咀嚼力降低对发育期大鼠髁突发育的组织学影响

目的：探讨咀嚼力降低对断乳后大鼠下颌骨髁突尺寸和髁突软骨厚度的影响。方法：16只刚断乳的雄性大鼠随机分为两组。一组喂以标准的硬食,另一组以粉末状软食喂养。4w后,取每只大鼠的一侧髁突进行长度和宽度的测量,另一侧髁突进行组织学观察。结果：软食组大鼠的髁突长度、宽度和软骨层厚度均明显小于硬食组。结论：咀嚼力降低会导致下颌骨髁突生长减慢和髁突软骨厚度的变化。     

咀嚼负荷对幼兔髁突软骨细胞BrdU标记影响的研究

目的 探讨咀嚼负荷对幼兔髁突软骨发育及软骨细胞中BrdU标记的影响。方法 出生10 d龄的32只幼兔,随机分为喂食固体硬饲料组和喂食粉末软饲料组。连续2周每天每只腹腔注射BrdU （50 mg/kg）,在2、4、6、8周时每组处死4只幼兔,HE观察组织形态学变化测量软硬组髁突厚度变化,免疫组化检测BrdU表达。结果 髁突软骨前部厚度硬食组明显高于软食组,在中部无明显差异,髁突软骨后部厚度软食组高于硬食组。BrdU免疫组化染色显示,发育期髁突软骨各层及软骨下骨骨髓腔内均有BrdU阳性标记,前部的增殖层表达最显著。第2周,硬食组在前、中、后3个部位的BrdU阳性率都高于软食组;第6周,软食组的中、后部位的阳性率高于硬食组;第8周,软硬组均检测不到阳性细胞。结论 软食组在低咀嚼力刺激下,幼兔髁突软骨细胞增殖启动较慢。适当的咀嚼压力能促进髁突软骨细胞的增殖,同时幼兔发育期的髁突生长还受到基因等内环境因素影响。     

不同咀嚼压力对幼兔髁突软骨成骨的影响研究

目的：分析生长发育期不同咀嚼模式下兔髁突软骨成骨形态学的差异,研究咀嚼压力对幼兔髁突生长发育的影响和作用机制。方法：选取15天龄的未断奶幼兔16只,随机分为硬食组和软食组各8只,硬食组提供颗粒饲料,软食组提供由同种颗粒饲料碾磨而成的粉末状饲料。幼兔28天龄断奶,分别于28天龄、33天龄肌肉注射四环素、钙黄绿素,38天龄处死幼兔。制作硬组织切片,荧光显微镜观察,独立t检验分析比较两组髁突软骨成骨厚度,骨形态学参数。结果：硬食组的软骨成骨厚度、骨小梁相对体积、骨小梁数量、骨小梁厚度均大于软食组（P<0.05）,硬食组骨小梁分离度小于软食组（P<0.05）。相同时间硬食组骨形成沉积率[（3.25±0.02）μm / d ]大于软食组[（2.37±0.02）μm / d ]。结论：生长期软食组幼兔的髁突软骨成骨能力较硬食组弱,咀嚼负荷力是影响髁突软骨成骨生长的重要因素。     

The effect of dietary consistency on bone mass and turnover in the growing rat mandible.

Hard and soft diets were fed to weanling rats for up to 8 weeks. Some animals were switched after 4 weeks to the opposite diet. A histomorphometric study of bone formation activity at the mandibular ramus, body, and condyle was made after in vivo fluorochrome labelling. Mineral apposition rates at the lateral and inferior periosteal surfaces of the ramus were lower in the soft diet than in the hard diet animals. The rate of bone formation at the lateral periosteal surface of the ramus was significantly lower in soft than in hard diet animals. The medial periosteal surface of the ramus sometimes changed to bone formation in the soft diet groups. Condylar cartilage zones were somewhat thinner in soft diet groups. In the mandibular body, differences due to dietary consistency were less marked than near the gonial angle. Adaptation of periosteal bone and condylar cartilage to a new dietary consistency occurred within 4 weeks of switching. These results suggest that lateral and inferior periosteal bone growth of the ramus and condylar elongation were slowed in rats consuming soft diets. Decreased functional force during rapid mandibular bone growth causes changes in shape. The changes are due to regional decreases in osteoblast function, realignment of bone formation surfaces in the ramus area, and slowed growth in the condylar cartilage.     

Effects of age on the ability of the rat temporomandibular joint to respond to changing functional demands

This investigation examined the ability of the tissues of the temporomandibular joint (TMJ) to adapt to changing functional demands in young, growing rats compared with mature rats. Functional demands on the TMJ were varied by feeding diets with different physical consistencies. The first group was fed a soft diet for the experimental period. The second group was fed a hard diet, and the third group was initially fed the soft diet, then switched to the hard diet at the mid-point of the experimental period. Gross dimensions of the condyle, mandible, and maxilla were measured with calipers. Thickness of the articular, proliferative, transitional, and hypertrophic zones of the condylar cartilage, and the amount of bone in the subcondylar region and condylar neck were measured on histological sections. Gross dimensions of the condyle were significantly smaller in the soft-diet group compared with the hard- and soft/hard-diet groups in both growing and mature rats. The individual zones of the condylar cartilage were also significantly narrower in the soft-diet group in both growing and mature rats. However, the soft/hard-diet group of mature rats showed only a significant reduction in the thickness of the articular zone of the condylar cartilage compared with the hard-diet group. There were also narrower proliferative and transitional zones in the mature rats fed a soft/hard diet. In contrast, all of these zones showed full recovery in the young rats fed a soft/hard diet. The data presented here suggest that increasing age may diminish the capacity of the TMJ to adapt to altered function and consequently may play a significant role in the development of degenerative joint disease.     

不同咀嚼负荷对幼兔髁突软骨内印度豪猪蛋白-人甲状旁腺激素相关蛋白通路表达的影响

目的 分析不同咀嚼负荷作用下,幼兔髁突软骨内印度豪猪蛋白（Ihh）-人甲状旁腺激素相关蛋白（PThrP）通路表达的差异性,探讨咀嚼应力负荷对髁突软骨Ihh-PThrP信号通路的影响。方法 选取10 d龄幼兔48只,随机分为硬食组和软食组,分别喂以同种颗粒状（硬食）和粉状（软食）,于饲养的第2、4、6、8周处死。采用苏木精-伊红（HE）染色、免疫组织化学、免疫印迹、实时定量荧光聚合酶链反应检测Ihh和PThrP mRNA和蛋白的动态变化。结果 HE染色显示硬食组髁突软骨厚度高于软食组的厚度;第2、4、6、8周,Ihh蛋白、PThrP蛋白和mRNA表达量在两组中呈递减趋势;软食组中Ihh和PThrP蛋白以及mRNA表达量显著低于硬食组。结论 较低的咀嚼负荷会造成髁突软骨生长因子Ihh和PThrP分泌减少,Ihh-PThrP通路表达迟缓,软骨发育受阻碍;适当的咀嚼负荷对髁突的正常发育有至关重要的作用。     

The effects of gradually induced backward movement of the mandible by a Twin Inclined Plane Device in rats

Abstract Objective: To develop a model of gradually induced backward movement of the mandible under normal masticatory action and to examine morphological changes in the mandible and condylar cartilage in rats. Materials and Methods: The newly developed Twin Inclined Plane Device (TIPD) was composed of upper and lower posterior metal crowns with a long inclined plane on both sides separately and was applied in experimental groups of 6-week-old male Wister rats. After 3, 14, 30, and 60?days, the rats were euthanatized and samples were collected. Various measurements and hematoxylin-and-eosin stains were performed. Results: From day 30 on, the length of the condylar process was shorter in the TIPD groups than in the control groups (P < .05). The angulation of the condylar process axis to the mandibular plane was greater in the TIPD groups (P < .01). The thickness of the condylar cartilage in the posterior part of the posterior region was thinner in the TIPD groups (P < .05) on day 30 and even thinner (P < .01) on day 60; from day 30 on, the thickness in the anterior part of the posterior region was thicker in the TIPD groups (P < .01). Conclusion: TIPD can successfully induce backward movement of the mandible under normal masticatory action. TIPD can cause region-specific changes in condylar cartilage and leads to a continuous remodeling.     

咀嚼力降低对发育期大鼠髁突软骨中细胞增殖和凋亡的影响

目的：研究咀嚼力降低对发育期大鼠下颌髁突软骨细胞增殖和细胞凋亡活性的影响。方法：40只18d雄性大鼠随机分为两组。一组喂以标准的硬食,另一组以粉末状软食喂养。于大鼠4周、5周、6周、7周龄时取其髁突软骨,分别采用免疫组织化学染色和TUNEL染色,观察髁突软骨中增殖细胞核抗原阳性细胞数和凋亡细胞数。结果：在4～7周,PCNA阳性细胞数在软、硬食组都逐步上升。但在各相同时间点,软、硬食组的PCNA阳性细胞数不存在统计学差异。在4～7周,硬食组大鼠髁突软骨中凋亡细胞数基本保持一致,而软食组凋亡细胞数随时间推移而降低,但在4周、5周、6周时,软食组的凋亡细胞数均明显多于同期硬食组（P〈0.05）。结论：咀嚼力降低对发育期大鼠髁突软骨中的细胞增殖活性影响不大,但却使细胞凋亡的数量增加,提示咀嚼力降低可能加速了髁突软骨中细胞的分化、成熟、凋亡,进而影响髁突的正常发育。     

Histologic examination of the temporomandibular joint after mandibular advancement with and without rigid fixation: An experimental investigation in adult Macaca mulatta

This study evaluated the histologic response of the temporomandibular joint (TMJ) following mandibular advancement using rigid and nonrigid fixation in monkeys. Twelve adult female rhesus monkeys underwent sagittal ramus osteotomies with advancement. Six of them were placed into maxillomandibular fixation (MMF); six underwent bicortical bone-screw fixation without MMF. Changes in condylar position were quantified using lateral cephalograms with the aid of bone markers. The animals were killed at 6 weeks and the TMJs were prepared for histologic analysis. Three measures of condylar cartilage thickness were obtained for each animal and were correlated to changes in position of the condyle. Animals who underwent MMF showed a tendency for anterior movement of the condyles; animals who underwent rigid fixation showed a tendency for posterior condylar position. Thicker cartilage layers were found in the MMF animals. Animals who had posterior displacement of the condyles showed evidence of resorption of the posterior surface of the condyle and anterior surface of the postglenoid spine. There was a significant correlation between a change in the horizontal position of the condyle and the thickness of the posterior aspect of the condylar cartilage. The results of this study indicate that alterations in condylar position may induce remodeling changes within the TMJ.     

咬合紊乱与去除咬合紊乱对大鼠髁突软骨中骨形成蛋白-2表达的影响

目的探讨咬合紊乱与去除咬合紊乱大鼠髁突软骨内骨形成蛋白- 2（BMP- 2）的变化。方法幼年和成年雌性大鼠各9只,等分为咬合紊乱组、去除咬合紊乱组和对照组。咬合紊乱组在建立咬合紊乱8周后处死,去除咬合紊乱组在建立咬合紊乱6周时拔除造成紊乱的双侧第一磨牙,2周后处死。对照组不作任何处理,同环境饲养、同期处死。测量各组髁突组织切片上软骨前、中、后部的厚度,SABC法检测软骨前、中、后部BMP- 2的表达。结果成年咬合紊乱组髁突软骨中部变薄,后部增厚;去除咬合紊乱后后部恢复正常,中部仍薄于对照组（P<0.05）。幼年髁突软骨的前、中、后部厚度三组间未见显著性差异（P>0.05）。幼年髁突软骨前、中、后部咬合紊乱组BMP-2表达高于去除咬合紊乱组和对照组（P<0.05）,成年髁突软骨中部咬合紊乱组和去除咬合紊乱组均高于对照组（P<0.05）,后部咬合紊乱组高于和去除咬合紊乱组,后者高于对照组（P<0.05）,前部无差异。结论咬合紊乱可导致幼年和成年大鼠髁突软骨BMP- 2高表达,成年大鼠髁突软骨对咬合紊乱的适应能力较幼年大鼠差,中部尤为明显。     

Changes in growth and morphology of the condyle following mandibular distraction in minipigs: overloading or underloading?

OBJECTIVE: Loading of temporomandibular tissues during mandibular distraction may cause changes in condylar growth and cartilage thickness. This study examines the effects of distraction on the condyle in a large animal model by explicitly measuring growth and in vivo loading. DESIGN: Unilateral mandibular distraction was carried out on 20 growing minipigs divided into three groups. One group underwent distraction but not consolidation, whereas the other two groups were allowed a period of consolidation of either 1 or 2 weeks. Animals received fluorochrome and 5-bromo-2'-deoxyuridine (BrdU) labelling and masticatory strain was measured from the condylar neck. Condylar strain was also recorded in an age-matched sample of eight animals that received no distraction surgery. Immunohistochemical procedures were used to identify dividing prechondroblasts and histological analysis was used to measure mineral apposition rate, count dividing cells, and measure the thickness of condylar cartilage. RESULTS: Strain magnitude, particularly compressive strain, was much larger on the non-distraction side compared to the distraction side condyle. Compared to normal loading levels, the distraction side condyle was underloaded whereas the condyle on the intact side was overloaded. Mineral apposition and cartilage thickness were greater on the distraction side condyle compared to the opposite side. Differences between the sides were most pronounced in the group with no consolidation and became progressively reduced with consolidation time. CONCLUSIONS: Increased mineralisation and cartilage thickness on the distraction side condyle are associated with reduced, not increased loading, perhaps because of disruption of the distraction side masseter muscle.     

Do masticatory functional changes influence the mandibular morphology in adult rats

Objective

This study aimed to investigate the effect of masticatory functional changes on the lateral view morphology of the mandible in adult rats.

Design

Sixty 21-day-old male Sprague–Dawley rats were divided into two groups. Sixteen received ordinary (hard) food during the whole experimental period (normal group). The remaining 44 received a soft diet during 21 weeks to develop a hypofunctional masticatory system. Then, the now adult animals were divided into two equal groups: one continued on a soft diet (hypofunctional group), and the other changed to an ordinary diet with the aim of functionally rehabilitating their masticatory system (rehabilitation group). After another 6 weeks all animals were sacrificed and their left mandible was dissected, photographed and customised cephalometric software was used to perform morphometric measurements.

Results

The area of the mandible was smaller in the hypofunctional compared to the normal group. Interestingly, the alveolar process was shorter in the normal group. Morphometric analysis revealed significant differences such as the area of the angular process and the inclination of the condylar process. The rehabilitation group was only marginally different compared to the hypofunctional group, although a general tendency to approach (catch-up) the normal group was observed, and one morphometric variable (condylar base inclination) was indeed significantly different.

Conclusions

Morphometric analysis revealed only marginal changes of the adult rat mandibular morphology during a 6-week period of masticatory function rehabilitation. However, the observed catch-up tendency might suggest that a longer rehabilitation period may have significant effect on mandibular morphology.     

Facial development disorders due to inhibition to endochondral ossification of mandibular condyle process caused by malnutrition

Objective:To analyze the effect of protein restriction on histomorphometric parameters of bone remodeling in mandibular condyle process and its possible influence in facial development in growing rats.Materials and Methods:Wistar rats weaned at the age of 21 days were assigned to one of the following groups: control (fed a regular hard diet ad libitum) and protein restricted (PR) (fed a hard diet lacking in protein ad libitum). The animals were euthanized on day 35 after the onset of the experiment. Mandibles were resected, fixed in 10% formalin, hemisected at the symphysis, and then radiographed in order to perform cephalometric studies of the condylar process length and the height of the lower alveolar process. Mandibles were then processed for light microscopy, and histomorphometric determinations were performed on histologic sections of the condylar process subchondral bone.Results:The PR group showed a significantly lower body weight than control group at the end of the experiment. The length of the condylar process was lower in the PR group; however, the diet used in this study did not affect the height of the lower alveolar process. The histomorphometric analysis showed that the PR group exhibited a statistically significant decrease in bone formation and bone volume in condylar process subchondral bone.Conclusion:Protein restriction inhibits bone formation and longitudinal growth in the mandibular condylar process. This result suggest that protein restriction can alter normal facial development.     

生长期山羊囊内骨折复位术中保留髁突软骨的重要性及组织学研究

目的：通过组织学观察,探讨髁突囊内骨折内固定术中保留和切除髁突软骨对髁突生长发育的影响。方法：6个月龄山羊12只,随机分为实验组（n=8）和对照组（n=4）。实验组双侧髁突造成囊内骨折并同期行手术复位固定．一侧保留髁突软骨,另一侧切除髁突软骨。术后3个月、6个月处死动物,切取髁突标本行石蜡切片和硬组织切片观察骨折愈合和髁突生长情况。结果：实验组髁突骨折愈合良好,钛板被新生骨组织覆盖;HE染色显示．保留髁突软骨组,髁突软骨结构清晰,与正常对照髁突相同,软骨成骨活跃,髁突生长发育正常;切除髁突软骨组,髁突软骨层消失,表面为成熟的骨细胞覆盖,直接与关节盘的纤维组织相连,新生骨组织少见：硬组织切片显示,钛板与骨组织直接结合．未见组织渗出和排异反应。结论：手术复位髁突囊内骨折时保留髁突软骨,不会影响髁突的生长发育;损伤髁突软骨．会造成髁突与关节盘黏连．引起髁突生长发育障碍。     

Effects of the masticatory demand on the rat mandibular development

The influence of masticatory loading stimulus on mandibular development is not fully clear. In this paper, experimental alterations in the daily muscle use, caused by a changed diet consistency, were continuously monitored, while adaptations in bone and cartilage were examined. It is hypothesised that decreased muscular loading will result in a decrease in the growth factor expression and mandible growth. Fourteen 21‐day‐old Wistar strain male rats were randomly divided into two groups and fed on either a hard or soft diet for 14 weeks. An implanted radio‐telemetric device recorded continuously muscle activity of the superficial masseter muscle. Chondroblast proliferation in the condylar cartilage was identified by insulin‐like growth factor‐1 receptor (IGF‐1r) immunostaining. Furthermore, an X‐ray was taken for cephalometric analysis. In the soft‐diet group, the duty time of the superficial masseter muscle at higher activity levels was significantly lower than that in the hard‐diet group. This decrease in muscular loading of the jaw system was accompanied by: a significant reduction in (i) articular cartilage thickness, (ii) expression of IGF‐1r immunopositive cells and (iii) mandible ramus height. In conclusion, a decrease in masticatory demand during the growth period leads to insufficient mandibular development.     

An impact of masticatory muscle function on IL-1beta and SOX9 expression in condyle

The importance of masticatory muscle function on the growth and differentiation of condyle was examined. The aim of this study was to investigate how reduced masticatory muscle function influences intrinsic regulatory factors which govern growth and differentiation of condylar cartilage. Masseter muscles of 3-week-old Wistar rats were resected bilaterally. Masseteric resected animals and corresponding control animals were sacrificed at 3, 6, 12 and 21 days post-resection. The condyles were then processed for histological and immunohistochemical analysis. The expression patterns of an inhibitory regulator (IL-1beta) and a master regulator (Sox9) of chondrogenesis in condylar cartilage of growing rats were investigated. Quantitative analysis shows that masseteric resection significantly increased the number of IL-1beta positive cells in proliferative layer. In contrast, the number of Sox9 positive cells was significantly decreased compared to the control animals. It can be concluded that the reduced articular function due to masseteric resection decreased condylar cartilage cell differentiation which led to the decrease in the thickness of condylar cartilage.     

功能性矫治器引导下颌后退对大鼠髁突软骨新骨形成影响的实验研究

目的探讨功能性矫治器引导下颌后退所致的髁突软骨新骨形成的影响。方法选用4周龄SD雄性大鼠45只,分为实验组及对照组,每组5只。实验组模拟临床功能性矫治器引导大鼠下颌后退。分别在0、3、7、14及21 d全麻下处死动物各5只。苏木精-伊红染色观察细胞形态反应,PAS染色观察新骨形成量。结果大鼠在下颌持续后退情况下,实验组髁突后份软骨组织增殖层和成熟层明显变薄,移行区可见破骨细胞增加,此区附近的骨小梁骨沉积减少;实验第3天及第7天,实验组和对照组髁突软骨新骨形成的量有明显区别,第14天和第21天,二者间的差别趋于减小。结论后退大鼠下颌后,髁突软骨后份新骨形成量减少。     

Micro X-ray computed tomography analysis for the evaluation of asymmetrical condylar growth in the rat

Authors– Nakano H, Watahiki J, Kubota M, Maki K, Shibasaki Y, Hatcher D, Miller AJ Objectives – To investigate the influence of forced lateral bite on mandibular growth, micro X‐ray computed tomography (CT) was used for the purpose evaluating condylar cartilage and cancellous bone formation in 10 male Wister rats (3 weeks of age). Settings and sample population – The rats were divided into two groups – experimental and control. In experimental group, an inclined crown was cemented onto the maxillary incisors to produce 2.5 mm shift toward the left side during mastication. Right–left differences in whole mandibular length, mandibular height, condylar size, trabecular structure of the condylar head and three‐dimensional (3‐D) finite element analysis were assessed using 3‐D images reconstructed from micro X‐ray CT scans when the mice had reached 21 weeks. Measurements and Results – Asymmetrical growth was found in the experimental group, in which the left condylar head became thicker and shorter than the right condylar head during development. When comparing the left and right condyles of the experimental animals, histomorphometric analysis from micro X‐ray CT showed that the bone volume (BV) of the cancellous bone, the surface area of the cancellous bone (BS), the BS/BV ratio, the BV fraction (BV/TV), and the trabecular thickness and trabecular number were less for the right condyle than for the left condyle. Conclusions – These findings suggested that artificial changes in the mastication do influence the growth of condylar head, condylar bone trabecular structure, and mineralization.     

渐进性咬合紊乱对大鼠颞下颌关节的影响

目的:探讨渐进性咬合紊乱对大鼠颞下颌关节的影响。方法:9只成年雄性SD大鼠,随机分为实验组(3只),操作对照组(3只)和空白对照组(3只)。实验组大鼠左侧上颌第一、第二磨牙间及右侧下颌第一、第二磨牙间分别嵌入正畸用皮圈推第一磨牙向近中,造成磨牙尖窝接触不吻合的渐进性咬合紊乱。术后6周取双侧颞下颌关节作HE染色观察。结果:操作对照组与空白对照组间未见差异,实验组与对照组相比髁突软骨中部增厚,后部变薄(P<0.05);关节盘中带和后带均明显变薄(P<0.05);实验组中部和后部软骨下骨小梁变得明显垂直于髁突表面排列。结论:渐进性咬合紊乱可以引起大鼠髁突软骨及软骨下骨小梁适应性改建。     

Effects of cyclosporin A on the mandibular condylar cartilage in rats.

Twenty, 5-week-old, male, Sprague-Dawley rats were divided into a control and a cyclosporin A (CSA) group for evaluating effects of the drug on condylar cartilage. Animals in the treatment group daily received CSA (15 mg/kg body wt) in mineral oil by gastric feeding over a 4-week observation interval. Control animals received mineral oil only. Five animals from each group were killed at weeks 2 and 4 of study. After histological processing, five tissue sections from the mid-region of the condyle were selected and examined. Three compositional zones (articular fibrous, proliferative, and hypertrophic) of the superior, posteriosuperior and posterior regions of the condylar cartilage were evaluated by light microscopy. At week 2, total condylar cartilage thickness was similar in the CSA and control groups, but the thickness of each zone was altered in CSA-treated animals, including a decrease of the fibrous and proliferative zones and an increase in hypertrophic zone compared to control (P<0.05). At week 4, CSA-treated animals exhibited overall decreased cartilage thickness, including decreased thickness of each zone compared to control (P<0.05). The results suggest that CSA has an inhibitory effect on the maturation of the mandibular condyle in rats.