Zvi Artzi

In vitro Effects of Enamel Matrix Proteins on Rat Bone Marrow Cells and Gingival Fibroblasts

Emdogain® (EMD), a formulation of Enamel Matrix Proteins (EMP), is used clinically for periodontal regeneration, where it stimulates cementum formation and promotes gingival healing. In this study, we investigated the in vitro effects of EMD on rat bone marrow stromal cells (BMSC) and gingival fibroblasts (GF). EMD (at 25 μg/mL) increased the osteogenic capacity of bone marrow, as evidenced by ~ three-fold increase in BMSC cell number and ~ two-fold increase in alkaline phosphatase (ALP) activity and mineralized nodule formation. The presence of EMD in the initial stages (first 48 hrs) of the culture was crucial for this effect. In contrast, EMD did not induce osteoblastic differentiation of GF (evidenced by lack of mineralization or ALP activity) but increased up to two-fold both their number and the amount of matrix produced. These in vitro data on BMSC and GF could explain the promotive effect of EMD on bone formation and connective tissue regeneration, respectively.

Histomorphometric assessment of bone formation in sinus augmentation utilizing a combination of autogenous and hydroxyapatite/biphasic tricalcium phosphate graft materials: at 6 and 9 months in humans

OBJECTIVE The aim of this study was to examine the efficacy of a new biphasic hydroxyapatite/tricalcium phosphate (HA/TCP) bone substitute in combination with particulate autogenous bone in sinus floor augmentation procedures. MATERIAL AND METHODS A simultaneous or a two-stage sinus augmentation and implant placement were conducted in 28 patients. A mixture of HA/TCP and autogenous bone chips in a 1 : 1 ratio was used as the grafting biomaterial. Cylindrical specimen bone retrieval was performed in all patients except one. Specimens were harvested either at 6 (n=14) or 9 (n=13) months post-augmentation. For histologic and histomorphometric evaluations, the non-decalcified tissue processing (Donaths technique) was performed. RESULTS Newly formed bone around the grafted particles was found in all samples. The encircling, highly cellular bone followed the outline of the grafted particles in direct contact. Both woven and lamellar types of bone were observed. Morphometrically, the total mean bone area fraction of all sections was 34.8+/-10.3%, increasing from 28.6+/-7.8% at 6 months to 41.6+/-8.3% at 9 months (P<0.001). Mean particle area fraction average was 25.5+/-11.6% and 23.5+/-9.3% at 6 and 9 months, respectively, with a total mean of 24.5+/-10.4%. The increase in bone area fraction was not significantly correlated to the decrease of the grafted particles area fraction. CONCLUSIONS The biphasic HA/TCP showed biocompatible and osteoconductive properties. This alloplast as a composite with autogenous bone chips promotes newly formed bone, which increases in its fraction along an extended healing period.

Mucosal considerations for osseointegrated implants

Tissue resistance is determined by the nature of cells and intercellular contacts irrespective of the presence or absence of keratinization, masticatory mucosa, or skin. However, these tissues are more easily maintained and less vulnerable to inflammation when in contact with dental implants. Lack of masticatory mucosa and the presence of alveolar mucosa embracing the implant are often associated with plaque, which can induce inflammation resulting in subsequent peri-implant destruction. To facilitate proper mechanical oral hygiene maintenance, transplantation of autogenous masticatory mucosal grafts at the implant sites was performed in patients without attached gingiva, unfavorable vestibulum with submucosal muscular activity, and uncontrolled peri-implant mucositis. The rationale for having attached mucosa around osseointegrated implants and illustration of possible methods of mucosal management in the different phases of implant rehabilitation are presented.

Long‐term bio‐degradation of cross‐linked and non‐cross‐linked collagen barriers in human guided bone regeneration

OBJECTIVE This double-blind study clinically and histologically evaluated long-term barrier bio-durability of cross-linked and non-cross-linked collagen membranes (CLM and NCLM) in sites treated by guided bone regeneration procedures. MATERIALS AND METHODS In 52 patients, 52 bony defects were randomly assigned to treatment with either a CLM or a NCLM. Post-surgical spontaneous membrane exposures were recorded. Before implant placement, full-thickness standard soft tissue discs were retrieved wherever suitable for histologic examination. RESULTS Spontaneous membrane exposure was observed in 13 (50%) CLM sites and in six (23.1%) NCLM sites (P<0.05). Clinical healing at exposed sites lasted 2-4 weeks. CLM were histologically intact in all non-perforated sites, were interrupted in five perforated sites, and undetected in four. NCLMs were undetected in all 18 specimens examined. In three non-perforated CLM sites, bone apposition and ossification at or within the membrane was observed. CONCLUSIONS CLMs were more resistant to tissue degradation than NCLMs, and maintained integrity during the study. Neither membrane was resistant to degradation when exposed to the oral environment. CLMs were associated with a higher incidence of tissue perforations. In non-perforated sites, CLM ossification at or within the membrane was occasionally observed.

Antral computerized tomography pre‐operative evaluation: relationship between mucosal thickening and maxillary sinus function

OBJECTIVES to assess the correlation between maxillary sinus inferior mucosal thickening and sinus outflow obstruction. MATERIAL AND METHODS the study included 280 computerized tomography (CT) scans (560 maxillary sinuses). CT aimed to assess sinusitis; trauma to the face and intubated patients were excluded. Mucosal thickening was graded as < 5 mm (1), < 10 mm (2), < 15 mm (3), < 20 mm (4) and > 20 mm (5), and classified by appearance as normal, rounded, circumferential, irregular, or complete. Maxillary sinus outflow was classified as patent or obstructed. RESULTS mucosal thickening was found in 36.1% of the maxillary sinuses, graded as 31.2% (1), 34.2% (2), 12.9% (3), 5.4% (4) and 16.3% (5), and classified as rounded (11.8%), irregular (10.4%), circumferential (8.8%) and complete (5.2%). Sinus outflow was obstructed in 15% of the scans. Mucosal thickening of < 5 mm (11.1%), < 10 mm (36.2%) and > 10 mm (74.3%) was associated with sinus obstruction (P<0.0001). Rounded (6.1%), circumferential (55.2%), irregular (38.8%) and complete (100%) mucosal appearances were associated with sinus obstruction (P<0.001). When statistically combined, a substantial risk for sinus obstruction was observed with irregular mucosal appearance of > 5 mm (56.5% for grade 2 up to 82.6% for grades 3-5) and circumferential appearance (21.4% for grade 1 up to 100% for grades 3-5). A low risk for obstruction was found with the rounded appearance (mean 6.1%). CONCLUSIONS irregular (> 5 mm), circumferential and complete mucosal appearance are associated with an increased risk for sinus outflow obstruction and an ENT consultation is recommended. A rounded mucosal appearance of any grade is associated with a low risk for sinus obstruction. Routine CT scans, including the maxillary sinus ostium, are recommended.

Clinical evaluation of particulate allogeneic with and without autogenous bone grafts and resorbable collagen membranes for bone augmentation of atrophic alveolar ridges

OBJECTIVES The evaluate the clinical outcome of bone augmentation with the use of particulate mineralized freeze-dried bone allograft (FDBA) with or without the addition of autogeneous bone chips, applied in a bi-layered (BL) technique, covered by a resorbable cross-linked collagen membrane. MATERIAL AND METHODS Fifty patients presenting with a vertical and/or lateral ridge deficiency of at least 3 mm were included: Group FDBA, N=27 patients, particulate FDBA was the only graft; and Group BL, N=23 patients, a BL bone grafting procedure where autogenous bone chips were the inner layer and FDBA the outer. Bone graft was covered with a ribose cross-linked collagen barrier membrane. Ridge dimensions were clinically or radiographically (computerized tomography scan) measured at the time of the bone augmentation procedure and at implant placement or uncovering and the maximum linear vertical or horizontal calcified tissue gain was calculated. Statistical analysis consisted of linear regression analysis, with maximum bone gain being the dependent variable. RESULTS In the FDBA group, mean vertical bone gain was 3.47 mm (SD 1.25) and the horizontal, 5 mm (SD 1.28), while in the BL values were 3.5 mm (SD 1.2) and 3.6 mm (SD 1.72), respectively. Addition of autogenous bone does not appear to statistically significantly enhance the results. Spontaneous membrane exposure occurred in 24% of the cases and was the only variant that significantly influenced results (P<0.001). CONCLUSIONS Large vertical and/or horizontal ridge deficiencies may be treated with FDBA and ribose cross-linked collagen barrier membranes with good clinical outcome. No added effect of the application of a layer of autogenous bone in these bone augmentation procedures could be demonstrated. Spontaneous membrane exposure was the only parameter to affect the degree of new calcified tissue formation.

Bio-degradation of a resorbable collagen membrane (Bio-Gide) applied in a double-layer technique in rats.

OBJECTIVE The aim of this study was to evaluate histologically the bio-degradation of two layers of Bio-Gide((R)) (BG) membrane, as compared with that of a single layer. MATERIAL AND METHODS Two circular calvarial bony defects, 5 mm in diameter, were made in 24 Wistar rats. BG membrane, labeled with biotin, was cut into 5-mm-diameter disks, and placed in defects either as a mono-layer membrane (MLM) or as a double-layer membrane (DLM). Rats were sacrificed after 4 or 9 weeks and histology was performed. Membranes were stained with horseradish peroxidase-conjugated streptavidin and aminoethyl carbazole as a substrate for detection of biotinylated collagen. The area of collagen and thickness of the residual membranes were measured by image analysis software. Statistical analysis was performed using the non-parametric Wilcoxons signed-ranks test. RESULTS At 4-week collagen area per measurement window within the DLM sites (0.09+/-0.05 mm(2)) was significantly greater (P<0.01) than that in the MLM sites (0.047+/-0.034 mm(2)). At 9 weeks, the collagen area was also greater in the DLM sites (0.037+/-0.026 mm(2)) compared with that of the MLM sites (0.025+/-0.016 mm(2)); however, this difference did not reach statistical significance. The rate of membrane degradation, calculated as percent membrane lost compared with baseline, was similar for the DLM and MLM at both time points ( approximately 60% at 4 weeks and approximately 80% at 9 weeks). In addition, the residual DLM thickness at 4 weeks (475.5+/-73.77 mum) was significantly (P<0.01) greater than that of MLM (262.38+/-48.01 mum). At 9 weeks, membrane thickness was also greater in the DLM sites (318.22+/-70.45 mum) compared with that of the MLM sites (183.32+/-26.72 mum); however, this difference did not reach statistical significance. The reduction in thickness between 4 and 9 weeks was 30% for MLM and 33% for DLM. DISCUSSION The use of a double layer of BG membrane results in a barrier of increased collagen area and thickness, compared with application of a single layer.

Effect of Guided Tissue Regeneration on Newly Formed Bone and Cementum in Periapical Tissue Healing after Endodontic Surgery: An In Vivo Study in the Cat

INTRODUCTION The purpose of this study was to evaluate the influence of anorganic bovine bone as a grafted biomaterial on newly formed bone and cementum in periapical regions after surgical endodontic treatment in cats. METHODS After inducing apical periodontitis in 9 cats, root canal and surgical endodontic treatment were performed on 72 roots of first and second maxillary premolars. Bone defects were treated with biomaterial particles + a membrane, biomaterial only, a membrane only, or left unfilled (control). Histomorphometry on nondecalcified sections were performed at 3 and 6 months after surgery. Analysis of variance with repeated measures was used within 2 and 3 subject factors to analyze newly formed bone, cementum, biomaterial conduction, and resorption. RESULTS At each time period, bone formation was greater at the grafted membrane-protected sites than in the grafted-unprotected sites. At 6 months, the bone area fraction at membrane nongrafted sites was greater than in the grafted-protected sites. The new cementum was significantly greater at 6 months than at 3 months and greater at the grafted membrane-protected sites over the unprotected ones at 6 months. Statistically, the grafted biomaterial, the membrane, and the time contributed significantly to the amount of new bone (P<.05) with no significant interaction. Biomaterial osteoconduction was significantly affected by the time. All 3 variables showed a significant interaction on new cementum. CONCLUSIONS There was significantly more bone formation after surgical endodontic treatment when membrane and bone grafts were used as compared with bone grafts only or unfilled control sites. However, it appears that the key factor to the enhanced tissue regeneration is the membrane and not the grafted biomaterial.

Simultaneous versus two-stage implant placement and guided bone regeneration in the canine: histomorphometry at 8 and 16 months.

AIM To compare the effect of timing of implant placement and guided bone regeneration (GBR) procedure on osseointegration and newly formed bone at 8 and 16 months. MATERIAL AND METHODS In seven dogs, four different sites were bilaterally established: (1) an implant placed in a 6-month healed (6m-GBR) bovine bone mineral (BBM) grafted site; (2) a simultaneously placed implant with the grafted BBM (Si-GBR) followed by a membrane coverage; (3) an implant placed in a membrane-protected non-grafted defect; and (4) an implant placement in a naturally healed site (Cont). Histomorphometry was obtained at 8 and 16 months post-implant placement. Bone-implant contact (BIC), crestal bone resorption (CBR), vertical intra-bony (VIB) defect, bone (BAF) and particle (PAF) area fractions, and osteoconductivity (CON) levels were measured. RESULTS In all sites, BIC ranged between 62% and 79% with no significant differences. PAF ranged from 17% to 27%, with no effect of time. At 8 and 16 months, BAF was significantly smaller at the Si-GBR site when compared with all other sites, CON was significantly greater at the 6m-GBR site, and CBR and VIB were significantly smaller at the 6m-GBR when compared with the Si-GBR sites. CONCLUSIONS The simultaneous and delayed techniques both showed a similar osseointegration level over time. However, the staged approach showed enhanced newly formed bone, higher osteoconduction around the grafted mineral, less CBR, and smaller vertical bone defect over time compared with the combined approach.

Deproteinized bovine bone in association with guided tissue regeneration or enamel matrix derivatives procedures in aggressive periodontitis patients: a 1-year retrospective study.

OBJECTIVES To retrospectively evaluate and compare two regenerative periodontal procedures in young individuals with aggressive periodontitis (AgP). METHODS Thirty-two patients aged 14-25 years (mean ± SD 19.3 ± 5.7) were diagnosed as having AgP with multiple intra-bony defects (IBDs) and treated by one of two regenerative modalities of periodontal therapy: guided tissue regeneration (GTR) using deproteinized bone xenograft (DBX) particles and a resorbable membrane (the GTR group), or an application of enamel matrix derivatives (EMD) combined with DBX (the EMD/DBX group). Periodic monitoring of treated sites included recording of probing depth (PD), clinical attachment level (CAL) and gingival recession. Pre-treatment and 1-year post-operative findings were statistically analysed within and between groups. RESULTS The PD and CAL values decreased significantly with time, but not those between study groups. The mean pre-treatment and 1-year post-treatment PDs of the IBDs of the GTR group (n = 16; sites = 67) were 8.93 ± 1.14 mm and 3.58 ± 0.50 mm, respectively, and the mean CALs were 9.03 ± 1.03 mm and 4.16 ± 0.53 mm respectively. The mean PDs of the EMD/DBX group (n = 16; sites = 73) were 8.77 ± 1.04 mm and 3.61 ± 0.36 mm, respectively, and the mean CALS were 8.79 ± 1.04 mm and 3.77 ± 0.22 mm respectively (p < 0.001 for all). CONCLUSION Surgical treatment of AgP patients by either GTR or by application of EMD/DBX yielded similarly successful clinical results at 1-year post-treatment.