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Proteomic insights into the immunomodulatory effects of Ca/Sr co-doped sol-gel coatings for titanium implants.

Ionic functionalization of biomaterial coatings has emerged as a powerful strategy to regulate early host responses at the implant interface. However, how combined Ca/Sr incorporation governs the adsorbed proteome and downstream immune signaling remains poorly understood. This study analyses, employing in vitro tests and proteomics, the effect of adding Sr and Ca to Si-based coatings designed to bioactivate Ti implants. Hybrid Si-based coatings were synthesized by the sol-gel route with a fixed Ca content (0.5 wt%) and increasing Sr contents (0.5, 1.0, 1.5 wt%), and their physicochemical properties, ion release kinetics, and hydrolytic stability were characterized. The coatings remained highly crosslinked despite Ca/Sr incorporation, whereas the highest Sr content increased hydrolytic degradation to around 70% after 56 days. Proteomic analysis identified 183 adsorbed proteins, of which 56 were differentially adsorbed on Ca/Sr-coatings, mainly associated with immune and coagulation pathways. In vitro, RAW 264.7 showed increased gene expression of TNF-α and TGF-β; with an enhanced TNF-α secretion by the addition of Ca and Sr. In parallel, MC3T3-E1 indicated that Ca/Sr-coatings were not cytotoxic and did not impair cell proliferation. However, ALP activity was reduced in the co-doped groups, indicating that the immunomodulatory effects induced by Ca/Sr incorporation were not accompanied by enhanced early osteogenic differentiation. The Ca/Sr combination induced alterations in the adsorption of immune-related proteins, which correlated with the in vitro findings. The deeper insight into how Ca/Sr mixtures modulate protein adsorption on biomaterial surfaces may be key to understanding the immunomodulatory capacity of these bioactive cations.

Animals↗

Surface spectroscopic characterization of titanium implants after separation from plastic-embedded tissue.

The method of plastic embedding of tissue and implant and subsequent separation of plastic and implant for preparing sections of tissue adjacent to solid metallic implants relies on a successful separation of the embedment and the implant. In this work, the surface of machined Ti implants has been analysed in order to investigate to what extent plastic remnants exist on the implant after separation. SEM and AES analyses show that at least 70% of the implant surface is free of plastic remnants to a proximity of 10 nm or less from the implant surface. The method is simple and suitable for both light and transmission electron microscopy of the interface tissue.

Bone Screws↗

Initial responses of human osteoblasts to sol-gel modified titanium with hydroxyapatite and titania composition.

Sol-gel thin films of hydroxyapatite (HA) and titania (TiO(2)) have received a great deal of attention in the area of bioactive surface modification of titanium (Ti) implants. Sol-gel coatings were developed on Ti substrates of pure HA and TiO(2) and two composite forms, HA+10% TiO(2) and HA+20% TiO(2), and the biological properties of the coatings were evaluated. All the coating layers exhibited thin and homogeneous structures and phase-pure compositions (either HA or TiO(2)). Primary human osteoblast cells showed good attachment, spreading and proliferation on all the sol-gel coated surfaces, with enhanced cell numbers on all the coated surfaces relative to uncoated Ti control at day 1, as observed by MTT assay and scanning electron microscopy. Cell attachment rates were also enhanced on the pure HA coating relative to control Ti. The pure HA and HA+10% TiO(2) composite coating furthermore enhanced proliferation of osteoblasts at 4 days. Moreover, the gene expression level of several osteogenic markers including bone sialoprotein and osteopontin, as measured by RT-PCR at 24h, was shown to vary according to coating composition. These findings suggest that human primary bone cells show marked and rapid early functional changes in response to HA and TiO(2) sol-gel coatings on Ti.

Cell Proliferation↗

The bovine bone protein lyophilisate Colloss improves fixation of allografted implants--an experimental study in dogs.

BACKGROUND: Impacted morselized bone allograft is a well-established way of giving joint arthroplasties additional support in situations where there is insufficient bone stock. For long-term survival of the implant, early implant fixation is important. We hypothesized that Col-loss, a bone protein lyophilisate, might improve early implant fixation of allografted implants. METHOD: We inserted 4 porous-coated Ti implants in the distal femurs of 16 dogs. All implants were surrounded by a 2.5-mm gap, which was impacted with morselized allograft with or without Colloss. In each dog, the implants were treated with no Collos or low-, middle- or high-dose (0, 10, 20 and 40 mg) Colloss per cm3 allograft. The observation time was 4 weeks. RESULTS: Mechanical implant fixation was improved for all 3 groups with Colloss-treated implants (p < 0.05). The best anchorage was seen in the middle-dose group, where fixation was improved by 100%. We saw a dramatic reduction in fibrous tissue on the surface of the Colloss-treated implants (p < 0.001). The Colloss groups showed increased ongrowth of new bone (p < 0.01) and accelerated gap remodeling (p < 0.05). INTERPRETATION: Colloss can improve early osseointegration and fixation of allografted implants.

Animals↗

Physical characteristics of Ti-6A1-4V implants fabricated by electrodischarge compaction.

Physical characteristics of a Ti-6A1-4V implant were evaluated following fabrication using a new electrodischarge compaction technique. Ti-6A1-4V atomized powders were loaded into Pyrex tubes (3.3 mm ID) and subjected to a high-voltage, high-current-density pulse in air for a period of less than 300 microseconds. Single pulses (1.0, 1.5, 2.0 and 2.5 KJ/grampowder) were applied from a capacitor bank (240, 480 or 720 microF) to produce solid core implants with porous surface layers. Implants were evaluated microscopically for core size, neck size, pore size, grain structure, and incorporated oxide film. Hardness was also evaluated. Implants were compared with Ti-6A1-4V commercial powders. Core size increased and pore size decreased with increases in energy and capacitance. The cores were composed of equiaxed grains which were free of oxide at the grain boundary. Porous layers, consisting of particles connected in three dimensions by necks, were free of oxide films at the connecting interfaces. Neck size increased with increases in input energy and capacitance. Hardness readings of the core, necks, and porous particles resulted in readings higher than or similar to control materials. Electrodischarge compaction did not alter the physical characteristics during compaction.

Alloys↗

Histomorphometrical and clinical comparison of submerged and nonsubmerged implants subjected to experimental peri-implantitis in dogs.

OBJECTIVES: Soft and hard tissue healing around submerged and nonsubmerged versions of one dental implant design was evaluated in an experimental canine peri-implantitis model. MATERIAL AND METHODS: Forty-eight c.p. Ti-implants with integrated (one-piece=OPI) or screw-on (two-piece=TPI) abutments were inserted in edentulous mandibles of eight beagle dogs, one OPI and one TPI with connected abutments for nonsubmerged and one TPI without abutment (SMI) for submerged healing. After 3 months, all implants were functionally loaded, and at 4 months peri-implantitis was ligature-induced in one jaw side. Intravital polyfluorochrome labeling, monthly conventional radiography and gingival probing of all 48 implants were performed until sacrifice 8 months postimplantation. Undecalcified ground sections in the bucco-lingual and mesio-distal planes of four dogs (23 implants, one implant lost) were evaluated by light and fluorescence microscopy. The immunohistochemical and SEM-vascular corrosion cast results of the four other dogs (24 implants) will be reported elsewhere. Levels of alveolar bone-to-implant contact (ABICL), alveolar crest (ACL) and junctional epithelium-to-implant contact were determined by computer-assisted histometry. Peri-implant alveolar bone loss (=saucerization) was assessed on the radiographs and calculated as ACL minus ABICL from histometric data. RESULTS: Around SMIs and OPIs without ligature less plaque adhesion and lower gingival indices were found when compared to TPIs. Radiologically, all ligatured, but also some nonligatured implants showed alveolar bone loss. Histometry demonstrated reduced ABICL around all these implants. Saucerization was more pronounced on the lingual and mesio-distal sides. Particularly around TPIs, bone resorption was still active or bone formation was impaired on fluorochrome labeling. Only around SMIs and one OPI without ligature continuing alveolar bone formation reflected by gains in ABICL were found. CONCLUSION: The clinical and histometric results of this study demonstrate that healing of submerged SMIs was not impaired by the two-stage procedure, resulting in equally good healing as around nonsubmerged OPIs. However, peri-implantitis plaque-induced by ligature and/or dilated abutment connection microgaps in TPIs affected alveolar bone-to-implant contacts more than transmucosal or submerged healing mechanisms.

Alveolar Bone Loss↗

Protective covering of surgical wounds with honey impedes tumor implantation.

HYPOTHESIS: Tumor implantation (TI) development at the surgical wound following cancer surgery is still an unresolved concern. Trocar site recurrence, which is likely a form of TI, has become one of the most controversial topics and, with the widespread acceptance of laparoscopic surgery, has caused renewed interest in questions about TI. Honey has positive effects on wound healing. Physiological and chemical properties of honey might prevent TI when applied locally. DESIGN, INTERVENTIONS, AND MAIN OUTCOME MEASURES: Sixty BALB/c strain mice, divided into 2 groups, were wounded in the posterior neck area. Group 1 mice formed the control group, and group 2 mice had wounds coated with honey before and after tumor inoculation. All wounds were inoculated with transplantable Ehrlich ascites tumor. The presence of TI was confirmed in the wounded area by histopathological examination on the 10th day. RESULTS: Tumor implantation was achieved in all group 1 animals and verified by palpable mass and histopathological examination. In group 2 mice, although TI could not be detected macroscopically, it was revealed by pathological examination in 8 cases. Tumor implantation was less likely in group 2 mice (8 of 30 vs 30 of 30; P<.001). CONCLUSIONS: Tumor implantation was markedly decreased by the application of honey pre- and postoperatively. It is possible that the physiological and chemical properties of honey protected wounds against TI. Honey could be used as a wound barrier against TI during pneumoperitoneum in laparoscopic oncological surgery and in other fields of oncological surgery.

Animals↗

In vitro stability of plasma-sprayed hydroxyapatite coatings on Ti-6Al-4V implants under cyclic loading.

The success of hydroxyapatite (HA)-coated Ti-6Al-4V implants relies on the long-term stability of HA coatings. In this study, the mechanical stability of plasma-sprayed HA coatings on Ti-6Al-4V implants under four-point cyclic bending was systematically investigated in both air and simulated body fluid (SBF) environments at room temperature. To have a clear view of the microscale damage evolution, the surface morphology change of HA coatings during cyclic loading was carefully examined by scanning electron microscopy at the same locations on the coating surfaces after four-point bending for 4, 6.5, 8.5, and 10 million cycles. Also, possible changes of other characteristics such as thickness, weight, crystallinity, and residual stress of HA coatings were measured as a function of loading cycles. Up to 10 million cycles of bending in air and SBF, we found no significant microcracking or coating spalling on the surface of coatings, and no significant changes in thickness, weight, crystallinity, or residual stress of the plasma-sprayed HA coatings. The experiment results indicate that thickness and crystallinity had no effects on the stability of the HA coatings. HA coating resistance to the cyclic four-point bending might result from the stress shielding effects of preexisting microcracks in the coatings.

Biocompatible Materials↗

In vitro evaluation of bio-functional performances of Ghimas titanium implants.

Titanium is the most widely used material for dental implants. The natural formation, in presence of oxygen, of different oxide films (passivation films) is correlated to titanium implant biocompatibility, resistance to corrosion and is responsible for implant bacteriostatic action. Surface roughness is another surface property of Ti-implants that, affecting implant-to-bone contact, improves integration. In the present study data concerning composition, surface roughness and biocompatibility of Ghimas implants and mini-implants undergoing sandblasting with Calcium Magnesium Carbonate (CaMg(CO3)2) are reported. AFM, SEM/EDX, XRD analyses and morpho-functional tests (MTT and ALP) were performed. Cell actin cytoskeletal modification (fluorescence phalloidin staining) was also observed with confocal laser microscopy (CLSM). Data related to surface geometry and chemical properties, associated with evidence of high purity of all the tested materials (XRD and EDX), highlighted the elevated biocompatibility of tested implants and mini-implants. CLSM investigation confirmed osteoblast features of an active cell behavior able to fit cell to chemico-mechanical stimuli present at the bone/implant interface and suggests an effective implant/alveolar bone integration in vivo.

Biocompatible Materials↗

Bone ingrowths to press-fit and loose-fit implants: comparisons between titanium and hydroxyapatite.

The aim of this study was to investigate whether the coating of titanium (Ti) implants with hydroxyapatite (HA) might create a better fixation when titanium implants are implanted into a gap. In each of 16 rats, the medullary cavity of both femurs was entered by an awl from the trochanteric area. With steel burrs it was successively reamed to a diameter of 1.5 mm. In a random manner the proximal part of the cavity in half of the bones was reamed once again to a diameter of 2.0 mm. Nails with a diameter of 1.5 mm and a length of 34 mm were then inserted into the medullary cavity of these bones with press fit at the distal half and a gap to the bone in the proximal half. In the remaining bones the whole medullary canal was reamed to a diameter of 2.0 mm, and nails with a diameter of 2.0 mm and a length of 34 mm were introduced. In all cases, either a pure Ti nail or a Ti nail entirely plasma sprayed with HA was used in a random manner. The surface roughness of the pure Ti was characterized by Ra 2.6 microm and Rt 22 microm. Ra of HA was 7.5 microm and Rt 52 microm. At sacrifice after 16 weeks, both femurs were dissected free from soft tissues and then immersed in fixative. A specimen slice of about 5 mm in thickness was prepared from the subtrochanteric region with a water-cooled band saw. Sample preparation for undecalcified tissue followed the internal guidelines at the laboratories of the Department of Biomaterials/Handicap Research. Generally, bone contact to the nails with HA coating was more predictable than was bone contact to the Ti nails. But due to rather large variations in bone contact between the samples, statistical analyses revealed non-significant differences between the 4 groups (p = 0.083). There were no significant differences between Ti and HA coated nails of 2.0 mm (p = 0.633), nor between Ti and HA coated nails of 1.5 mm (p = 0.924). The pooled values for the 2.0 mm nails showed significantly higher bone bonding contact than the pooled values of the 1.5 mm nails (p = 0.011). Our results, then, indicate that bone bonding contact to implants with a loose fit insertion is less predictable than in press fit insertion, and HA coating seemed to be more predictable than pure Ti. However, due to large variations between the samples, the differences did not reach significant levels.

Animals↗

Effect of cpTi surface roughness on human bone marrow cell attachment, proliferation, and differentiation.

There is general agreement that rough surfaces improve both biologic and biomechanical responses to titanium (Ti) implants. The aim of this investigation was to study the effect of Ti surface roughness on the response of human bone marrow cell culture evaluating: cell attachment, cell proliferation, total protein content, alkaline phosphatase (ALP) activity, and bone-like nodule formation. Cells were cultured on commercially pure titanium (cpTi) discs with fourdifferent average roughnesses (Ra). For attachment evaluation, cells were cultured for 4 h. After 21 days, cell proliferation, total protein content, and ALP activity were evaluated. For bone-like nodule formation, cells were cultured for 28 days. Data were compared by ANOVA and Duncan's multiple range test. Cell attachment was not affected by surface roughness. For cells cultured on Ti with Ra ranging from 0.80 microm to 1.90 microm, proliferation was reduced while total protein content, and ALP activity were increased. There was a non-statistically significant increase of bone-like nodule formation on a surface with Ra near 0.80 microm. These results suggest that for Ti an Ra ranging from 0.80 microm to 1.90 microm would optimize both intermediary and final cellular responses but not affect the initial response, and a smoother surface would not favor any evaluated response.

Alkaline Phosphatase↗

[A biomechanical investigation of Ha-coated ceramic implant]

Two samples of Ha-coated ceramic implants of which sintering temperatures were 850 degrees centigrade,1050 degrees centigrade respectively,were prepared.Pure titanium implant of the same size was accomplished using a transcortical implant model in adult Mongrel dogs.Follow-up periods were 1,3 and 6 months.The results in vivo were evaluated using push-out tests and scanning electron microscope.There were significant differences in push-coated ceramic implants and pure titanium implants,especially for the 1 and 3 months follow-up period.SEM investigation of the interface after push-out test showed for the HA-coated ceramic implants to be fractured in the coating layer and for the Ti-implant to be fractured at the implant-bone interface.

Journal Article↗

Bone reactions to oxidized titanium implants with electrochemical anion sulphuric acid and phosphoric acid incorporation.

BACKGROUND: The importance of the surface properties of implants for a successful osseointegration has been emphasized. It is generally known that bone response to implant surfaces is considerably related to the various surface properties. PURPOSE: The purpose of this study was to investigate bone tissue reactions to multifactorial biocompatibility of the surface oxide of electrochemically oxidized titanium implants. The ultimate objective was to improve surface quality, resulting in enhancement of clinical outcomes of osseointegrated implants. MATERIALS AND METHODS: Three different surface types of commercially pure titanium (c.p. Ti) implants were prepared. Turned implants were used for controls and test implants were prepared by the micro arc oxidation (MAO) method, either in sulphuric acid (S implants) or in phosphoric acid (P implants). Implants were inserted in the femur and tibia of 10 mature New Zealand White rabbits. The bone response was evaluated by biomechanical tests, histology, and histomorphometry. The follow-up time was 6 weeks. RESULTS: The mean peak values of the removal torque showed significant differences between control and test S implants (p =.022) but showed no significant differences between control and test P implants (p =.195) or between test S and test P implants (p =.457). In addition, the histomorphometric comparisons of the bone-to-metal contact around entire implants demonstrated 186% increase in S implants (p =.028) and 232% increase in P implants (p =.028) compared with the paired control groups. Quantification of the bone area in the threads did not show any significant differences. CONCLUSIONS: The present results suggest that the primary mode of action in strong bone response to S implants is mechanical interlocking, and to P implants, it is biochemical interaction. It is possible that the phosphate groups in the titanium oxide of P implants provide potential chemical bonding sites for calcium ions and hydroxyapatite of the bone matrix during biologic mineralization. key words: bone responses, histomorphometry, oxidized implants, removal torque test, surface oxide properties

Animals↗

Surface modification of titanium through amino group implantation.

We modified Ti surfaces by implantation of amino (NH(2+)) groups at 10(16) and 10(17) cm(-2). The implanted surfaces were characterized by X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), scanning Auger electron spectroscopy (AES), and second ion mass spectroscopy (SIMS). The experimental results showed that the implanted Ti specimens were covered by a dominant hydrocarbon overlayer due to contamination and the surface oxide layer of implanted specimens became thicker. XPS, AES, and SIMS depth profiles showed that implanted elements had a typical ion implantation distribution and that titanium nitride (TiN) was formed.

Amines↗

Systemically administered human growth hormone improves initial implant stability: an experimental study in the rabbit.

PURPOSE: This study was an investigation to determine whether human growth hormone (hGH) continuously administered to rabbits may improve implant integration in bone. MATERIALS AND METHODS: Thirty-two commercially pure titanium (c.p. Ti) implants were inserted in the tibiae of 16 rabbits. Human growth hormone (0.3 U/kg/d) or sodium chloride (NaCl) was administered by subcutaneous pumps. Insulin-like growth factor-1 (IGF-1) levels in blood were measured. Two biomechanical tests were performed: (1) every second week resonance frequency analysis (RFA) was used to investigate implant stability or stiffness at the interface and, after 8 weeks of follow-up, (2) removal torque (a measure of implant integration and stability) was registered. Further evaluation was performed by dual energy x-ray analysis (DEXA), to evaluate bone mineral density, and histomorphometric analysis of tissue-to-implant integration on undecalcified cut and ground sections. RESULTS: A difference in implant stability was detected with the RFA technique after 2 weeks and 8 weeks in favor of the hGH-treated rabbits. No significant differences were detected with removal torque, DEXA, and histomorphometric measurements. The blood test demonstrated antibody development in the rabbits treated with hGH after 4 weeks. CONCLUSION: Growth hormone has an initial beneficial effect on implant integration; however, owing to rapid antibody formation, this study did not demonstrate whether this effect remains in the long term.

Absorptiometry, Photon↗

A new implant designed to maximize contact with trabecular bone: survival to 18 months.

PROBLEM: A roughened, commercially pure titanium (CP-Ti) implant design has been developed that features a different length and pitch for each screw thread to direct functional stresses away from cortical bone and to the more resilient trabecular bone. Abutment-implant connection is made using a conical taper to provide a seal against invasion by microorganisms. PURPOSE: To assess short-term (18 months) clinical performance of this innovative implant design. METHODS: A total of 1,419 implants were placed in 313 patients to support 419 prostheses in a multidisciplinary, multicentered, prospective clinical study conducted by the Ankylos Implant Clinical Research Group (AICRG). More than 100 dentists at 32 centers in the United States, 1 in Korea, and 1 in Taiwan are involved in the study. Failure was defined as implant removal for any reason. The influence of mobility at placement, implant length and diameter, incision type, augmentation, crestal bone reduction, bone density, and the use of the operating room or dental clinic on survival were evaluated over 18 months. Crestal bone loss between placement and uncovering was also determined. RESULTS: Crestal bone loss ranged from 0.2 to 0.5 mm. The overall success rate from placement to 18 months was 96.6%. Implants mobile at placement failed more frequently (16.9%) compared with stable implants (3.1%). Wide-diameter implants and longer implants exhibited higher survival rates. Incision design and surgery location did not influence survival. Bone density was important to clinical survival.

Adult↗

Effect of platelet-rich plasma on the peri-implant bone response: an experimental study.

Platelet-rich plasma (PRP) is used as a source of growth factors to stimulate and accelerate bone formation and soft tissue healing. The use of PRP in bone regeneration, both around dental implants and in periodontic treatments, has become particularly appealing. The aim of this study was to evaluate the effect of PRP in an experimental model of osteogenesis around laminar implants. Fifteen male Wistar rats, weighing 90 +/- 10 g, were used in this study. One milliliter of blood was obtained from each animal by intracardiac puncture and transferred into Eppendorf tubes containing 10% sodium citrate. The tubes were centrifuged at 1500 rpm for 15 minutes and PRP was prepared. The laminar test was used to evaluate the bone peri-implant response. PRP and a titanium laminar implant were introduced into the right tibia (Ti/PRP group), whereas the left tibia (control) received only a laminar implant (Ti group). Thirty days postimplantation, the tibiae were resected, radiographed, and processed for embedding in acrylic resin. Ground sections (50 microm) were stained with toluidine blue. The peri-implant bone volume was evaluated histomorphometrically. Statistical analysis of the data was performed. The amount of newly formed bone in the Ti/PRP group (30 +/- 7 cm) was significantly greater than in the Ti group (16 +/- 3 cm). A greater volume of peri-implant bone was observed when PRP was used in the laminar implant test model.

Animals↗

Mechanical and histological evaluation of amorphous calcium phosphate and poorly crystallized hydroxyapatite coatings on titanium implants.

The effect of amorphous calcium phosphate (Ca/P) and poorly crystallized (60% crystalline) hydroxyapatite (HA) coatings on bone fixation to "smooth" and "rough" (Ti-6A1-4V powder sprayed) titanium-6Al-4V (Ti) implants was investigated. Implants were evaluated histologically, mechanically, and by scanning electron microscopy (SEM) after 4 and 12 weeks of implantation in a rabbit transcortical femoral model. Histological evaluation of amorphous vs. poorly crystallized HA coatings showed significant differences in bone apposition (for rough-coated implants only) and coating resorption (for smooth- and rough-coated implants) that were increased within cortical compared to cancellous bone. The poorly crystallized HA coatings showed most degradation and least bone apposition. Mechanical evaluation, however, showed no significant differences in push-out shear strengths between the two types of coatings evaluated. Differences between 4 and 12 weeks were significant for coating resorption and push-out shear strength but not for bone apposition. Significant enhancement in interfacial shear strengths for bioceramic coated as compared to uncoated implants were seen for smooth-surfaced implants (3.5-5 times greater) but not for rough-surfaced implants at 4 and 12 weeks. Rough implants showed greater mean interfacial strengths than uncoated smooth implants at 4 and 12 weeks (seven times greater) and to coated smooth implants at 12 weeks only (two times greater). Mechanical failure of the bone/coating/implant interface consistently occurred within the bone, even in the case of the poorly crystallized HA coatings, which had almost completely resorbed on rough implants. These results suggest that once early osteointegration is achieved biodegradation of a bioactive coating should not be detrimental to the bone/coating/implant fixation.

Alloys↗