复合珊瑚羟基磷灰石人工骨的研制和临床应用

目的　自行研制复合人工骨并评价其临床应用价值。方法　将天然海珊瑚碳酸钙在特定条件下经过“热液交换反应”先制成一种珊瑚羟基磷灰石 (CorallingHydroxyAptite,CHA)人工骨。再将此CHA人工骨与重组人BMP2 (rhBMP2 )和 2 %几丁糖复合 ,制备成rhBMP2 /CHA复合人工骨。将此新型复合人工骨分别植入 2 0只SD大白鼠体内 ,观察其生物相容性和成骨能力。并应用临床治疗骨缺损和椎间融合共 30例 ,观察术后局部伤口情况 ,全身反应和骨愈合情况。结果　随访观察平均 14个月 ,无全身性异常反应 ,仅 1例伤口有渗液 ,3周内伤口愈合 ,其余无不良反应 ,伤口均在 2周内一期愈合。骨愈合情况 :骨囊肿和胫骨干骺端骨折骨缺损植骨后 2个月愈合 ;脊柱椎间融合术后 3个月融合 ;骨不连术后 4～ 6个月愈合 ,无并发症发生。人工骨在观察期内未能完全降解。结论　自行研制的rhBMP2 /CHA复合人工骨生物相容性好 ,无毒性 ,成骨诱导作用优于单纯CHA和异体脱钙骨 ,临床应用满意 ,是一种比较理想的骨移植替代物。     

The use of coral for cervical interbody arthrodesis (IBA) and vertebral body excision

Summary The value of using biomaterials and especially coral in cervical spinal surgery is linked to a desire to eliminate the disadvantages associated with the taking of bone grafts in arthrodeses. A chemical, microscopic, biomechanical and clinical study of coral from 1970 has shown the value of this material, whose structure resembles that of bone, and which degrades completely to be replaced by newly-formed bone. Our initial results in 20 cases confirmed these findings by studying the progress of the coral: its good clinical tolerance, and its excellent radiologic incorporation without modification of spinal statics. Subsequently 83 cases have ben treated with this biomaterial, of which 45 were followed up. The results are satisfactory from both the clinical viewpoint (technques of transdiscal osteophytectomy) and the biomechanical aspect.     

珊瑚复合人骨髓基质细胞构建组织工程骨

目的：观察人骨髓基质细胞在角孔珊瑚上立体培养后的生长情况及其在体内的成骨能力。方法：穿刺抽吸人髂骨区骨髓基质细胞，体外培养扩增、诱导后，接种于角孔珊瑚中立体培养5d，观察细胞在角孔珊瑚表面的贴附及伸展情况；将上述细胞/角孔珊瑚复合物植入体内，观察植入物在体内的成骨情况。结果：细胞可在角孔珊瑚中立体培养成活。体内植入后4周，复合物中有少量新骨形成；体内植入后8周，复合物中出现大量较成熟骨组织；而对照组4、8周均无骨组织形成。结论：穿刺抽吸的人骨髓基质细胞可作为骨组织工程的种子细胞，角孔珊瑚可作为骨组织工程的支架材料。     

载药珊瑚人工骨修复骨缺损及预防感染的实验研究

目的：探讨载药珊瑚人工骨修复骨缺损及预防感染的可能性。方法：将珊瑚与克林霉素复合，制成载药珊瑚人工骨；将其置于接种有金葡菌的培养基中检测其体外缓释特性及抗菌活性；将其植入金葡菌污染的兔颅骨缺损区，通过细菌培养和组织病理检查评价其骨缺损修复能力及抗感染能力。结果：载药珊瑚人工骨在体外持续释放高浓度抗菌素；植入金葡菌污染的骨缺损后可有效预防骨感染，骨感染的发生率明显低于单纯珊瑚人工骨植入组。结论：载药珊瑚人工骨是伴有污染或感染骨缺损较理想的修复材料。     

生物珊瑚、胶原和rhBMP-2合成人工骨修复兔下颌骨缺损的实验研究

目的：研究生物珊瑚、胶原和ｒｈＢＭＰ－２复合人工骨修复兔下颌骨缺损的效果。方法：选择３６只兔下颌骨缺损模型，分别采用新鲜自体骨，生物珊瑚和生物珊瑚、胶原和ｒｈＢＭＰ－２复合人工骨修复，大体、组织切片、Ｘ线、统计学等方法进行分析。结果：修复兔下颌骨缺损，合成人工骨形成新骨的时间比生物珊瑚早，２个月时修复效果优于生物珊瑚；３个月时，复合骨修复效果与自体骨无明显差别。结论：生物珊瑚、胶原和ｒｈＢＭＰ－２复合人工骨能够引导和诱导新骨形成。     

Repair of cranial bone defects with adipose derived stem cells and coral scaffold in a canine model

Adipose derived stem cells (ASCs) with osteogenic differentiation potential have been documented as an alternative cell source for bone regeneration. However, most of previous in vivo studies were carried out on small animals along with relatively short-term follow-up. In this study, we investigated the feasibility of using ASCs and coral scaffolds to repair a cranial bone defect in a canine model, and followed up the outcome for up to 6 month. Autologous ASCs isolated from canine subcutaneous fat were expanded, osteogenically induced, and seeded on coral scaffolds. Bilateral full-thickness defects (20 mm×20 mm) of parietal bone were created. The defects were either repaired with ASC-coral constructs (experimental group) or with coral alone (control group). Three-dimensional CT scan showed that new bones were formed in the experimental group at 12 weeks post-implantation, while coral scaffolds were partially degraded in the control group. By radiographic analysis at 24 weeks post-transplantation, it was shown that an average of 84.19±6.45% of each defect volume had been repaired in experimental side, while the control side had only 25.04±18.82% of its volume filled. Histological examination revealed that the defect was repaired by typical bone tissue in experimental side, while only minimal bone formation with fibrous connection was observed in the control group. The successful repair of critical-sized bone defects via the current approach substantiates the potentiality of using ASCs with coral scaffolds for bone regeneration.     

富血小板血浆对珊瑚支架上骨髓基质细胞异位成骨的影响

目的：评价富血小板血浆（platelet rich plasma,PRP）对珊瑚支架上骨髓基质细胞（marrow stromal cells,MSCs）异位成骨的影响。方法：将MSCs悬液与同一供体来源的PRP混合后滴加到珊瑚圆片上,形成珊瑚/MSCs/PRP复合物。以同样数量MSCs或PRP制备珊瑚/MSCs和珊瑚/PRP复合物。将3种复合物植入同一裸鼠的背部皮下。术后4周和8周取材,通过组织学观察和组织形态测量分析评价其异位成骨情况。结果：植入后4周或8周,珊瑚/MSCs/PRP组珊瑚表面及其孔洞内有大量的软骨或骨质形成;珊瑚/MSCs组珊瑚表面及其孔洞内也有软骨或骨质形成,并有纤维结缔组织长入;珊瑚/MSCs/PRP组形成的软骨或骨质的量明显多于珊瑚/MSCs组（P〈0.01）;珊瑚/PRP组珊瑚周围及其孔洞内有大量纤维组织,未见骨或软骨形成。结论：PRP促进了珊瑚支架上MSCs的异位成骨。     

二十五昧珊瑚胶囊对继发性脊髓损伤中腺苷的影响

目的探讨二十五味珊瑚胶囊对脊髓损伤大鼠腺苷的影响及在脊髓继发性损伤中的作用。方法选择Wistar大鼠40只,随机分为．Y-常对照组、模型对照组、模型服药l组、模型服药2组,除正常对照组外,其他3组建立大鼠胸椎脊髓腹侧压迫模型,收集脊髓损伤后脊髓组织细胞外液中的腺苷,用高效液相色谱仪紫外检测法检测细胞外液腺苷的含量。结果模型对照组脊髓损伤后腺苷浓度明显高于正常对照组,服用二十五味珊瑚胶囊2周后模型服药1组、模型服药2组脊髓损伤后腺苷浓度高于模型对照组,二者比较有显著性差异。结论二十五味珊瑚胶囊在继发性脊髓损伤中具有提高腺苷水平的作用,从而增加了神经细胞的自我修复能力,这可能是其治疗继发性脊髓损伤的作用机制之一。     

Dynamics and diversity of phosphate mineralizing bacteria in the coral reefs of Gulf of Mannar

Phosphatase Producing Bacteria (PPB) and Inorganic Phosphate Solubilizing Bacteria (IPSB) are important to reef nutrition. These microbes and phosphate concentration play a significant role in the productivity of coral reef ecosystems. A study was conducted in Gulf of Mannar coral reef ecosystem to understand the diversity of these groups of bacteria and their competence in mineralizing the phosphate. The PPB isolates were identified under six genera i.e. Bacillus, Pseudomonas, Micrococcus, Vibrio, Arthrobacter and Brevibacterium. Likewise, the IPSB isolates were also identified that belong to six genera i.e. Bacillus, Arthrobacter, Pseudomonas, Flavobacterium, Flavomonas and Micrococcus. Among the PPB and IPSB strains, Bacillus represented more number of species than others. PPB exhibited maximum activity between pH 8 and 9 and the lowest was at pH 6. Among the phosphatase producers Bacillus cereus (546) showed maximum activity (0.333 mmol/l P) at pH 8. In general, the phosphatase activity in most of the Bacillus species varied with reference to different pH. The species Bacillus megaterium (573) showed highest phosphate solubilizing activity (0.906 mmol/l P) by producing 2-ketogluconic acid. The production of organic acids and phosphatase enzymes by these bacterial groups are responsible for the conversion of insoluble inorganic and organic phosphates into soluble forms which are available for the reef organisms.     

Reproductive effects of the water-accommodated fraction of a natural gas condensate in the Indo-Pacific reef-building coral Pocillopora damicornis

Toxic effects of the water-accommodated fraction (WAF) of a natural gas condensate on the reproduction of the brooding coral Pocillopora damicornis were studied in short-term (24 h) laboratory experiments. Coral fragments were exposed to varying concentrations of condensate WAF during different reproductive phases: gametogenesis, early embryogenesis, and late embryogenesis (when nighttime planulation occurs). During gametogenesis, exposure to condensate WAF did not inhibit subsequent production of larvae. On the other hand, exposure to >25% WAF of gravid corals, at early and late embryogenesis, resulted in abortion and early release of larvae, respectively, with higher percentages of larvae expelled in fragments treated with higher concentrations of condensate WAF at least 3 h after onset of exposure. Aborted larvae during early embryogenesis were ‘premature’, as they are of small size (0.06±0.03 mm³), low metamorphic competency (54%), and white in coloration, with a pale brown oral end (indicating low density of zooxanthellae). Those larvae released at the latter part of embryogenesis are bigger in size (0.22±0.08 mm³), possess 100% metamorphic competency, and are brown in coloration (high density of zooxanthellae). Aside from direct effects on reproduction, fragment mortality index was higher in samples exposed to higher concentrations of condensate WAF (>25%), hence lowering the number of potentially reproducing polyps. Altogether, exposure to >25% natural gas condensate WAF for at least 3 h can potentially disrupt the replenishment of coral populations due to negative effects on reproduction and early life processes.