Search PubMed⌕ Search

Biomedical subjects

D L Cochran

Publications and source records attributed to D L Cochran.

At least 73 records · Page 4Linked to original sources
PubMed1999

Biological mediators for periodontal regeneration.

A review of the literature on the use of growth-regulatory molecules in the oral cavity permits a model in which to consider approaches to oral tissue engineering. These concepts apply to periodontal regeneration and to regeneration of alveolar bone. In either case, the formation of tissues is complex but proceeds in a deliberate and orderly sequence. In these sequence of events resulting in either bone or cementum formation, periodontal ligament and bone can be stimulated at various points. Different signals can apparently be used to stimulate tissue formation including mitogenic signals and differentiation factors. Additionally, both hard and soft tissue stimulatory molecules appear to be permissive. Classic receptor-mediated peptides or extracellular matrix molecules for soft and hard tissues appear to allow stimulation of tissue formation cascades. Importantly, it also appears that the stimulatory event is transitory (that is, short-lived) and leads itself to a sequence of cellular events. These cellular events in turn stimulate a number of subsequent events (such as chemotaxis, proliferation, differentiation or angiogenesis), which lead to further progression of tissue formation. While a solid scientific rationale exists for the use of a variety of growth and attachment factors in regeneration of oral tissues, only a small number are being pursued clinically. Many therapeutic regimens have failed in preclinical testing or have resulted in limited regenerative capacity. The mitogenic polypeptides that stimulate soft tissue growth (such as platelet-derived growth factor) and both hard and soft tissue growth (such as transforming growth factor-beta) appear to have not led to successful enough outcomes to facilitate further work towards regulatory approval. The demonstrated ability of bone morphogenetic proteins to generate substantial quantities of bone suggest many applications in the oral cavity where this is the only tissue desired. Another therapeutic candidate is enamel matrix derivative, a set of matrix proteins. Enamel matrix derivative appears to stimulate first acellular cementum formation, which may allow for functional periodontal ligament formation. It will be of interest in the future to determine whether the protein matrix contains classic mitogenic or differentiation factors as well as the amelogenins. It is also evident that the bone morphogenetic proteins permit periodontal ligament formation. The conditions for stimulating predictable periodontal ligament tissues with bone morphogenetic proteins however are not known. It is clear that the bone morphogenetic proteins are excellent molecules for stimulating oral bone formation. The results of all these studies will determine the future therapeutic potential for these growth molecules such that they may be used to optimally stimulate and direct specific points along tissue formation cascades.

Alveolar Bone Loss↗
PubMed1999

The accuracy of radiographic methods in assessing the outcome of periodontal regenerative therapy.

BACKGROUND: The study of regenerative therapy in the periodontal intrabony defect has relied upon surgical re-entry as the gold standard of outcome assessment. The search for a non-invasive method has led to the application of various radiographic techniques in evaluating post-treatment bone fill. METHODS: The purpose of this study was to determine the ability of 2 forms of radiographic analyses (linear measurement and computer assisted densitometric image analysis, CADIA) to assess postsurgical bone fill as measured at a re-entry procedure. A method that incorporates linear measurements and CADIA (linear-CADIA) was developed and tested as well. Forty-five intrabony defects in 15 patients were treated with open flap debridement, demineralized freeze-dried bone allograft (DFDBA), or a combination of DFDBA and tetracycline. Standardized radiographs were obtained at baseline and at 1-year postsurgery. RESULTS: A 12-month surgical re-entry provided clinical measurements for post-treatment bone fill. All radiographs were digitally scanned and analyzed on a computer. Fifty-three percent of the defects were excluded from the study due to poor standardization or poor defect quality. Forty percent of all pairs of radiographs were judged to have poor standardization. In the first analysis, standardized images were subtracted and quantitatively analyzed utilizing CADIA. It was found that CADIA had the highest correlation with clinical bone fill when a region of interest (ROI) was examined in the middle portion of the defect. This quantitative evaluation provided very little clinically relevant information regarding actual bone fill. For the second analysis, pre- and post-treatment linear radiographic measurements were obtained. In only 43% of the sites, did linear radiographic measurements determine post-treatment bone fill within 1.0 mm of the clinical measurements. Overall, linear measurements underestimated bone fill by 0.96 mm (+/-1.2). These differences were statistically significant (paired Student t-test, P = 0.0023). A method, which incorporates the use of both CADIA and linear radiographic measurements (linear-CADIA), was tested. The linear-CADIA method underestimated bone fill by 0.26 mm (+/-1.4), but these differences were not statistically significant (paired Student t-test, P = 0.41). CONCLUSION: Linear radiographic measurements significantly underestimate post-treatment bone fill when compared to re-entry data. The linear-CADIA method provided the highest level of accuracy of the 3 methods tested. This study also emphasizes the importance of developing a consistent method of radiographic standardization.

Adult↗
PubMed1999

A comparison of endosseous dental implant surfaces.

Endosseous dental implants are available with various surface characteristics ranging from relatively smooth machined surfaces to more roughened surfaces created by coatings, blasting by various substances, by acid treatments, or by combinations of the treatments. Studies characterizing these implants and surfaces include in vitro experimentation, animal studies, and human clinical trials. Both descriptive and functional testing of the bone-implant interface includes histomorphometrics and biomechanical testing such as torque removal values and push out/pull out strength. Using these assays to evaluate and compare different surfaces, the data demonstrate that rough implant surfaces have increased bone-to-implant contact and require greater forces to break the bone-implant interface compared to more smooth surfaces. The objective of this report was to evaluate publications of human clinical experiences evaluating implant use in patients and to determine if differences existed in success rates of implants with relatively smooth surfaces compared to implants having roughened implant surfaces. Human trials were reviewed to determine the clinical efficacy of implants under various clinical indications. Synopsis tables were constructed and the experiences segregated by implant surface characteristics. Meta-analyses were performed on all implants in all locations, on implants placed only in the maxilla or the mandible, and, finally, on implants placed in the maxilla compared to implants placed in the mandible. Evaluation of the data revealed that predictably high success rates can be achieved for implants with both rough and smooth titanium surfaces and for hydroxyapatite-coated implants. When studies were clustered by specific indications or patient populations, rough surfaced implants had significantly higher success rates compared to implants with more smooth surfaces except in the case of single tooth replacements where the success rates were comparable. In general, implants placed in the mandible had significantly higher success rates than implants placed in the maxilla. However, in the partially edentulous patient group, titanium implants with a rough surface had significantly higher success rates in the maxilla compared to the mandible and, in cases of single tooth replacement, success rates were similar in the maxilla and in the mandible as was the case for hydroxyapatite-coated implants. The documented advantage of implants with a roughened surface in animal and in vitro experiments has been demonstrated in clinical cases when studies were compared in which specific indications or patients were treated. Additionally, implants placed in the mandible have, in general, higher success rates than implants placed in the maxilla, with only a few exceptions noted. These data from human clinical experiences support the documented advantage of implants with a roughened surface in animal and in vitro experimentation and indicate that the magnitude of the advantage is significant for patient care.

Animals↗
PubMed1999

Recombinant human bone morphogenetic protein-2 stimulation of bone formation around endosseous dental implants.

BACKGROUND: Successful endosseous implant placement requires that the implant be stable in alveolar bone. In certain cases, the implant can be stabilized in native bone but some part of the implant is not covered by bone tissue. This often occurs during placement of implants into extraction sites or in areas where bone resorption has occurred and the ridge width is not sufficient to completely surround the implant. In those cases, the clinician usually employs a procedure to encourage bone formation. These procedures typically include a bone graft and/or membrane therapy. Recent advances have led to the isolation, cloning, and production of recombinant human proteins that stimulate bone formation. One of these bone morphogenetic proteins (rhBMP-2) has been extensively studied in animal models and is currently being tested in human clinical trials. METHODS: In this study, rhBMP-2 was tested using a collagen sponge carrier to stimulate bone formation in defects in the canine mandible around endosseous dental implants. Six animals had a total of 48 implants placed. rhBMP-2 with the collagen carrier was implanted around 24 of these, the remainder having only the collagen carrier placed. Half the sites were covered with a nonresorbable expanded polytetrafluoroethylene membrane. Histologic analysis was performed after 4 and 12 weeks. The area of new bone formed, percentage of bone-to-implant contact in the defect area, and percentage fill of the defect was calculated. RESULTS: The addition of rhBMP-2 resulted in significantly greater amounts of new bone area and percentage of bone-to-implant contact and with more percentage fill after 4 and 12 weeks of healing. The area of new bone formed was reduced after 4 weeks when a membrane was present but after 12 weeks, there was no significant difference between membrane and non-membrane treated sites. In some specimens, new bone was found coronal to the membranes, with rhBMP-2-treated sites having greater amounts than non-rhBMP-2-treated sites. CONCLUSIONS: These data demonstrate that a bone differentiation factor significantly stimulates bone formation in peri-implant bone defects in the canine mandible. In addition, bone-to-implant contact was significantly enhanced along the rough implant surface. Membrane-treated sites had less new bone formation after 4 weeks of healing but were similar to non-membrane sites after 12 weeks. These results demonstrate that rhBMP-2 can be used to stimulate bone growth both around and onto the surface of endosseous dental implants placed in sites with extended peri-implant osseous defects.

Animals↗
PubMed1999

Evaluation of 2 novel approaches for assessing the ability of demineralized freeze-dried bone allograft to induce new bone formation.

BACKGROUND: Because of the wide variation in the ability of human demineralized freeze-dried bone allograft (DFDBA) to reproducibly induce new bone formation, there is a need for a reliable measure of bone induction activity. In this study we examined an immature osteoprogenitor cell line for its potential utility in measuring the activity of DFDBA in vitro. METHODS: We characterized the response of 2T9 cells, an immature osteoprogenitor cell line derived from the calvariae of transgenic mice containing the SV40 T-antigen driven by the mouse bone morphogenetic protein (BMP)-2 promoter, to recombinant human BMP-2 by measuring alkaline phosphatase specific activity, osteocalcin production, and matrix mineralization. Responses were compared to those obtained with 1,25-(OH)2D3. In addition, 2T9 cells were cultured with active or inactive human DFDBA in the presence or absence of BMP-2. We also tested the hypothesis that radio-opacity of tissue following implantation of DFDBA in vivo correlates with the ability of human DFDBA to induce new bone. DFDBA from 9 different donors, stratified by age, were implanted subcutaneously in the thorax of 18 nude (nu/nu) mice. Tissue was harvested at 36 days postoperatively and examined histologically and biochemically for calcium and phosphorus uptake. RESULTS: 2T9 cells exhibited a dose- and time-dependent response to soluble BMP-2. Proliferation was decreased and alkaline phosphatase activity, osteocalcin production, and mineralized nodule formation were increased. The effects were dose- and time-dependent. Peak effects on alkaline phosphatase and osteocalcin were noted on day 8, whereas mineral deposition did not begin to occur until day 12. 1,25-(OH)2D3 did not regulate these effects unless used with BMP-2. When the cells were exposed to active or inactive DFDBA in the presence or absence of BMP-2, no effect on 2T9 cell differentiation was observed. This indicated that DFDBA released no soluble factors with bone inductive ability and that if any active factors were adsorbed to the DFDBA, they were inactivated. When DFDBA was implanted subcutaneously in the thorax of nude mice, there was no histologic evidence of new bone formation. However, there was a donor age-dependent decrease in Ca and P uptake of the implanted tissue, reflecting a donor age-dependent decrease in remineralization of DFDBA. CONCLUSIONS: These data indicate that cell culture assays like the one used in this study may not be appropriate indicators of bone induction ability by DFDBA since soluble factors may not be responsible for bone induction in vivo. Nonetheless, in vitro assays are still needed. While Ca and P uptake by DFDBA-implanted tissue in the present study correlated with the age-dependent decrease in bone induction at intramuscular sites in a previously reported study, these data show that early x-rays may actually detect remineralization and not new bone formation. Thus, assessment of bone induction ability may still depend on histologic analysis of animal models.

Age Factors↗
PubMed1999

Comparison of bioactive glass to demineralized freeze-dried bone allograft in the treatment of intrabony defects around implants in the canine mandible.

BACKGROUND: The purpose of this study was to evaluate and compare the healing of different bone grafting materials adjacent to titanium plasma-sprayed (TPS) endosseous dental implants. METHODS: Implant osteotomy sites were prepared and standardized 3-walled intrabony defects (3 mm x 5 mm x 5 mm) were created at the mesial of each implant site. Thirty-two TPS implants were placed in edentulous mandibular ridges of the 4 dogs. Periodontal dressings were placed in the defect sites so as to create a defect simulating bone loss around an implant. After 3 months, the periodontal dressing was removed, the defect sites debrided and evaluated for size, and intramarrow penetration performed. The graft materials tested were 1) canine demineralized freeze-dried bone allograft (cDFDBA); 2) bioactive glass granules of a broad size range 90 to 710 microns (BRG); and 3) bioactive glass granules of narrow size range 300 to 355 microns (NRG). One site on each side of the mandible was not filled and served as a control. Dogs were sacrificed 4 months after graft placement. RESULTS: Histologically, differences in percent bone-to-implant contact in the defect area were observed between the treatment groups. cDFDBA>control=BRG=NRG with statistical significance found between cDFDBA and control (P = 0.0379), but no statistically significant difference between control or either bioactive glass material. When comparing percent bone height fill of the defect in the grafted area, cDFDBA (65.7%) was significantly better than the control (48.9%; P < or = 0.05) with no statistically significant difference between control, broad range bioactive glass (57.3%) and narrow range bioactive glass (56.6%). When total bone area was measured, the percentage of new bone in the grafted area was cDFDBA (42.1%), broad range glass (33.1%) and narrow range glass (22.6%) with significance found between cDFDBA and NRG (P = 0.0102). The content of residual graft particles in soft tissue was significant (P = 0.0304) between cDFDBA (1.4%) and NRG (11.4%) with no significant difference between graft material for residual particle content in bone tissue. CONCLUSIONS: The results of this study indicate that percent bone-to-implant contact and percent bone height fill in an intrabony defect around titanium plasma-sprayed implants are statistically significantly higher with the use of DFDBA when compared to bioactive glass material.

Alveolar Bone Loss↗
PubMed1998

Prostaglandins mediate the effects of titanium surface roughness on MG63 osteoblast-like cells and alter cell responsiveness to 1 alpha,25-(OH)2D3.

Surface roughness affects proliferation, differentiation (alkaline phosphatase and osteocalcin), local factor production (transforming growth factor (TGF beta) and prostaglandin E2 (PGE2)], and response to 1,25-(OH)2D3 (1,25) of MG63 osteoblast-like cells. In this study, we examined whether the effect of surface roughness on MG63 cells is mediated by prostaglandins produced by the cells. Unalloyed titanium (Ti) disks were pretreated with HF/HNO3 (PT) and then machined and acid-etched (MA). Disks were also coarse grit-sandblasted (SB), coarse grit-sandblasted and acid-etched (CA), or plasma-sprayed with Ti particles (PS). The surfaces, from smoothest to roughest, were PT, MA, CA, SB, and PS. MG63 cells were cultured to confluence on the Ti disks in the presence or absence of 10(-7) M indomethacin (Indo), a specific inhibitor of cyclooxygenase activity, resulting in decreased prostaglandin production. When the cells reached confluence, cell number, cell layer alkaline phosphatase specific activity (ALPase), and osteocalcin (OC) and latent TGF beta (LTGF beta) production were determined. In addition, confluent cultures which had been grown in the absence of Indo were exposed to 10(-7) M 1,25, 10(-7) M Indo, or a combination of the two for 24 h. On the rougher surfaces, cell number was decreased and ALPase, OC, and LTGF beta were increased. When indomethacin was present throughout the culture period, the effect of surface roughness on cell number, OC, and LTGF beta was abolished. ALPase was reduced, but surface roughness-dependent effects were still observed. Addition of indomethacin to confluent cultures for 24 h had no effect on any of the parameters examined, with one exception: Cells cultured on MA surfaces exhibited a more differentiated phenotype. 1,25 increased all parameters examined on SB, CA, and PS surfaces. When indomethacin was added with 1,25, the 1,25-dependent effects on cell number and OC and LTGF beta production were abolished; however, ALPase was unaffected. This indicates that bone cell response to systemic hormones may be modified by implant surface roughness. This effect may be mediated, at least in part, by prostaglandins produced by the same cells.

Alkaline Phosphatase↗
PubMed1998

Titanium surface roughness alters responsiveness of MG63 osteoblast-like cells to 1 alpha,25-(OH)2D3.

Surface roughness has been shown to affect differentiation and local factor production of MG63 osteoblast-like cells. This study examined whether surface roughness alters cellular response to circulating hormones such as 1 alpha,25-(OH)2D3. Unalloyed titanium (Ti) disks were pretreated with HF/HNO3 (PT) and then were machined and acid-etched (MA). Ti disks also were sandblasted (SB), sandblasted and acid etched (CA), or plasma sprayed with Ti particles (PS). The surfaces, from smoothest to roughest, were: PT, MA, CA, SB, and PS. MG63 cells were cultured to confluence on standard tissue culture polystyrene (plastic) or the Ti surfaces and then treated for 24 h with either 10(-8) M or 10(-7) M 1 alpha,25-(OH)2D3 or vehicle (control). Cellular response was measured by assaying cell number, cell layer alkaline phosphatase specific-activity, and the production of osteocalcin, latent (L) TGF beta, and PGE2. Alkaline phosphatase activity was affected by surface roughness; as the surface became rougher, the cells showed a significant increase in alkaline phosphatase activity. Addition of 1 alpha,25-(OH)2D3 to the cultures caused a dose-dependent stimulation of alkaline phosphatase activity that was synergistic with the effect caused by surface roughness alone. 1 alpha,25-(OH)2D3 also caused a synergistic increase in osteocalcin production as well as local factor (LTGF beta and PGE2) production on the rougher CA, SB, and PS surfaces, but it had no effect on the production on smooth surfaces. The inhibitory effect of surface roughness on cell number was not affected by 1 alpha,25-(OH)2D3 except on the SB surface. 1 alpha,25-(OH)2D3 decreased cell number, increased alkaline phosphatase activity and osteocalcin production, and had no effect on LTGF beta or PGE2 production by MG63 cells grown on tissue culture polystyrene. These data suggest that bone cell response to systemic hormones is modified by surface roughness and that surface roughness increases the responsiveness of MG63 cells to 1 alpha,25-(OH)2D3. They also suggest that the endocrine system is actively involved in normal bone healing around implants.

Alkaline Phosphatase↗
PubMed1998

Bone response to unloaded and loaded titanium implants with a sandblasted and acid-etched surface: a histometric study in the canine mandible.

Many dental clinical implant studies have focused on the success of endosseous implants with a variety of surface characteristics. Most of the surface alterations have been aimed at achieving greater bone-to-implant contact as determined histometrically at the light microscopic level. A previous investigation in non-oral bone under short-term healing periods (3 and 6 weeks) indicated that a sandblasted and acid-etched titanium (SLA) implant had a greater bone-to-implant contact than did a comparably-shaped implant with a titanium plasma-sprayed (TPS) surface. In this canine mandible study, nonsubmerged implants with a SLA surface were compared to TPS-coated implants under loaded and nonloaded conditions for up to 15 months. Six foxhound dogs had 69 implants placed in an alternating pattern with six implants placed bilaterally in each dog. Gold crowns that mimicked the natural occlusion were fabricated for four dogs. Histometric analysis of bone contact with the implants was made for two dogs after 3 months of healing (unloaded group), 6 months of healing (3 months loaded), and after 15 months of healing (12 months loaded). The SLA implants had a significantly higher (p < 0.001) percentage of bone-to-implant contact than did the TPS implants after 3 months of healing (72.33 +/- 7.16 versus 52.15 +/- 9.19; mean +/- SD). After 3 months of loading (6 months of healing) no significant difference was found between the SLA and TPS surfaced implants (68.21 +/- 10.44 and 78.18 +/- 6.81, respectively). After 12 months of loading (15 months of healing) the SLA implants had a significantly greater percentage (p < 0.001) of bone-to-implant contact than did the TPS implants (71.68 +/- 6.64 and 58.88 +/- 4.62, respectively). No qualitative differences in bone tissue were observed between the two groups of implants nor was there any difference between the implants at the clinical level. These results are consistent with earlier studies on SLA implants and suggest that this surface promotes greater osseous contact at earlier time points compared to TPS-coated implants.

Acid Etching, Dental↗
PubMed1998

The soft tissue response to osseointegrated dental implants.

The use of dental implants in the treatment of fully edentulous patients has become an important addition in oral/dental rehabilitation. The fact that these implants penetrate the oral mucosa can lead to the assumption that peri-implant tissues, similar to the periodontal tissues, are fulfilling an important function as a barrier to protect the bony anchorage underneath. It has been shown that insufficient plaque removal may lead to peri-implant tissue disease with bone loss similar to teeth. However, it is unclear how important this cause is as a source of implant failure compared with other factors, such as inadequate bone healing, unfavorable quantity and quality of bone, or (bio)mechanical and functional problems. It is also not understood if peri-implant epithelium and connective tissue are equally needed and/or qualified to slow down or prevent tissue breakdown as their periodontal counterparts. The scientific work focusing on peri-implant soft tissues has dramatically increased in the past few years. Most studies to date have examined and described their structure but little data exist on their true biologic function. This review analyzes the current understanding of morphologic and clinical features of the peri-implant soft tissues. Furthermore, evidence shall be provided that peri-implant soft tissues do not interfere with the current favorable results obtained when treating the edentulous patient with osseointegrated implants.

Alveolar Bone Loss↗
PubMed1998

Response of MG63 osteoblast-like cells to titanium and titanium alloy is dependent on surface roughness and composition.

The success of an implant is determined by its integration into the tissue surrounding the biomaterial. Surface roughness and composition are considered to influence the properties of adherent cells. The aim of this study was to determine the effect of chemical composition and surface roughness of commercially pure titanium (Ti) and Ti-6A1-4V alloy (Ti-A) on MG63 osteoblast-like cells. Unalloyed and alloyed Ti disks were machined and either fine-polished or wet-ground, resulting in smooth (S) and rough (R) finishes, respectively. Standard tissue culture plastic was used as a control. Surface topography and profile were evaluated by cold field emission scanning electron microscopy and profilometry, while chemical composition was determined using Auger electron spectroscopy and Fourier transform infrared spectroscopy. The effect on the cells was evaluated 24 h postconfluence by measuring cell number, [3H]-thymidine incorporation into DNA, cell and cell layer alkaline phosphatase specific activity (ALPase), osteocalcin and collagen production, [35S]-sulfate incorporation into proteoglycan, and prostaglandin E2 (PGE2) and transforming growth factor-beta (TGF-beta) production. When compared to plastic, the number of cells was reduced on the pure Ti surfaces, while it was equivalent on the Ti-A surfaces; [3H]-thymidine incorporation was reduced on all surfaces. The stimulatory effect of surface roughness on ALPase in isolated cells and the cell layer was more pronounced on the rougher surfaces, with enzyme activity on Ti-R being greater than on Ti-A-R. Osteocalcin production was increased only on the Ti-R surface. Collagen production was decreased on Ti surfaces except Ti-R; [35S]-sulfate incorporation was reduced on all surfaces. Surface roughness affected local factor production (TGF-beta, PGE2). The stimulatory effect of the rougher surfaces on PGE2 and TGF-beta was greater on Ti than Ti-A. In summary, cell proliferation, differentiation, protein synthesis and local factor production were affected by surface roughness and composition. Enhanced differentiation of cells grown on rough vs. smooth surfaces for both Ti and Ti-A surfaces was indicated by decreased proliferation and increased ALPase and osteocalcin production. Local factor production was also enhanced on rough surfaces, supporting the contention that these cells are more differentiated. Surface composition also played a role in cell differentiation, since cells cultured on Ti-R surfaces produced more ALPase than those cultured on Ti-A-R. While it is still unknown which material properties induce which cellular responses, this study suggests that surface roughness and composition may play a major role and that the best design for an orthopaedic implant is a pure titanium surface with a rough microtopography.

Alkaline Phosphatase↗
PubMed1998

PDGF-alpha receptor subunit expression down-regulated by IL-1beta in human periodontal ligament cells.

The responses of cells to the distinct PDGF isoforms have been correlated directly to the relative numbers of specific PDGF receptor subunits on the cell surface. The modulation of PDGF-alpha receptor subunits, the major subunit expressed in human periodontal ligament (PDL) cells, by cytokines present in the periodontal wound site, such as interleukin-1 (IL-1), may be an important factor influencing regenerative outcomes. The purpose of the present study was to examine the effects of IL-1 beta on PDGF-alpha receptor subunit expression in human PDL cells. Primary cultures of human PDL cells were treated with IL-1 beta over a range of concentrations. We assessed PDGF-alpha receptor subunits by examining the mitogenic responses of cells to PDGF-AA, specific binding of 125I-labeled PDGF-AA, immunofluorescent analysis of PDGF-alpha receptor subunits, and PDGF-alpha receptor subunit mRNA levels using Northern blot analysis. The results demonstrate a significant concentration-dependent decrease in 3H-thymidine incorporation in response to PDGF-AA following IL-1 beta treatment (p < 0.001). This decreased response correlated directly with IL-1-induced decreases in 125I-labeled PDGF-AA binding (p < 0.01), the numbers of immunolabeled PDGF-alpha receptor subunits, and in PDGF-alpha receptor subunit mRNA levels. However, when combined with TGF-beta, IL-1 beta did not show additional down-regulation in proliferative response to PDGF-AA or PDGF-alpha receptor subunits beyond that achieved with these factors individually. These experiments identify IL-1 beta, along with TGF-beta, as significant inhibitors of PDGF stimulation in human PDL cells, acting through the down-regulation of PDGF-alpha receptor subunit expression.

Analysis of Variance↗
PubMed1998

Addition of human recombinant bone morphogenetic protein-2 to inactive commercial human demineralized freeze-dried bone allograft makes an effective composite bone inductive implant material.

COMMERCIAL PREPARATIONS OF HUMAN DEMINERALIZED freeze-dried bone allograft (DFDBA) vary in their ability to induce new bone formation. This study tested the hypothesis that inactive DFDBA can be used as an effective carrier of recombinant human bone morphogenetic protein-2 (rhBMP-2). Two batches of active DFDBA were used as controls. Two batches of DFDBA, previously shown to be inactive, were treated with vehicle or with 5 or 20 microg rhBMP-2 and implanted into the calf muscle of male Nu/Nu (nude) mice. Each mouse received one implant in each hind limb, both of which were of the same formulation, resulting in 8 groups of 4 mice per group: active DFDBA batch A, active DFDBA batch B, inactive DFDBA batch A, inactive DFDBA batch B, inactive DFDBA batch A plus 5 microg rhBMP-2, inactive DFDBA batch A plus 20 microg rhBMP-2, inactive DFDBA batch B plus 5 microg rhBMP-2, and inactive DFDBA batch B plus 20 microg rhBMP-2. After 56 days, the implants were removed and histologically examined. A semiquantitative bone induction index was calculated based on the amount of new bone covering each histological section. Histomorphometry was also used to evaluate the area of new bone formed and the area of residual implant material. The results showed that active DFDBA induces new bone formation, whereas inactive DFDBA does not. Addition of rhBMP-2 to inactive DFDBA results in new bone formation with a bone induction index comparable to that of active DFDBA. Histomorphometric analysis, however, revealed that the rhBMP-2-containing implants caused a dose-dependent increase in new bone area that exceeded that induced by active DFDBA. At the highest concentration of rhBMP-2, bone formation was exuberant. rhBMP-2 also caused the resorption of residual implant material to levels comparable to that seen in sites treated with active DFDBA, suggesting that this growth factor may regulate resorptive cells either directly or indirectly. This study shows that addition of rhBMP-2 to inactive DFDBA provides reproducible, consistent bone induction, and suggests that inactive commercial preparations may contain inadequate amounts of BMP to cause bone induction compared to active preparations.

Animals↗
PubMed1998

Ability of commercial demineralized freeze-dried bone allograft to induce new bone formation is dependent on donor age but not gender.

Demineralized freeze-dried bone allografts (DFDBA) have been used extensively in periodontal therapy. DFDBA is used because it contains bone morphogenetic protein (BMP), which induces new bone formation during the healing process. Most commercial bone banks do not verify the presence or activity of BMP in DFDBA nor the ability of DFDBA to induce new bone. Recently, we showed that different bone bank preparations of DFDBA, even from the same bank, varied considerably in their ability to induce new bone, suggesting inherent differences in the quality of the material. Therefore, we examined whether donor age or gender contributed to the variability seen with these preparations. Twenty-seven batches of DFDBA from different donors were donated by one bone bank which had been shown previously to supply DFDBA that was consistently able to induce new bone formation. Each batch was implanted bilaterally in the thigh muscle of nude mice. After 56 days, the implants were excised and examined by light microscopy and histomorphometry. Seventy percent of the preparations tested induced new bone formation. Most of these preparations produced ossicles containing cortical bone surrounding bone marrow-like tissue. The ability to induce bone appears to be age-dependent, with DFDBA from older donors being less likely to have strong bone-inducing activity. By contrast, no difference in ability to induce new bone was noticed between male or female donors. The results of this study confirm that commercial preparations of DFDBA differ in their ability to induce new bone formation. In fact, some of the batches had no activity at all. The ability of DFDBA to induce new bone formation is suggested to be age-dependent, but not gender-dependent by our study. These results indicate that commercial bone banks need to verify the ability of DFDBA to induce new bone formation and should reconsider the advisability of using bone from older donors.

Adolescent↗
PubMed1998

Clinical evaluation of bioactive glass in the treatment of periodontal osseous defects in humans.

The purpose of this study was to compare the use of bioactive glass to demineralized freeze-dried bone allograft (DFDBA) in the treatment of human periodontal osseous defects. Fifteen systemically healthy patients (6 males and 9 females, aged 30 to 63) with moderate to advanced adult periodontitis were selected for the study. All patients underwent initial therapy, which included scaling and root planing, oral hygiene instruction, and an occlusal adjustment when indicated, followed by re-evaluation 4 to 6 weeks later. Paired osseous defects in each subject were randomly selected to receive grafts of bioactive glass or DFDBA. Both soft and hard tissue measurements were taken the day of surgery (baseline) and at the 6-month re-entry surgery. The clinical examiner was calibrated and blinded to the surgical procedures, while the surgeon was masked to the clinical measurements. Statistical analysis was performed by using the paired Student's t test. The results indicated that probing depths were reduced by 3.07 +/- 0.80 mm with the bioactive glass and 2.60 +/- 1.40 mm with DFDBA. Sites grafted with bioactive glass resulted in 2.27 +/- 0.88 mm attachment level gain, while sites grafted with DFDBA had a 1.93 +/- 1.33 mm gain in attachment. Bioactive glass sites displayed 0.53 +/- 0.64 mm of crestal resorption and 2.73 mm bone fill. DFDBA-grafted sites experienced 0.80 +/- 0.56 mm of crestal resorption and 2.80 mm defect fill. The use of bioactive glass resulted in 61.8% bone fill and 73.33% defect resolution. DFDBA-grafted defects showed similar results, with 62.5% bone fill and 80.87% defect resolution. Both treatments provided soft and hard tissue improvements when compared to baseline (P < or = 0.0001). No statistical difference was found when comparing bioactive glass to DFDBA; however, studies with larger sample sizes may reveal true differences between the materials. This study suggests that bioactive glass is capable of producing results in the short term (6 months) similar to that of DFDBA when used in moderate to deep intrabony periodontal defects.

Adult↗
PubMed1997

Development and characterization of a transformed human periodontal ligament cell line.

Periodontal ligament (PDL) cells are thought to be important for establishing and maintaining a stable interface between bone and teeth. In addition, PDL cells are thought to play critical roles in both the pathogenesis of periodontal disease and the regeneration of periodontal ligament tissues. The purpose of this study was to develop a continuous or stable human PDL cell line as an in vitro model for the investigation of cellular mechanisms involved in periodontal regeneration and destruction. Human PDL cells, derived from a primary cell culture, were transfected with simian virus 40 (SV40) T antigen-containing virus with a neomycin resistance gene. The transformed cells expressed the SV40 T antigen mRNA as assayed by reverse transcription polymerase chain reaction (RT-PCR). This cell line was also characterized for morphological changes and growth characteristics compared to primary PDL cell cultures. The transformed cells were shown to form a multilayer pattern and distinct colonies on tissue culture surfaces. However, no colony formation was found in soft agar. The transformed PDL cell line was found to have a greater rate of proliferation in 10% fetal bovine serum than primary culture, and continued to proliferate in low serum concentrations capable of producing quiescence in primary cells. Interleukin-1 beta (IL-1 beta) was shown to produce a 7-fold elevation in collagenase (MMP-1) mRNA levels, consistent with primary PDL cells. In addition, IL-1 beta was shown to produce a decrease in alkaline phosphatase activity in a concentration-dependent manner. The transformed cell line has been maintained for over 30 generations of cell culture. In conclusion, a stable human PDL cell line has been established to serve as a model for future in vitro investigations into periodontal pathogenic mechanisms and to evaluate therapies directed at the regeneration of periodontal ligament.

Adult↗
PubMed1997

Crestal bone changes around titanium implants. A radiographic evaluation of unloaded nonsubmerged and submerged implants in the canine mandible.

Current implant placement utilizes both nonsubmerged and submerged techniques. However, the implications of the location of a rough/smooth implant interface as well as the location of a microgap between implant and abutment on crestal bone changes are not well understood. The purpose of this study was to radiographically evaluate crestal bone changes around unloaded nonsubmerged and submerged titanium implants in a side-by-side comparison. Fifty-nine (59) implants were placed at different levels to the alveolar crest in 5 foxhounds. Standardized radiographs were taken at baseline and at monthly intervals until sacrifice at 6 months. Radiographic assessment was carried out by measuring the distance between the top of the implant/abutment and the most coronal bone-to-implant contact (DIB), and by evaluation of bone density changes using computer-assisted densitometric image analysis (CADIA). DIB measurements revealed that in 1-part, nonsubmerged implants, the most coronal bone-to-implant contact followed at all time points the rough/smooth implant interface. In all 2-part implants, nonsubmerged and submerged, the most coronal bone-to-implant contact was consistently located approximately 2 mm below the microgap. In addition, CADIA values for all 2-part implants were decreased in the most coronal area-of-interest (AOI). All bone changes were statistically significant and detectable 1 month after implant placement in nonsubmerged implants or 1 month after abutment connection in submerged implants. Neither implant position nor individual dog effects were statistically significant. These results demonstrate that the rough/smooth implant interface as well as the location of the microgap have a significant effect on marginal bone formation as evaluated by standardized longitudinal radiography. Bone remodeling occurs rapidly during the early healing phase after implant placement for non-submerged implants and after abutment connection for submerged implants.

Alveolar Process↗