Immunohistochemical detection of matrix metalloproteinase-1 in the periodontal ligament of equine cheek teeth

The hypsodont equine cheek tooth erupts continuously throughout life. The collagen fibers of the periodontal ligament (PDL) have to remodel constantly to allow the tooth to move in an occlusal direction. Remodeling of the collagen fiber bundles needs to be well-coordinated in order to maintain functional tooth support. The aim of this study was to examine the role of matrix metalloproteinase-1 (MMP-1) in the collagen remodeling of the equine PDL under physiological conditions. Specimens containing the PDL interposed between the dental cementum and the alveolar bone were taken from nine Warmblood horses at three designated horizontal levels: subgingival, middle, and apical. The expression of MMP-1 was detected immunohistochemically. MMP-1 was found to be present in the specimens of all horses. Immunopositive fibroblasts/fibrocytes were accumulated within individual single collagen fascicles. Our results suggest that MMP-1 induced collagen degradation plays a central role in the physiological remodeling of the equine PDL. The distribution of MMP-1 positive fascicles indicates well-directed remodeling which occurs as an asynchronous process, so that only single collagen fascicles are remodeled at the same time. Due to this remodeling of one fascicle at a time, the overall anchorage of the tooth is preserved at all times.     

Role of regulator of G-protein signaling 2 (RGS2) in periodontal ligament cells under mechanical stress

Mechanical stress is thought to regulate the expression of genes in the periodontal ligament (PDL) cells. Using a microarray approach, we recently identified a regulator of G-protein signaling 2 (RGS2) as an up-regulated gene in the PDL cells under compressive force. The RGS protein family is known to turn off G-protein signaling. G-protein signaling involves the production of cAMP, which is thought to be one of the biological mediators in response to mechanical stress. Here, we investigated the role of RGS2 in the PDL cells under mechanical stress. PDL cells derived from the ligament tissues of human premolar teeth were cultured in collagen gels and subjected to static compressive force. Compressive force application time-dependently enhanced RGS2 expression and intracellular cAMP levels. To examine the interrelationship between RGS2 and cAMP, the PDL cells were treated with 2',5'-dideoxyadenosine (DDA), an inhibitor of adenyl cyclase, or antisense S-oligonucleotide (S-ODN) to RGS2 under compressive force. DDA dose-dependently inhibited RGS2 stimulated by compressive force. Blockage of RGS2 by antisense S-ODN elevated the cAMP levels compared with controls. These results indicate that cAMP stimulates RGS2 expression, which in turn leads to a decrease in the cAMP production by inactivating the G-protein signaling in the mechanically stressed PDL cells.     

A simple cell motility assay demonstrates differential motility of human periodontal ligament fibroblasts, gingival fibroblasts, and pre-osteoblasts

During periodontal regeneration, multiple cell types can invade the wound site, thereby leading to repair. Cell motility requires interactions mediated by integrin receptors for the extracellular matrix (ECM), which might be useful in guiding specific cell populations into the periodontal defect. Our data demonstrate that fibroblasts exhibit differential motility when grown on ECM proteins. Specifically, gingival fibroblasts are twice as motile as periodontal ligament fibroblasts, whereas osteoblasts are essentially non-motile. Collagens promote the greatest motility of gingival fibroblasts in the following order: collagen III>collagen V>collagen I. Differences in motility do not correlate with cell proliferation or integrin expression. Osteoblasts display greater attachment to collagens than does either fibroblast population, but lower motility. Gingival fibroblast motility on collagen I is generally mediated by α2 integrins, whereas motility on collagen III involves α1 integrins. Other integrins (α10 or α11) may also contribute to gingival fibroblast motility. Thus, ECM proteins do indeed differentially promote the cell motility of periodontal cells. Because of their greater motility, gingival fibroblasts have more of a potential to invade periodontal wound sites and to contribute to regeneration. This finding may explain the formation of disorganized connective tissue masses rather than the occurrence of the true regeneration of the periodontium. This research was supported by the Louisiana Board of Regents through the Millennium Trust Health Excellence Fund, HEF-(2000-05)-04.     

关节镜下多种移植物联合重建膝关节前、后交叉韧带

目的：评估关节镜下膝关节前交叉韧带（ACL）与后交叉韧带（PCL）同时重建的技术和临床效果。方法：自2003年6月～2009年10月,27例病人（28膝）经MRJ检查及关节镜检查证实ACL和PCL均断裂,其中9膝伴内侧副韧带损伤（MCL）,8膝伴后外侧角损伤（PLC）,5膝伴内侧半月板破裂,4膝伴外侧半月板损伤。27例患者于伤后3～10周在关节镜下行膝关节前、后交叉韧带联合重建。结果：本组术后早期均未发生严重并发症。术后随访12-88个月,平均（42．67±3．34）个月,Lysholm膝关节功能评分为78-93分,平均（86．67±5．21）分。国际膝关节文件编制委员会（mDC）综合评定由术前显著异常（D级）28膝,改进为随访时正常（A级）9膝、接近正常（B级）16膝、异常（C级）3膝。结论：关节镜下膝关节前交叉韧带（ACL）与后交叉韧带（PCL）同时重建创伤小、手术操作精细,术后膝关节功能恢复满意。     

关节镜下多种移植物联合重建膝关节前、后交叉韧带

目的:评估关节镜下膝关节前交叉韧带(ACL)与后交叉韧带(PCL)同时重建的技术和临床效果。方法:自2003年6月～2009年10月,27例病人(28膝)经MRI检查及关节镜检查证实ACL和PCL均断裂,其中9膝伴内侧副韧带损伤(MCL),8膝伴后外侧角损伤(PLC),5膝伴内侧半月板破裂,4膝伴外侧半月板损伤。27例患者于伤后3～10周在关节镜下行膝关节前、后交叉韧带联合重建。结果:本组术后早期均未发生严重并发症。术后随访12～88个月,平均(42.67±3.34)个月,Lysholm膝关节功能评分为78～93分,平均(86.67±5.21)分。国际膝关节文件编制委员会(IKDC)综合评定由术前显著异常(D级)28膝,改进为随访时正常(A级)9膝、接近正常(B级)16膝、异常(C级)3膝。结论:关节镜下膝关节前交叉韧带(ACL)与后交叉韧带(PCL)同时重建创伤小、手术操作精细,术后膝关节功能恢复满意。     

Bold 螺钉治疗内踝骨折的疗效分析

目的:对比Bold螺钉和普通空心螺钉内固定治疗单纯内踝骨折的疗效。方法:空心螺钉治疗单纯内踝撕脱骨折57例,分为A组Bold螺钉内固定治疗内踝骨折25例,B组使用普通空心螺钉内固定32例。结果:两组57例均获得随访,两组病例远期均能得到较坚强的固定和良好的功能恢复,但Bold螺钉组相对普通螺钉组愈合时间更快(P<0.05),下床活动时间更早(P<0.05),早期踝关节功能评分高(P<0.05),但六个月后没有明显差异。结论:Bold螺钉内固定有助于内踝骨折早期愈合和早期功能锻炼,是一种比较好的内固定材料,值得推广。     

双侧自体腘绳肌腱一期重建前后交叉韧带损伤的临床疗效

目的:探讨关节镜辅助下使用双侧自体腘绳肌腱一期修复膝关节前后交叉韧带损伤的方法和临床疗效。方法:内窥镜微创双侧自体腘绳肌腱修复膝关节内韧带,术后用IKDC分级、影像学IKDC分级、Lysholm功能评分和KT2000TM测量进行关节机能打分。结果:11例患者获得3-5年随访,平均随访3.8年。术前Lysholm功能评分平均(46.8±5.7)分,终末随访时平均(81.3±10.5)分,差异有显著性(P<0.05)。术后关节稳定性测量,在20磅时、30磅和最大拉力时健膝和患膝分别是:6.1±0.3和6.8±0.8;6.3±0.5和7.7±1.3;7.5±0.6和9.6±2.4,统计学上差异无显著性(P>0.05)。主观IKDC分级:A级4例,B级6例,C级1例;影像学IKDC分级:A级8例,B级2例,C级1例。结论:关节镜辅助下使用双侧自体腘绳肌腱一期修复膝关节前后交叉韧带损伤是重建膝关节稳定性的良好有效方法。     

Platelet-rich plasma provides nucleus for mineralization in cultures of partially differentiated periodontal ligament cells

Summary Platelet-rich plasma (PRP) has been used to promote periodontal regeneration following the premise that constituent transforming growth factor-β1 (TGF-β1) and platelet-derived growth factor-AB will stimulate cell proliferation at the site of application. In previous studies, we demonstrated that PRP mimics TGF-β1 to modulate proliferation in a cell type-specific manner, that fibrin clot formation by PRP upregulates type I collagen, and that an unidentified factor(s) in PRP increases alkaline phosphatase (ALP) activity in human periodontal ligament (PDL) cell cultures. We have now examined the effects of PRP on in vitro mineralization. Platelet-rich plasma and PDL cells were prepared from human adult volunteers or rats. After 20 d of continuous treatment with PRP in dexamethazone (Dex)-containing osteogenic medium, PRP time dependently promoted mineralization by rat PDL cells but failed to fully induce the osteoblastic phenotype. Furthermore, when human PDL cells were induced to increase ALP activity in osteogenic medium that lacked Dex, a condition that should delay (or suppress) osteoblastic differentiation, transmission electron microscopy revealed that mineralized spicules were initially deposited onto PRP-derived platelet aggregates. Taken together with our previous data, these findings suggest that PRP provides platelet aggregates as nuclei to initiate mineralization while stimulating PDL cell proliferation, differentiation, and collagen production. The combination of these effects may effectively mediate PRP's ability to promote regeneration of periodontal tissue, including skeletal tissue, at the site of injury.     

Proliferation and tube formation of periodontal endothelial cells

Angiogenesis is indispensable to guide a regeneration of good periodontal tissue in the wound healing after periodontal surgery. Hepatocyte growth factor is well known for a strong angiogenic factor and it may play important roles in the periodontal tissue during periodontal wound healing. In exploring the promotion of angiogenesis in the periodontal ligament, proliferative and tubulogenic responses of endothelial cells to hepatocyte growth factor and to soluble factors secreted by fibroblasts were investigated. Pavement-shaped cells isolated from a human periodontal ligament were identified as the endothelial cell by their granular immunoreactivity for factor VIII. The proliferation of the endothelial cells was accelerated by the addition of hepatocyte growth factor or fibroblast-conditioned medium, and far more by adding both than either. The endothelial cells seeded on the agar containing both hepatocyte growth factor and fibroblast products formed a dense network in a shorter time than on the agar containing either. The endothelial cells in the dense network took a tube-like structure with lumen and were covered with laminin. These results suggest that hepatocyte growth factor administered into the regenerating periodontal tissue may promote, synergistically with local factors produced by the activated fibroblast, the proliferation and tubulogenesis of the remaining endothelial cells.