanical usage even with a deficiency of gonadal hormones. © 1994 Wiley-Liss, Inc.     

Sensitivity of bone histomorphometry in the diagnosis of metabolic bone diseases

Diagnostic sensitivity of bone histomorphometry was assessed in different metabolic bone diseases, after fixing the specificity at 75%, 90% and 95% reference levels. Sensitivity was particularly high in cases with greatly increased osteoid and/or resorption features, as in renal osteodystrophy (ROD). All the remodeling indicators were highly sensitive toward advanced or severe forms of mixed ROD (mROD). Osteoid indicators were the most sensitive parameters in ROD with predominant osteomalacia (oROD). Osteoclastic and several osteoid indicators were very sensitive in all grades of ROD with predominant hyperparathyroidism (hROD). Sensitivity was generally low in uremic patients without bone changes (wROD) and also in patients with idiopathic osteoporosis (OP). It is our recommendation, however, that for each individual patient the definite diagnosis should be based on both morphological, clinical and metabolic parameters.     

Transmission electron microscopic visualization of the degradation and phagocytosis of a poly-L-lactide screw in cancellous bone: a long-term experimental study

The increasing clinical use of biodegradable implants in orthopedic surgery makes it necessary to determine their long-term behavior in tissues. In this study, a biodegradable screw made of poly-L-lactide (PLLA) was inserted axially into the right distal femur in 18 rabbits. The degradation and phagocytosis process of PLLA was assessed histologically and by transmission electron microscopy (TEM). The follow-up times were 3 and 4.5 years for groups of nine and eight animals, respectively. Abundant birefringent polymeric material was still present in the center of the implant channel in all specimens in both follow-up groups. The PLLA material studied appeared to be a biologically relatively inert material, with only sparse reactive cellular activity at the tissue-implant boundary. In the TEM specimens, polymeric particles of an average area of 2 microm2 were seen to be located intracellularly within phagocytic cells. The spheric and polygonal particles were membrane-bound and to a great extent filled up each phagocyte. In the 4.5-year specimens, the size of the polymeric particles, measured as area and perimeter, was significantly smaller (p < 0.02) than that of the 3-year specimens. The findings indicate that the ultimate degradation process of PLLA is much longer than it previously was thought to be. Complete degradation probably still would have taken years after the 4.5-year span of this study.     

腰椎爆裂骨折椎体松质骨内力学分布特点的有限元研究

目的运用有限元方法模拟腰椎爆裂骨折的过程,观察腰椎在轴向压缩载荷作用下松质骨内的应力分布情况。方法建立正常人体胸腰段(T12~L2)运动节段的三维有限元模型,在T12椎体上表面施加不同等级的压力(0.4、0.6、0.8、1.0、1.2 k N),模拟腰椎爆裂骨折发生时椎体承受的不同等级的轴向压缩载荷。将连接椎体上下终板凹面顶点的连线7等分,在此基础上将L1椎体中的松质骨划分为7个具有统计节点的层面,每个统计层面划分成6个统计区。分别测量L1椎体松质骨内3个层面(第1、4、7层面)18个统计区的平均应力。在同一等级载荷下对3个层面内的平均应力进行单因素方差分析,分析腰椎椎体松质骨内不同载荷作用下的应力分布情况。结果在5个不同等级载荷下,第1、7层松质骨平均应力分别与第4层比较有统计学意义(P0.05),而第1、7层平均应力比较无统计学意义(P0.05)。轴向加载时,相比第1、7层应力,椎体松质骨中间层面(第4层)应力最小。结论腰椎在轴向压缩载荷作用下,椎体松质骨内存在应力集中的现象,接近椎体上下软骨板的松质骨应力较大,而椎体松质骨中间层面应力较小,椎体内应力集中分布在上下软骨板的特点与腰椎爆裂骨折所致终板破裂的生物力学机制相一致,提示腰椎椎体骨结构损伤可能与椎体内应力集中有关。     

Orthogonal cutting of cancellous bone with application to the harvesting of bone autograft

Autogenous bone graft harvesting results in cell death within the graft and trauma at the donor site. The latter can be mitigated by using minimally invasive tools and techniques, while cell morbidity may be reduced by improving cutter design and cutting parameters. We have performed orthogonal cutting experiments on bovine cancellous bone samples, to gain a basic understanding of the cutting mechanism and to determine design guidelines for tooling. Measurements were performed at cutting speeds from 11.2 to 5000mm/min, with tool rake angles of 23 degrees , 45 degrees and 60 degrees , and depths of cut in the range of 0.1-3.0mm. Horizontal and vertical cutting forces were measured, and the chip formation process video recorded. Continuous chip formation was observed for rake angles of 45 degrees and 60 degrees , and depths of cut greater than 0.8mm. Chip formation for depths of cut greater than 1.0mm was accompanied by bone marrow extruding out of the free surfaces and away from the rake face. Specific cutting energies decreased with increasing rake angle, increasing depth of cut and increasing cutting speed. Our orthogonal cutting experiments showed that a rake angle of 60 degrees and a depth of cut of 1mm, will avoid excessive fragmentation, keep specific cutting energy low and promote bone marrow extrusion, which may be beneficial for cell survival. We demonstrate how drill bit clearance angle and feed rate can be calculated facilitating a 1mm depth of cut.     

骨组织形态计量学在临床骨病应用中的研究与进展

背景：骨组织形态计量学是一种定量的不脱钙骨组织学切片检查,能取得骨重建和结构的定量信息。近年来已广泛应用于代谢性骨病发生发展机制、抗骨质疏松症药物等研究。 目的：从骨组织形态计量学被认识到临床研究、临床应用以及展望进行综述。 方法：由第一作者分别以“骨组织形态计量学,骨活检,代谢性骨病,骨重建,骨结构,人”和“Bone histomorphometry,metabolic bone diseases,bone biopsy,bone remodeling,humans”为关键词进行检索,CNKI 数据库的检索时限为2000/2010,PubMed 数据库的检索时限为1965/2010。筛选骨组织形态计量学方面的文献,最终纳入30 篇文献进行分析。 结果与结论：骨计量学可得到三维的骨结构和重建的参数,在评估、提供骨代谢信息方面是一个功能强大的工具,其他任何研究方法不可代替。对于各种各样的骨代谢疾病,可从骨计量学方面更深层了解骨病的发生发展机制,为治疗提供更可靠的依据。此外,在临床新的药物安全干预、研究试验中,它提供了宝贵的骨结构和重建的参数资料。     

The effects of cracks on the quantification of the cancellous bone fabric tensor in fossil and archaeological specimens: a simulation study

Cancellous bone is very sensitive to its prevailing mechanical environment, and study of its architecture has previously aided interpretations of locomotor biomechanics in extinct animals or archaeological populations. However, quantification of architectural features may be compromised by poor preservation in fossil and archaeological specimens, such as post mortem cracking or fracturing. In this study, the effects of post mortem cracks on the quantification of cancellous bone fabric were investigated through the simulation of cracks in otherwise undamaged modern bone samples. The effect on both scalar (degree of fabric anisotropy, fabric elongation index) and vector (principal fabric directions) variables was assessed through comparing the results of architectural analyses of cracked vs. non‐cracked samples. Error was found to decrease as the relative size of the crack decreased, and as the orientation of the crack approached the orientation of the primary fabric direction. However, even in the best‐case scenario simulated, error remained substantial, with at least 18% of simulations showing a > 10% error when scalar variables were considered, and at least 6.7% of simulations showing a > 10° error when vector variables were considered. As a 10% (scalar) or 10° (vector) difference is probably too large for reliable interpretation of a fossil or archaeological specimen, these results suggest that cracks should be avoided if possible when analysing cancellous bone architecture in such specimens.