Subcutaneous implantation of hydroxylapatite/collagen in induced diabetic and non-diabetic rats

To evaluate tissue reaction to hydroxylapatite (HA) and HA/collagen mixtures in rats with uncontrolled induced diabetes, 48 males were divided: 24 with induced diabetes (ID) from Streptozotocin (70 mg/kg) and 24 non-diabetic (ND) controls. Three subcutaneous sites in each chest were randomly implanted with non-porous HA, or non-porous HA and bovine collagen, or non-porous HA and purified fibrillar collagen. Subgroups of 6 ID and 6 ND rats were killed at 4, 6, 12, and 24 weeks post-implantation. Histologic specimens showed that all materials elicited greater inflammatory response in ID than in ND at all intervals. Each specimen had HA particles encapsulated by host fibrous tissue. Compared to ND, ID specimens had markedly reduced ingrowth and maturity of collagen at each time interval. There was no osteogenesis, but there was dystrophic mineralization within the implant sites in both ID and ND. Mixed HA/collagen exceeded HA alone in maintaining implant contour. In soft tissue, no materials were osteoinductive. Adding collagen did not increase or decrease inflammatory reaction nor improve density and maturity of tissue synthesized around implants.     

Tissue response to hydroxylapatite in induced diabetic and nondiabetic rats: histologic evaluation

This study evaluated the tissue response to the subcutaneous implantation of nonporous hydroxylapatite (HA) in 24 induced-diabetic (ID) and 24 nondiabetic (ND) rats. One cubic centimeter of HA was implanted subcutaneously in each rat's chest. Subgroups of 6 rats from the ID and ND groups were killed at 3, 6, 12, and 24 weeks postimplantation. The implants were removed with the surrounding soft tissues and processed for histologic evaluation. This revealed that soft tissue inflammation was mild at each time interval. There was a decreased response at 6 months in ND rats and a persistent inflammatory reaction in ID rats. Collagen maturity and fibroplasia increased within ND rats, whereas the ID rats showed a marked delay in collagen maturity and density. No osteogenesis was observed in any specimen. Dystrophic calcification was observed at the HA-tissue interface in 37% of ND and 59% of ID specimens. It was concluded that HA elicited a greater inflammatory response in ID than in ND rats when implated subcutaneously.     

Loaded hydroxylapatite-coated implants and uncoated titanium-threaded implants in distracted dog alveolar ridges

AIMS: The aim of this study was to determine the response of alveolar bone after it was augmented vertically with distraction osteogenesis, implanted with hydroxylapatite (HA)-coated implants and noncoated titanium-threaded implants, and subsequently loaded for 1 year. METHODS: Eight dogs each had 4 implants placed horizontally into an edentulous mandibular quadrant. After integration, a distraction osteogenesis device was fabricated in the laboratory. An osteotomy was made to allow the crest of the alveolar ridge to be distracted vertically. After 10 mm of vertical distraction, the distraction devices were stabilized with light cured resin. After bone fill of the distraction gap was radiographically confirmed in all dogs at 10 weeks, 2 implants were placed into the ridges. Four dogs had threaded titanium implants placed, and 4 dogs had threaded HA-coated implants placed, with 1 implant in the distracted bone and 1 implant in adjacent nondistracted bone, for both groups. After 4 months for implant integration, bridges were fabricated and secured to the implants with screws. Crestal bone levels were evaluated by radiographs through 1 year of function. Animals were killed after 1 year of loading for histologic evaluation. RESULTS: The vertical ridge augmentation averaged 8.8 +/- 1.0 mm after 10 weeks of healing after distraction. Analysis of variance indicated a significantly greater change from baseline for HA-coated implants and for distracted bone sites. Histologic examination showed that bone had formed between the distracted segments creating an augmented ridge. The average thickness of the labial cortex in the distraction gap was significantly thinner than the lingual cortex in distracted bone or the lingual and labial nondistracted cortical bone. The presence of a dental implant did not significantly affect cortical bone thickness. Serial sections showed that implants remained integrated and functional without soft tissue inflammation. CONCLUSION: Dental implants placed into alveolar ridges augmented with the technique of distraction osteogenesis were functional for the length of this study.     

Bone augmentation osteogenesis using hydroxyapatite and beta-tricalcium phosphate blocks.

PURPOSE: In this study, we investigated the differences in osteogenesis and resorption between hydroxyapatite (HA) and beta-tricalcium phosphate (beta-TCP) implanted on the parietal bone of rats. MATERIALS AND METHODS: HA and beta-TCP were used in blocks with macropores and micropores. They were implanted between the parietal bone and the cranial periosteum in rats. Osteogenesis around the implanted materials was investigated histopathologically and histomorphometrically at 1, 2, 4, 8, and 24 weeks after surgery. RESULTS: At 2 weeks, osteogenesis from the parietal bone was observed around both materials, and new bone had attached directly to the surfaces of both materials. New bone grew into the pores of the upper regions of both materials with time. The beta-TCP block had a characteristic basophilic reticular structure in which the dissolution of the materials was observed close to the new bone. The HA blocks were stable for 24 weeks, whereas parts of the beta-TCP blocks were fractured and resorbed at 24 weeks. Histomorphometrically, the volume of new bone around HA was larger than that around beta-TCP. There was no remarkable change in the amount of remaining HA, but that of beta-TCP was decreased. CONCLUSION: HA blocks in this model are suitable for onlay grafts because of its stability and osteogenesis, beta-TCP is not stable. Therefore, when beta-TCP blocks are used for onlay grafts, the mechanical stress on the recipient site should be taken into consideration because of resorption and fracture.     

种植体周骨缺损时羟基磷灰石颗粒植入的实验研究

目的为研究羟基磷灰石颗粒在即刻人工种植体的愈合过程中的作用。方法作者应用国产ＨＡ层钛种植体植入犬股骨中进行观察，术后１、２、３、４个月处死动物，通过Ｘ线检查及扫描电镜观察标本。结果即刻种植组中，用ＨＡ充填空隙后，其愈合情况在术后３个月与延期种植无明显差异，种植体和骨组织结合紧密；无ＨＡ充填的标本中与延期种植存在差异，４个月时颈部尚有部分骨组织缺损。结论即刻种植术中，在植入种植体的同时植入ＨＡ颗粒。有助于种植体的稳定，对种植体与周围骨组织早期愈合起到较大作用。     

The effect of therapeutic radiation on canine alveolar ridges augmented with hydroxylapatite.

The purpose of this investigation was to evaluate the effect of radiation on hydroxylapatite (HA) implanted subperiosteally for alveolar ridge augmentation in dogs. All bicuspids and molars were extracted from 16 dogs. After 6 weeks, nonporous HA granules were implanted subperiosteally on the alveolar ridge. Following 4 months of healing, 12 dogs (experimental group) underwent therapeutic radiation therapy (Co60, 4,000 rad [40 Gy]) to the head and neck region. Four dogs were not irradiated and served as controls. Four animals (three experimental and one control) were killed at 5,6,7, and 8 months after HA augmentation. Light microscopic evaluation showed that approximately 25% of HA granules were encased by bone while the others were surrounded by fibrous connective tissue. Dissolution of the HA was observed. Microparticles of HA were phagocytized as part of a granulomatous inflammatory reaction. This reaction decreased significantly as time elapsed after implantation. Osteoclastic activity was seen at the junction of HA and periosteum and as part of bone remodeling. Dissolution of the HA granules and the granulomatous inflammatory reaction were not significantly increased by therapeutic radiation. The radiation did not cause development of dehiscence or osteonecrosis.     

Bone augmentation by onlay implant using recombinant human BMP-2 and collagen on adult rat skull without periosteum

The purpose of this study was to determine whether bone augmentation could be obtained by the composite of recombinant human bone morphogenetic protein-2 (rhBMP-2) and bioabsorbable atelocollagen when the periosteum was resected, and to compare the efficacy of the rhBMP-2/collagen implant and the collagen alone implant. The onlay implant was inserted into the space between the elevated galea aponeurotica and the skull without the periosteum of 10-month-old rats. The rhBMP-2/collagen implant resulted in osteoblasts differentiation under the galea at 1 week and active bone formation without a prior formation of cartilage. At 4 weeks, the bony trabeculae were interconnected and connected directly with the compact bone of the skull. Histomorphometric analysis at 4 weeks demonstrated that the rhBMP-2/collagen implant showed 92.5% in the volume of bone tissue, whereas the collagen alone showed 0%. The implanted collagen was gradually replaced by bone tissue in the presence of rhBMP-2. Our present results indicate that rhBMP-2 stimulates undifferentiated mesenchymal cells in the galea overlying the implant to proliferate and differentiate directly into osteoblasts on the carrier collagen fibers. The collagen matrix was stably placed on the skull and suitable as a substitute for rhBMP-2. The rhBMP-2/collagen onlay implant might be clinically applicable for bone augmentation even under the condition without the periosteum.     

The influence of radiation therapy on subperiosteal hydroxyapatite implants in rabbits.

Granular hydroxyapatite (HA) was implanted into subperiosteal pockets along both the right and left proximal tibias of 12 adult New Zealand white rabbits. The left extremities of 10 rabbits served as controls, whereas the right sides were irradiated with 2,250 rad in three doses over 5 days. The animals were killed at 1-week intervals starting at 2 weeks postirradiation. One half of each site was evaluated using standard histologic techniques while the other half was examined as a nondecalcified section. The quantity and the quality of new bone formation was determined using a rating scale and histomorphometric digitization. The results of this 4-month study showed that the amount of new bone formation around the HA granules was significantly greater in the irradiated sites. The irradiation produced no deleterious effects on the implant or the surrounding tissue areas.     

Effect of recombinant human bone morphogenetic protein-2 on mandibular distraction osteogenesis

The purpose of this investigation was to study the effects of recombinant human bone morphogenetic protein-2 (rhBMP-2) on bone formation of mandibular distraction osteogenesis. Six skeletally mature sheep underwent 10 mm of bilateral mandibular distraction osteogenesis via a custom-made distractor. Three micrograms of rhBMP-2 with a collagen carrier was implanted in the osteotomy site of one side of the mandible during the osteotomy phase. The contralateral side was used as the control group, and no material was implanted into the distracted area. At 10 days after the end of distraction, all animals were killed, and the distracted calluses were harvested for radiologic and histologic analysis. New bone was generated in the distracted zone in all groups. Histologic and radiologic examination showed that the new bone formation was greater in the rhBMP-2 group than in the control group. Quantitative computed tomography evaluation, however, did not demonstrate a significantly different mean bone density of the regenerates between the 2 groups. The results indicate that application of a rhBMP-2/collagen implant during the osteotomy phase of distraction osteogenesis increased bone formation but did not have a significant effect on bone density of the regenerates.     

Alveolar ridge augmentation in rats by combined hydroxylapatite and osteoinductive material

Abstract— The purpose of this study was to examine if increased bony ingrowth developed when hydroxylapatite was implanted together with an osteoinductive substrate. Dense hydroxylapatite granules (HA) (Calcitite, Calcitek, San Diego, CA, USA) were mixed with equal volumes of allogenic, demineralized, lyophilized dentin or bone and implanted subperiosteally for alveolar ridge augmentation purposes and heterotopically in the abdominal muscles in rats. Light microscopic evaluation revealed that HA was surrounded by fibrous connective tissue containing foreign body giant cells and it had neither an osteoinductive nor an osteoconductive effect. The newly formed bone induced from the implanted allogenic, demineralized, lyophilized dentin or bone was never found in close contact with the HA and did not incorporate the implant.     

Alveolar ridge augmentation in rats by combined hydroxylapatite and osteoinductive material.

The purpose of this study was to examine if increased bony ingrowth developed when hydroxylapatite was implanted together with an osteoinductive substrate. Dense hydroxylapatite granules (HA) (Calcitite, Calcitek, San Diego, CA, USA) were mixed with equal volumes of allogenic, demineralized, lyophilized dentin or bone and implanted subperiosteally for alveolar ridge augmentation purposes and heterotopically in the abdominal muscles in rats. Light microscopic evaluation revealed that HA was surrounded by fibrous connective tissue containing foreign body giant cells and it had neither an osteoinductive nor an osteoconductive effect. The newly formed bone induced from the implanted allogenic, demineralized, lyophilized dentin or bone was never found in close contact with the HA and did not incorporate the implant.     

Hydroxylapatite granules and blocks as an extracranial augmenting material in rhesus monkeys

This study evaluates the use of the nonporous hydroxylapatite (HA) granules and blocks as an extracranial augmentation material in monkeys. Augmentation was performed in 12 Rhesus monkeys killed at three, six, and 12 months postoperatively. Clinical evaluation revealed that the HA granules become stable within three to four weeks, but there is loss of some augmented height due to settling of the material within the subpericranial pocket. The HA blocks maintained the augmented cranial contours, but 25% of them exhibited mobility. Histologic evaluation revealed no evidence of inflammation or bone resorption with either the HA granular or block form. The HA granules were separated from each other and from the bone interface by a layer of collagen fibers, and the blocks were also surrounded by a fibrous capsule. There was no evidence of bonding between the HA granules, and the bone-HA interface was separated by an intervening fibrous layer in all the HA granular specimens except for one in the one year group which showed evidence of bone formation in part of the HA bone interface. It was concluded that when used as an extracranial augmentation material, the nonporous HA granules were more stable than blocks; however, the blocks maintain a better contour.     

兔骨髓基质干细胞自体嚼肌内移植异位成骨实验研究

目的：观察骨髓基质干细胞成骨向分化诱导后植入嚼肌内异位成骨的过程。方法：从兔髂骨分离骨髓基质干细胞,经体外诱导分化,与磷酸三钙／羟基磷灰石（tricalcium phosphate／hydroxyapatite,TCP／HA）支架复合,植入兔自体嚼肌内,形态学、免疫细胞化学、组织学观察其成骨特征。结果：体外培养的兔骨髓基质干细胞性状稳定,经矿化诱导培养的骨髓基质干细胞表现为成骨细胞分化,表达Ⅰ型胶原蛋白,与支架材料复合植入自体嚼肌内可见成熟骨组织形成。结论：利用骨髓基质干细胞为种子细胞植入嚼肌内开展功能性组织工程化颌骨移植方法可行。     

Enhancement of bone ingrowth into collagen/HA composite implants using e-PTFE membranes

The objective of this investigation was to study the effectiveness of expanded polytetrafluoroethylene (e-PTFE) membranes for enhancement of bone ingrowth through subperiosteally implanted collagen/HA composite blocks. Twelve rabbits aged 12-15 months served as the experimental animals in this study. Two compressed Collagen/HA composite blocks in the shape of two attached cylinders of different diameters were inserted into two defects of each rabbit calvarium, the smaller cylinder being intrabony, the larger subperiosteal in location. One of the two implants was covered with non-resorbable e-PTFE membrane. The other implant was left uncovered. Specimens were obtained at 4, 8, and 12 weeks. While the implant specimens on the membrane side showed progressive bone formation between and around HA particles at the subperiosteal extrabony locations, the HA particles on the non-membrane side were surrounded and separated by dense fibrous tissue. At intraosseous sites, HA particles were surrounded by new bone throughout the defect on the membrane side, but new bone formation occurred only along the periphery on the non-membrane side. It appears that guided tissue regeneration may be used to enhance new bone formation around and between subperiosteally implanted HA particles.     

A new approach to enhancement of bone formation by electrically polarized hydroxyapatite.

An electrical field may affect osteogenesis. Since we found that hydroxyapatite (HA) ceramics may be polarizable, we hypothesized that electrically polarized HA may foster production of new bone in vivo. Both polarized and non-polarized HA ceramics were inserted into the subperiosteum spaces at the parietal bone area of rats. After 2, 4, and 8 weeks, the implant sites were examined histologically. Morphometric analysis revealed that new bone formation was accelerated on the negatively charged surface of the polarized HA (N-surface) at 2 weeks. The newly formed bone approached maturation at 4 weeks and was thicker on the N-surface than in the controls. By 8 weeks, newly formed bone in the controls was almost the same as that on the N-surface. These findings suggest that polarized HA is biocompatible and that bone formation on the N-surface is enhanced in the early stage of bone healing.     

Effect of recombinant human tissue inhibitor of matrix metalloproteinase-1 in mandibular distraction osteogenesis in rabbits: a computed tomographic study

Matrix metalloproteinases (MMPs), together with their tissue inhibitors (TIMPs), are responsible for the controlled degradation of collagen in bone. We studied the long-term stability of the regenerated bone formed by distraction osteogenesis, and the effect of recombinant human TIMP-1 on the remodelling of bone formed by mandibular distraction osteogenesis in rabbits. Nine rabbits were subjected to distraction osteogenesis of the mandible and divided into three groups: a control group with no collagen implanted; a sham-control group with a collagen sheet implanted; and an experimental group with a collagen sheet impregnated with rhTIMP-1 implanted. Computed tomograms were taken at weeks 4, 12, and 24 after distraction, and micro-computed tomograms and histological examinations were made at week 24. There was no significant resorption of regenerated bone at the site of distraction in any group after 6 months of consolidation, suggesting that the regenerated bone formed by distraction osteogenesis is stable. We found no obvious influence of rhTIMP-1 in the collagen sheet on the bony regenerate after 6 months of consolidation.     

Bone changes around hydroxyapatite and titanium implants after abutment placement in rabbits-observations using histological and three-dimensional examinations

We have previously developed a computer-aided system for examination of the three-dimensional bone structure around implants and observed the bone changes in the healing period after implant placement. This paper describes the bone changes around hydroxyapatite (HA) and titanium (Ti) implants after abutment placement using histological and three-dimensional examinations. Twenty-four HA and Ti implants were embedded in the tibias of adult male New Zealand white rabbits. After 8 weeks, the abutment had passed through periosteum and was placed under the skin. Rabbits were sacrificed 4 and 8 weeks following abutment placement. In conclusion, histological examination showed that, at 4 weeks after abutment placement, bone resorption around the implant neck was seen in both HA and Ti implants, and at 8 weeks, excessive bone formation was seen around the implant neck. Three-dimensional bone examination showed that abutment placement may affect bone formation and cause additional bone hypertrophy in the bone marrow area.     

HA—涂层种植体不同种植间隙的组织学评价

目的 :对国产羟基磷灰石涂层的种植体在狗股骨上模拟临床上即刻种植与延期种植的两种种植方法 ,进行组织学评价。方法 :在狗股骨上分别形成 1～ 4mm宽的种植间隙 ,用软X线、偏光显微镜和甲美兰染色检查等方法 ,比较了其组织学的反应。结果 :所有种植孔内皮质骨区新骨生长充满间隙 ,与种植体HA涂层结合 ,但髓质骨区仅间隙内放置HA颗粒者 ,骨小梁呈网状生长 ,HA颗粒、新生骨质和HA涂层结合紧密 ,类似骨皮质结构。结论 :种植间隙大时 ,应用HA颗粒有利于新骨生长。     

选择性激光烧结多孔钛种植体及其性能研究

目的 分析选择性激光烧结（SLS）多孔钛种植体的机械性能及生物相容性,探讨其与壳聚糖（CS）/羟磷灰石（HA）复合涂层结合后的促骨结合作用。方法 制备Ti6Al4V试件,部分试件表面进行CS/HA涂层处理;对试件进行扫描电子显微镜观察和机械性能检测;体外培养MC3T3-E1细胞,进行活/死细胞染色、甲基噻唑基四唑（MTT）及碱性磷酸酶（ALP）水平检测;将柱状螺纹种植体植入兔股骨髁部,分析其体内生物学性能。结果 试件准弹性梯度随孔隙率增大而减小,孔隙率为30%时与皮质骨接近,70%时与松质骨接近;试件具有良好的生物相容性。复合CS/HA涂层后可促进MC3T3-E1细胞增殖、分化,有利于骨组织长入孔隙,形成良好的骨结合。结论 多孔钛种植体具有良好的机械性能和生物相容性,与CS/HA涂层结合后具有骨诱导性,利于形成稳定的骨结合。     

The effect of electrical stimulation on bone formation around hydroxyapatite implants placed on the rabbit mandible.

Nonresorbable, nonporous, particulate hydroxyapatite (HA) was implanted on the mandible in rabbits and stimulated electrically, 4 hours per day, during the first postoperative week. Stimulated and control implant sites were recovered 8 weeks postoperatively and examined histologically. The HA migrated into the mandible in the electrically treated specimens, and was routinely found in intimate association with preexisting mandibular bone. In the controls, the HA remained superior to the mandibular surface. In further studies (without electrical stimulation) in which the implant site was recovered 26 weeks postoperatively, HA was observed in the mandible; some HA particles migrated completely through the mandible and were found in the adjacent soft tissue. It was concluded that, under the conditions studied, electrical stimulation does not promote bone growth into HA, but rather produces the opposite result--it promotes more rapid movement of HA particles into the mandibular bone. The HA particle migration into the mandible observed (longer postoperative times) in the absence of electrical stimulation suggests that migration is a general property of HA particles when placed over bone under muscle.