Osteogenic potential of cultured bone/ceramic construct: comparison with marrow mesenchymal cell/ceramic composite

Osteogenesis occurs in porous hydroxyapatite (HA) when porous HA blocks combined with marrow mesenchymal cells are grafted in vivo. In vitro bone formation occurs in HA pores when HA combined with marrow cells is cultured in osteogenic medium containing dexamethasone. This cultured bone/HA construct possesses higher osteogenic ability when it is grafted in vivo. In the present study, we compared the osteogenic potential of a cultured bone/HA construct with that of a marrow mesenchymal cell/HA composite. Marrow cells were obtained from the femoral bone shaft of 7-week-old, male Fischer 344 rats and were cultured in T-75 flasks. Cells were concentrated, then frozen and stored in liquid nitrogen for 6 months. The cryopreserved cells were then thawed and prepared for subculture in porous HA (5 x 5 x 5 mm, Interpore 500) and for implantation with porous HA. After 2 weeks of subculture, three cultured bone/HA constructs were separately implanted in the right side of the back of each syngeneic 7-week-old male Fischer rat, and three thawed cell/HA composites (without subculture) were separately implanted in the left side. These implants were harvested at 2 or 4 weeks postimplantation, and prepared for histological, biochemical, and genetic analysis. Alkaline phosphatase activity and osteocalcin content of cultured bone/HA constructs were much higher than those of the cell/HA composites at 2 and 4 weeks postimplantation. Histological examination and gene expression data agreed with these findings. The culture technique discussed herein should facilitate the development of biosynthetic bone implants with higher osteogenic capacity.     

Locally delivered rhBMP-2 enhances bone ingrowth and gap healing in a canine model.

The purpose of the present study was to determine if recombinant human bone morphogenetic protein-2 (rhBMP-2) enhances bone ingrowth into porous-coated implants and gap healing around the implants. In the presence of a 3-mm gap between the implant and host bone, porous-coated implants were placed bilaterally for four weeks in the proximal humeri of skeletally mature, adult male dogs. In three treatment groups, the test implant was treated with HA/TCP and rhBMP-2 in buffer at a dose of 100 microg/implant (n=5), 400 microg/implant (n=6), or 800 microg/implant (n=5) and placed in the left humerus. In these same animals, an internal control implant was treated only with HA/TCP and buffer and placed in the right humerus. These groups were compared with a previously reported external control group of seven animals in which no growth factor was delivered [J. Orthop. Res. 19 (2001) 85]. The BMP treated implants in the two lower dose groups had significantly more bone ingrowth than the external controls with the greatest effect in the 100 g/implant group (a 3.5-fold increase over the external control, p=0.008). All three dose groups had significantly more bone formation in the 3-mm gap surrounding the BMP treated implants than the external controls with the greatest effect in the 800 microg group (2.9-fold increase, p<0.001). Thus, application of rhBMP-2 to a porous-coated implant stimulated local bone ingrowth and gap healing. The enhancement of bone formation within the implant (bone ingrowth) was inversely related to dose.     

Demineralized bone matrix and hydroxyapatite/tri-calcium phosphate mixture for bone healing in rats

Purpose: Hydroxyapatite/tri-calcium phosphate (HA/TCP) mixture is an osteoconductive material used as a bone graft substitute, and demineralised bone matrix (DBM) is an osteoinductive material. A combination of DBM and HA/TCP mixture would probably create a composite with both osteoconductive and osteoinductive properties. The purpose of this study was to determine the effect of the combination of DBM and HA/TCP mixture on healing of rat radius segmental defects. Methods: Twenty-four adult male Wistar rats were used. Bilateral radial defects were created in each animal. Radial defects were implanted with DBM, HA/TCP mixture and a combination of both substances. Control defects were left unfilled. Ten weeks after implantation, the animals were sacrificed, and the radii were evaluated by radiograhic and histopathological studies. Results: The use of DBM alone demonstrated improved healing on radiographic and histological studies compared to other groups and the control group. There were no differences between the other two groups and the control group. Conclusion: The DBM group showed the best healing response. Combined use of DBM and HA/TCP mixture did not improve bone healing, and the osteoinductive properties of DBM were inhibited by HA/TCP mixture.     

Bone tissue engineering using novel interconnected porous hydroxyapatite ceramics combined with marrow mesenchymal cells: quantitative and three-dimensional image analysis

We developed fully opened interconnected porous calcium hydroxyapatite ceramics having two different pore sizes. One has pores with an average size of 150 microm in diameter, an average 40-microm interconnecting pore diameter, and 75% porosity (HA150). The other has pores with an average size of 300 microm in diameter, an average 60-100-microm interconnecting pore diameter, and 75% porosity (HA300). Because of its smaller pore diameter, HA150 has greater mechanical strength than that of HA300. These ceramics were combined with rat marrow mesenchymal cells and cultured for 2 weeks in the presence of dexamethasone. The cultured ceramics were then implanted into subcutaneous sites in syngeneic rats and harvested 2-8 weeks after implantation. All the implants showed bone formation inside the pore areas as evidenced by decalcified histological sections and microcomputed tomography images, which enabled three-dimensional analysis of the newly formed bone and calculation of the bone volume in the implants. The bone volume increased over time. At 8 weeks after implantation, extensive bone volume was detected not only in the surface pore areas but also in the center pore areas of the implants. A high degree of alkaline phosphatase activity with a peak at 2 weeks and a high level of osteocalcin with a gradual increase over time were detected in the implants. The levels of these biochemical parameters were higher in HA150 than in HA300. The results indicate that a combination of HA150 and mesenchymal cells could be used as an excellent bone graft substitute because of its mechanical properties and capability of inducing bone formation.     

In vivo osteogenic durability of cultured bone in porous ceramics: a novel method for autogenous bone graft substitution

BACKGROUND: Bone marrow cells differentiate into bone-forming osteoblasts when cultured in medium supplemented with 15% fetal bovine serum, ascorbic acid, beta-glycerophosphate, and dexamethasone. METHODS: To investigate in vivo osteoblastic activity and bone matrix formation by cultured bone marrow cells, Fischer rat marrow cells were cultured for 2 weeks in porous hydroxyapatite (HA) and then subcutaneously implanted into 7-week-old male syngeneic rats. The implants were harvested after 8 and 52 weeks for biochemical and histological analyses. RESULTS: At both times, formation of lamellar bone accompanied by regeneration of marrow were seen in many of the HA pores. When a fluorochrome (calcein) was administered at 50 weeks after implantation, it was detected in the pores of implants harvested at 52 weeks. Osteoclastic resorption followed by new bone formation was seen in some pores at 52 weeks, indicating that bone remodeling was continuing. The alkaline phosphatase activity of implants harvested at 52 weeks was comparable to that at 8 weeks, whereas the osteocalcin content of the implants harvested at 52 weeks was about twice that at 8 weeks. CONCLUSION: These results demonstrated that there was persistent in vivo osteogenic and hematopoietic activity in the prefabricated bone/HA constructs, and indicated that normal bone tissue was regenerated after grafting of the constructs, which were brittle before implantation. Tissue engineering using HA and cultured marrow cells culture may provide an alternative method of bone transplantation for patients with skeletal disorders, although further in vivo and in vitro experiments are needed.     

Effects of hydroxyapatite tricalcium phosphate coating and intracancellous placement on bone ingrowth in titanium fibermetal implants

The purpose of this study was to compare the host—bone response to hydroxyapatite/tricalcium phosphate (HA/TCP)-coated and noncoated titanium fibermetal implants placed in a load-sharing cancellous bone environment of the distal femurs of rabbits. The influence of implantation site was also investigated by comparing these intracancellous implants with intramedullary implants evaluated in a previous study. Three parameters were measured: percentage implant perimeter surface length in contact with new bone, percentage internal fibermetal surface length in contact with ingrown bone, and percentage of available pore space filled with bone. The HA/TCP coating significantly accelerated and increased bone ongrowth, new bone formation on the perimeter and internal surface of the implants. This effect was evident as early as 2 weeks after implantation. In contrast, there was no difference between HA/TCP-coated and noncoated implants in the bone ingrowth parameter, percentage of available pore space filled with bone, or pull-out strength. Scanning electron microscopy in the backscatter mode demonstrated that new bone formed directly onto the HA/TCP-coated fibers and did not usually form directly on noncoated fibers. Analysis of fluorochrome labeling revealed that bone formation in weeks 1 through 4 was primarily woven and there-after lamellar. Compared with intramedullary placement, intracancellous placement significantly accelerated the apposition of bone to the perimeter and internal surface of HA/TCP-coated implants and both accelerated and increased bone ingrowth as a percentage of available pore volume. These data show that the host response to titanium fibermetal implants is influenced both by HA/TCP coating and by the implantation site.     

Effect of Hydroxyapatite porous characteristics on healing outcomes in rabbit posterolateral spinal fusion model

Hydroxyapatite (HA) has been commonly used as a bone graft substitute in various kinds of clinical fields. To improve the healing capability of HA, many studies have been performed to reveal its optimal structural characteristics for better healing outcomes. In spinal reconstruction surgery, non-interconnected porous HAs have already been applied as a bone graft extender in order to avoid autogenous bone harvesting. However, there have been few experimental studies regarding the effects of the structural characteristics of HA in posterolateral lumbar intertransverse process spine fusion (PLF). The aims of this study were to investigate the effect of HA porous characteristics on healing outcomes in a rabbit PLF model in order to elucidate appropriate structural characteristics of HA as a bone graft extender. Thirty-six adult female Japanese White rabbits underwent bilateral intertransverse process fusion at the level of L5–6 without internal fixation. We prepared three types of HA with different porosities: HA with 15% porosity (HA15%), HA with 50% porosity (HA50%), and HA with 85% porosity (HA85%), all of which were clinically available materials. The HA15% and HA50% had few interconnecting pores, whereas the HA85%, which was a recently developed material, had abundant interconnecting pores. All rabbits were randomly divided into the following four groups according to the grafted materials: (1) HA15% + autogenous bone, (2) HA50% + autogenous bone, (3) HA85% + autogenous bone, (4) pure autogenous bone graft. The animals were euthanized at 5 weeks after surgery, and post-mortem analyses including biomechanical testing, radiographical and histological evaluations were performed. There was no statistically significant difference in either fusion rate and/or bending stiffness among the three HA groups. However, in histological and radiological analyses, both bone ingrowth rate and direct bone bonding rate in the HA85% group were significantly higher than those in the HA15% and HA50% groups, despite the similar value of bone volume rate in fusion mass among the three HA groups. In the HA85% group, bone ingrowth was achieved throughout the implanted HAs via interconnecting pores and there was excellent unification between the HA granules and the newly mineralized bone. On the other hand, in the non-interconnected porous HA groups, only a little bone ingrowth could be seen at the peripheral pores of the implanted HA, and its surface was mostly covered with fibrous tissue or empty space. The current study demonstrated that the HA porous characteristics had an effect on the histological outcomes in a rabbit PLF model. We would like to conclude that the interconnected high porous structure seems to be promising for the environment of PLF in the point of producing fusion mass with higher cellular viability. This is because the HA85% is superior in terms of integration with the newly formed bone in fusion mass compared to the non-interconnected porous HAs. However, the porous modifications of HA have little influence on fusion rate and mechanical strength because primary stabilization of the fusion segment is mainly achieved by bridging bone between the adjacent transverse processes outside the implanted materials, rather than the degree of integration between the newly formed bone and the HA granules in PLF.     

Bone ingrowth into two porous ceramics with different pore sizes: an experimental study

Many properties of porous calcium phosphate ceramics have been described, but how pore size influences bony integration of various porous ceramics remains unclear. This study was performed to quantify the bony ingrowth and biodegradability of two porous calcium phosphate ceramics with four different pore size ranges (45-80 microm, 80-140 microm, 140-200 microm, and 200-250 microm). Hydroxyapatite (HA) and beta-tricalcium phosphate (TCP) cylinders were implanted into the femoral condyles of rabbits and were left in situ for up to 12 months. The percentage of bone ingrowth and the depth of ingrowth within the pores were determined. Biodegradability of the implants was also evaluated. Bone ingrowth occurred at a higher rate into the TCP than into the HA ceramics with the same pore size ranges. The amount of newly formed bone was statistically smaller (p < 0.05) into ceramics with 45-80 microm pore size than with larger pore size, whatever the implantation time for HA and until four months for TCP. No statistical difference was noted between the three highest pore size ranges. No implant degradation was noted up to four months. Our results suggest that a pore size above 80 microm improves bony ingrowth in both HA and TCP ceramics. Bone formation was higher in the TCP than in the HA implants.     

Locally delivered rhTGF-beta2 enhances bone ingrowth and bone regeneration at local and remote sites of skeletal injury.

The purposes of the present study were to determine if recombinant human transforming growth factor-beta-2 (rhTGF-beta2) enhances bone ingrowth into porous-coated implants and bone regeneration in gaps between the implant and surrounding host bone. The implants were placed bilaterally for four weeks in the proximal humeri of skeletally mature, adult male dogs in the presence of a 3-mm gap. In three treatment groups of animals, the test implant was treated with hydroxyapatite/tricalcium phosphate (HA/TCP) and rhTGF-beta2 in buffer at a dose per implant of 1.2 microg (n = 6), 12 microg (n = 7), or 120 microg (n = 7) and placed in the left humerus. In these same animals, an internal control implant treated only with HA/TCP and buffer was placed in the right humerus. In a non-TGF-beta treated external control group of animals (n = 7), one implant was treated with HA/TCP while the contralateral implant was not treated with the ceramic. In vitro analyses showed that approximately 15%, of the applied dose was released within 120 h with most of the release occurring in the first 24 h. The TGF-beta treated implants had significantly more bone ingrowth than the controls with the greatest effect in the 12 microg/implant group (a 2.2-fold increase over the paired internal control (P = 0.004) and a 4-fold increase over the external control (P < 0.001)). The TGF-beta treated implants had significantly more bone formation in the gap than the controls with the greatest effect in the 12 and 120 microg groups (1.8-fold increases over the paired internal controls (P = 0.003 and P = 0.012, respectively) and 2.8-fold increases over the external controls (P < 0.001 and P = 0.001, respectively)). Compared to the external controls, the internal control implants tended to have more bone ingrowth (1.9-fold increase, P = 0.066) and had significantly more bone formation in the gap (1.7-fold increase. P = 0.008). Thus, application of rhTGF-beta2 to a porous-coated implant-stimulated local bone ingrowth and gap healing in a weakly dose-dependent manner and stimulated bone regeneration in the 3-mm gap surrounding the contralateral control implant, a site remote from the local treatment with the growth factor.     

三种钙磷陶瓷材料复合重组人骨形成蛋白-2体内成骨的研究

目的 探讨和比较3种钙磷陶瓷材料HA、TCP、HA／TCP复合重组入骨形成蛋白。2(rhBMP—2)体内异位成骨效果，为临床应用提供依据。方法 取35只3月龄Wistar大鼠，将复合rhBMP—2的3种钙磷陶瓷材料(1：1)植于鼠背部肌袋内，未复合rhBMP-2的上述3种单纯陶瓷材料为对照组。术后2、4和8周取材，测定植入物碱性磷酸酶(ALP)活性，通过HE染色和计算机图像分析进行组织学和组织计量学观察，比较新生骨组织的形成。结果 术后2、4周复合植入物ALP活性测定从高到低依次为HA、HA／TCP、TCP，但在相同rhBMP—2剂量下，其差异无统计学意义(P＞0．05)，与相对应单纯支架材料比较有统计学意义(P＜0．05)；组织学和组织计量学检测结果显示各复合材料组均有新骨形成，成骨量随时间推移而增加，2周时以HA/rhBMP—2成骨量较多，但3组间差异无统计学意义(P＞0．05)；8周时新骨形成以双相陶瓷HA／TCP最佳，相关参数和图像分析有统计学意义(P＜0．05)，成骨量8周较2、4周多，有统计学意义(P＜0．01)；3种单纯支架材料各观察期均无骨样组织形成。结论 双相陶瓷材料HA／TCP是携带rhBMP—2的钙磷陶瓷良好支架材料。     

Electromagnetic effects on bone formation at implants in the medullary canal in rabbits

This experiment was aimed at illuminating the relationship between electromagnetic and mechanical stimuli of bone formation when present simultaneously. Movable and stationary intramedullary wire implants were studied in rabbits treated with a pulsing electromagnetic field (PEMF) 4 h/day for 3 weeks, and were compared with identical control animals without PEMF. Trabecular bone formed routinely at spontaneously movable implants, but not at stationary ones. On average, PEMF-treated movable implants in the femur induced 44% more bone than untreated movable implants. Also, in the PEMF-treated femora, a 22% enlargement of the area of the medullary canal was observed compared with no-field controls. In the tibia, these effects were weak or nonexistent. The PEMF used did not induce bone at stationary implants, suggesting that under these conditions it is not a primary trigger in vivo.     

Comparison of hydroxyapatite and hydroxyapatite tricalcium-phosphate coatings

This study compared the effects of hydroxyapatite (HA) coating and biphasic HA/tricalcium-phosphate (HA/TCP) coating on the osseointegration of grit-blasted titanium-alloy implants. Each coated implant was compared with uncoated grit-blasted implants as well. The implants were press-fit into the medullary canal of rabbit femora, and their osseointegration was evaluated 3 to 24 weeks after surgery. The coated implants had significantly (P<.05) greater new bone ongrowth than the uncoated implants (HA, 56.1 +/- 3.1%; HA/TCP, 53.8 +/- 2.6%; uncoated, 32.2 +/- 1.4% of the implant perimeter, 12 weeks). Unmineralized tissue (cartilage and osteoid) was seen on the uncoated implants but never on the coated implants. The coated implants had significantly (P<.05) greater interfacial shear strength than the uncoated implants (HA, 4.1 +/- 0.4 MPa; HA/TCP, 4.8 +/- 0.5 MPa; uncoated, 2.6 +/- 0.2 MPa, 12 weeks). There was no difference between HA and HA/TCP coating in regard to new bone growth or interfacial shear strength. These data show a comparable enhancement effect of HA and HA/TCP coatings on the osseointegration of titanium-alloy implants.     

新型生物活性陶瓷复合人工骨成骨效应的实验研究

目的　探讨新型生物活性陶瓷复合材料成骨效应 ,为人工骨替代材料临床应用提供依据。　方法　小鼠 96只 ,随机分为 4组 ,每组 2 4只。采用具有诱导活性的骨形成蛋白 (bone morphogenetic protein,BMP)分别与羟基磷灰石 (hydroxyapatite,HA)、磷酸三钙 (tricalcium phosphate,TCP)、胶原复合羟基磷灰石 (collagen HA,CHA)及氟化羟基磷灰石 (fluoridated HA,FHA)复合 ,将 4种复合材料 (HA/ BMP,TCP/ BMP,CHA/ BMP及 FHA/ BMP)分别植入 4组小鼠左侧股部肌肉内为实验侧 ,右侧分别植入 HA、TCP、CHA及脱钙牙基质 (decalcified dentin matrix,DDM)作为对照 ,在 1、3、5及 7周取材作大体观察、组织形态学、扫描电镜观察及生化测定。　结果　各组实验侧及第 4组对照侧植入后 1周软骨形成 ,第 1～ 3组对照侧为纤维结缔组织 ;3周时各组实验侧均有较多的成熟骨组织 ,组织碱性磷酸酶 (alkalinephosphatase,AL P)染色均为阳性。各组对照侧材料被结缔组织包囊 ,AL P染色阴性 ,未见骨组织形成。各组实验侧材料AL P活性及磷 (phosphrus,P)检测水平明显高于相应的对照侧材料 ,实验侧与对照侧比较具有统计学意义 (P<0 .0 5 ) ,而 TCP/ BMP复合材料明显高于另 3种复合材料 ,有统计学意义 (P<0 .0 5 )。5、7周各实验侧及对照     

Comparative study of porous hydroxyapatite and tricalcium phosphate as bone substitute

This study was performed to quantitate the bone ingrowth and biodegradability of porous calcium phosphate. Two kinds of coralline hydroxyapatite (CHA), along with sintered tricalcium phosphate (TCP), were evaluated. All implants had totally interconnecting pores. These pores measured 260-600 mu for CHA-Goniopora (CHAG), 190-230 microns for CHA-Porites (CHAP), and 100-300 microns for TCP. Cylindrical implants (3 mm in diameter by 8 mm in length) were implanted into the diaphyses of rabbit tibias for 3 to 24 weeks. The quantity of regenerated bone, remaining implant, nonmineralized space, and the middle two quarters of the cortical area were measured by a computerized operator-assisted analysis for bone histomorphometry. At 3 weeks after implantation, 17.3% of CHAG, 11.3% of CHAP, and 7.0% of TCP were infiltrated by regenerated bone. At 24 weeks, 56.1% of CHAG, 52.7% of CHAP, and 44.7% of TCP were occupied by lamellar-type bone. Implant degradation was noted to be 46.4% for TCP and 27.5% for CHAP. In contrast, CHAG did not show appreciable degradation until 24 weeks.     

Zoledronic acid causes enhancement of bone growth into porous implants

The effect of zoledronic acid on bone ingrowth was examined in an animal model in which porous tantalum implants were placed bilaterally within the ulnae of seven dogs. Zoledronic acid in saline was administered via a single post-operative intravenous injection at a dose of 0.1 mg/kg. The ulnae were harvested six weeks after surgery. Undecalcified transverse histological sections of the implant-bone interfaces were imaged with backscattered scanning electron microscopy and the percentage of available pore space that was filled with new bone was calculated. The mean extent of bone ingrowth was 6.6% for the control implants and 12.2% for the zoledronic acid-treated implants, an absolute difference of 5.6% (95% confidence interval, 1.2 to 10.1) and a relative difference of 85% which was statistically significant. Individual islands of new bone formation within the implant pores were similar in number in both groups but were 69% larger in the zoledronic acid-treated group. The bisphosphonate zoledronic acid should be further investigated for use in accelerating or enhancing the biological fixation of implants to bone.     

Enhanced Bone Bonding of the Hydroxyapatite/β‐Tricalcium Phosphate Composite by Electrical Polarization in Rabbit Long Bone

A review of the osteogenic cell activity and new bone growth in the regions bordering negatively charged surfaces of polarized Hydroxyapatite/β‐tricalcium phosphate (HA/TCP) composites implanted in the long bone in rabbits was conducted. Polarized and non‐polarized HA/TCP specimens were implanted into the right and left femoral condyle, respectively (each n = 10). After 3 and 6 weeks, five rabbits were sacrificed in each group, and histological analysis was administered. Large cuboidal‐shaped osteoblastic cells were predominantly observed lining the newly formed bone on the negatively charged surface (N‐surface) in the polarized HA/TCP implants. The TRAP‐positive multinucleated cells were observed extensively in the newly formed bone on the N‐surfaces compared with the 0‐surface and adhered directly to the HA/TCP composite. The bone area (B.Ar) value, newly formed bone area contacting the implant, and contact length (C.Le) value, percentage length of newly formed bone directly attaching to the implant, on both the 0‐ and N‐surface increased significantly with time in each group. Both the B.Ar and C.Le value on the N‐surface were significantly greater than those on the 0‐surface after 3 and 6 weeks. The number of TRAP‐positive cells/total length value on the N‐surface was significantly greater than that on the 0‐surface after 3 and 6 weeks postoperatively. It is hypothesized that electrical charge acquired by electrical polarization treatment may modify the biochemical and biophysical processes of the osteogenic cells, resulting in enhanced new bone formation and direct bonding between the recipient bone and implants.     

新型双相陶瓷材料复合成骨细胞构建人工骨及其体内成骨的观察

目的 观察骨髓来源的成骨细胞与新型双相陶瓷材料复合培养后细胞生长状况及其体内异位成骨能力。方法 将来源于兔骨髓的成骨细胞与新法制备的HA/TCP共培养2周,通过相差显微镜、扫描电镜观察体外细胞生长情况;然后将复合物自体异位植入体内,6周后观察体内成骨情况。结果 扫描电镜显示材料表面和孔隙内均有成骨细胞生长,有良好增殖活动性和稳定细胞表型,材料中心区未见细胞生长;植入体内6周后取材见内植物有新骨形成、多位于内植物表面,可见成骨细胞、骨细胞、髓腔样结构、板层样骨基质等正常骨组织结构。结论 新型双相陶瓷支架可用作组织工程骨的细胞外基质材料,骨髓来源的成骨细胞可用作骨组织工程的种子细胞。     

Bioactive glass enhances bone ingrowth into the porous titanium coating on orthopaedic implants

Purpose

The aim of the study was to verify the ability of nanoparticulate bioactive glass (BAG) to infiltrate into the porous titanium (Ti) layer on Ti-based implants to promote osseointegration.

Methods

The porous titanium layer on Ti-based implants was impregnated with nanoparticulate BAG. The implants without or with BAG were implanted bilaterally in tibial holes of ten New Zealand white rabbits. The rabbits were sacrificed after ten weeks for examinations. Beside histological examination, EDXS analysis of polished cross-sections of explanted implants was also performed with the aim to quantitatively evaluate the bone-to-pore contact and bone-in-pore ratio.

Results

After ten weeks, EDXS analyses of cross-sections of the explanted implants confirmed that bioactive glass was fully resorbed and that the pores throughout the thickness of the porous titanium layer were to a large extent filled with a new bone. In the absence of bioactive glass, only the outer part of the porous layer was filled with bone. The implants without BAG in the porous Ti-layer exhibited similar bone-to-pore contact, while significant improvement of bone ingrowth into the pores was observed for the implants with BAG (38%), as opposed to those without it (22%).

Conclusion

This study confirmed that the nanoparticulate bioactive glass within the porous titanium surface layer on implants promotes osseointegration and stimulates the formation of bone within the pores.     

Fresh bone marrow introduction into porous scaffolds using a simple low‐pressure loading method for effective osteogenesis in a rabbit model

Recent advances in tissue engineering techniques have allowed porous biomaterials to be combined with osteogenic cells for effective bone regeneration. We developed a simple low‐pressure cell‐loading method using only syringes and stopcocks, and examined the effect of this method on osteogenesis when applied to the combination of highly porous β‐tricalcium phosphate (β‐TCP) and fresh autologous bone marrow. Both block and granule β‐TCP scaffolds were used to prepare implants in three different ways: without bone marrow as a control, with bone marrow that was allowed to penetrate spontaneously under atmospheric pressure (AP group), and with bone marrow that was seeded under low pressure (ULP group). These implants were transplanted into rabbit intramuscular sites, and the samples were examined biologically and histologically. The penetration efficiency of the block implants after marrow introduction was significantly higher in the ULP group than in the AP group. In the transplanted block samples, alkaline phosphatase activity was significantly higher in the ULP group at 2 weeks after implantation, and significantly more newly formed bone was observed in the ULP group at both 5 and 10 weeks compared with the AP group. Similar results were observed even in the experiment using β‐TCP granules, which are smaller than the blocks and frequently used clinically. Because of its convenience and safety, this low‐pressure method might be a novel, effective treatment to promote osteogenesis with bone marrow in clinical bone reconstruction surgeries. © 2008 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 27:1–7, 2009     

A long-term study on defect filling and bone ingrowth using a canine fiber metal total hip model.

When performing primary and revision total hip arthroplasty (THA), bone defects are often encountered. At present, grafting osseous defects with autogeneic bone is a common means of treatment. In this study, defects in bone were created in the femora and acetabula of dogs being treated with cementless THA with a fiber metal implant (Group A) or a hydroxyapatite tricalcium phosphate (HA/TCP) sprayed implant (Group B). The following methods of defect filling were compared: (1) leaving defects unfilled, (2) filling with autogeneic bone graft, (3) filling with a 50:50 mixture of autograft and a biphasic ceramic composed of HA/TCP, and (4) filling with a collagen-HA/TCP-bone marrow mixture. Analysis of defect healing and the extent of ingrowth into the overlying fiber metal, at defect sites and sites distant from defects, was made at six, 12, and 24 weeks postimplantation. Defect healing was enhanced at six and 12 weeks in all grafted groups when compared with ungrafted controls. Bone ingrowth into the porous fiber metal overlying the defects was not significantly affected by grafting the defects, compared with the ungrafted defects. The extent of bone ingrowth into the fiber metal acetabular implant at sites away from the defects increased during the entire study. In contrast, the extent of bone ingrowth on the femoral side was maximal at 12 weeks. The HA/TCP coating enhanced ingrowth into the acetabular component at 12 weeks, compared with the uncoated prosthesis, but did not enhance ingrowth on the femoral side. The data from this study demonstrate that defect healing is enhanced with graft materials. However, this does not necessarily result in increased ingrowth into porous surfaces overlying osseous defects. General bone ingrowth and ingrowth at defect sites at 12 weeks postimplantation can be enhanced on the acetabular side with the use of HA/TCP-sprayed implants. However, no positive effect is seen with the use of an HA/TCP-sprayed femoral implant.