Mohamed Sharawy

Osteoarthrosis of the temporomandibular joint following experimental disc perforation in Macaca fascicularis.

The aim of this experiment was to study the sequela of experimental temporomandibular joint (TMJ) disc perforation. Each TMJ of four Macaca fascicularis adult monkeys was surgically exposed, and a 4- to 6-mm perforation at the posterolateral portion of the avascular disc was produced by electrosurgery. Four monkeys were used as controls. The animals were killed 11 weeks (two experimental and two controls) or 12 weeks (two experimental and two controls) after disc perforation. The perforations were increased in size in five joints, and healed in one joint. In addition, two joints of one animal showed complete loss of the disc, denudation of articular surfaces, and bone-to-bone contact. In contrast to control joints, the experimental joints exhibited the following changes histopathologically: thick, highly cellular and fibrillated fibrous coverings of articular surfaces (five joints); marked hyperplasia of synovial membrane; migration of synovial cells on the surfaces of the disc and margins of perforation; multiple adhesions of disc to articular surfaces; increase in cellularity and vascularity of discs; and chondrocytic clustering in temporal fibrous covering; and osteophytes of condylar and temporal components and focal or complete denudation of articular surfaces (2 joints). Most of these changes were consistent with the diagnosis of osteoarthritis. From this study, one can conclude that disc perforation can lead to osteoarthritis.

Effect of Ovariectomy and Alendronate on Implant Osseointegration in Rat Maxillary Bone

Bisphosphonates such as alendronate (ALD), although controversial, are worthy of investigation for the enhancement of implant osseointegration in patients with low bone mass who are already taking bisphosphonates for osteoporosis. These patients may receive additional benefits and be acceptable candidates for dental implants without needing to change their medication regimen and possibly as a result of their medication regimen. The purpose of this study was to compare implant osseointegration in maxillary bone of normal rats with a rat model of postmenopausal estrogen deficiency (ovariectomized [OVX]), with and without ALD. An experimental group of 32 rats was divided in 4 groups: ALD-OVX (n=8 OVX with ALD), OVX (n=8 OVX without ALD), ALD (n=8 normal rats with ALD), and control (n=8 normal rats). All rats received one titanium microscrew implant in the left edentulous region of the maxillary arch. The ALD-OVX and ALD groups received subcutaneous injections of ALD 3 times a week. On the fourth week after ALD administration, an implant was placed in all 32 rats. The maxilla of each rat was radiographed 4 times: at 0, 7, 14, and 28 days. On day 28 after implant placement, all rats were killed, and the peri-implant tissue was embedded in plastic or paraffin for histological examination. The X rays were used for a chronologic calculation of the contact ratio between implant and bone surfaces. Radiographic bone density was determined at 3 points: mesial, apical, and distal. The results show that osseointegration of the implants was impaired in the estrogen-deficient OVX rats compared with the ALD-OVX rats. Fifty percent of the implants were lost at 2 weeks in the OVX group. Radiographic evidence suggested that none of the implants in the OVX group osseointegrated. In the histologic examination more bone was observed around implants from the ALD-OVX and ALD groups than around implants from the OVX group. The OVX group presented a dramatic reduction in implant bone contact at 2 weeks and a significant 13% reduction at 4 weeks vs day of implant (P = .006). The ALD-OVX group presented 50% more bone density than the OVX group (P = .0003). Both ALD groups (ALD and ALD-OVX) had significantly higher radiographic bone density than the other groups (P < .01 for each comparison). In conclusion, osseointegration of implants was enhanced by ALD. Radiographic bone density and contact ratio improved with ALD administration. Implant osseointegration was impaired by estrogen deficiency in the OVX group.

Workshop Guidelines on Immediate Loading in Implant Dentistry

P redictable formation of a direct bone-toimplant interface is a treatment goal in implant dentistry. The 2-stage surgical protocol established by Branemark et al to accomplish osseointegration consisted of several prerequisites, including (1) countersinking the implant below the crestal bone, (2) obtaining and maintaining a soft-tissue covering over the implant for 3 to 6 months, and (3) maintaining a minimally loaded implant environment for 3 to 6 months. The primary reasons cited for the submerged, countersunk, surgical approach to implant placement were (1) to reduce and minimize the risk of bacterial infection, (2) to prevent apical migration of the oral epithelium along the body of the implant, and (3) to minimize the risk of early implant loading during bone remodeling. After this procedure, a second-stage surgery was necessary to uncover these implants and place a prosthetic abutment. Predictable, long-term, clinical rigid fixation has been reported after this protocol in patients who were either completely or partially edentulous. During the past 15 years, several authors have reported that root-form implants may osseointegrate, even though the implants extend above the bone and through the soft tissues during early bone remodeling. This surgical approach has been called a 1-stage or nonsubmerged implant procedure because it eliminates the second-stage implant uncovery surgery. As a result, the discomfort, inconvenience, and appointments of the surgery and suture removal are eliminated. In addition, the soft tissue is more mature before fabricating a final prosthesis.

Osteoinduction in Rhesus Monkeys using demineralized bone powder allografts

This experiment tested the osteoinductivity of demineralized bone powder (DBP) in nonhuman primates. Six DBP implants were implanted subcutaneously close to the pectoralis major muscle in each of four rhesus monkeys. Excisional biopsies were obtained 20, 40, and 72 days after implantation and were processed for light and electron microscopy. Decalcified and undecalcified sections of the implants were studied. Large numbers of undifferentiated mesenchymal and fibroblast-like cells were observed around and within the DBP matrix particles on day 20. Cartilage formation was also evident at that time and had increased by day 40, when chondroid bone also appeared. By day 72, the implants showed mature (lamellar) and immature (woven and chondroid) bone and bone marrow formation. Areas of DBP that were incorporated within the induced bone contained empty lacunae and stained similarly to mineralized bone. It was concluded that allogenic DBP can induce bone formation in monkeys. The results justify the use of DBP as a bone-banked material to induce bone formation in humans.

Green tea polyphenols reduce autoimmune symptoms in a murine model for human Sjogren's syndrome and protect human salivary acinar cells from TNF-a-induced cytotoxicity

Sjogrens syndrome (SS) is a relatively common autoimmune disorder. A key feature of SS is lymphocytic infiltration of the salivary and lacrimal glands, associated with the destruction of secretory functions of these glands. Current treatment of SS targets the symptoms but is unable to reduce or prevent the damage to the glands. We reported previously that the major green tea polyphenol (GTP) epigallocatechin-3-gallate (EGCG) inhibits autoantigen expression in normal human keratinocytes and immortalized normal human salivary acinar cells (Hsu et al. 2005). However, it is not known whether GTPs have this effect in vivo, if they can reduce lymphocytic infiltration, or protect salivary acinar cells from tumor necrosis factor-α (TNF-α)-induced cytotoxicity. Here, we demonstrate that in the NOD mouse, a model for human SS, oral administration of green tea extract reduced the serum total autoantibody levels and the autoimmune-induced lymphocytic infiltration of the submandibular glands. Further, we show that EGCG protected normal human salivary acinar cells from TNF-α-induced cytotoxicity. This protection was associated with specific phosphorylation of p38 MAPK, and inhibitors of the p38 MAPK pathway blocked the protective effect. In conclusion, GTPs may provide a degree of protection against autoimmune-induced tissue damage in SS, mediated in part through activation of MAPK elements.

Effects of oral consumption of the green tea polyphenol EGCG in a murine model for human Sjogren's syndrome, an autoimmune disease.

SIGNIFICANCE Protection of glandular cells from autoimmune-induced damage would be of significant clinical benefit to Sjogrens syndrome (SS) patients. Epigallocatechin-3-gallate (EGCG) possesses anti-apoptotic, anti-inflammatory, and autoantigen-inhibitory properties. AIMS To investigate if EGCG protects against certain autoimmune-induced pathological changes in the salivary glands of the non-obese diabetic (NOD) mouse model for SS. MAIN METHODS Animals were provided with either water or water containing 0.2% EGCG. At the age of 8, 16 and 22 weeks, submandibular salivary gland tissue and serum samples were collected for pathological and serological analysis. KEY FINDINGS Significant lymphocyte infiltration was observed in the salivary glands of the water-fed group at the age of 16 weeks, while the EGCG group showed reduced lymphocyte infiltration. By 22 weeks of age, water-fed animals demonstrated elevated levels of apoptotic activity within the lymphocytic infiltrates, and high levels of serum total anti-nuclear antibody, compared to EGCG-fed animals. Remarkably, proliferating cell nuclear antigen (PCNA) and Ki-67 levels in the salivary glands of water-fed NOD mice were significantly elevated in comparison to BALB/c control mice; in contrast, PCNA and Ki-67 levels in EGCG-fed NOD animals were similar to BALB/c mice. These results indicate that EGCG protects the NOD mouse submandibular glands from autoimmune-induced inflammation, and reduces serum autoantibody levels. Abnormal proliferation, rather than apoptosis, appears to be a characteristic of the NOD mouse gland that is normalized by EGCG. The evidence suggests that EGCG could be useful in delaying or managing SS-like autoimmune disorders.

Enhancement of osteoblast proliferation in vitro by selective enrichment of demineralized freeze-dried bone allograft with specific growth factors.

Decalcified freeze-dried bone allograft (DFDBA), believed to serve as a matrix for new bone growth and to contain various bone-inducing growth factors, is currently used to regenerate periodontal defects and to restore and maintain dental alveolar ridges. Growth factors within DFDBA are extracted during the demineralization process, thus rendering the allograft incapable of spontaneous osteogenesis; however, exogenous growth factor addition to DFDBA may enhance the osteogenic capacity of native osteoblasts. This studys purpose is to evaluate murine osteoblast proliferation in the presence of various exogenous soluble growth factors as measured by fluorescence units. Osteoblasts harvested from mouse pup calvaria were cultured with 2% residual calcium-DFDBA and supplemented by one of the following growth factors or combinations of these factors: transforming growth factor-beta (TGF-beta), insulin-like growth factor-I (IGF-I), platelet-derived growth factor (PDGF), fibroblast growth factors basic (bFGF), or vascular endothelial growth factors (VEGF). Osteoblast proliferation rates indicate that the in vitro supplementation of 2% residual calcium-DFDBA with the combination of IGF and TGF-beta, IGF and PDGF, and PDGF and TGF-beta significantly (P < or = .05) enhance murine osteoblast activity and proliferation at 7 days compared with the control containing no exogenous growth factors.

Morphological Alterations in the Elastic Fibers of the Rabbit Craniomandibular Joint following Experimentally Induced Anterior Disk Displacement

Elastic fibers are important components of the connective tissue that attaches the articular disk of the craniomandibular joint (CMJ) to the skull and mandible. Biopsies of the articular disk proper and bilaminar zone (BZ) tissues from patients with anterior disk displacement (ADD) have shown previously that there is a marked loss of elastic fibers. In the present study, the effects of inducing ADD on the elastic fibers in the rabbit CMJ disk proper, BZ and condylar cartilage were investigated. The right CMJ was exposed surgically and the discal attachments were severed except for the BZ attachments. Then, the disk was displaced anteriorly and sutured to the zygomatic arch. The CMJs were removed after 1, 2 or 6 weeks and processed for histochemical demonstration of elastic fibers. The results showed osteoarthritic changes following ADD, and a significant decrease in the number of the elastic fibers in the disk proper and BZ. The remaining elastic fibers were abnormal in their appearance and orientation. In addition, ADD led to the appearance of fine elastic fibers among the chondrocytes in the hyaline cartilage of the condyle that were not present in the cartilage of the control condyle. We conclude that induced ADD can lead to a significant loss of elastic fibers in the articular disk, and result in the appearance of elastic fibers within the cartilage of the mandibular condyle.

Osteoinduction in young and old rats using demineralized bone powder allografts

The process of inducing differentiated and undifferentiated cells to become osteogenic using demineralized bone powder (DBP) is a well-known phenomenon in developmental biology. The aim of this study was to examine whether age has an effect on the process of bone induction. DBP was implanted in the subcutaneous thoracic tissue of young rats (28-56 days) and old rats (14 months or older), and the animals were examined seven, 12, 20, and 60 days after implantation. The amount of newly induced bone in the implant was quantitatively measured using histomorphometry and 45Ca uptake. Undecalcified and decalcified specimens were processed for histologic examination using several stains that demonstrate osteoid. Both the young rats and the old rats formed bone in response to the DBP implants. In old animals the induced bone appeared to be less in quantity, it formed at a slower rate, and it exhibited less bone marrow cellularity than did the bone in young animals.

Periodontal ligament fibroblasts sustain destructive immune modulators of chronic periodontitis.

BACKGROUND In healthy periodontal tissue, innate immune responses effectively confine and suppress a bacterial insult. However, a disruption of the host-bacterial equilibrium may produce an overexpression of cytokines and lead to permanent, host-mediated tissue damage. Although such periodontal destruction primarily results from activated immune mechanisms, the site-specific damage suggests that local tissues participate in these pathologic changes. Periodontal ligament fibroblasts (PDLFs) are prominent in the periodontium and are critical in homeostasis and regeneration because they have the ability to produce multiple cytokines in response to a bacterial insult. These cells could play a role in the local pathogenesis of periodontal disease. METHODS We studied alkaline phosphatase (ALP) activity, interleukin (IL)-6 production, and morphologic characteristics of cultured PDLFs that were isolated from periodontally healthy sites (H-PDLFs) and diseased sites (D-PDLFs) in humans. Quantitative analyses of 84 genes that are related to inflammation were performed using real-time polymerase chain reaction arrays. RESULTS A mineralizing medium induced a significant increase of ALP in H-PDLFs, but no significant enzymatic changes were detected in D-PDLFs after such treatment. The protein and gene expression of IL6 showed a significant upregulation in D-PDLFs, which also demonstrated a significant upregulation of 54% of genes in the inflammatory gene arrays. CONCLUSIONS To our knowledge, these results represent the first biologic evidence that D-PDLFs retain uniquely inflammatory phenotypes that could maintain localized destructive signals in periodontitis. The overexpression of proinflammatory cytokines by PDLFs could amplify local inflammation by the continuous triggering of immune responses. In addition, the location of these cells could be critical in the progression of the inflammatory front into the deeper tissues.