人异体骨软骨移植物保存方法研究

[目的]探讨六种保存方法对人关节软骨组织结构和细胞活性的影响,寻求一种保存效果较好的方法,为临床提供一种有活性的异体骨软骨移植物.[方法]自捐献新鲜尸体膝关节利用专用手术器械获取4.5 mm ×4.5 mm大小的人骨软骨块,分别采用梯度降温法、Co60射线照射+梯度降温法、玻璃化法、连续降温法、直接液氮法和酒精浸泡法对软骨块进行保存处理,分别于保存第8、15、30、60d时,采用蕃红-O组织染色、扫描电镜、软骨细胞胎盼蓝染色、MTT法等,观察并比较以上6种方法保存后关节软骨细胞存活率及其代谢功能变化.[结果]除了酒精保存方法外,其余保存方法随着时间延长软骨组织的细胞成活率和细胞代谢活性逐渐降低;保存60d时,采用玻璃化法保存软骨组织的细胞存活率为62.47％,软骨基质成分丢失较少,胶原纤维断裂不明显;采用慢速梯度降温法保存软骨组织的细胞存活率为59.75％,软骨基质成分丢失较多,胶原纤维断裂明显;其他保存方法软骨细胞存活率不足40％,软骨基质成分大量丢失,胶原纤维杂乱.[结论]六种保存方法中,玻璃化保存法能够较好的保存关节软骨,活性最好,其次是梯度降温保存法,两者均具有一定临床价值.Co60射线照射对软骨细胞有一定的损伤作用;酒精浸泡不能保存软骨细胞活性.     

自体软骨移植矫正鼻小柱退缩

目的 探索一种通过调整鼻小柱与鼻翼位置的关系来矫正鼻小柱退缩的方法,形成正面观"海鸥翼"状的鼻下段美学曲线.方法 采取自体软骨移植支撑鼻小柱,同时行鼻中隔延伸及韧带分离技术,使鼻小柱向下延伸,并对鼻小柱短小及鼻尖突出度不足进行矫正.结果 对18例鼻小柱内陷缺陷的患者进行治疗,术后随访6～24个月,满意率为83%.结论 联合应用自体软骨移植和鼻尖区软骨重构,可以有效矫正鼻小柱退缩.     

组织工程软骨移植修复兔膝关节软骨缺损

研究用组织工程的方法进行差了软骨缺损修复的基本原理。方法取新生兔关节－干骺复后体软骨,胶原酶消化,将所获软骨细胞种入９６孔板内几丁质纤维无纺布上。结果培养２１天时形成“膜状软骨”,至１１０天时形成直径为４．４ｍｍ的“圆盘状软骨”。经Ｓａｆｒａｎ“Ｏ”ｉｖｓ ｑｃ ｙｇｈ ｐｕ 其基质富含蛋白多糖,原位杂交法 产其软骨细胞表达Ⅱ型胶原ｍＲＮＡ。将培养２１天的“膜状软骨”移植于成年兔膝关节圆形全厚缺损     

自体骨软骨移植修复软骨缺损的研究

把非负重区正常软骨移植到负重区病损软骨部位，让其在受区发挥作用、改善关节功能、减轻患者痛苦是目前研究的热点。拟在关节镜监视下，取非负重区软骨自体移植替代负重区病变软骨．探讨自体骨软骨移植修复软骨缺损的可行性、效果及并发症。     

自体镶嵌式骨软骨移植修复关节软骨损伤

关节软骨的修复能力极其有限，一旦发生损伤，很难自行修复．最终会发生不可逆的病理改变，演变为创伤性关节炎或骨性关节炎，严重影响患者的生活质量：目前，治疗关节软骨损伤的方法主要有软骨下骨板钻孔术、微骨折术、关节削磨成形术、骨膜软骨膜移植术、自体软骨细胞移植术，并且有学者运用电流、电磁波、激光、超声、微波等刺激软骨细胞增殖，虽然取得了一定的疗效，但这些方法均不能产生形态、功能、力学性质和正常软骨一样的修复组织。     

关节镜下自体骨软骨移植修复股骨关节面软骨缺损

目的探讨关节镜下自体骨软骨镶嵌式移植术治疗股骨负重面关节软骨缺损的可行性。方法2001年6月起,共17例股骨关节面软骨缺损患者,男12例,女5例;年龄18～45岁,平均29岁;左膝10例,右膝7例。按照Brittberg-Peterson功能评定标准,膝关节的功能为65～105分,平均(80.65±9.69)分。3例无明显外伤史,但有风湿病史;14例有外伤史,均有膝关节疼痛,大腿肌肉萎缩。3例伴绞锁,2例伴弹响。14例外伤患者为股骨外髁负重面局灶性软骨损伤,均为单个创面,损伤范围2.5～3.0cm2;3例无外伤患者为股骨内髁负重面局灶性软骨软化或软骨剥脱,损伤范围2.0～2.5cm2。采用镶嵌式骨软骨移植器,在关节镜下取膝关节非负重关节面骨软骨条,将之移植修复膝关节负重面的局灶性软骨缺损。结果术后随访10～20个月,平均15个月,患者临床症状消失,关节活动度正常。术后Brit-tberg-Peterson评分,14例为0分,3例因活动时轻微疼痛分别评为3分和2分。MRI显示软骨缺损区软骨表面平整,移植的骨软骨柱位置良好。结论自体骨软骨镶嵌式移植术对关节负重面局灶性软骨缺损有较好、确切的治疗效果。     

跖趾关节软骨移植修复掌指关节面缺损

目的　利用自体跖趾关节软骨移植修复掌指关节缺损的软骨面,为掌指关节创伤性关节炎提供简便、可靠的方法。方法　1990 年以来,应用改良处理后的自体跖趾关节面软骨移植修复9 例11 侧掌指关节面,通过内固定、侧副韧带修复并提供适宜的生物力学和营养环境。结果　经6 个月7 年随访,关节活动度平均增加35-1°,供趾骨融合良好,成形术后关节活动度减少甚至强直,但无疼痛,对行走无明显影响。结论　软骨面匹配良好,可靠的内固定、重建韧带、供区薄的软骨面和少的骨量及适宜的生物力学环境是手术成功的关键     

同种异体软骨移植修复关节软骨缺损实验研究

目的 采用兔膝关节软骨标本经打孔梯度降温冻存后行同种异体移植,研究打孔梯度降温冻存对兔关节软骨的影响及其修复关节软骨缺损的效果.方法 自16只2月龄新西兰白兔膝关节股骨髌面取分别取3块骨软骨移植物,随机分为3组.Ⅰ组为实验组,在软骨上以3 mm×3 mm矩阵打孔,Ⅱ、Ⅲ组为对照组,不打孔.分别将软骨标本置于二甲基亚砜冷冻保护溶液中,并经梯度降温至-80℃(Ⅰ、Ⅱ组)或直接置于-80℃(Ⅲ组)保存1周,复温后移植到成年兔相应膝关节部位.术后分批处死动物,通过对移植物的大体形态学、组织学、免疫组化染色光镜观察,研究各组移植物保存效果的差异.结果 Ⅰ、Ⅱ组光镜观察结果明显优于Ⅲ组;Ⅰ组与Ⅱ组结果差异不明显,但Ⅰ组对中间层软骨组织的保护明显加强.结论 关节软骨的梯度降温冷冻保存效果明显优于快速降温冷冻保存,且关节软骨打孔冷冻保存对深层软骨细胞有一定的保护作用,可提高软骨细胞存活率,延缓移植软骨组织的退变过程.     

Allogenous Cartilage Graft Versus Autogenous Cartilage Graft in Augmentation Rhinoplasty: A Decade of Clinical Experience

Cartilage grafts have great value in augmentation rhinoplasty. For most surgeons, an autogenous cartilage graft is the first choice in rhinoplasty because of its resistance to infection and resorption. On the other hand, an allogenous cartilage graft might be preferred over an autogenous graft to avoid additional morbidity and lengthened operating time. Allogenous cartilage grafts not only have the advantage of averting donor site morbidity but also are resistant to infection, resembling autogenous cartilage grafts. The authors present their experience with 41 patients who underwent augmentation rhinoplasty using 22 autogenous and 19 allogenous cartilage grafts between June 1994 and August 2004. For evaluation of adequate augmentation rates, photographic analyses were performed on preoperative, early postoperative, and late postoperative photographs from all the patients. To assess patient satisfaction, the Facial Appearance Sorting Test (FAST) was applied preoperatively and late postoperatively in both groups. These results were compared, and it was concluded that in terms of resorption, there was no difference in the early and late postoperative follow-up data between allogenous and autogenous cartilage grafts. Evaluation of the preoperative and early postoperative photographic outcomes showed statistically significant differences with respect to adequate augmentation rates between the two groups. The FAST scores showed statistically significant differences between preoperative and late postoperative outcomes. There were no infections in the two groups of patients. Commentary to DOI: .     

Autologous Cartilage Graft Rhinoplasties

Background: The authors report their experience with autologous graft rhinoplasties. Methods: Data were collected retrospectively, with selection of only autologous grafts from 2,000 rhinoplasties performed at the Plastic and Reconstructive Department of the University of Rome Tor Vergata. Results: A total of 62 patients from January 1995 to January 2005 were selected. Most of the patients were treated with the “open tip” technique, whereas 9.7% had a classic endonasal approach. Follow-up evaluation was performed with outpatient visits at 2 and 6 weeks, then at 3, 6, and 12 months. Good aesthetic results were obtained for 93.5% of the patients, and 83.7% had complete satisfaction. Conclusion: Autologuos cartilage graft rhinoplasty is an affordable technique easy to learn that widens possibilities of interventions for nasal pyramid reconstruction.     

软骨组织工程的研究进展

目前，软骨组织工程已在基础研究方面取得了令人瞩目的成果，并在临床应用方面进行了初步尝试，为I临床软骨缺损的修复提供了新的思路与途径，已经成为该领域主要的研究方向，但其大规模临床应用尚存在诸多困难。现结合近年的研究成果，就组织工程化软骨的种子细胞、生物支架材料、体内外构建以及临床应用等方面进行综述。     

Cartilage chips synthesized with fibrin glue in rhinoplasty

A new type of cartilage graft that is synthesized from cartilage fragments minced by our instrument and mixed with fibrin glue is described. This new material is useful when particular grafts are required in primary or secondary rhinoplasty.     

Bone Chips and Diced Cartilage: An Anatomically Adopted Graft for the Nasal Dorsum

Since 1996, cranial bone chips or septal bone chips harvested during septal deviation surgery and small chips of ear or septal cartilage have been used in 67 patients for dorsal nasal augmentation or for smoothing dorsal nasal irregularities. In this study, 59 overresections of ostecartilaginous nose structures during previous aesthetic nose surgeries and 8 primary rhinoplasties occasioned the use of bone or cartilage grafts. For 57 patients both bone and ear cartilage grafts were used for the reconstruction. Bone grafts were used for seven cases and cartilage grafts for three cases.The results from 7 years, of experience with this method of nasal dorsum reconstruction were satisfactory and durable. The most important advantage of this method is that the bony side of the nose is reconstructed with bone and the cartilage side with cartilage. Another advantage is that the bone chips are incorporated with both nasal bones, building a strong dorsal nasal bony monoblack.This technique also is useful for augmenting mild saddle nose deformity and dorsal nasal projection deficiency on the bony part, cartilage part, or both parts.Presented at 16th Congress of ISAPS, Istanbul, Turkey, 26–29 May 2002 and 24th National Meeting of the Turkish Plastic Reconstructive Aesthetic Surgery Society, Ankara, Turkey, 18–20 October 2002.     

Radiologic and Histologic Assessment of Diced Cartilage Grafts for Cranial Bone Defects of Rabbits: An Experimental Study

This study was performed to experimentally evaluate the viability and coverage of diced cartilage grafts for cranial defects. Biparietal bone defects were prepared in each of 20 rabbits. Otogenous bone grafts were fixed to one side, whereas cartilage grafts taken from the right ear, diced, and wrapped with oxidized regenerated cellulose (Surgicel) were placed on other side. Parenchymal impression, contour, and ossification of all grafts in the 16 rabbits surviving after 8 weeks were evaluated with computed tomography and magnetic resonance imaging. The existence of ossification was examined pathologically. Parenchymal impression was significantly more frequent in bone grafts than in cartilage grafts (p < 0.05). This difference probably is attributable to the application technique. There was no significant radiographic difference in other parameters between the two groups. No significant difference in ossification was found. In conclusion, ossification was seen radiologically and pathologically in diced cartilage grafts. Because of autogenicity, ease of shaping, absence of postoperative warping, reduced resorption rates, and inherent viability and ossification properties, the authors suggest that diced cartilage grafts may be an alternative material for cranial defects, especially for frontal cranial reconstruction in which deformities may cause aesthetic and functional disabilities. The authors believe that further long-term studies also are needed.     

Ex Vivo Perfusion for Accurate Assessment of Liver Graft Viability in Dogs

A New type of ex vivo liver perfusion model (E VPM) was developed. The system includes a physiological porto-portal connection, using a centrifugal pump and an aorto-aortal connection, through a heparin-coated tube. Ten pairs of mongrel dogs, weighing 13–20 kg, were used. Grafts of the liver procured from donors were perfused on the EVPM for 3 h. These grafts were divided into two groups: group A (n = 5), nonpreserved graft; group B (n = 5), 8-h graft preserved with lactated Ringer's solution (4°C). The orthotopic liver transplantation (OLTx) series with the nonpreserved graft was reviewed, and dogs surviving for over 7 days following OL Tx, group C (n = 5), were selected for liver grafts in the actual OL Tx. In comparison with groups A and C, no difference was noted among enzyme levels (sGOT sGPT, LDH) or in the recovery rate of A TP content in the graft liver tissue after revascularization. There were significant differences between group A and group B after revascularization. This simple and physiological EVPM accurately reflects graft function and hemodynamics in the actual OLTx. Graft viability, as a substitute for OLTx, in large animals can be reliably assessed. This EVPM is expected to contribute to research on events related to liver transplantation.     

Egr-1在自体移植静脉中的表达及意义

目的研究自体静脉移植后早期生长反应基因-1(Egr-1)表达的动态变化,探讨其在内膜增生中的作用。方法Wistar大鼠90只,将大鼠右颈总静脉端-端吻合于肾下腹主动脉建立自体静脉移植模型。术后随机分别于1、2、6和24h,3、7、14、28及42d相应时间处死动物取移植静脉,同时取正常静脉作对照。应用原位杂交和RT-PCR方法检测Egr-1mRNA的表达,联合应用Western蛋白印迹和免疫组织化学方法检测Egr-1蛋白表达情况,同时进行组织形态学观察。结果自体静脉移植后,Egr-1mRNA和Egr-1蛋白的表达呈双相变化,即移植后1h,Egr-1mRNA表达迅速升高,阳性率为(35±7)%,6h、24h及3d时下降到较低水平,阳性率分别为(8±2)%、(8±6)%和(8±4)%,7d时又再升高,28d时达高峰,阳性率为(45±6)%,此与其余各时点比较差异均有统计学意义(P<0.01),42d时,Egr-1mRNA的表达再次下降;移植早期(2h)即有Egr-1蛋白的表达,阳性率为(30±5)%,并持续至6h,24h~3d表达下降到较低水平,阳性率分别为(7±3)%和(7±8)%,7d时又再升高,至移植后28d,Egr-1蛋白的表达阳性率达到高峰,为(40±9)%,此与其余各时点比较差异有统计学意义(P<0.01)。移植后7d,免疫组化结果显示,Egr-1蛋白表达主要位于中膜血管平滑肌细胞(VSMCs)和单核细胞/巨噬细胞,移植后期28d,Egr-1蛋白表达主要位于新生内膜和中膜的VSMCs,同时部分内皮细胞也有Egr-1蛋白的表达。结论移植静脉内膜增生与Egr-1的激活及表达关系密切,Egr-1可能成为防治移植静脉内膜增生、狭窄及闭塞的一个新的干预靶点。