多在磷灰石陶瓷诱导成骨的作用

孔羟基磷灰石陶瓷种体埋值于狗背部肌肉内，分别于植入后１、２、４和８月处死动物，取出种植体，将其固定，脱钙、切片、ＨＥ染色，在光镜下组织形态学观察发现：一月的种植体孔隙内充满了血管丰富的结缔组织；二、四和八月的种植体孔隙内有较多量的新骨组织形成，种植体周有完整的纤维结缔组织包膜。本研究从组织形态学说明了多羟基磷灰石陶瓷具有诱导骨形成的能力。     

国外磷灰石陶瓷-机体界面研究近况

磷灰石陶瓷是一种生物种植材料，磷灰石陶瓷一机体界面的研究是这种种植材料的基础性研究内容。本文综述了９０年代以来国外在这方面的研究状况，包括组织形态学、生物力学和分析学研究内容，特别是胶原／羟基磷灰石与机体界面、羟基磷灰石与细胞界面、等离子体喷涂ＨＡ种植体和ＨＡ复合材料与骨界面的研究。     

国外磷灰石陶瓷—机体界面研究近况

磷灰石陶瓷是一种生物种植材料，磷灰石陶瓷－机体界面的研究是这种种植材料的基础性研究内容。本文综述了９０年代以来国外在这方面的研究状况，包括组织形态学、生物力学和分析学研究内容，特别是胶原／羟基磷灰石与机体界面、羟基磷灰石与细胞界面、等离子体喷涂ＨＡ种植体和ＨＡ复合材料与骨界面的研究。     

多孔磷酸钙陶瓷在动态SBF中类骨磷灰石形成的研究

本实验在模拟体液（SBF）以静止、等于或大于骨骼肌组织内体液正常生理流率（2ml/100ml.min)的流动条件下，研究多孔磷酸钙陶瓷上类骨磷灰石的形成，结果表明，SBF在生理流率（2ml/100ml.min)情况下，材料仅在多孔磷酸钙陶瓷孔隙内部形成类骨磷灰石，静态（0ml/100ml.min)情况下，多孔磷酸钙陶瓷材料表面有类骨磷灰石形成；高于生理流率（10ml/100ml/min)时，表面和断面均无类骨磷灰石形成，但如果增加SBF中的Ca^2 ,HPO4^2-的浓度，则在孔隙内部有类骨磷灰石形成，生理流条件下的结果与多数肌肉内植入磷酸钙陶瓷试验的结果一致。仅在多孔材料内部成骨，这个结果表明，比起通常使用的静态浸泡试验，SBF以生理流率流动的动态体外试验能够更好地模拟类骨 磷灰石生长的体内环境，动态SBF对了解类骨磷灰石形成，进而了解磷酸钙陶瓷在体内诱导成骨机理是十分有用的。     

多孔碳酸化羟基磷灰石骨水泥的生物相容性研究

目的探讨一种新型的代骨材料--多孔碳酸化羟基磷灰石骨水泥的组织相容性.方法合成碳酸化羟基磷灰石骨水泥,添加成孔剂,制备能原位固化形成多孔结构的碳酸化羟基磷灰石代骨材料,并通过细胞毒性实验和肌内埋植实验检测其组织相容性.结果多孔碳酸化羟基磷灰石骨水泥的浸提液对骨髓基质细胞的生长无影响.细胞于材料表面的生长速度、形态与空白对照组无差别.肌内植入实验发现,材料周围纤维组织包膜的最大厚度为22.5μm,未发现炎性细胞反应.结论多孔碳酸化羟基磷灰石骨水泥具有良好的组织相容性,材料测试结果符合和标准.     

在不同动物肌肉中磷酸钙陶瓷表面类骨磷灰石的形成研究

磷酸钙陶瓷材料植入动物体内后其表面类骨磷灰石层的形成对骨的形成有非常重要的作用，并被认为是骨诱导发生的先决条件。我们将相同大小的孔壁有微孔的多孔材料和致密磷酸钙陶瓷材料植入猪，狗、兔和鼠的背肌或腿肌内，研究陶瓷表面类骨磷灰石的形成，以了解类骨磷灰石层的形成与骨诱导的联系。结果表明：磷酸钙陶瓷材料植入动物的肌肉内14d后，狗，兔和鼠体内的多孔材料孔隙内表面（包括陶瓷表面较深孔隙）有一层类骨磷灰石层形成；植入猪体内的多孔材料内外表面都形成了一层类骨磷灰石，致密材料在几种动物体内都未观察到类骨磷灰石层形成，类骨磷灰石层形成的快慢次序与动物组织学观察到的在不同动物的肌内骨诱导性高低的次序不一致。证实了类骨磷灰石层的形成的确是骨诱导的先决条件。但还有其它因素影响骨诱导的发生。     

多孔碳酸钙陶瓷与干细胞源性成骨细胞的相容性

背景：国内外的研究证实普通碳酸钙陶瓷作为骨替代材料时具有细胞支架作用。 目的：观察多孔碳酸钙陶瓷与成骨细胞的相容性,及作为骨组织工程支架的可能性。 方法：SD大鼠骨髓基质干细胞经矿化诱导培养、扩增并检测证实其已具成骨细胞表型后,分别与多孔碳酸钙陶瓷支架、普通羟基磷灰石陶瓷支架体外复合培养。 结果与结论：骨髓基质干细胞经体外诱导形成成骨细胞,钙结节、Ⅰ型胶原和碱性磷酸酶免疫染色结果阳性。多孔碳酸钙陶瓷支架材料与羟基磷灰石陶瓷材料皆有细胞附着生长,但多孔碳酸钙陶瓷支架材料细胞的黏附能力、增殖活力及成骨活性均强于羟基磷灰石陶瓷材料。提示多孔碳酸钙陶瓷支架材料与SD大鼠骨髓基质干细胞源性成骨细胞有良好相容性。     

新型羟基磷灰石多孔支架的制备与性状

背景：目前几种钙羟磷灰石作为骨的替代品已经在临床上使用,但是由于缺少内部交互的连通孔,使得骨形成过程中经常发生病理性的骨折。 目的：采用改良的技术制备互联多孔羟磷灰石陶瓷支架材料,并对其物理化学特征进行检测。 方法：将羟基磷灰石(质量分数60%)与聚乙烯亚胺质量分数40%混合,采用改良的泡沫凝胶技术技术(聚氧乙烯十二烷基醚质量分数1%)制备多孔羟磷灰石陶瓷。扫面电镜检测材料内部结构,将制备的材料移植动物体骨缺损区观察成骨情况。 结果与结论：多孔羟磷灰石陶瓷具有三维结构,内部充满大小较均一的球形孔隙,直径平均为150 μm,孔隙率75%,孔隙上充满窗孔样相互交通的小洞,平均40 μm, 具有足够的抗压度(10~12 MPa)。动物实验表明,多孔羟磷灰石陶瓷表现出较好的成骨诱导性,可见孔隙里形成了新生骨。结果提示改良的羟基磷灰石支架能够在骨组织工程上应用,有望成为新型的改良品。     

多孔碳酸化羟基磷灰石水泥植入修复兔包容性骨缺损的力学分析

背景:采用发泡剂成孔技术,制成了有知识产权的新型骨修复材料多孔碳酸化羟基磷灰石,既保留了碳酸化羟基磷灰石骨水泥原位固化性能等所有的优点,同时又形成多孔结构。目的:通过动物实验观察新型的骨修复材料多孔碳酸化羟基磷灰石水泥修复骨缺损的力学效果。方法:30只新西兰大白兔,手术组25只在双侧股骨髁制备直径为5.5mm、深12mm的骨缺损动物模型,左侧植入多孔碳酸化羟基磷灰石骨水泥为实验组,右侧植入碳酸化羟基磷灰石骨水泥为对照组。非手术组5只,用于正常力学对照。将多孔碳酸化羟基磷灰石骨水泥和碳酸化羟基磷灰石骨水泥试件经模仿体液浸泡,检测力学强度。同时在手术组背肌内分别植入多孔碳酸化羟基磷灰石骨水泥和碳酸化羟基磷灰石骨水泥标准试件。分别于术后2,4,8,12,16周分批处死动物,进行试件骨内和肌内植入的力学实验分析和试件在模仿体液中浸泡后的抗压强度测试。结果与结论:多孔碳酸化羟基磷灰石骨水泥:2周时的骨内力学强度较低,4周时降到最低,8周时接近正常松质骨强度,12周时超过正常松质骨强度,16周时恢复到正常松质骨水平。碳酸化羟基磷灰石骨水泥:2周时骨内植入强度较多孔碳酸化羟基磷灰石骨水泥略高,4周时有所降低,8,12,16周时略升高,但是始终低于正常松质骨的强度。多孔碳酸化羟基磷灰石骨水泥和碳酸化羟基磷灰石骨水泥在SBF中浸泡的抗压强度变化不大。试件植入肌内后抗压强度变化非常显著。结果表明,多孔碳酸化羟基磷灰石水泥具有原位固化性能和一定的力学支撑作用,能作为自体骨移植的一种替代物修复骨缺损。     

纳米双相陶瓷人工骨的成骨及降解

背景：烧结后的纳米羟基磷灰石结晶度很高,在体内很难降解;纳米β-磷酸三钙的降解速度太快,不利于体内生物组织在材料上附着,不利于引导成骨。 目的：观察纳米羟基磷灰石/纳米β-磷酸三钙双相陶瓷人工骨的成骨及降解性能。 方法：将36只青紫蓝兔随机分为实验组、对照组及空白组,制作左侧挠骨缺损模型,实验组与对照组分别植入纳米羟基磷灰石/纳米β-磷酸三钙双相陶瓷人工骨、纳米羟基磷灰石人工骨,空白组不植入任何材料。术后4,8,12周观察成骨和材料降解情况。 结果与结论：①术后12周时X射线：实验组可见材料基本降解,连续性骨痂通过骨缺损部位。对照组材料未见明显降解,骨缺损处有骨痂修复。空白组骨缺损未见修复。②术后12周时组织学观察：实验组材料孔隙内以骨细胞和成骨细胞为主,有少量软骨细胞,出现散乱的骨松质,材料完全降解。对照组材料孔隙内以骨细胞为主,有少量成骨细胞和软骨细胞,材料未见明显降解。空白组可见纤维结缔组织及胶原纤维。③术后12周时扫描电镜观察：实验组材料降解,骨缺损部位被新生骨松质取代。对照组材料未见降解,骨缺损部位大都被新生骨松质取代。空白组无明显骨重建。表明纳米羟基磷灰石/纳米β-磷酸三钙双相陶瓷人工骨具有良好成骨能力及降解性能。     

Permucosal implantation pilot study with HA-coated dental implant in dogs.

The histological response of transmucosal one-stage titanium dental implants coated with hydroxyapatite is described. The gingival adhesion to the implant was examined with regard to coated, partially coated or non-coated surfaces in the cervical region. From each coating type, 9 implants were inserted into dogs. Six months after the insertion, 19 implants could be evaluated, but 8 implants were lost. From these 19 implants, 6 implants showed severe pockets with inflammation up to the bony tissue. The 13 successful implants showed direct bone bonding with the hydroxyapatite coating and adhesion of submucosal connective tissue to the implant surface, with inflammation. The marginal gingiva showed slight inflammation. A totally coated implant will probably introduce inflammation by debris formation against the rough implant surface more easily. The hydroxyapatite coating often disappeared in the soft tissue or in the oral cavity. Bone which directly adapted to the coating seemed to prevent it from resorption.     

Integration of metallic endoprotheses in dog femur studied by near-infrared Fourier-transform Raman microscopy.

The integration of hydroxyapatite-coated implants in dog femur was studied by near-infrared Fourier-transform Raman microscopy. Raman spectra were taken in lateral scans in step widths of 10-40 microm from the implant surface up to a distance of 320 microm into the bone tissue. The spectra were subjected to a component analysis for the quantitative determination of the protein and the inorganic components. This quantitative analysis is shown to be more reliable than conventional band fitting procedures and allows, for the first time, the quantitative distinction between the hydroxyapatite form of mature bone tissue and synthetic hydroxyapatite introduced by the implant coating. It is demonstrated that full mineralization of the ongrowing bone is not achieved after 6 months. In contrast, after a residence time of 18 months in the body, the Raman spectra reveal a complete calcification of the new bone tissue as indicated by content of biological hydroxyapatite that is the same as in mature bone tissue throughout the whole implant/bone interface. On the other hand, the content of synthetic hydroxyapatite is strongly reduced in the sample prepared after eighteen months implantation whereas for the shorter implantation time. substantial contributions of synthetic hydroxyapatite are found even at positions beyond the thickness of the implant coating. These results indicate that the coating material is actively involved in the mineralization of ongrowing bone. Possible mechanisms for the underlying transport processes in the implant/bone interface are discussed.     

Healing of osteochondral osteotomies after fixation with a hydroxyapatite-buffered polylactide. A histomorphometric and radiographic study in rabbits

The tissue response of subchondral bone to a biodegradable fixation device manufactured in the shape of a screw and made of polylactide with a hydroxyapatite buffer were implanted through the articular surface of the intercondylar portion of the distal rabbit femur. One screw was implanted per animal. The screws had a core diameter of 3.2 mm and an outer diameter of 4.5 mm. At insertion, the implants were cut flush with the articular surface. After follow-up times of 8 and 16 weeks, the specimens were examined radiographically and histomorphometrically. The intact contralateral femur served as a control for comparison. Only minimal signs of degradation of the polymer could be seen in the histologic specimens. These implant degradation sites were commonly areas of new bone formation adjacent to the screw implant. A brim of repair tissue was formed at the entrance and exit of the implant channel. The width of the repair tissue from the tissue-implant boundary towards the center of the entrance hole varied greatly between the specimens, from 80 to 750 microm. In most specimens this bridging tissue consisted of newly formed bone and undifferentiated mesenchymal tissue. Degenerative chondrocyte clustering occurred in the pre-existing cartilage within a 400 microm wide zone from the tissue-implant interface into the recipient tissues. Some new-bone formation was seen to envelop the implant in all specimens, but the fractional osteoid formation surface of the trabeculae was only significantly higher in the screw-implanted 16-week specimens, when compared to the non-operated contralateral controls. Although the bony osteotomy was invariably healed in all specimens with good implant integration, the quality and quantity of the reparative tissue of the articular cartilage near the screw hole was variable. This study showed that large polylactide implants, which are buffered with hydroxyapatite show benign tissue responses and good implant osteointegration when implanted in bone. They may be suitable for fixation of small bone fractures. However, insertion through intra-articular surfaces may require further improvement of the implant material to avoid the degenerative repair processes seen in this study.     

Threaded versus porous-surfaced designs for implant stabilization in bone-endodontic implant model

An endodontic implant model system was used to compare the effect of implant design on stabilization in bone. Specifically a porous-surfaced design was compared to conventional threaded and smooth-tapered endodontic implant designs. All implants were placed in immediate function thereby assessing the effect of early limited movement on the fixation achieved. A total of eighty-three endodontic implants were inserted in the mandibles of six adult mongrel dogs. Animals were sacrificed immediately after implantation and after 3, 6, and 12 months. Implants were evaluated by clinical and radiographic examination and after animal sacrifice by pull-out tests of the implant from the tissues, SEM examination of the pulled-out implants and, finally, histology. The pull-out test results indicated increasing shear strength with implantation time for the porous-surfaced implants in contrast to the gradual loss of fixation for the threaded implants and the continuous low shear strength for the smooth implants. Histological studies and SEM examination indicated the reason for these changes. Smooth implants became encapsulated by fibrous connective tissue from early post-implantation time periods. Threaded implants, although initially mechanically interlocked with bone, developed a fibrous connective tissue capsule that gradually thickened with time until, by 6 months, little mechanical interlock of bone and implant was present. It was assumed that this fibrous capsule thickening was caused by implant movement. The porous-surfaced implants, however, became stabilized by bone ingrowth and showed more extensive bone formation within the surface pores with time. It is concluded that for implants that are made functional immediately after implantation, as in this study, porous-surfaced implants can become strongly fixed by bone ingrowth, in contrast to conventional threaded or smooth-surfaced designs, thus presenting a more favourable long term prognosis.     

Assessment of the effect of a biphasic ceramic on bone response in a rat calvarial defect model

Calcium phosphate ceramics are being extensively used for orthopedic, periodontal, and dental applications. This study aimed to assess the effect of a biphasic ceramic such as Ceraform on the osteogenesis in a rat calvarial defect model. 20 Wistar rats were enrolled in the study. Two symmetrical, circular, and 5-mm-wide full thickness defects were created in the parietal bones of each animal. The left defect was left empty as a control and the right defect was filled with the particular implant material. Animals were divided into two groups, and 10 animals were sacrificed at month 3 and the rest were sacrificed at month 6. The calvarial specimens were harvested for histological examinations. Defect area samples were stained with hematoxylin-eosin and Masson Thrichrom. A semiquantitative method was used to quantify the bone regeneration. The defects were mostly filled with fibrous connective tissue (3-6 months) in the control site. A loose, fibrovascular tissue was observed at the side of ceraform implantation at month 3. By 6 month, a dense collagenous tissue was observed at the same area. Multinuclear giant cells (MNGC) were detected around the implant bed at month 3 and month 6. No necrosis, tumorigenesis, or infection was observed at the implantation site at any time. There was no statistically meaningful difference regarding bone regeneration between the two defects at each observation period (p>0.05). This study showed that Ceraform is biocompatible. However, this study indicates that biphasic ceramic do not offer any advantage over hydroxyapatite ceramics. It was also revealed that it had no effects on bone regeneration and that it seemed to be a space maintainer.     

Quantitative bone remodelling resulting from the use of porous dental implants

Histomorphometric analyses were used to quantitatively determine the patterns of bony ingrowth which resulted from the placement of porous-surfaced dental implants into the mandibles of Rhesus monkeys for up to 74 months utilizing a two-stage approach. Quantitative histopathologic evaluations were made using ground section microscopy. Implant stability resulting from bone remodelling and ingrowth occurred to varying degrees with all implants. Bone ingrowth occurred from medullary trabeculae and contact with the adjacent cortical plates. Quantitative histomorphometric analyses revealed that in only one case was the bone ingrowth into the available internal pores less than 45%. Minimal fibrous connective tissue ingrowth was observed in the implant crypts and was not thought to be due to micro-motion. The observed bone remodelling indicated a favorable prognosis for long-term implant performance.     

HA/TCP双相陶瓷肌内长期植入诱导成骨的实验观察

通过肌内长期植入研究了Ca-P生物材料骨诱导性及诱生新骨的生长状况。HA／TCP多孔陶瓷植入狗背部肌内15月后，植入体为一层很薄的纤维结缔组织膜包裹。3种形态的组织，即间充质组织，骨和骨髓在植入体内部孔中有规律地分布：植入体内部所有的孔中都有骨组织存在；但中部内孔中的骨为较成熟的板状骨，有明显的骨陷窝，已形成髓腔和骨髓，边缘外孔中的骨组织为未成熟的编织骨，与间充质组织相连，无髓腔和骨髓形成，这表明HA／TCP具有骨诱导性，在非骨环境中确实能够诱导成骨，而且，各种组织有规律地分布在植入材料孔中，表明植入体内部骨组织形态的发生和发展遵循特定的时空顺序。     

Bone bonding in sintered hydroxyapatite combined with a new synthesized agent, TAK-778.

We studied the stimulatory effects of TAK-778, a new synthetic 3-benzothiepin derivative that promotes osteoblast differentiation, in bone bonding to sintered hydroxyapatite implants in rabbit tibiae. Smooth-surfaced rectangular plates (15 x 10 x 2 mm) made of sintered hydroxyapatite were implanted into the proximal metaphyses of bilateral rabbit tibiae, with TAK-778-containing sustained-release microcapsules packed into the medullary cavity in one limb and untreated microcapsules packed in the contralateral limb to serve as a paired control. At 4, 8, and 16 weeks after implantation, bone bonding at the bone-implant interfaces was evaluated by a detaching test and undecalcified histological examination. The tensile failure load increased from 4 to 16 weeks for both groups; however, the tensile failure load of the TAK-778-treated group was significantly greater than that of the control group at each interval after implantation. Histologically, the TAK-778-treated specimens showed greater active new bone formation mainly in the medullary cavity and more extensive bonding between the implant and bone than the untreated specimens. The results of this study suggest that adding osteoinductive TAK-778 to hydroxyapatite implants may significantly accelerate bone apposition to the implants and improve the bonding process at the interface. This would help to establish an earlier and stronger bonding of orthopedic ceramic implants between the surrounding bone tissue.     

组织工程骨-假体界面微观结构与元素分析的意义

目的研究假体-组织工程骨界面的微观结构和钙磷元素及构成比动态变化及意义。方法取新西兰大白兔15只,将兔骨髓间充质干细胞培养、扩增及诱导后,与珊瑚羟基磷灰石复合构建组织工程骨;双侧股骨髁分别制作一0.5cm×1.2cm骨缺损,骨缺损中央植入0.2cmx1.0cm钛合金植入体,左侧植入体周围植入组织工程骨,右侧仅植入珊瑚羟基磷灰石为对照,于术后4周、8周和l2周分别行X线检查、扫描电镜及能谱分析观察植入体表面微观结构和钙磷元素百分含量及比值的变化。结果 X线检查示,实验组术后8-12周可见大量模糊的高密度骨痂影,缺损区与周围骨质间分界变模糊。对照组各时间点无明显变化;扫描电镜示,实验组术后8、12周植入体表面孔隙内有大量无定形物质,而对照组未见有无定形物质。能谱分析示,不同时间点实验组较对照组内钙、磷元素百分含量高,有显著性差异（〈0.05）,随着时间的变化实验组和对照组Ca、P百分含量都呈增高的趋势,但12周和8周相比无显著性差异（〉0.05）。实验组内钙磷比值随时间变化有逐渐增大趋势,8周时达峰值,8周后缓慢下降。结论假体-组织工程骨界面钙、磷元素及构成比的动态变化表明骨整合的形成,并随着时间的延长骨改建逐步增加。     

Contractile behavior of smooth muscle actin-containing osteoblasts in collagen-GAG matrices in vitro: implant-related cell contraction

The contraction of connective tissue cells may facilitate their production and maintenance of extracellular matrix architecture and can benefit healing through wound closure. However, aberrant cell contraction can result in pathological contracture. Implants may stimulate this process leading to contracture of the surrounding fibrous capsule. In the case of compliant porous matrices used for tissue engineering the cell contraction may cause the constriction of pores and the distortion of the implant. The objective of this study was to determine if osteoblasts expressed a specific muscle actin, alpha-smooth muscle actin (SMA), that could provide for their contraction. Immunohistochemistry revealed the presence of SMA in some cells in all of three human and three canine trabecular bone specimens. The majority of the cells of an osteoblastic cell line, MC3T3-E 1, also expressed this actin isoform in two-dimensional culture and when seeded into a collagen-glycosaminoglycan (GAG) matrix. These SMA-containing cells were found to cause contraction of the collagen-GAG analog of extracellular matrix. These findings demonstrate that osteoblasts can display contractile behavior that might help to explain the mechanism by which they impart architecture to bone matrix, including that at implant interfaces. An understanding of this process could also guide the development of matrices for bone tissue engineering.