目的 研究海螵蛸-骨形态发生蛋白(BMP)复合材料对骨缺损大鼠成骨及再血管化的影响. 方法 将海螵蛸去外壳,制成圆柱体(直径约5 mm,高约2mm),浸泡于重组人骨形态发生蛋白2溶液中制作海螵蛸-BMP复合材料.将30只SD大鼠制成颅骨缺损模型,按照随机数字表法分成海螵蛸-BMP组与单纯海螵蛸组,每组15只,分别植入相应材料修复颅骨缺损.移植术后4、6、8周,2组各取5只大鼠,分批放血处死同时行墨汁灌注.ld后取出全部植入物和周围部分颅骨同时进行植入区大体观察,制作切片行HE及Masson染色,通过组织病理学技术、图像采集系统,观察、分析微血管面积和新生骨面积.对数据行两因素析因设计方差分析和LSD检验,并对海螵蛸-BMP组大鼠微血管面积和新生骨面积的相关性行Pearson相关分析. 结果 (1)植入区大体观察可见植入物被纤维结缔组织包膜包裹,移植术后4周包膜质地柔软、较厚;移植术后6、8周包膜质韧、薄,但较致密.植入物移植术后4周时转为胶冻样物质,移植术后6、8周时与软骨组织类似.(2)组织学观察显示,移植术后4周,2组大鼠植入物结构均遭到破坏,并可见中等数量的炎性细胞浸润;海螵蛸-BMP组初级骨小梁、微血管数量明显多于单纯海螵蛸组,破骨细胞数量则少于单纯海螵蛸组.移植术后6周,2组大鼠植入物炎性细胞明显减少,海螵蛸进一步降解,新生微血管更加丰富,植入物周边均形成明显的成熟骨小梁,但海螵蛸-BMP组在降解后的海螵蛸-BMP中也出现了成熟骨小梁.移植术后8周,2组大鼠植入物炎症反应消失,成熟骨小梁显著增多,海螵蛸的网格状结构基本被破坏,单纯海螵蛸组的成熟骨小梁仍分布在海螵蛸周边;而海螵蛸-BMP组中,海螵蛸-BMP中央、周围均可见成熟骨小梁,微血管数量仍多于单纯海螵蛸组.(3)移植术后4～8周,海螵蛸-BMP组微血管面积[(63±4)、(136 ±36)、(347±31) μm2]大于单纯海螵蛸组[(44±7)、(73±4)、(268±42) μm2,P＜0.05或P＜0.01];新生骨面积[(236±26)、(339 ±42) 、(553 ±40)μm2]大于单纯海螵蛸组[(137±15)、(243 ±21)、(445±29) μm2,P值均小于0.01].(4)海螵蛸-BMP组微血管面积与新生骨面积呈显著正相关(r =0.948,P=0.00l) 结论 海螵蛸-BMP复合材料对骨缺损大鼠具有较好的成骨及再血管化作用,是骨组织工程优良的三维支架和载体材料
目的 研究海螵蛸-骨形態髮生蛋白(BMP)複閤材料對骨缺損大鼠成骨及再血管化的影響. 方法 將海螵蛸去外殼,製成圓柱體(直徑約5 mm,高約2mm),浸泡于重組人骨形態髮生蛋白2溶液中製作海螵蛸-BMP複閤材料.將30隻SD大鼠製成顱骨缺損模型,按照隨機數字錶法分成海螵蛸-BMP組與單純海螵蛸組,每組15隻,分彆植入相應材料脩複顱骨缺損.移植術後4、6、8週,2組各取5隻大鼠,分批放血處死同時行墨汁灌註.ld後取齣全部植入物和週圍部分顱骨同時進行植入區大體觀察,製作切片行HE及Masson染色,通過組織病理學技術、圖像採集繫統,觀察、分析微血管麵積和新生骨麵積.對數據行兩因素析因設計方差分析和LSD檢驗,併對海螵蛸-BMP組大鼠微血管麵積和新生骨麵積的相關性行Pearson相關分析. 結果 (1)植入區大體觀察可見植入物被纖維結締組織包膜包裹,移植術後4週包膜質地柔軟、較厚;移植術後6、8週包膜質韌、薄,但較緻密.植入物移植術後4週時轉為膠凍樣物質,移植術後6、8週時與軟骨組織類似.(2)組織學觀察顯示,移植術後4週,2組大鼠植入物結構均遭到破壞,併可見中等數量的炎性細胞浸潤;海螵蛸-BMP組初級骨小樑、微血管數量明顯多于單純海螵蛸組,破骨細胞數量則少于單純海螵蛸組.移植術後6週,2組大鼠植入物炎性細胞明顯減少,海螵蛸進一步降解,新生微血管更加豐富,植入物週邊均形成明顯的成熟骨小樑,但海螵蛸-BMP組在降解後的海螵蛸-BMP中也齣現瞭成熟骨小樑.移植術後8週,2組大鼠植入物炎癥反應消失,成熟骨小樑顯著增多,海螵蛸的網格狀結構基本被破壞,單純海螵蛸組的成熟骨小樑仍分佈在海螵蛸週邊;而海螵蛸-BMP組中,海螵蛸-BMP中央、週圍均可見成熟骨小樑,微血管數量仍多于單純海螵蛸組.(3)移植術後4～8週,海螵蛸-BMP組微血管麵積[(63±4)、(136 ±36)、(347±31) μm2]大于單純海螵蛸組[(44±7)、(73±4)、(268±42) μm2,P＜0.05或P＜0.01];新生骨麵積[(236±26)、(339 ±42) 、(553 ±40)μm2]大于單純海螵蛸組[(137±15)、(243 ±21)、(445±29) μm2,P值均小于0.01].(4)海螵蛸-BMP組微血管麵積與新生骨麵積呈顯著正相關(r =0.948,P=0.00l) 結論 海螵蛸-BMP複閤材料對骨缺損大鼠具有較好的成骨及再血管化作用,是骨組織工程優良的三維支架和載體材料
목적 연구해표소-골형태발생단백(BMP)복합재료대골결손대서성골급재혈관화적영향. 방법 장해표소거외각,제성원주체(직경약5 mm,고약2mm),침포우중조인골형태발생단백2용액중제작해표소-BMP복합재료.장30지SD대서제성로골결손모형,안조수궤수자표법분성해표소-BMP조여단순해표소조,매조15지,분별식입상응재료수복로골결손.이식술후4、6、8주,2조각취5지대서,분비방혈처사동시행묵즙관주.ld후취출전부식입물화주위부분로골동시진행식입구대체관찰,제작절편행HE급Masson염색,통과조직병이학기술、도상채집계통,관찰、분석미혈관면적화신생골면적.대수거행량인소석인설계방차분석화LSD검험,병대해표소-BMP조대서미혈관면적화신생골면적적상관성행Pearson상관분석. 결과 (1)식입구대체관찰가견식입물피섬유결체조직포막포과,이식술후4주포막질지유연、교후;이식술후6、8주포막질인、박,단교치밀.식입물이식술후4주시전위효동양물질,이식술후6、8주시여연골조직유사.(2)조직학관찰현시,이식술후4주,2조대서식입물결구균조도파배,병가견중등수량적염성세포침윤;해표소-BMP조초급골소량、미혈관수량명현다우단순해표소조,파골세포수량칙소우단순해표소조.이식술후6주,2조대서식입물염성세포명현감소,해표소진일보강해,신생미혈관경가봉부,식입물주변균형성명현적성숙골소량,단해표소-BMP조재강해후적해표소-BMP중야출현료성숙골소량.이식술후8주,2조대서식입물염증반응소실,성숙골소량현저증다,해표소적망격상결구기본피파배,단순해표소조적성숙골소량잉분포재해표소주변;이해표소-BMP조중,해표소-BMP중앙、주위균가견성숙골소량,미혈관수량잉다우단순해표소조.(3)이식술후4～8주,해표소-BMP조미혈관면적[(63±4)、(136 ±36)、(347±31) μm2]대우단순해표소조[(44±7)、(73±4)、(268±42) μm2,P＜0.05혹P＜0.01];신생골면적[(236±26)、(339 ±42) 、(553 ±40)μm2]대우단순해표소조[(137±15)、(243 ±21)、(445±29) μm2,P치균소우0.01].(4)해표소-BMP조미혈관면적여신생골면적정현저정상관(r =0.948,P=0.00l) 결론 해표소-BMP복합재료대골결손대서구유교호적성골급재혈관화작용,시골조직공정우량적삼유지가화재체재료
Objective To study the effects of cuttlefish bone-bone morphogenetic protein (BMP)composite material on osteogenesis and revascularization of bone defect in rats.Methods The cuttlefish bone was formed into cylinder with the dianeter of about 5 mm and height of about 2 mm after the shell was removed,and then it was soaked in the recombinant human BMP 2 to make a cuttlefish bone-BMP (CBB)composite material.Thirty SD rats,with a defect of skull in every rat,were divided into the CBB and pure cuttlefish bone (PCB) groups according to the random numbertable,with 15 rats in each group.The rats in the group CBB and group PCB were transplanted with the corresponding material to repair the skull defect.At post transplantation week (PTW) 4,6,and 8,5 rats from every group were sacrificed by exsanguination,and ink perfusion was performed.One day later,all the transplants and part of the skull surrounding the defect were harvested,and general observation was conducted at the same time.The specimens were paraffin sectioned for HE staining and Masson staining.The area of microvessel and the area of newborn bone were observed and analyzed through histopathological techniques and image collection system.Data were processed with the analysis of variance of factorial design and LSD test.The correlation between the area of microvessel and the area of newborn bone of the group CBB was analyzed with Pearson correlation analysis.Results (1) The general observation of the transplant region showed that the transplants were encapsulated by a capsule of fibrous connective tissue.The texture of capsule was soft and relatively thick at PTW 4.The texture was tenacious and thin,but rather compact at PTW 6 and 8.The transplants became gelatinous at PTW 4,and similar to the cartilage tissue at PTW 6 and 8.(2) Histological observation showed that the structure of the transplants in two groups was damaged at PTW 4.A moderate quantity of inflammatory cell infiltration could be observed.The amounts of the primary bone trabeculae and microvessels in group CBB were more abundant than those of group PCB,while the number of osteoclasts was less than those of group PCB.At PTW 6,the inflammatory cell infiltration in the transplants in both groups decreased obviously,the cuttlefish bone was found to be further degraded,and the number of newborn microvessels was increased.There were mature bone trabeculae around the transplants in both groups.And there were also mature bone trabeculae in the degraded CBB in group CBB.At PTW 8,the inflammatory reaction in the transplants in both groups disappeared; there were more mature bone trabeculae; the structure of the cuttlefish bone was found to be damaged basically.Bone trabeculae in group PCB were found around the transplant,while the bone trabeculae could be observed not only around the transplant but also in the degraded CBB in group CBB.The amount of the microvessels in group CBB was still larger than that of group PCB.(3) From PTW 4 to 8,the area of microvessel in group CBB [(63 ±4),(136 ±36),(347 ±31) μm2] was larger than that in groupPCB [(44±7),(73 ±4),(268 ±42) μm2,P ＜0.05 orP ＜0.01].FromPTW4to8,the area of newborn bone in group CBB [(236 ±26),(339 ±42),(553 ±40) μm2] was larger than that in group PCB [(137±15),(243 ±21),(445 ±29) μm2,with P values all below 0.01].(4) The relation between the area of microvessel and the area of newborn bone was significantly positive (r =0.948,P =0.001).Conclusions The CBB may exert good effect on osteogenesis and vascularization of rats with bone defect.It is a good three dimensional scaffold in bone tissue engineering.
