Antimicrobial peptide KSL-W promotes gingival fibroblast healing properties in vitro

We investigated the effect of synthetic antimicrobial decapeptide KSL-W (KKVVFWVKFK) on normal human gingival fibroblast growth, migration, collagen gel contraction, and α-smooth muscle actin protein expression. Results show that in addition to promoting fibroblast adhesion by increasing F-actin production, peptide KSL-W promoted cell growth by increasing the S and G2/M cell cycle phases, and enhanced the secretion of metalloproteinase (MMP)-1 and MMP-2 by upregulating MMP inhibitors, such as tissue inhibitors of metalloproteinase (TIMP)-1 and TIMP-2 in fibroblasts. An in vitro wound healing assay confirmed that peptide KSL-W promoted fibroblast migration and contraction of a collagen gel matrix. We also demonstrated a high expression of α-smooth muscle actin by gingival fibroblasts being exposed to KSL-W. This work shows that peptide KSL-W enhances gingival fibroblast growth, migration, and metalloproteinase secretion, and the expression of α-smooth muscle actin, thus promoting wound healing.     

Genetic control of variation in human gingival fibroblast proliferation rate

Summary Logarithmic proliferation rate (Days 1 to 6) of gingival fibroblasts derived from 15 pairs of monozygotic (MZ) and 9 pairs of dizygotic (DZ) human twins was compared under optimal and suboptimal growth conditions. Cell proliferation rates exhibited considerable variability among strains. For Caucasian donors (13 MZ, 6 DZ pairs) DZ twins demonstrated significantly greater (P<0.01) within-pair variance in cell proliferation rate compared to MZ twins when evaluated under optimal growth conditions. Heritability analysis indicated strong genetic control of proliferation rate of human gingival fibroblasts (HGF) under optimal growth conditions (1.0±0.67), whereas proliferation rate of HGF under suboptimal growth conditions revealed less genetic control (0.42±0.61). These findings emphasize the importance of carefully matching control and test HGF in assays dependent on cellular proliferation. This work was supported by grants DE-06671 and DE-07841 from the National Institutes of Health.     

Aggregatibacter actinomycetemcomitans lipopolysaccharide affects human gingival fibroblast cytoskeletal organization

The cytoskeleton is a dynamic structure that plays a key role in maintaining cell morphology and function. This study investigates the effect of bacterial wall lipopolysaccharide (LPS), a strong inflammatory agent, on the dynamics and organization of actin, tubulin, vimentin, and vinculin proteins in human gingival fibroblasts (HGF). A time-dependent study showed a noticeable change in actin architecture after 1.5 h of incubation with LPS (1 microg/ml) with the formation of orthogonal fibers and further accumulation of actin filament at the cell periphery by 24 h. When 0.01-10 microg/ml of LPS was added to human gingival fibroblast cultures, cells acquired a round, flat shape and gradually developed cytoplasmic ruffling. Lipopolysaccharides extracted from Aggregatibacter actinomycetemcomitans periodontopathogenic bacteria promoted alterations in F-actin stress fibres of human gingival cells. Normally, human gingival cells have F-actin fibres that are organized in linear distribution throughout the cells, extending along the cell's length. LPS-treated cells exhibited changes in cytoskeletal protein organization, and F-actin was reorganized by the formation of bundles underneath and parallel to the cell membrane. We also found the reorganization of the vimentin network into vimentin bundling after 1.5 h of treatment. HGF cells exhibited diffuse and granular gamma-tubulin stain. There was no change in LPS-treated HGF. However, vinculin plaques distributed in the cell body diminished after LPS treatment. We conclude that the dynamic and structured organization of cytoskeletal filaments and actin assembly in human gingival fibroblasts is altered by LPS treatment and is accompanied by a decrease in F-actin pools.     

Mandibular bone remodeling induced by dental implant

The ability to assess the effects of an implant on bone remodeling is of particular importance to prosthesis placement planning and associated treatment assurance. Prediction of on-going bone responses will enable us to improve the performance of a restoration. Although the bone remodeling for long bones had been extensively studied, there have been relatively few reports for dental scenarios despite its increasing significance with more and more dental implant placements. This paper aimed to develop a systematic protocol to assess mandibular bone remodeling induced by dental implantation, which extends the remodeling algorithms established for the long bones into dental settings. In this study, a 3D model for a segment of a human mandible was generated from in vivo CT scan images, together with a titanium implant embedded to the mandible. The results examined the changes in bone density and stiffness as a result of bone remodeling over a period of 48 months. Resonance frequency analysis was also performed to relate natural frequencies to bone remodeling. The density contours are qualitatively compared with clinical follow-up X-ray images, thereby providing validity for the bone remodeling algorithm presented in dental bone analysis.     

Predictions of bone remodeling around dental implant systems

This study presents the implementation of a mathematical bone remodeling algorithm to bone adaptation in the premolar area of the mandible around various dental implant systems, and thus sheds a new perspective to the complex interactions in dental implant mechanics. A two-dimensional, plane strain model of the bone was built from a CT-scan. The effect of implant contour on internal bone remodeling was investigated by considering four dental implant systems with contours similar to commercially available ones and another four with cylindrical and conical cross-sections. The remodeling algorithm predicts non-homogeneous density/elastic modulus distribution; and, implant contour has some effect on how this is distributed. Bone density is predicted to increase on the tips of the threads of the implants, but to decrease inside the grooves. Threadless implants favor to develop a softer bone around their periphery, compared to implant systems that have threads. The overall contour (dimensions and the shape) of an implant affect the bone density redistribution, but the differences between different implant systems are relatively small.     

Application of the finite element method in dental implant research

This article provides a review of the achievements and advancements in dental technology brought about by computer-aided design and the all powerful finite element method (FEM) of analysis. The scope of the review covers dental implants, jawbone surrounding the implant and the biomechanical implant and jawbone interaction. Prevailing assumptions made in the published finite element analysis (FEA) and their limitations are discussed in some detail which helps identify the gaps in research as well as future research direction.     

Growth of bones as revealed by implant markers in animals

Implantation of screws, pegs, wires, or other material as markers has been of value during the last 200 years in the serial study of growth of bones. With this direct method, for a given period, the increase in distance between implants on either side of an endochondral growth center or suture can be readily determined. In contrast, the relatively constant position of two or more implants within a single bone can be utilized to establish the pattern of peripheral apposition and resorption. When radiopaque implants are used in combination with serial roentgenography, not only the amount but also the rate, periods of activity and relative direction of growth may be determined. This indirect method obviates reoperating or killing the animal to measure the distance between implants. Some of the problems in the use of these methods are that (1) growth occurs in more than one plane, (2) the implants may not remain where inserted into the bone, (3) the implants may not be in a plane parallel to the X -ray film, and (4) the distance between the implants and the film may not be constant. (Abstract previously published Am. J. Phys. Anthrop., 27: 236, September, 1967).     

Proliferative effects of angiotensin II and endothelin-1 on guinea pig gingival fibroblast cells in culture

We investigated whether phenytoin (PHT) and nifedipine (NIF) induce angiotensin II (Ang II) and endothelin-1 (ET-1) generation by cultured gingival fibroblasts derived from guinea pigs and whether Ang II and ET-1 induce proliferation of these cells. Immunohistochemical experiments showed that PHT (250 nM) and NIF (250 nM) increased the immunostaining intensities of immunoreactive Ang II and ET-1 (IRET-1) in these cells. Captopril (3 microM), an angiotensin-converting enzyme inhibitor, reduced these enhanced intensities to control levels. Ang II (100 nM) enhanced the immunostaining intensity of IRET-1. PHT (250 nM) and NIF (250 nM)-induced cell proliferation. Both PHT- and NIF-induced proliferation was inhibited by captopril (3 microM). Ang II (100 nM) and ET-1 (100 nM) also induced cell proliferation. Ang II-induced proliferation was inhibited by CV11974 (1 microM), an AT(1) receptor antagonist and saralasin (1 microM), an AT(1)/AT(2) receptor antagonist, but not by PD123,319 (1 microM), an AT(2) receptor antagonist. ET-1-induced proliferation was inhibited by BQ123 (10 microM), an ET(A) receptor antagonist, but not by BQ788 (1 microM), an ET(B) receptor antagonist. These findings suggest that PHT- and NIF-induced gingival fibroblast proliferation is mediated indirectly through the induction of Ang II and ET-1 and probably mediated through AT(1) and ET(A) receptors present in or on gingival fibroblasts.     

Histological and biomechanical studies of hydroxylapatite implant.

Bony defects measuring 12x12x12 millimeters in size were created bilaterally over the proximal end of the femurs of 10 Macaca cyclopis monkeys and implanted with particulate hydroxylapatite. The animals were sacrificed according to a predetermined sequence of post-operative 1, 2, 3, 6, and 8 months. The specimens taken from the implant sites in the two femurs of each animal were chosen randomly either for histological observation or for compressive strength tests under vertical loads. Histological pictures showed only loose fibrous tissue over the interparticular space of the implant site at the first month after surgery. Woven bone could be observed over the basal part of the implant site at the second month. New bone formed over the central part of the implant site at the third month after surgery. Continuing maturation of the bone tissue could be observed over the interparticular space of the implant site until the late stages at the 6th and 8th month after surgery. The results of compressive strength tests were compatible with the histological observations. The value of the elastic modulus was low initially at the first month, but increased gradually with time due to organization of fibrous tissue, condensation of HA particles, and ossification as well as maturation of bone tissue. The value of elastic modulus at the third month was equal to that of cortical bone tissue and even two-fold higher than that of the cortical bone at the eighth month after surgery. These results provide valuable information about the compressive strength of the HA implant in host tissue.     

Comparison of short-latency trigeminal evoked potentials elicited by painful dental and gingival stimulation

Painful stimulation of tooth pulp and of the maxillary gingiva was undertaken in 16 volunteers. Short-latency evoked potentials (15–20 msec) were recorded over 800 trials in each case at F3-P3 of F4-P4, and the resultant averaged wave forms were compared. The gingival wave was distinct in all subjects and could be averaged across subjects while the dental waves were either noise or very inconsistent over subjects. Averaging of the dental wave forms across subjects yielded an uninterpretable result. It was clear that dental evoked potentials could not be recorded at these sites. These findings could be explained by either or both of two hypotheses: (1) dental afferents are predominantly small fiber, nociceptive end organs that conduct more slowly than soft tissue afferents whereas gingival stimulation activates both large and small fiber populations; and (2) dental representation in somatosensory cortex is different and phylogenetically more primitive than that of neighboring soft tissue. Therefore, the location of the generator sites in cortex and the orientation of the dipole may be different for dental than for gingival wave forms.     

Comparative proteomics analysis of human stem cells from dental gingival and periodontal ligament

Dental stem cells isolated from oral tissues have been shown to provide with high proliferation ability and multilineage differentiation potential. Gingival mesenchymal stem cells (GMSCs) and periodontal ligament stem cells (PDLSCs), kinds of dental stem cells, can be used as substitutes for tissue repair materials because of their similar regenerative functions. In this study, we aim to explore the similarities and differences between the protein profiles of GMSCs and PDLSCs through quantitative proteomics. A total of 2821 proteins were identified and retrieved, of which 271 were upregulated and 57 were downregulated in GMSCs compared to PDLSCs. Gene Ontology (GO) analysis demonstrated that the 328 differentially abundant proteins (DAPs) were involved in the regulation of gene expression, metabolism, and signal transduction in biological process, mainly distributed in organelles related to vesicle transport, and involved in the molecular function of binding protein. And Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that the DAPs were committed to regulating the synthesis of proteasome and spliceosome. Real-time quantitative polymerase chain reaction (RT-qPCR) results showed that ARPC1B, PDAP1, and SEC61B can be used as special markers to distinguish GMSCs from PDLSCs. This research contributes to explaining the molecular mechanism and promoting the clinical application of tissue regeneration of GMSCs and PDLSCs.     

Computational simulation of dental implant osseointegration through resonance frequency analysis

The aim of this study is to predict the evolution of the resonance frequency of the bone-implant interface in a dental implant by means of finite element simulation. A phenomenological interface model able to simulate the mechanical effects of the osseointegration process at the bone-implant interface is applied and compared with some experimental results in rabbits. An early stage of slow bone ingrowth, followed by a faster osseointegration phase until final stability is predicted by the simulations. The evolution of the resonance frequency of the implant and surrounding tissues along the simulation period was also obtained, observing a 3-fold increase in the first principal frequency. These findings are in quantitative agreement with the experimental measurements and suggest that the model can be useful to evaluate the influence of mechanical factors such as implant geometry or implant loading on the indirect evaluation of the process of implant osseintegration.     

Characterization of Actinomyces species isolated from failed dental implant fixtures

In the oral cavity, Actinomyces form a fundamental component of the indigenous microflora, being among initial colonizers in polymicrobial biofilms. However, some differences may exist between different species in terms of their attachment not only to teeth but also to biomaterials. In this study we investigated the distribution of Actinomyces in 33 dental implant fixtures explanted from 17 patients. The identification was based on comprehensive biochemical testing and gas-liquid chromatography and when needed, 16S rRNA sequencing. Actinomyces was the most prevalent bacterial genus in these failed implants, colonizing 31/33 (94%) of the fixtures. Proportions of Actinomyces growth of the total bacterial growth in the Actinomyces-positive fixtures varied from 0.01% up to 75%. A. odontolyticus was the most common Actinomyces finding, present in 26/31 (84%) Actinomyces-positive fixtures. Actinomyces naeslundii and A. viscosus were both detected in 10/31 (32%) and A. israelii in 7/31 (23%) fixtures. Other Actinomyces species, including A. georgiae, A. gerencseriae and A. graevenitzii, were detected less frequently. Our results suggest that Actinomyces species are frequent colonizers on failed implant surfaces, where A. odontolyticus was the far most prominent Actinomyces species.     

Plasminogen activator inhibitor type 1 expression induced by lipopolysaccharide of Porphyromonas gingivalis in human gingival fibroblast

In the gingival tissues of patients with periodontitis, inflammatory responses are mediated by a wide variety of genes. In our previous screening study, plasminogen activator inhibitor type 1 (PAI-1) mRNA binding protein expression was increased in gingiva from periodontitis patients. In this study, we further investigated the signaling pathway involved in PAI-1 expression induced by Porphyromonas gingivalis LPS (Pg LPS) in human gingival fibroblasts (HGF). When HGFs were treated with Pg LPS, both PAI-1 mRNA expression and PAI-1 protein were induced in a dose-dependent manner. Pg LPS induced NF-κB activation and the expressions of PAI-1 mRNA and protein were suppressed by pretreating with a NF-κB inhibitor. Pg LPS also induced ERK, p38, and JNK activation, and Pg LPS-induced PAI-1 expression was inhibited by ERK/p38/JNK inhibitor pretreatment. In conclusion, Pg LPS induced PAI-1 expression through NF-κB and MAP kinases activation in HGF.     

A three dimensional finite element study on dental implant design

Implant diameter and length are the most effective parameters affecting stress distribution in surrounding bones. In order to extract simplified design equations to better understand implants behavior, 25 different implant designs with gradual increase in diameter and length were analyzed in 3D using Finite Element Method. Four types of loadings were applied on each design: tension of 50 N, compression of 100 N, bending of 20 N, and torque of 2 Nm to derive design curves.Analysis of results showed that increasing implant diameter and length generate better stress distribution on spongy and cortical bones. Approximate design equations and curves were obtained as a result of this study.