In vivo effects of coating loaded and unloaded Ti implants with collagen, chondroitin sulfate, and hydroxyapatite in the sheep tibia.

The in vivo effects of coating titanium implants with organic extracellular matrix molecules were examined in the sheep tibia. Titanium screws (5.0 mm) were coated with type I collagen (Ti/Coll) or type I collagen and chondroitin sulfate (Ti/Coll/CS) by biomimetic fibrillogenesis. Uncoated screws (Ti) and screws coated with hydroxyapatite (Ti/HA) served as control. Six adult female sheep received one screw of each type to stabilize a midshaft tibial fracture with external fixation. Four cylindrical implants of 4-mm outer diameter and 3.3-mm inner diameter with the same coatings were inserted into the tibial head. No pin track infections were seen at the time of implant retrieval 6 weeks after implantation. Extraction torque was greater for Ti/HA (1181 Nmm) and Ti/Coll/CS (1088 Nmm) compared to Ti/Coll (900 Nmm) and Ti (904 Nmm) [N.S.]. Newly formed bone was noted around all coated screws within the medullary cavity. Macrophage and osteoclast activity was significantly reduced around Ti/Coll/CS in both types of implants compared to uncoated controls (p < 0.05). Osteoblast activity was significantly increased around loaded Ti/Coll and Ti/Coll/CS screws compared to uncoated Ti screws (p < 0.05). Microtomographic evaluation (SRmicroCT) revealed no significant differences in new bone formation around the unloaded tibial head implants.Coating of external fixation devices with of type I collagen and chondroitin sulfate appears to have similar effects with respect to stability and bone healing as HA but with less osteoclast activity. These findings were more pronounced under loaded than unloaded conditions in the sheeptibia.     

Effect of modification of hydroxyapatite/collagen composites with sodium citrate, phosphoserine, phosphoserine/RGD-peptide and calcium carbonate on bone remodelling

This study describes the early interface reaction of cancellous bone to a nanocrystalline hydroxyapatite cement containing type I collagen (HA/Coll) and its modifications with sodium citrate (CI), calcium carbonate (CA), phosphoserine (P) and phosphoserine plus RGD-peptide (RGD). Cylindrical implants of HA/Coll and its modifications were inserted into the tibia of Wistar rats. We analysed 6 specimens per group at days 2, 4, 7, 14 and 28. CI, P and RGD modifications showed improved material properties (finer microstructure and higher compressive strength) compared to CA and HA/Coll implants. The powder X-ray diffraction (XRD) showed that the addition of P and CI led to an increase of alpha-TCP peaks while the diffraction patterns of the non-modified cement (HA/Coll) were quite similar with that of natural bone. All of the implants healed without adverse reactions. A significantly higher number of TRAP-positive osteoclasts were observed around CI, RGD and P on day 7 compared to CA and HA/Coll. Around CI, P and RGD a significantly delayed increase of ED1-positive mononuclear cells was detected. The amount of direct bone contact after 28 days was significantly higher around CI, P and RGD compared to CA and HA/Coll implants. The addition of CI, P and RGD appears to enhance bone remodelling at the early stages of bone healing, leading to increased bone formation around HA/Coll composite cements.     

Development of a hydroxyapatite/collagen nanocomposite as a medical device

The effect of cross-linking of a hydroxyapatite/collagen (HA/Col) nanocomposite, in which HA nanocrystals and collagen fibers are aligned like natural bone by a self-organization mechanism between HA and collagen in vitro, on mechanical properties was examined. The influence of degree of cross-linking, as well as rhBMP-2 preadsorption to the composite on the substitution pattern and rate with bone, was examined. In Experiment 1, anterior fusion was carried out at the C3-C4 vertebrae on 10 dogs and they were implanted as follows: without cross-linking and without adsorbed rhBMP-2 (three dogs), with cross-linking and without adsorbed rhBMP-2 (three dogs), without cross-linking and with adsorbed rhBMP-2 (two dogs), and with cross-linking and adsorbed rhBMP-2 (two dogs). Implants were removed from each dog for histology determinations after 12, 16, and 24 weeks in the non-rhBMP-treated groups, and after 16 and 24 weeks in the rhBMP-treated groups. In Experiment 2, the HA/Col composites with cross-linking and both with and without rhBMP-2 pretreatment were implanted into a bone defect of 20 mm made in the central part of tibiae in dogs (N = 3 in each group). As a control, bone defects of 20 mm remained without implantation (N = 3). The dogs were allowed to walk using an Ilizarov extra skeletal fixator. The implants were removed after 12, 16, and 24 weeks from one dog in each group. The cross-linking of the HA/Col composite was effective in controlling both the mechanical strength and bioresorbability. A "self-organization process" on the HA/Col implant surface resulted in the formation of bone remodeling units in and around the implant. Radiographic and histological findings suggest that a combined treatment of cross-linking of the HA/Col composite with preadsorption of rhBMP-2 molecules may be a very suitable replacement of existing ceramic systems in the anterior fusion of the cervical spine, as well as inlay grafting of bone defects in weight-bearing sites.     

Reconstruction of Extensive Long-Bone Defects in Sheep Using Porous Hydroxyapatite Sponges

The capacity of hydroxyapatite (HA) implants to support large defect repair in weight-bearing long bones of large size animals was investigated. Diaphyseal resections 3.5 cm of the tibia were performed in five adult sheep. They were substituted with HA macroporous ceramic cylinders anatomically shaped, and an external fixator was assembled. The sheep were sacrificed at 20, 40, 60, 120, and 270 days after surgery, respectively. Histology and micro X-ray study of resected implants and adjacent tissues showed proper integration of ceramic with newly formed periosteal bone as early as 20 days after surgery. In one sheep, the external fixator was removed 5 months after surgery. The animal gained the ability to walk with no functional impairment until it was sacrificed 4 months later. At this time, extensive integration of ceramic with bone was detected radiographically and confirmed by a morphological study of the resected sample. Our data indicate that large defects in a weight-bearing long bone can be repaired to the extent necessary for full functional recovery in large animals. These data set the stage for further intervention on material properties as well as for preliminary attempts to use ceramic prostheses for reconstruction of large bone defects in humans. Received: 15 July 1997 / Accepted: 9 July 1998     

Effects of the systemic administration of alendronate on bone formation in a porous hydroxyapatite/collagen composite and resorption by osteoclasts in a bone defect model in rabbits

Several bisphosphonates are now available for the treatment of osteoporosis. Porous hydroxyapatite/collagen (HA/Col) composite is an osteoconductive bone substitute which is resorbed by osteoclasts. The effects of the bisphosphonate alendronate on the formation of bone in porous HA/Col and its resorption by osteoclasts were evaluated using a rabbit model. Porous HA/Col cylinders measuring 6 mm in diameter and 8 mm in length, with a pore size of 100 μm to 500 μm and 95% porosity, were inserted into a defect produced in the lateral femoral condyles of 72 rabbits. The rabbits were divided into four groups based on the protocol of alendronate administration: the control group did not receive any alendronate, the pre group had alendronate treatment for three weeks prior to the implantation of the HA/Col, the post group had alendronate treatment following implantation until euthanasia, and the pre+post group had continuous alendronate treatment from three weeks prior to surgery until euthanasia. All rabbits were injected intravenously with either saline or alendronate (7.5 μg/kg) once a week. Each group had 18 rabbits, six in each group being killed at three, six and 12 weeks post-operatively. Alendronate administration suppressed the resorption of the implants. Additionally, the mineral densities of newly formed bone in the alendronate-treated groups were lower than those in the control group at 12 weeks post-operatively. Interestingly, the number of osteoclasts attached to the implant correlated with the extent of bone formation at three weeks. In conclusion, the systemic administration of alendronate in our rabbit model at a dose-for-weight equivalent to the clinical dose used in the treatment of osteoporosis in Japan affected the mineral density and remodelling of bone tissue in implanted porous HA/Col composites.     

胶原支架增强自固化磷酸钙骨水泥的力学及成骨性能研究

[目的]研究胶原支架（CS）对磷酸钙骨水泥（CPC）的力学及其在体内成骨的影响。[方法]试验分CPC／CS及CPC两组，三点弯曲试验测试材料的强度和弹性模量；组织学观察材料植入兔股骨22及54周的成骨状况。[结果]CPC／CS比CPC的弯曲强度、韧性强度分别提高了64．2％、3933．3％，弹性模量降低了45．7％；组织学显示22周CPC／CS内的胶原支架已完全被新骨替代，CPC只在边缘有少量成骨及材料降解而内部无成骨；54周CPC／CS已大部分降解孔化，孔内充满大量新骨及髓样组织，而CPC边缘区的成骨及材料降解虽比22周时明显，但其内部仍未见成骨。[结论]胶原支架既可改善CPC的力学性能，又能促进新骨长入CPC／CS复合材料内部，因此，CPC／胶原支架是较好的骨缺损修复材料。     

Hydroxyapatite coating modifies implant membrane formation. Controlled micromotion studied in dogs.

We studied the influence of controlled micromovements between bone and porous titanium alloy implants with and without hydroxyapatite coating. A dynamically loaded unstable device producing approximately 150-microns axial translation of knee implants during each gait cycle was developed. Stable implants served as controls. Matched stable and unstable implants with either porous titanium (Ti) or hydroxyapatite (HA) coating surrounded by a gap of 0.75 mm were inserted into the weight-bearing regions of the medial femoral condyles in 14 mature dogs. Histologic analysis after 4 weeks showed a fibrous membrane surrounding both types of implants subjected to micromovements, whereas various amounts of bone ingrowth was obtained in the stable implants. The membrane around unstable HA implants was thinner than that around unstable Ti implants. Islands of fibrocartilaginous tissue characterized the membrane around unstable HA implants, whereas fibrous connective tissue surrounded unstable Ti implants. The collagen concentration of the fibrous membranes was higher around unstable HA implants compared with Ti implants. Instability reduced the shear strength of the implants. However, the shear strength of unstable HA implants exceeded that of the Ti implants, both unstable and stable. The greatest shear strength was obtained by stable HA implants, i.e., tenfold greater than that of stable Ti implants. The gap-healing capacity around stable HA implants increased toward the HA surface, and was greater than that around Ti implants. Our study demonstrates that micromovements between bone and implant inhibit bone ingrowth and lead to the development of a fibrous membrane. The superior fixation of unstable HA implants compared with unstable Ti implants may be ascribed to the presence of fibrocartilage, a higher collagen concentration, and radiating orientation of collagen fibers in the membrane. The strongest mechanical anchorage and the greatest amount of bone ingrowth was obtained by stable implants coated with hydroxyapatite.     

Novel application of HA‐TCP biomaterials in distraction osteogenesis shortened the lengthening time and promoted bone consolidation

This study tested the hypothesis that use of biomaterials in distraction osteogenesis (DO) would reduce the treatment time and enhance bone formation quality. A 1.0‐cm tibial shaft was removed in the left tibia of 36 rabbits. Rabbits were randomly divided into three groups: group A, the defect gap was reduced with the tibia shortened for 1.0‐cm; group B, the defect gap was filled with 1.0‐cm restorable porous hydroxyapatite and Tri‐calcium phosphates cylindrical block (HA/TCP block, diameter is 0.5‐cm); group C, The 1.0‐cm defect gap was reduced 0.5 cm and the remaining 0.5‐cm defect gap was filled with the 0.5‐cm HA/TCP block. The tibia was then fixed with unilateral lengthener; for groups A and C; lengthening started 7 days after surgery at a rate of 1.0 mm/day, in two steps. Group A received lengthening for 10 days and group C for 5 days, there was no lengthening for group B. All animals were terminated at day 37 following surgery. The excised bone specimens were subject to microcomputed tomography (micro‐CT), mechanical testing, and histological examinations. Bone mineral density and content and tissue mineral density and content, as well as the mechanical properties of the regenerates were significantly higher in group C compared to groups A and B. Micro‐CT and histological examinations also confirmed that the regenerates in Group C had most advanced bone formation, consolidation, and remodeling compared to other groups. In conclusion, the combined use of biomaterials and DO technique can reduce the treatment time and enhance bone consolidation in bone defect management. © 2008 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 27: 477–482, 2009     

胶原/羟基磷灰石支架负载软骨细胞修复兔膝关节骨软骨缺损

目的 探讨胶原复合梯度羟基磷灰石(Col/HA)双相支架负载软骨细胞修复兔膝关节骨软骨缺损的可行性及疗效.方法 构建Col/HA双相支架,将软骨细胞种植于支架培养1周,再将软骨细胞-支架复合体移植修复兔膝关节股骨髁的骨软骨缺损,并对骨软骨缺损的修复进行检测.结果 光镜及扫描电镜观察显示软骨细胞在Col/HA支架中贴附良好,表型维持稳定,分泌胞外基质.大体观察和组织学检测显示,植入体内16周后实验组软骨层呈透明软骨样修复,软骨下骨缺损有新骨构建;对照组骨软骨缺损修复不良,组织学检测以纤维性组织或纤维软骨组织形成.Wakitani评分显示实验组修复组织优于对照组,差异有统计学意义(P＜0.05).结论 双相Col/HA复合支架可作为骨软骨组织工程支架,负载软骨细胞可修复兔膝关节骨软骨缺损,重建关节软骨的结构和功能.     

The effect of different mineral frames on ectopic bone formation in mouse hind leg muscles induced by native reindeer bone morphogenetic protein

Introduction Bone morphogenetic proteins (BMPs) require carrier material for slow release and framing material for osteoconduction.Materials and methods The effect of a frame on early bone formation induced by partially purified native reindeer BMP in composite implants containing 3 mg of BMP, type IV collagen and tricalcium phosphate (TCP/Col/BMP) or hydroxyapatite (HA/Col/BMP) or biphasic tricalcium phosphate-hydroxyapatite (TCP/HA/Col/BMP) or biocoral (NC/Col/BMP) was evaluated using a mouse hind leg muscle pouch model. Collagen with native reindeer BMP (Col/BMP) and corresponding implants without native reindeer BMP served as controls. Evaluation was done by incorporation of ⁴⁵Ca, radiographically and histologically 3 weeks after the implantation.Results None of the implants without native reindeer BMP were able to induce new bone visible on radiographs. The area of new bone formation in the Col/BMP (p=0.026) and TCP/HA/Col/BMP (p=0.012) groups was significantly greater than in the TCP/Col/BMP group. The optical density of the new bone area was significantly greater in the TCP/HA/Col/BMP group than in the TCP/Col/BMP (p=0.036) or Col/BMP (p=0.02) groups. ⁴⁵Ca incorporation was many times greater in all the groups containing native reindeer BMP than in the corresponding groups without BMP. In the Col/BMP (p=0.046) and TCP/HA/Col/BMP (p=0.046) groups, ⁴⁵Ca incorporation was significantly greater than in the TCP/Col/BMP group. No significant differences were found in any parameters between HA/Col/BMP and NC/Col/BMP groups and the other BMP-containing groups.Conclusions Hydroxyapatite, biocoral and biphasic tricalciumphosphate-hydroxyapatite are equally good as framing material for native reindeer BMP, while tricalciumphosphate is somewhat worse. Osteoinduction of native reindeer BMP works well with collagen alone.     

The influence of hydroxyapatite granules on the healing of a segmental defect filled with autologous bone marrow.

Hydroxyapatite(HA) ceramics are frequently used as a bone graft substitutes for the filling of bony defects. The addition of autologous bone marrow to HA ceramics does improve defect healing. There is conflicting evidence in the literature whether autologous bone marrow transplantation alone is as effective as the combination of HA ceramics and bone marrow combined. It was the purpose of this study to identify the role of additional HA ceramic granules on the healing of a sheep tibia segmental defect filled with autologous bone marrow. After permission of the local animal rights committee was obtained, a 3 cm segmental defect in the midshaft of 31 adult sheep was stabilized with an unreamed tibia nail. The animals were divided into 4 groups according to the mode of defect filling: HA plus autologous bone marrow (HA + MAR) (n = 8), autologous bone marrow (MAR) (n = 9), empty defect (DEF) (n = 6), cancellous bone graft (CAN) (n = 8). After three months follow up animals were sacrificed and analysed for the key parameters of union and maximum torque at failure. One nonunion was present in each of the HA + MAR, MAR, and CAN groups. Four of the six animals in the DEF group developed a nonunion. Maximum torque at failure was reported as percentage of the intact contralateral tibia: HA + MAR 39% +/- 24%, MAR 26% +/- 17%, DEF 22% +/- 13%, CAN 41% +/- 20%. The difference between the groups was statistically significant, but appeared to be relevant. We conclude from our data, that HA ceramics do improve healing of a segmental defect in the sheep tibia filled with autologous bone marrow. The results of this combination are comparable to cancellous autograft.     

Hydroxyapatite/Calcium Carbonate (HA/CC) vs. Plaster of Paris: A Histomorphometric and Radiographic Study in a Rabbit Tibial Defect Model

The search for an ideal bone substitute is ongoing. Multiple osteoconductive bone substitutes are available today. Plaster of Paris (POP) (calcium sulfate) has been used for more than 100 years for treatment of skeletal defects. This implant is compared to a new material, hydroxyapatite/calcium carbonate (HA/CC), in a rabbit tibia model. HA/CC is made from partial conversion of coralline calcium carbonate to hydroxyapatite and has an outer hydroxyapatite layer and an inner calcium carbonate core, a combination that leads to faster resorption than that of pure hydroxyapatite. This study compares the histomorphometric and radiographic properties of POP and HA/CC in a rabbit tibial defect. Both implants preferentially restore bone to the cortex relative to the canal. Plaster of Paris was fully resorbed by 6 weeks both radiographically and histometrically and HA/CC was substantially resorbed by 42 weeks. No significant difference was noted in volume fraction of bone between the two implants at 42 weeks postimplantation. Hydroxyapatite/calcium carbonate is a biocompatible bone graft substitute with a rate of resorption significantly slower than plaster of Paris.     

Osteopontin and bone repair in rabbit tibial defect

Human osteopontin (OPN) was produced by a recombinant technique. A circular defect was created in each tibia of 30 adult rabbits. The animals were divided in six equal groups. Four animals of each group were randomly chosen as experimental group in which OPN coated hydroxyapatite granules (OPN-HA) and non-coated hydroxyapatite (HA) granules were inserted alternatively in created defects. One animal of each group was used as control animal to observe the spontaneous healing process of the defects. The created defects in these animals did not receive any implants. The animals were sacrificed after 1, 2, 6, 12, 18 and 30 weeks. The histological sections were magnified and scanned digitally. In the first measurement, the total amount of bone formation in the entire defect was calculated. In a second measurement, new bone formation was quantified at the edges and at the centre of the defects. The total amount of bone formation in OPN treated defects was significantly higher, whereas no significant difference of bone formation was observed at the edges or at the centre of the defects. A careful interpretation of these results suggests that human OPN might stimulate bone formation in the rabbit tibia.     

羟基磷灰石晶体纤维增强磷酸钙骨水泥力学性能及生物相容性研究

目的将高长径比的羟基磷灰石(HA)晶体纤维添加入磷酸钙骨水泥(CPC)中,研究HA晶体纤维对CPC抗压力学性能增强的最佳配方及在体内的生物相容性。方法采用水热合成法制备高长径比HA晶体纤维,并验证其细胞毒安全性。将2.5%、5%及10%(wt%)HA晶体纤维添加入CPC中,分析并获取HA晶体纤维对CPC力学性能增强的最佳配方。使用最佳配方制备CPC+HA晶体纤维复合材料,植入大鼠胫骨近端骨缺损模型,4、8周组织学观察HA晶体纤维添加入CPC的成骨性能及生物相容性。对照组为单纯CPC实验组。结果成功制备高长径比HA晶体纤维,其细胞毒安全性为1级(RGR79%)。与对照组相比,2.5%及5%HA晶体纤维添加入CPC可以增强材料的抗压力学性能(P<0.05),其中5%HA晶体纤维对CPC抗压性能的增强效果最佳,10%HA晶体纤维略降低CPC的抗压性能,但无统计学差异(P>0.05)。大鼠体内研究结果显示:与对照组相比,5%HA晶体纤维添加入CPC复合材料植入后材料周围骨体积分数(BV/TV)无显著性差异(P>0.05),HA晶体纤维添加入CPC后材料周围成骨与材料接触好,生物相容性良好。结论水热合成法制备的HA晶体纤维作为添加剂,具有增强CPC抗压力学性能,减低CPC脆性,生物相容性良好的特点,可用作骨植入生物材料的添加剂。     

Bridging the gap: Bone marrow aspiration concentrate reduces autologous bone grafting in osseous defects

Although autologous bone grafting represents an effective tool to induce osteogenic regeneration in local bone defects or pseudarthroses, it is associated with significant donor site morbidity and limited by the amount available for grafting. We investigate the potency of bone marrow aspiration concentrate (BMAC) to augment bone grafting and support bone healing. The functional and radiographic outcome of 39 patients with volumetric bone deficiencies treated with BMAC are presented and evaluated in a prospective clinical trial. A collagen sponge (Col) served as scaffold in 12 patients and a bovine hydroxyapatite (HA) was applied in the other 27 individuals. The minimal follow‐up was 6 months. Clinical and radiographic findings were completed by in vitro data. All patients showed new bone formation in radiographs during follow‐up. However, two patients underwent revision surgery due to a lack in bone healing. In contrast to the Col group, the postoperative bone formation appeared earlier in the HA group (HA group: 6.8 weeks vs. Col group 13.6 weeks). Complete bone healing was achieved in the HA group after 17.3 weeks compared to 22.4 weeks in the Col group. The average concentration factor of BMAC was 5.2 (SD 1.3). Flow cytometry confirmed the mesenchymal nature of the cells. Cells from BMAC created earlier and larger colonies of forming units fibroblasts (CFU‐F) compared to cells from bone marrow aspirate. BMAC combined with HA can reduce the time needed for healing of bone defects when compared to BMAC in combination with collagen sponge. © 2010 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 29:173–180, 2011     

Repair of large osteochondral defects in rabbits using porous hydroxyapatite/collagen (HAp/Col) and fibroblast growth factor‐2 (FGF‐2)

Articular cartilage has a limited capacity for self‐renewal. This article reports the development of a porous hydroxyapatite/collagen (HAp/Col) scaffold as a bone void filler and a vehicle for drug administration. The scaffold consists of HAp nanocrystals and type I atelocollagen. The purpose of this study was to investigate the efficacy of porous HAp/Col impregnated with FGF‐2 to repair large osteochondral defects in a rabbit model. Ninety‐six cylindrical osteochondral defects 5 mm in diameter and 5 mm in depth were created in the femoral trochlear groove of the right knee. Animals were assigned to one of four treatment groups: porous HAp/Col impregnated with 50 µl of FGF‐2 at a concentration of 10 or 100 µg/ml (FGF10 or FGF100 group); porous HAp/Col with 50 µl of PBS (HAp/Col group); and no implantation (defect group). The defect areas were examined grossly and histologically. Subchondral bone regeneration was quantified 3, 6, 12, and 24 weeks after surgery. Abundant bone formation was observed in the HAp/Col implanted groups as compared to the defect group. The FGF10 group displayed not only the most abundant bone regeneration but also the most satisfactory cartilage regeneration, with cartilage presenting a hyaline‐like appearance. These findings suggest that porous HAp/Col with FGF‐2 augments the cartilage repair process. © 2009 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 28:677–686, 2010     

Recombinant human bone morphogenetic protein-2 potentiates the in vivo osteogenic ability of marrow/hydroxyapatite composites

A composite of marrow mesenchymal stem cells (MSCs) and porous hydroxyapatite (HA) has bone-forming capability. To promote the capability, we added recombinant human bone morphogenetic protein-2 (BMP) to the composite. The bone formation was assessed by rat subcutaneous implantation of 4 different kinds of implants, i.e., HA alone, BMP/HA composites, MSCs/HA composites, and the composites containing BMP (MSCs/BMP/HA). Both HA and the BMP/HA composites did not show bone formation at any time after implantation. The MSCs/HA composites showed moderate bone formation at 4 weeks and extensive bone formation at 8 weeks. The MSCs/BMP/HA composites showed obvious bone formation together with active osteoblasts at 2 weeks and more bone formation at 4 and 8 weeks. The MSCs/BMP/HA composites demonstrated high alkaline phosphatase and osteocalcin expression at both the protein and gene levels. These results indicate that the combination of MSCs, porous HA, and BMP synergistically enhances osteogenic potential and provides a rational basis for their clinical application in bone reconstruction surgery.     

Osteogenic protein 1 device stimulates bone healing to hydroxyapaptite-coated and titanium implants

This study evaluated the effects of osteogenic protein 1 (OP-1) placed in a gap around uncoated and hydroxyapatite (HA)-coated implants. Unloaded cylindrical titanium alloy implants were inserted in the femoral condyles of 16 skeletally mature dogs surrounded by a 3-mm gap. The gap around the implants was filled with 325 microg OP-1 in 130 mg bovine collagen type I as carrier (OP-1 device) or collagen carrier alone or left empty. All groups were tested with both HA-coated and uncoated implants, and the animals were sacrificed after 6 weeks. Implant fixation was determined by push-out test. Bone ongrowth and bone formation were evaluated by quantitative histomorphometry. OP-1 device enhanced mechanical fixation of uncoated and HA-coated implants, resulting in a higher shear strength than implants with collagen matrix and control implants. Bone ingrowth and bone formation in the gap were also stimulated 3-fold for OP-1 groups when compared with empty controls, but no difference was found between OP-1 groups and collagen matrix groups. This study suggests that the OP-1 device placed in a gap around uncoated and HA-coated implants is capable of enhancing mechanical fixation and periimplant bone formation. The collagen carrier alone also enhanced bone ongrowth and fixation significantly.     

羟基磷灰石材料结合牵拉成骨技术修复骨缺损的实验研究

 目的 探讨羟基磷灰石（hydroxyapatite/tri-calcium phosphase,HA/TCP）材料结合牵拉成骨技术是否能减少治疗骨缺损所需的时间并促进骨的矿化过程。方法 将36只健康雄性新西兰大白兔（20~24周,体重2.2~2.8 kg）随机分为3组,每组12只。于左胫腓关节下方做1 cm的胫骨缺损。A组：将骨缺损两端靠拢,外固定架加压固定。B组：将1 cm长的HA/TCP材料填入1 cm的骨缺损中,外固定架加压固定。C组：将骨缺损两端拉近至间距0.5 cm,将0.5 cm长的HA/TCP材料填入剩余的0.5 cm骨缺损中,外固定架加压固定。除B组外,A、C组均于术后7 d开始延长胫骨,A组延长10 d,C组延长5 d。在术后即刻、12、17、27、37 d用“C”型臂X线机观察各组新生骨的矿化过程。所有动物于术后37 d处死,取新生骨样本测量骨矿物质含量（BMC）和组织矿物质含量（TMC）,行胫骨扭转实验及组织病理学分析。结果 C组BMC为（454.44±89.98） mg和TMC为（454.40±89.97） mg显著高于A、B两组。C组新生骨的最大扭矩、新生骨的成熟度均明显高于A、B两组,骨的矿化和重塑也较快。C组在观察期总共37 d内骨愈合良好,但A、B两组在37 d内并未达到骨愈合。结论 在兔胫骨缺损延长模型中,与单一疗法相比,联合应用羟基磷灰石材料和牵拉成骨技术可减少治疗时间并促进骨的矿化。     

Coating of titanium implants with type-I collagen.

PURPOSE: Type-I collagen, the major structural protein in bone, has beneficial properties regarding bone regeneration. Little is known about the potential effects of collagen coating on orthopedic implants. METHODS: 3 to 6 microg/cm2 of lyophilized type-I collagen was absorbed on titanium rods. Six coated and uncoated pins of 0.9 mm diameter were inserted into the tibia of adult male Wistar rats for 1, 2, 4, 7, 14, and 28 days. Specimens were embedded in methacrylate-based Technovit 9100N resin. From one portion cutting and grinding sections were obtained. The implant was removed from the other half that was depolymerized, sectioned, and mounted for immunohistochemistry. RESULTS: At day 4, the interface around the collagen-coated implants displayed a granulation tissue with higher numbers of cathepsin D-positive mononucleated cells compared to the uncoated implants (p<0.05). Active osteoblasts, reactive for osteopontin, were increased around the collagen-coated pins at day 4 and 7 (p<0.01). After 28 days of implantation, direct bone contact averaged 74.9% around the collagen-coated implants and 62.1% around uncoated implants (NS). The amount of newly formed bone averaged 76.3% around the collagen-coated pins and 67.8% around the uncoated pins (NS). The histomorphometric findings were confirmed by SRmicroCT in two specimens. CONCLUSIONS: The earlier observation of mononuclear phagocytozing cells and the earlier and higher expression of bone-specific matrix proteins suggest an increased early bone remodeling around titanium pins through collagen coating. A tendency towards increased bone formation was observed around the coated implants.