男科学组织工程研究前景

利用组织工程技术进行细胞移植的设想 ,已经为男科领域的组织再建工作提供了许多可能。为改善、修复或替代现有组织的功能 ,组织工程的应用研究 ,已经在睾丸间质细胞、睾丸假体、阴茎海绵体、阴茎假体等方面开展。虽然大多数再建工作仍然停留在实验阶段 ,但是有些技术也被用于临床 ,并取得满意的结果。本文简要地综述了组织工程在男科学的应用     

The use of tissue expanders in syndactyly repair

The syndactyly repairs of 11 hands in seven patients (average age 20.4) who had simple complete syndactyly between the third and fourth fingers were done by the use of tissue expanders in order to obtain adequate skin closure. At the first stage, a wedge type, 7 cc tissue expander was placed beneath the dorsal skin of the syndactylous digits via a vertical dorsal hand incision under local anesthesia. Inflation was begun on the 15th postoperative day. After the inflation period (average 32 days), syndactyly repair similar to the Littler technique was performed using axillary block anesthesia. During the expansion period one tissue expander became exposed and this hand was repaired by the classical skin grafting technique. In the remaining ten hands the expanded dorsal skin ensured skin closure of the digits and web reconstruction without any need for skin grafting. No complications were encountered during the postoperative period. Active and passive abduction angles, tip-to-tip length, active range of motion of the digits, and two point discrimination tests were done at follow-up (average six months). The web appearances were normal in all patients. Active abduction angles and active range of motion of the digits was always normal at the third postoperative month. Two point discrimination was 8.7 mm on the interdigital sides of the digits and there was no tip-to-tip length differences of the digits. No contractures were seen.Presented at the International Congress of Surgery of the Hand and Upper Extremity, Izmir, Turkey, September 25, 1994.     

Precise and accurate measurement of proton T1 in human brain in vivo: Validation and preliminary clinical application

Precise and accurate inversion-recovery (PAIR) magnetic resonance (MR) measurements of T1 were obtained in eight brain regions and cerebrospinal fluid of 26 healthy volunteers. Accuracy of the technique was assessed by measuring T1 in small fluid volumes with the PAIR technique and with two independent spectroscopic techniques. The mean difference between T1 measured with PAIR and with the two spectroscopic techniques was 3.1% ± 1.3. The precision (reproducibility) of measurements with the PAIR technique was excellent. The coefficient of variation (CV) across 16 measurements in a head phantom was 2.0%, compared with a CV of 2.7% across 45 separate measurements in a single subject. The within-subject CV was 1.8% ± 0.6 in white matter and 1.4% ± 1.0 in basal ganglia. The between-subject CV in 26 healthy volunteers was 3.6% ± 0.6 in white matter and 4.1% ± 1.9 in basal ganglia. Comparison between a patient with an active recurrent brain tumor and an agematched patient with an inactive brain tumor showed that T1 was significantly elevated throughout the brain of the active-tumor patient, especially in white matter tracts, even though no tumor or edema was detected in the white matter on standard MR images. Comparisons between five brain tumor patients and four healthy volunteers of similar age showed that T1 was significantly and substantially elevated throughout the white matter tracts and in the caudate nucleus, putamen, and thalamus. These results are consistent with the hypothesis that white matter tracts are selectively vulnerable to edema and that T1 increases in white matter are a sensitive indicator of patient status or tumor aggressiveness.     

组织型纤溶酶原激活物双单抗夹心ELISA建立及临床应用

本文动用自制ｔ－ＰＡ单克隆抗体建立ｔ－ＰＡ：Ａａ夹心ＥＬＩＳＡ法。结合ｔ－ＰＡ：Ａ、ＰＡＩ：Ａ测定，对５０例正常人，８７例肝病，４７例冠心病和１９例深静脉血栓 成患者的血浆ｔ－ＰＡ：Ａｇ、ｔ－ＰＡ：Ａ、ＰＡＩ：Ａ水平进行了研究。     

采用基因配型行同种异体肾移植术

目的 探讨基因配型技术在同种异体肾移植中的应用。方法采用顺序特异引物聚合酶链反应法(PCR-SSP)分别进行供、受体的基因分型，为3例配型适当的终末性肾病患者行同种异体肾移植术，并观察其术后肾功能恢复情况。结果所有接受肾移植的患者于术后2d肾功能恢复至正常水平，无急性和超急性排异发生。随访11-15个月，各项检查指标均正常。结论 基因配型的准确率较之传统的血清学配型显著增加，显著提高同种异体肾移植的成功率。     

生物衍生骨复合骨髓基质干细胞修复山羊胫骨缺损的血管化研究

目的研究生物衍生骨与骨髓基质干细胞(marrow stromal stem cells, MSCs)复合修复山羊胫骨缺损的血管化过程,了解其修复长段管状负重骨缺损的血管化情况. 方法制备生物衍生骨作为支架材料,培养、诱导MSCs作为种子细胞,二者在体外复合构建组织工程骨.20只山羊双侧胫骨中段制备成20 mm长的骨-骨膜缺损模型,采取自身左右侧对照,实验侧(右侧)缺损处植入组织工程骨,对照侧(左侧)植入单纯支架材料,采用钢板内固定.术后2、4、6及8周用墨汁灌注透明标本及血管面积图像分析方法观察血管化过程,组织学观察血管形成及成骨情况. 结果术后2、4周,实验侧血管形成较对照侧少(P<0.05);术后8周,两侧均完全血管化,差异无统计学意义(P>0.05).实验侧于术后6、8周新骨形成逐渐增加,材料降解吸收较对照组快;对照侧术后8周材料孔隙内仍无明显新骨形成. 结论生物衍生骨作为骨组织工程的支架材料,能够较快发生血管化;组织工程骨成骨能力较单纯支架材料强.     

Ⅱ型糖尿病血浆PAI-1、t-PA检测的意义和结果评价

目的:通过对Ⅱ型糖尿病患者分组(伴血管病变组、胰岛素抵抗组、胰岛素抵抗伴血管病变组、Ⅱ型糖尿病组)检测PAI-1、t-PA抗原及活性,并计算抗原比和活性比,以评价各检测指标的临床价值。方法:采用ELISA法检测PAI-1及t-PA抗原,采用发色底物法检测PAl-1、t-PA活性。结果:189例标本总体患病组及分组后各组的PAI-1活性、t-PA活性及t-PA活性/PAI-1活性比值与正常对照组比较均有显著性差异(P<0.05)。患病组间除t-PA抗原,其余检测显示胰岛素抵抗组、胰岛素抵抗组伴血管病变组与Ⅱ型糖尿病组存在显著性差异(P<0.05)。结论:在上述检测指标中t-PA活性、PAI-1活性反映血管病变的发生较敏感,t PA/PAI-1的活性比值更具临床应用价值。     

Time after excision and temperature alter ex vivo tissue relaxation time measurements

Previously unreported effects of tissue storage were recently observed in the authors' experimental magnetic resonance (MR) studies. To evaluate the effect of elapsed time after excision and storage temperature on tissue relaxation time measurements, tissue samples from the liver, pancreas, kidney, testis, spleen, and brain were obtained in rats. T1 and T2 were first measured within 5 minutes of excision, and between subsequent measurements, tubes were kept in a water bath at 40°C, at room temperature (28°C), or in an ice bath (4°C). Cellular and organellar integrity was assessed with electron microscopy and correlated with the MR findings. At 40°C (20-MHz spectrometer), the T1 of liver decreased from 280 msec ± 8 to 212 msec ± 10 during the first 60 minutes; the T1 of pancreas decreased from 276 msec ± 3 to 208 msec ± 2. Other tissues showed less than a 5% decrease in T1. T2 changes were smaller than T1 changes in all tissues. Electron microscopy of pancreatic acinar cells showed postmortem changes in mitochondria evolving over the first 60 minutes after death. Manganese loading experiments implicated mitochondrial manganese stores in the observed enhanced postmortem decrease in T1. This study calls into question reported relaxation time data for liver and pancreas. MR studies of excised tissues must account for time and temperature to prevent systematic experimental errors.     

藻酸钙凝胶-成骨细胞-骨粉构建工程化骨修复兔颅骨缺损的实验研究

目的探讨藻酸钙凝胶、成骨细胞、骨粉复合构建的可塑形组织工程骨修补兔颅骨缺损后的形态学变化和成骨效果.方法28只日本大耳白兔,随机分为A(n=20)、B(n=8)两组,手术在兔颅顶骨矢状缝两侧分别各建立一个直径1cm的圆形全层缺损,用两种方法藻酸钙凝胶、成骨细胞、骨粉和藻酸钙凝胶、骨粉分别构建组成可塑形的组织工程骨复合材料,分别填补修复A组兔颅骨左右两侧的缺损,B组为空白对照组,通过大体、组织学、X线观察材料的形态变化、成骨情况,并对X线片和组织学切片进行评分.结果材料植入后,局部未见红肿、积液、渗出等异常反应.①藻酸钙凝胶-成骨细胞-骨粉组修补后12周颅骨缺损基本被硬性组织所修复,镜下见修复材料大多被骨组织替代,骨粉基本被吸收,有块状凝胶残留其中,组织学评分为(5.50±1.00).X线片见兔颅骨缺损处有高密度骨痂影存在,布满整个缺损区,X线片评分为(3.25±0.95).②藻酸钙凝胶-骨粉组修补后12周部分颅骨缺损被硬性组织所修复,镜下见修复材料部分转变成骨组织,骨粉基本被吸收,有凝胶残留其中,组织学评分为(3.25±1.50).X线片见兔颅骨缺损处有高密度骨痂影存在,主要分布在缺损区的边缘部位,X线片评分(2.25±0.25).③空白对照组骨缺损主要被膜样纤维组织修复,在紧邻骨缺损边缘处有硬性组织形成,镜下见修复组织边缘有骨组织存在,中央大部为膜状致密纤维组织,组织学评分为(1.50±0.50),X线片仅见靠近骨缺损边缘的部位存在致密骨痂影,X线片评分为(1.00±0.57).结论藻酸钙凝胶、成骨细胞、骨粉构建的可塑形组织工程骨可根据颅骨缺损的形态进行塑形填补,在体内有良好的成骨能力,可达到对兔颅骨缺损的骨性修复,但部分藻酸盐凝胶吸收缓慢,手术后12周仍不能满意吸收.     

骺板软骨细胞复合三维支架体外构建组织工程软骨的研究

目的探讨将骺板软骨细胞复合三维支架经体外培养,构建组织工程软骨的效果及其生物学特点. 方法将3周龄幼兔第1代骺板软骨细胞与液态的生物凝胶混合,接种于聚磷酸钙纤维/L-聚乳酸(CPPF/PLLA)三维支架材料,构建组织工程软骨组织块,连续培养4周.行大体、倒置显微镜及组织学、Ⅰ型和Ⅱ型胶原免疫组织化学光镜观察,定量检测硫酸糖胺多糖(GAG)含量. 结果构建的组织工程软骨块在培养过程中能保持其初始外形,种子细胞呈稳定的三维均相分布,外观逐渐呈乳白色、半透明,硬度亦不断增加.培养1周有软骨细胞陷窝形成,2周后形成富含Ⅱ型胶原和蛋白聚糖、具有典型软骨组织结构的工程化软骨,且Ⅰ型胶原逐渐转为阴性.4周时构建软骨的组织结构与天然骺板软骨相类似,硫酸GAG含量平均为天然骺板软骨的34%以上. 结论骺板软骨细胞复合三维支架体外培养可生成典型软骨,且可形成类似天然骺板软骨的组织结构,能满足修复骺板缺损的基本要求.体外培养1～2周可能是植入体内修复骺板缺损的较佳时机.