Bone formation at rhBMP-2-coated titanium implants in the rat ectopic model

BACKGROUND: The objective of this study was to evaluate local bone formation at titanium porous oxide (TPO) implant surfaces adsorbed with recombinant human bone morphogenetic protein-2 (rhBMP-2). METHODS: In vitro studies were used to estimate the kinetics of I125-labeled rhBMP-2 released from TPO surfaces with narrow (N) or open (O) pores. Machined/turned titanium (MT) surfaces served as control. The rat ectopic model was used to assess local bone formation. Briefly, TPO-N, TPO-O, and MT disc implants adsorbed with 5, 10, or 20 microg rhBMP-2, respectively, were implanted subcutaneously into the ventral thoracic region in 5-week-old male Long Evans rats. The animals were euthanized at day 14 postsurgery when implants with surrounding tissues were removed, radiographed, and gross observations recorded. The specimens were processed for histologic evaluation using conventional cut-and-grind techniques. TPO implants without rhBMP-2 included in a preliminary evaluation revealed no evidence of bone formation, tissue encapsulation, or vascularity, thus such controls were not further used. RESULTS: TPO and MT implant surfaces adsorbed with 5 microg rhBMP-2 retained 2.3-5.4% rhBMP-2 following immersion and rinse in buffer, and 1.1-2.2% rhBMP-2 following repeated immersions and rinses over 27 days. TPO implants retained the most rhBMP-2 and MT implants retained the least. Explants revealed increased hard tissue formation, tissue encapsulation, and vascularity at TPO compared with MT implants. Radiographic observations were consistent with the explant observations. The histologic analysis showed greater amounts of bone formation, osteoblastic cells, osteoid, marrow, tissue encapsulation, vascularity, and bone voids for implants adsorbed with 10 and 20 microg rhBMP-2, and for TPO implants at the 5-microg rhBMP-2 dose. The histometric analysis revealed significantly greater bone formation at TPO-O than at MT implants at the 5-microg rhBMP-2 dose. All surfaces showed significant bone formation at the 10- and 20-microg dose. CONCLUSIONS: rhBMP-2 adsorbed onto TPO implant surfaces executes an osteoinductive effect including bone contacting the implant surface. This effect is surface- and dose-dependent; the TPO-O surface yielding the most bone at the low discriminating rhBMP-2 dose.     

Bone formation at titanium porous oxide (TiUnite) oral implants in type IV bone

BACKGROUND: Several oral implant design advances have been suggested to overcome poor bone quality, an impediment for successful implant treatment. A novel titanium porous oxide (TPO) surface has been shown to offer favorable results in several settings. The objective of this study was to evaluate the local bone formation and osseointegration at TPO-modified implants in type IV bone. METHOD: Three TPO surface-modified implants (TiUnite) were installed into the edentulated posterior maxilla in each of 8 Cynomolgus monkeys. The animals were injected with fluorescent bone labels at 2, 3, 4 and 16 weeks post-surgery and were euthanized at week 16 when block biopsies were collected for histologic analysis. RESULTS: The predominant observation of the TPO implant surface was a thin layer of new bone covering most of the implant threads. Mean (+/-SE) bone-implant contact for the whole study group was 74.1 +/- 4.8%. There was a significant variability in bone-implant contact between animals (P = 0.0003) and between sites of the same animal (P < 0.0001). The variance in bone-implant contact was 30% larger among sites of the same animal than between different animals (187.5 vs. 144.8, respectively). There was a small but significant difference in bone density immediately outside, compared to within the threaded area of the implants (37.1 +/- 3.2% vs. 32.1 +/- 3.2%, P < 0.0001). Bone density outside the implant threads was significantly correlated (beta = 0.682, P < 0.0001) with the bone density within the threaded area. Bone density within the threaded area was significantly correlated (beta = 0.493, P = 0.0002) with bone-implant contact, whereas bone density outside the implant threads did not have a significant effect (beta = 0.232, P = 0.1). CONCLUSIONS: The results suggest that the TPO surface possesses a considerable osteoconductive potential promoting a high level of implant osseointegration in type IV bone in the posterior maxilla.     

A mechanical evaluation of TiO2-gritblasted and Ca-P magnetron sputter coated implants placed into the trabecular bone of the goat: Part 1

The influence of Ca-P magnetron sputter coated implants on the mechanical anchorage was evaluated in a goat model. Therefore, uncoated and coated screw designed commercially pure titanium TiO2-blasted implants were inserted into the trabecular bone of the femoral condyles of 12 goats. The thicknesses of the coatings were 0.1 micron (CaP-0.1), 1.0 micron (CaP-1) and 4.0 microns (CaP-4). In addition, uncoated TiO2-blasted implants (Ti) were used as control. Evaluation of the interface strength and appearance, using torque test and scanning electron microscopy, was done at implantation periods of 6 and 12 weeks. Although, especially at 6 weeks of implantation, the Ca-P coated implants showed higher failure torque values than the TiO2 blasted implants, the observed differences for type of implant and evaluation period were not significant (P > 0.1). SEM evaluation showed that all implants with failure values of more than 100 N conducted bone growth into their screw threads. In addition, we observed that the fracture plane for the CaP-4 implants was situated at the coating-implant interface or inside the coating. For the CaP-1 and CaP-0.1 the fracture line could not be determined definitely. For the Ti implants, the fracture torque testing resulted in failure at the bone-implant interface. Therefore, we conclude that all implants resulted in a good bonding strength with the surrounding bone. The sputtered Ca-P coatings seemed to improve the initial fixation of the TiO2 blasted implants.     

Bone formation at recombinant human bone morphogenetic protein-2-coated titanium implants in the posterior mandible (Type II bone) in dogs

Background: Conventional oral/maxillofacial implants reach osseointegration over several months during which the titanium fixtures interact with alveolar bone. The objective of this study was to determine if adsorbing recombinant human bone morphogenetic protein‐2 (rhBMP‐2) onto a titanium porous oxide (TPO) implant surface might enhance or accelerate local bone formation and support osseointegration in a large animal oral/maxillofacial orthotopic model. Material and Methods: Endosseous implants with a TPO surface were installed into the edentulated posterior mandible in eight adult Hound Labrador mongrel dogs. The implant surface had been adsorbed with rhBMP‐2 at 0.2 or 4.0 mg/ml. TPO implants without rhBMP‐2 served as control. Treatments were randomized between jaw quadrants. Mucosal flaps were advanced and sutured leaving the implants submerged. Clinical and radiographic evaluations were made immediately post‐surgery, at day 10 (suture removal), and week 4 and 8 post‐surgery. The animals received fluorescent bone markers at week 3, 4, and at week 8 post‐surgery, when they were euthanized for histologic analysis. Results: TPO implants coated with rhBMP‐2 exhibited dose‐dependent bone remodelling including immediate resorption and formation of implant adjacent bone, and early establishment of clinically relevant osseointegration. The resulting bone–implant contact, although clinically respectable, appeared significantly lower for rhBMP‐2‐coated implants compared with the control [rhBMP‐2 (0.2 mg/ml) 43.3±10.8%versus 71.7±7.8%, p<0.02; rhBMP‐2 (4.0 mg/ml) 35.4±10.6%versus 68.2±11.0%, p<0.03]. Conclusions: rhBMP‐2 adsorbed onto TPO implant surfaces initiates dose‐dependent peri‐implant bone re‐modelling resulting in the formation of normal, physiologic bone and clinically relevant osseointegration within 8 weeks.     

Bone formation at recombinant human bone morphogenetic protein-2-coated titanium implants in the posterior maxilla (Type IV bone) in non-human primates

Background: Studies using ectopic rodent and orthotopic canine models (Type II bone) have shown that titanium porous oxide (TPO) surface implants adsorbed with recombinant human bone morphogenetic protein‐2 (rhBMP‐2) induce local bone formation including osseointegration. The objective of this study was to evaluate local bone formation and osseointegration at such implants placed into Type IV bone. Material and Methods: rhBMP‐2‐coated implants were installed into the edentulated posterior maxilla in eight young adult Cynomolgus monkeys: four animals each received three TPO implants adsorbed with rhBMP‐2 (2.0 mg/ml) and four animals each received three TPO implants adsorbed with rhBMP‐2 (0.2 mg/ml). Contra‐lateral jaw quadrants received three TPO implants without rhBMP‐2 (control). Treatments were alternated between left and right jaw quadrants. Mucosal flaps were advanced and sutured to submerge the implants. The animals received fluorescent bone markers at weeks 2, 3, 4, and at week 16 when they were euthanized for histologic analysis. Results: Clinical healing was uneventful. Extensive local bone formation was observed in animals receiving implants adsorbed with rhBMP‐2 (2.0 mg/ml). The newly formed bone exhibited a specific pinpoint bone–implant contact pattern regardless of rhBMP‐2 concentration resulting in significant osseointegration; rhBMP‐2 (2.0 mg/ml): 43% and rhBMP‐2 (0.2 mg/ml): 37%. Control implants exhibited a thin layer of bone covering a relatively larger portion of the implant threads. Thus, TPO control implants bone exhibited significantly greater bone–implant contact (～75%; p<0.05). There were no statistically significant differences between rhBMP‐2‐coated and control implants relative to any other parameter including peri‐implant and intra‐thread bone density. Conclusion: rhBMP‐2‐coated TPO implants enhanced/accelerated local bone formation in Type IV bone in a dose‐dependent fashion in non‐human primates resulting in significant osseointegration. rhBMP‐2‐induced de novo bone formation did not reach the level of osseointegration observed in native resident bone within the 16‐week interval.     

Ceramic hydroxyapatite coating on titanium implants drives selective bone marrow stromal cell adhesion

The aim of this study was to determine the cell characteristics that regulate implant osseointegration. The heterogeneity of bone marrow stromal cells obtained from 11 donors was assessed by measuring the expression of a large panel of adhesion molecules. Large differences in expression of adhesion molecules were detected depending on the culture conditions used. Cells cultured in fetal bovine serum induced the expression of different adhesion molecules from cells cultured in human serum. Donor-to-donor variation was determined by measuring the expression of adhesion molecules for stromal cells obtained from different donors that were processed identically. Fat adherent cells but also loose bone marrow cells showed large differences in expression of some but not all adhesion molecules. The flow cytometric data demonstrated large heterogeneity in expression of adhesion molecules, and this heterogeneity was influenced by culture conditions and varied from donor to donor. This demonstrates that the implant encounters different cell types, which could lead to different levels of integration. Surprisingly, in vitro only a subfraction of bone marrow stromal cells attached to titanium coated with ceramic hydroxyapatite. Adaptation of all cell types present in heterogeneous bone marrow to a coated surface is apparently not possible. Differential binding was not caused by aberrant staining of the stromal cells as the results were confirmed with bone marrow cells obtained from transgenic GFP mice. These results demonstrate that hydroxyapatite ceramics are selective in cell recruitment from the bone marrow, explaining the differences found in vivo for these coatings compared with titanium.     

Bone healing around titanium and titanium nitride-coated dental implants with three surfaces: an experimental study in rats

Background: Titanium nitride (TiN) has been used in many fields as a coating of surgical instruments, with the purpose of creating materials more resistant to wear and corrosion and also reducing adhesion. Purpose: The aim of this study was to evaluate the biocompatibility of TiN‐coated dental implants. Materials and Methods: Forty‐five rats were used in this study. One hundred eighty 2 mm 2 mm implants (P.H.I., San Vittore Olona, Milano, Italy) were used. The implants were divided into the following three groups: Group 1 (n= 60):30 machined and 30 machined coated with TiN Group 2(n= 60):30 sandblasted and 30 sandblasted coated with TiN Group 3 (n= 60):30 titanium plasma sprayed, 30 titanium plasma sprayed and coated with TiN Four implants were placed in each rat, two implants coated with TiN on the right tibia and two uncoated implants on the left. The animals were killed after 5, 10, 20, 30, or 60 days. Another 18 implants were used for surface roughness analysis. Results: The present study showed that the healing around the TiN‐coated implants was similar to that observed around the uncoated surfaces. Conclusions: TiN coating demonstrated a good biocompatibility, did not have untoward effects on the periimplant bone formation, and did not change the surface roughness values.     

Evaluation of stability of titanium and hydroxyapatite-coated osseointegrated dental implants: a pilot study

An endosseous implant is described as osseointegrated when it is immobile in function. Objective measures of stability testing have been described. The Periotest is a commercially available device that is used for this purpose. This study was designed to measure stability of endosseous implants placed in the mandible. Implants were placed in the mandibular canine or first premolar area to support an overdenture prosthesis. Stability was evaluated through the use of a Periotest device at the time of implant placement and following one year of functional loading. Implant designs were either a screw-shaped titanium alloy or a hydroxyapatite-coated cylinder. A total of 54 implants were placed, 37 were titanium screw-shaped implants, while the remaining 17 were hydroxyapatite cylinders. Initial measurements of stability showed no difference due to implant type. Following one year of functional loading, titanium screw-shaped implants were more stable than hydroxyapatite implants (P < 0.05). The difference in implant rigidity following a period of functional loading may be an indication of a difference in osseointegration between the two implants used in this study.     

钛种植体表面新型抗菌涂层预防种植术后早期感染的动物实验研究

目的：通过在犬下颌骨内植入种植体,观察新型抗菌涂层预防种植术后早期感染的实际效果。方法：选择2岁龄牙周健康的雄性Beagle犬4只,拔除双侧下颌第三前磨牙,并在近远中根拔牙窝内预备种植窝行即刻种植术。随机选取2只植入载有新型抗菌涂层的种植体作为实验组,另外2只植入仅微弧氧化处理的种植体作为对照组,并用丝线在所有种植体的颈部结扎,术后给予高糖软食,诱导种植体周围发生早期感染。结扎2个月后对种植体周进行临床指标检查和X线摄片,通过扫描电镜观察种植体／骨组织界面。结果：2个月后,实验组种植体周的临床检查指标明显低于对照组护〈0．05）。X线检查可见：实验组种植体周围无明显的牙槽骨吸收和透射影像,对照组种植体周围可见明显的骨吸收和裂隙状透射影像。扫描电镜观察可见：实验组种植体／骨界面结合较好,未见明显间隙,而对照组种植体与骨组织之间存在明显的间隙。结论：在钛种植体表面构建的新型抗菌涂层具有显著的预防种植术后早期感染的效果。     

大鼠正畸牙移动过程中牙周组织内转化生长因子β1的变化

目的 了解大鼠正畸牙移动过程中转化生长因子β１（ｔｒａｎｓｆｏｒｍｉｎｇ ｇｒｏｗｔｈ ｆａｃｔｏｒβ１,ＴＧＦ－β１）在牙周组织内的变化,探讨ＴＧＦ－β１在正畸牙移动过程中的作用。方法 对大习正畸牙移动不同阶段的牙及牙周组织联合切片,进行免疫组织化学研究。结果 正畸牙移动过程中,牙周膜内ＴＧＦ－β１表达增加。强力第５～１０天,张力区ＴＧＦ－β１表达增加的幅度明显大于压力区。结论 ＴＧＦ－β１在正     

Effects of Simvastatin on bone healing around titanium implants in osteoporotic rats

Objectives: Osteoporosis is known to impair the process of implant osseointegration. The recent discovery that statins (HMG‐CoA reductase inhibitors) act as bone anabolic agents suggests that statins can be used as potential agents in the treatment of osteoporosis. Therefore, we hypothesized that statins will promote osteogenesis around titanium implants in subjects with osteoporosis. Material and methods: Fifty‐four female Sprague Dawley rats, aged 3 months old, were randomly divided into three groups: Sham‐operated group (SHAM; n=18), ovariectomized group (OVX; n=18), and ovariectomized with Simvastatin treatment group (OVX+SIM; n=18). Fifty‐six days after being ovariectomized (OVX), screw‐shaped titanium implants were inserted into the tibiae. Simvastatin was administered orally at 5 mg/kg each day after the placement of the implant in the OVX+SIM group. The animals were sacrificed at either 28 or 84 days after implantation and the undecalcified tissue sections were obtained. Bone‐to‐implant contact (BIC) and bone area (BA) within the limits of implant threads were measured around the cortical (zone A) and cancellous (zone B) bone regions. Furthermore, bone density (BD) of zone B in a 500 μm wide zone lateral to the implants was also measured. Results: There were no significant differences in BIC and BA measurements in zone A in any of the three groups at either 28 or 84 days after implantation (P>0.05). By contrast, in zone B, significant differences in the measurement of BIC, BA, and BD were observed at 28 and 84 days between all three groups. Bone healing decreased with lower BIC, BA, and BD around implant in OVX group compared with other two groups, and Simvastatin reversed the negative effect of OVX on bone healing around implants with the improvement of BIC, BA, and BD in zone B. Conclusion: Osteoporosis can significantly influence bone healing in the cancellous bone around titanium implants and Simvastatin was shown to significantly improve the osseointegration of pure titanium implants in osteoporotic rats.     

Bone-tissue formation and integration of titanium implants: an evaluation with newly developed enzyme and immunohistochemical techniques

Background: Examination of the tissue surrounding retrieved implants involve routine investigations on cut and ground sections. Undecalcified sections with implants in situ are histologically stained followed by qualitative and quantitative observations of the tissue response to the implants by light microscopy. Purpose: A novel technique that allows for the accurate definition and quantification of enzymes involved in bone formation (alkaline phosphatase) and resorption (acid phosphatase) in the tissue is presented. Materials and Methods: Commercially pure titanium and titanium alloy (Ti6A14V) implants were retrieved after 6 and 12 weeks of healing in rabbit bone. In addition, 4-week specimens from commercially pure titanium bone harvest chambers placed in rabbit bone were used. Undecalcified cut and ground sections were produced and evaluated with enzyme and immunohistochemical staining techniques. Results: The titanium implants retrieved after 6 weeks of insertion in rabbit bone revealed a higher activity of both alkaline phosphatase and acid phosphatase activity compared to the implants followed for 12 months. The former samples revealed ongoing bone-tissue remodeling in the interface, whereas the latter ones showed steady-state bone conditions. Applying the new technique allowed for investigation of various bone proteins present in the tissue that had formed inside titanium canals of harvest chambers at various times of follow-up. Conclusions: The combination of routine histologic stainings with enzyme and immunohistochemical technique of cut and ground specimens is a valuable tool in the investigations of retrieved implants from humans and animals. This novel technique now may be used to describe the state of bone regeneration in the interface zone associated with implant research.     

多孔海绵状纯钛种植材料对成骨细胞生长的影响

目的：探讨具有连续多孔立体结构纯钛种植材料对成骨细胞生长的影响。方法：在体外对3种不同孔隙直径的多孔钛材料进行成骨细胞复合培养,在相差倒置显微镜下观察细胞在材料表面及孔隙中的生长情况,利用MTT法评价不同孔隙直径对细胞生长的影响。结果：成骨细胞在试件表面生长良好,成功长入连续孔隙之中,并以多突触的方式生长,在钛珠之间形成细胞桥;细胞增殖度检测显示多孔立体结构对成骨细胞的生长有一定的促进作用,200μTI孔径的作用最明显。结论：多孔立体结构能促进成骨细胞在纯钛种植材料表面和孔隙中的生长。     

放疗对狗下颌纯钛种植体与骨接触作用的影响

目的:观察放疗对狗下颌纯钛种植体与骨接触作用的影响。方法:8只成年雄性杂种狗,拔除双侧下颌第3、4前磨牙和第1磨牙,形成无牙区。拔牙后3个月,用电子直线加速器照射一侧下颌无牙区,单一剂量15Gy。另一侧不接受照射,作为对照。放疗后3个月,狗双侧下颌无牙区各植入纯钛种植体4枚,其中两枚种植体无任何辅助治疗措施。种植术后1.5个月和3个月分别处死2只动物取材。结果:放疗侧种植体的种植体-骨接触率与对照侧相比,有明显下降,在统计学上有显著差异(P=0.0001<0.01);放疗后种植体周围骨小梁体积百分比具体数值下降,统计学上差异不明显(P=0.071>0.05)。结论:放疗会影响狗下颌纯钛种植体与骨的接触作用。     

Prognostic factors for alveolar regeneration: bone formation at teeth and titanium implants

OBJECTIVES: There is a limited understanding of the effect of defect characteristics on alveolar bone healing. The objectives of this study were to assess the effect of alveolar bone width and space provision on bone regeneration at teeth and titanium implants, and to test the hypothesis that the regenerative potentials at teeth and implants are not significantly different. METHODS: Critical size, 5-6-mm, supra-alveolar, periodontal defects were surgically created in 10 young adult dogs. Similarly, critical size, 5-mm, supra-alveolar, peri-implant defects were created in four dogs. A space-providing expanded polytetrafluoroethylene device was implanted for guided tissue regeneration/guided bone regeneration. The animals were euthanized at 8 weeks postsurgery. Histometric analysis assessed alveolar bone regeneration (height) relative to space provision by the device and the width of the alveolar crest at the base of the defect. Statistical analysis used the linear mixed models. RESULTS: A significant correlation was found between bone width and wound area (r=0.55892, p<0.0001). Generally, bone width and wound area had statistically significant effects on the extent of bone regeneration (p<0.0005 and p<0.0001, respectively). Bone regeneration was linearly correlated with the bone width at periodontal (p<0.001) and implant (p=0.04) sites, and with the wound area at periodontal (p<0.0001) and implant (p=0.03) sites. The relationships of bone regeneration with these two variables were not significantly different between teeth and implants (bone width: p=0.83; wound area: p=0.09). When adjusted for wound area, bone regeneration was significantly greater at periodontal than at implant sites (p=0.047). CONCLUSIONS: The horizontal dimension of the alveolar bone influences space provision. Space provision and horizontal dimension of the alveolar bone appear to be important determinants of bone regeneration at teeth and implants. The extent of alveolar bone formation at implant sites is limited compared with that at periodontal sites.     

钛种植体表面改性方法

钛种植体具有美观舒适、不损伤邻牙、临床效果显著等优点,广泛应用于口腔种植修复领域;但应用中存在骨结合失败、种植体周围骨吸收、种植体周围炎等问题,使其应用受到一定的限制。对钛种植体进行表面改性,在其表面制备不同的理化涂层和生物活性涂层,可以提高种植体的种植成功率,满足临床应用需求。本文从物理、化学、生物三方面对各种钛种植体表面改性方法的特点进行总结,为牙种植材料的研究和临床应用提供参考。