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1.
Postmortem examination of human specimens is an extremely important aspect of evaluating the relative compatibility and long-term success of endosseous implant surfaces. The bone-implant interface of 5 commercially pure titanium screw-type mandibular implants after 85 months of service and 2 hydroxyapatite- (HA) coated maxillary implants after 38 months of service were examined. All implants were stable at the time of the patient's death. The mandibular implants had an average of 65% contact with bone and the maxillary implants had an average of 47% contact. The HA coating had separated from the maxillary implants in some areas and was free within surrounding connective tissue or surrounded by invaginating sulcular epithelium. The arrangement and pattern of bone contact appeared different between HA-coated and titanium implant surfaces.  相似文献   

2.
BACKGROUND: Controversy over the long-term clinical effectiveness of hydroxyapatite (HA)-coated dental implants still persists, despite numerous clinical studies documenting high survival rates. Concerns about the degradation of the coating over the years have been raised; it has been speculated that resorption of the HA could produce a space between the implant and the bone with a resultant mechanical instability. METHODS: Two HA-coated implants were retrieved due to a fracture of the abutment screws after a loading period of 14 years and were treated to obtain thin ground sections for histologic evaluation. RESULTS: At low-power magnification, it was possible to observe that the HA coating was in contact with mature bone. No gaps or connective fibrous tissue was found at the implant-bone interface. No epithelial downgrowth was present. No acute or chronic inflammatory cell infiltrate was present at the implant-bone interface. No foreign body reaction was present in the peri-implant tissues. Some osteocytes were in direct contact with the coating. For implant 1, the percentage of bone-titanium contact was 25% +/- 2.1%, and the percentage of bone-HA contact was 35% +/- 1.4%. The total bone-implant contact was approximately 60%. The HA coating appeared to be resorbed in 46% +/- 3.2% of the implant perimeter, especially in the coronal portions of the implant. For implant 2, the mean percentage of bone-HA contact was 13% +/- 1.8%, and the mean percentage of bone-titanium contact was 15% +/- 2.3%. The total bone-implant contact was approximately 28%. The HA coating appeared to be resorbed for a mean of 68% +/- 4.1% of the implant perimeter, especially in the coronal portion of the implant. CONCLUSIONS: No acute or chronic inflammatory cell infiltrate was present in the peri-implant tissues. No signs of coating infection, fatigue, or failure were observed in two specimens. The HA coating may not be susceptible to degradation or dissolution under long-term function.  相似文献   

3.
Long-term support of dental implants requires adequate bone thickness in the area surrounding the implant. A three-dimensional examination, including calculations of percent bone-implant contact and percent bone volume, was conducted to clarify the bone structure around pure titanium (Ti) and dense hydroxyapatite (HA) implants. The implants were installed in the tibiae of rabbits, and the bone structure was examined after 2, 4, 6 and 8 weeks. The bone formation following implantation differed in the cortical bone and cancellous bone areas. In the cortical bone area, the percent bone-implant contact and percent bone volume were comparatively consistent for both Ti and HA implants during the observation period. In the cancellous bone area, both findings were influenced by the implantation period, and the chronological bone structure in the cancellous bone area also differed between Ti and HA implants. The percent bone-implant contact and percent bone volume in the Ti implant increased over 8 weeks, whereas the dense HA implant increased for the first 4 weeks and then decreased. The implant materials, Ti and HA, affected the bone remodeling in the cancellous bone area.  相似文献   

4.
PURPOSE: The different implant systems available today present several types of surface treatment, with the aim of optimization of bone-implant contact. This study compared 4 different types of implant surfaces. MATERIALS AND METHODS: The first, second, third, and fourth mandibular premolars were extracted from 5 young adult mongrel male dogs. Ninety days after removal, four 3.75-mm-diameter, 10-mm-long screw-type implants (Paragon) were placed with different surface treatments in mandibular hemiarches. The dogs received 2 implants of each of the following surface treatments: smooth (machined), titanium plasma spray (TPS), hydroxyapatite coating (HA), and sandblasting with soluble particles (SBM). The implants were maintained unloaded for 90 days. After this period, the animals were sacrificed, and the hemimandibles were extracted and histologically processed to obtain non-decalcified sections. Two longitudinal ground sections were made for each implant and analyzed under light microscopy coupled to a computerized system for histomorphometry. RESULTS: The following means were obtained for bone-implant contact percentage: machined = 41.7%, TPS = 48.9%, HA = 57.9%, and SBM = 68.5%. DISCUSSION: The means for all treatments that added roughness to the implant surface were numerically superior to the mean found for the machined surface. However, this difference was statistically significant only between groups SBM and machined (Tukey test, P < .05). CONCLUSIONS: The SBM-treated surface provided a greater bone-implant contact than a machined surface after 90 days without loading in this model.  相似文献   

5.
The histologic examination of dental implants retrieved from humans provides a unique opportunity to evaluate the bone-implant interface. This case report presents a clinical, radiographic, and histologic evaluation of a cylindrical hydroxyapatite- (HA) coated implant retrieved from the posterior maxillary area of a patient after 9 years after placement. The implant had been placed in conjunction with a subantral augmentation procedure with HA as the graft material. Clinical examination revealed an immobile implant with no sign of pathosis. Radiographic examination indicated close proximity of the bone to the implant surface without evidence of radiolucency. Histologically, because of tissue destruction during implant retrieval, only the apical portion of the implant was available for examination under light microscopy, and it appeared to be integrated with the surrounding bone; 45.9% of the surface of the implant had close bone apposition at the interface. There was no evidence of dissolution of the HA coating and the bone appeared to be in immediate contact with the coating. Residual graft particles were present and in close proximity with the implant surface. These observations suggest that the subantral augmentation procedure performed simultaneously with the placement of an HA-coated implant with HA as the graft material apparently resulted in osseointegration between the implant and the surrounding bone. The implant was maintained without complication for 9 years.  相似文献   

6.
Hydroxylapatite (HA)-coated and grit-blasted (non-HA-coated) titanium dental implants were inserted into healed extraction sites of canine mandibles. After six weeks, the animals were killed and the implants mechanically tested in torsion to failure. Interface attachment strength, implant/tissue compatibility, integrity of the HA coating, and the location of interface failure were evaluated. Mechanical testing demonstrated an interface torsional strength of 3.98 +/- 0.93 MPa for the HA-coated implants and 2.25 +/- 0.65 MPa for the grit-blasted implants. This represents a 76.9% improvement in the maximum torsional interface strength, and is statistically-significant (p = 0.0004). On qualitative histologic analysis, interface failure was seen to occur primarily at the HA/implant interface, although failure through the HA coating and regions of bone/HA interface failure were observed. The HA-coated implants had bone in direct apposition to their surface with no fibrous tissue interposition. The grit-blasted implants also had regions of direct bone-implant apposition, but these areas were limited to a smaller proportion of the total interface area. There was no evidence of breakdown or change in thickness of the HA coating.  相似文献   

7.
PURPOSE: The aim of the present study was to test the hypothesis that calcium phosphate coating of titanium screw-type implants enhances peri-implant bone formation in the jaw. MATERIALS AND METHODS: Ten adult female foxhounds received experimental titanium screw-type implants in the mandible 3 months after removal of all premolar teeth. Four types of implants were evaluated in each animal: implants with machined titanium surface (the control group), implants coated with collagen I (the collagen-only group), implants with a composite coating of calcium phosphate and mineralized collagen I (the composite group), and implants with calcium phosphate (hydroxyapatite [HA]) coating (the HA-only group). Peri-implant bone regeneration was assessed histomorphometrically after 1 and 3 months in 5 dogs each by measuring bone-implant contact (BIC) and the volume density of the newly formed peri-implant bone (BVD). RESULTS: After 1 month, BIC was significantly enhanced only in the group of implants with composite coating of calcium phosphate and mineralized collagen (P = .038). Volume density of the newly formed peri-implant bone was significantly higher in all coated implants after 1 month. No significant difference from baseline was found in BIC for the collagen-only and HA-only groups, but BVD was significantly higher in implants with composite coating (P = .041). After 3 months, BIC and BVD were significantly higher in all coated implants than in the controls with machined surfaces. CONCLUSION: It was concluded that composite coating of dental screw-type implant surfaces using calcium phosphate and collagen can enhance BIC and peri-implant bone formation.  相似文献   

8.
Objective: Human retrieved implants with an intact bone-implant interface play a pivotal role in validating data obtained from in vitro studies and animal experiments. This study presents a histologic and histomorphometric analysis of peri-implant tissue reactions and of the bone-titanium interface in three machined and sandblasted dental implants retrieved after a 5-year loading period. Method and Materials: Three implants, with an intact bone-implant interface, were found in the Archives of the Implant Retrieval Center of the Dental School of the University of Chieti-Pescara, Chieti, Italy. The three implants had been used in a two-stage submerged procedure and loaded as part of a small prosthetic restoration. One implant had been retrieved because of an abutment fracture, while there was a fracture of the connecting screw in the other two. One implant was in the maxilla (sandblasted surface), and two were in the mandible (one with a machined surface and the other with a sandblasted surface). All implants had been processed for histology. Results: All three implants presented mature, compact, lamellar bone at the interface. Many remodeling areas were present in the peri-implant bone, especially inside the implant threads. The bone was always in close contact with the implant surface. The bone-implant contact percentage of the machined implant was 92.7%, while the two sandblasted implants showed bone-implant contact percentages of 85.9% and 76.6%. Conclusion: The present histologic results confirmed that these implants with different surfaces maintained a good level of osseointegration over a 5-year loading period, with continuous remodeling at the interface, and showed high bone-implant contact percentages. (Quintessence Int 2012;43:287?292).  相似文献   

9.
PURPOSE: This experimental study in adult female sheep examined the value of nonresorbable porous hydroxyapatite (HA) as a grafting material in a single-stage sinus-lift procedure. MATERIALS AND METHODS: Two titanium plasma-flame-sprayed cylindric implants were placed bilaterally in each of 54 sinuses in 27 adult female sheep. In 2 groups of 18 sinuses each, the subantral hollow space was filled with porous HA or autogenous cancellous bone harvested from the iliac crest, respectively. Eighteen sinuses were not augmented and served as controls. The time course of new bone formation and bone remodeling was evaluated by sequential polyfluorochrome labeling. Observation periods were 12, 16, and 26 weeks after the surgical procedure. Six sinuses per observation period and test group were available for histologic evaluation. RESULTS: All implants were osseointegrated in the local host bone. New bone formation was observed in a triangular area bounded by the implant surface, local buccal antral wall, and submucous connective tissue around all implants. The mean length of bone-implant contact was 3.9 +/- 0.3 mm in the control group, 5.7 +/- 0.3 mm in the autogenous bone group, and 5.9 +/- 0.3 mm in the group augmented with porous HA. During the observation period, the relative length of direct bone-implant contact increased from 20% to 25.1% in the control group, from 30.4% to 35.5% in the autogenous bone group, and from 29.8% to 41.7% in the HA group. At a distance of 1 mm from the implant, the mean bone volume was 29.7 +/- 15.7% in the autogenous bone group. In the group augmented with HA the mean bone volume was 11.2 +/- 13.0%. DISCUSSION: There was no significant difference between HA and autogenous bone regarding bone-implant contact (P = .89). CONCLUSIONS: Both groups showed a significantly greater bone-implant contact (HA: P = .002; autogenous bone: P = .0005) than the empty control group. However, since the results varied widely, the use of HA alone for sinus grafting should be used with discretion in sinus-lift procedures.  相似文献   

10.
The influence of surface quality, in particular surface topography and implant material, was evaluated by inserting titanium‐ and hydroxylapatite plasma‐sprayed coated implants into the maxilla of 10 goats. Three types of plasma‐spray coatings were applied to tapered, screw shaped implants; titanium plasma‐spray coating (TPS), titanium plasma‐spray coating with additional acid passivation (TPSA) and a bilayered coating (TPS/HA) consisting of titanium plasma‐spray coating (TPS) and a hydroxylapatite part (HA). In addition, as machined implants (TIM) were used as control. A total of 40 implants were inserted according a balanced split plot design. At the end of a 3‐month healing period, it appeared that 5 implants (2 TPS, 1 TPSA, 1 TPS/HA and 1 TiM) were lost. Histological examination revealed a stronger bone response to TPS/HA coated implants. Even the TPS/HA coated implants induced bone formation on the part of the implant inserted into the sinus. No signs of delamination of the TPS coatings were visible. The HA part of the dual coating showed signs of degradation. Histomorphometrical analysis confirmed these findings. A significant difference in bone contact ( P <0.05) was measured between the TPS/HA coated implants and the other types of implants. Linear regression ( r >0.2) showed no correlation between the inscrew values at the base line and the bone contact measurements 3 months after healing. On the basis of these results, we can conclude that the chemical composition of the HA coating has a positive influence on the bone reaction. The influence of roughness is less evident.  相似文献   

11.
This article presents a histologic evaluation of three hydroxyapatite (HA)-coated root-form implants retrieved from humans after being in function for 3.5 to 11 years. If the coronal portion, where bone loss was observed clinically and radiographically, is excluded, all implants appeared to be well osseointegrated, with intimate contact between the surrounding bone and the coating. There was no sign of resorption or dissolution of the HA coating. The coating had a uniform thickness (50 microm) equal to the thickness originally provided by the manufacturer In the few areas where there was no bone contact, the HA coating appeared to line the implant with no sign of dissolution. The few detached particles had tight contact with the bone, demonstrating the biocompatibility of the HA. The observations from the three reported cases suggest that the HA coating of dental implants may not be susceptible to resorption or dissolution under long-term function.  相似文献   

12.
A histomorphometric study was performed on six retrieved loaded hydroxyapatite (HA)-coated titanium implants. The implants had been clinically functioning in a 50-year-old patient. She originally presented a severely atrophic maxilla that was reconstructed by sinus floor augmentation using autogenous bone from the iliac crest. In spite of good clinical function, because of psychiatric problems, all implants, including some bordering bone, were removed using a trephine bur. Thin ground sections were prepared for histology and used for histomorphometry. The aim of this study was to observe the condition of the calcium phosphate coating after 2.5 years of functional loading, the nature of the bone-to-implant interface, as well as the density of the bone graft around the implants. Intimate and abundant bone-to-implant contact was observed, ranging from 90.4% to 99.8% along the implant surface. Active bone remodeling occurred within all threads, as demonstrated by secondary osteons close to the implant surface. The thickness of the calcium phosphate coating varied from 51 to 88 pm for the loaded retrieved implants, versus 53 to 89 microm for the nonused control implant. All implants showed bone contact including the first thread and up to the smooth titanium neck. Both the nature and thickness of the coating had hardly changed after 2.5 years of loading. The HA-coated implants used achieved excellent osseointegration and must be considered clinically safe and effective in maxillary grafted bone.  相似文献   

13.
The surface characteristics of dental implants play an important role in the osseointegration process. Over the years implant surfaces have been subjected to different treatments, including turning, plasma spraying, coating, sand blasting, acid etching, and anodization. FBR coating is a fully resorbable calcium phosphate (CaP) coating made of brushite, obtained by electrochemical deposition on titanium plasma-sprayed (TPS) implants; this bioactive layer may be totally resorbable in 6-12 weeks and once the FBR coating has been resorbed, the newly formed bone is in contact with the roughness of the TPS surface. Human biopsy of immediately-loaded implants is certainly the most definitive means of determining the occurrence of osseointegration. In this case series the histologic and histomorphometric features of the bone-implant interface are analyzed and discussed in 3 immediately restored implants, retrieved from human subjects at 8, 10 and 12 weeks, respectively. All 3 implants were osseointegrated, with a bone to implant contact (BIC) ranging from 54.4% to 70.1%. The FBR coating was resorbed and replaced by new bone. Osteoconduction was especially noticeable between the implant threads, where the pristine bone was removed during implant bed preparation. The results suggest that the resorption window of 6-12 weeks for the CaP coating seems to be confirmed at least in the human mandible, and that immediately loaded FBR-coated implants placed in the posterior mandible can achieve osseointegration within 6-12 weeks of loading.  相似文献   

14.
The influence of surface roughness and calcium phosphate (Ca-P) coating on the bone response of titanium implants was investigated. Four types of titanium implants, i.e. as-machined, grit blasted, as-machined with Ca-P sputter coating, and grit blasted with Ca-P sputter coating, were prepared. The Ca-P sputter-coating, produced by using the RF magnetron sputter technique, was rapid heat-treated with infrared radiation at 600 degrees C. These implants were inserted into the left and right femoral condyles and the left and right tibial diaphyses of the rabbits. After implantation periods of 2 and 12 weeks, the bone-implant interface was evaluated histologically and histomorphometrically. Histological evaluation revealed no new bone formation around different implant materials after 2 weeks of implantation. After 12 weeks, bone healing was almost completed. For both tibial and femoral implants, Ca-P coated implants always showed a higher amount of bone contact than either of the non-coated implants. On the other hand, surface roughness improved only the response to implants inserted into the tibial diaphysis. On the basis of these findings, we concluded that 1) deposition of a sputtered Ca-P coating on an implant has a beneficial effect on the bone response to this implant during the healing phase, and 2) besides implant surface conditions the bone response is also determined by local implant site conditions.  相似文献   

15.
PURPOSE: The objective of this study was to evaluate the effect of radiofrequency- (RF) sputtered calcium phosphate (CaP) coating of titanium implants on bond strength at the bone-implant interface and percent bone contact length. MATERIALS AND METHODS: Cylindric sputtered CaP-coated and plasma-sprayed hydroxyapatite- (HA) coated implants (4.0 mm diameter and 8 mm length) were implanted in dog mandibles. Half the sputtered CaP-coated implants were heat-treated. RESULTS: Twelve weeks after implant placement, no statistical differences in the mean ultimate interfacial strengths were observed between as-sputtered CaP-coated, sputtered CaP-coated heat-treated, and control plasma-sprayed HA-coated implants. Histomorphometric evaluation indicated that the percent bone contact lengths for the plasma-sprayed HA-coated implants and the as-sputtered CaP-coated implants were similar and significantly greater than that for the sputtered CaP-coated heat-treated implants. Differences in the ultimate interfacial strength and percent bone contact length between different implant sites in the mandible were not observed. DISCUSSION: The results of this study, considered together with the results of previous studies, suggest that once early osseointegration is achieved, biodegradation of the thin CaP coatings is not detrimental to bone-coating-implant fixation, and does not compromise bone responses to the coated implant surfaces. CONCLUSION: The interfacial strength and histomorphometric data suggest that the CaP coatings applied using the sputtering process produce bone responses similar to those of HA coatings applied using plasma spraying.  相似文献   

16.
Dental implant failures are still a common occurrence, especially in areas of poor bone quality. The purpose of this study was to observe whether the application of a growth factor, recombinant human transforming growth factor beta-1 (rhTGF-beta1), would improve the healing of bone adjacent to titanium dental implants. Four adult male Yucatan minipigs had four titanium dental implants placed into the inferior border of each hemimandible. On the control side, the implants were placed in standard fashion. On the experimental side, rhTGF-beta1 within a carrier gel was placed into the recipient site before implant placement. After a 6-week healing period, two implants from each side were evaluated by pull-out testing and histomorphometric analysis. The mean pull-out force for experimental implants was 1,124.01 N and 818.12 N (P = 0.5015) for controls. The mean percentage of bone between adjacent threads on the implant surface was 49.74% for the experimental group and 36.50% for controls (P = 0.0783). The mean percentage of bone-implant surface contact was 41.86% for the experimental implant sites and 24.60% for the control implant sites (P = 0.0452). The application of rhTGF-beta1 to implant sites appears to increase the amount of bone healing adjacent to a titanium dental implants in minipig mandibles at the 6-week period. Further studies are required to quantify better the amount of growth factor required and to study its effects over a broader period of time to see whether these differences are maintained.  相似文献   

17.
目的:观察不同微弧氧化时间处理对纯钛表面形成的掺锶羟基磷灰石涂层表面形貌的影响,以及不同表面形貌特征对其表面成骨特性的影响。方法:经5、10、15 min 3种微弧氧化时间在钛表形成3组掺锶羟基磷灰石涂层,分别采用扫描电镜观察表面形貌;采用表面粗糙度仪测量涂层表面粗糙度数值。然后再将3种钛种植体植入新西兰兔体内,术后4、12周取材,采用组织染色法观察植入体表面骨形成情况和骨接触率(Bone Implant Contact,BIC)。结果:随着微弧氧化时间的延长,涂层表面形貌成多孔状且越加不规则,粗糙度增加;丽春红染色显示4周时植入体表面有新骨形成,12周时转化为成熟的骨组织并与涂层形成紧密的骨结合。随植入时间的延长种植体表面骨接触率逐渐增加,而且15 min组和10 min组的骨接触率在第4周和12周均高于5 min组。结论:不同微弧氧化时间可以改变掺锶羟基磷灰石涂层的表面特性,而粗化的的涂层表面结构有利于骨组织的形成。  相似文献   

18.
PURPOSE: The influence of calcium phosphate (CaP) and hydroxyapatite (HA) crystallinity on bone-implant osseointegration is not well established. In this study, the effect of HA crystallinity and coating method on bone-implant osseointegration was investigated using a rat tibia model. MATERIALS AND METHODS: HA coatings 1 to 5 microm thick were produced using a supersonic particle acceleration (SPA) technology. The HA crystallinities used for this study were weight ratios of 30%, 50%, 70%, and 90%. A total of 128 HA-coated implants were placed into the tibiae of 64 male Sprague-Dawley rats. Bone-implant interfaces were evaluated using histology and push-out strength testing at 3 and 9 weeks after implantation. RESULTS: The 70% crystalline coatings exhibited significantly greater interfacial strength (5 implants/time point/treatment) than the 30%, 50%, and 90% crystalline coatings at 3 and 9 weeks following implantation. The implants with coatings of 70% crystallinity also had the greatest bone contact length. In addition, the HA coatings produced with SPA demonstrated greater interfacial strength and bone contact length than plasma-sprayed HA coatings (except for the HA coating with 30% crystallinity). DISCUSSION: HA coatings of different crystallinities exhibited different dissolution and re-precipitation properties which may enhance early bone formation and bone bonding. CONCLUSIONS: This study suggested that coating crystallinity and coating methods can influence the bone-implant interface.  相似文献   

19.
The aim of the present study was to test the hypothesis that immobilization of bone morphogenic protein (BMP2) on the surface of titanium implants can enhance peri-implant bone formation. Ten adult female foxhounds received experimental titanium screw implants in the mandible 3 months after removal of all premolar teeth. Three types of implant surfaces were evaluated in each animal: (i) implants with machined titanium surface, (ii) implants coated with collagen I, (iii) implants coated with collagen I, chondroitin sulphate (CS) and BMP2. Peri-implant bone regeneration was assessed using histomorphometry after 1 and 3 months in five dogs each by measuring bone-implant contact (BIC) and the volume density of the newly formed peri-implant bone (BVD). After 1 month, there was no significant enhancement in BIC values but volume density of the newly formed peri-implant bone was significantly higher in the two groups of coated implants. No significant difference was found between collagen and BMP2 coating. After 3 months, BIC was significantly higher in both collagen and BMP2-coated implants compared with implants with machined surfaces. Peri-implant BVD was also significantly increased in coated implants in comparison with machined surfaces. It was concluded that collagen coating of dental screw implants can enhance BIC and peri-implant bone formation. Addition of BMP2 does not increase peri-implant bone formation in the present application.  相似文献   

20.
The aim of the present study was to analyse the effect of organic coating of titanium implants on periimplant bone formation and bone/implant contact. Three types of implants were used: (i) Ti6Al4V implants with polished surface (control 1) (ii) Ti6Al4V implants with collagen coating (control 2) (iii) Ti6Al4V implants with collagen coating and covalently bound RGD peptides. All implants had square cross-sections with an oblique diameter of 4.6 mm and were inserted press fit into trephine burr holes of 4.6 mm in the mandibles of 10 beagle dogs. The implants of five animals each were evaluated after a healing period of 1 month and 3 months, during which sequential fluorochrome labelling of bone formation was performed. Bone formation was evaluated by morphometric measurement of the newly formed bone around the implant and the percentage of implant bone contact. After 1 month there was only little bone/implant contact, varying between 2.6 and 6.7% in the cortical bone and 4.4 and 5.7% in the cancellous bone, with no significant differences between the three types of implants. After 3 months, implants with polished surfaces exhibited 26.5 and 31.2% contact in the cortical and cancellous bone, respectively, while collagen-coated implants had 19.5 and 28.4% bone contact in these areas. Implants with RGD coating showed the highest values with 42.1% and 49.7%, respectively. Differences between the surface types as such were not significant, but the increase in bone/implant contact from 1 to 3 months postoperatively was significant only in the group of RGD-coated implants (P = 0.008 and P = 0.000). The results of this pilot study thus provide only weak evidence that coating of titanium implants with RGD peptides in the present form and dosage may increase periimplant bone formation in the alveolar process. The results therefore require further verification in a modified experimental setting.  相似文献   

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