The Studies towards Prevention of Osteoarthritis of the knee joint Using Adenovirus Vector Encording Hyaluronic Acid Synthe tase Type-2 Gene

1、通过长寿医学科学中心提供编码透明质酸合成酶2型基因的腺病毒载体（HAS 2病毒），并通过直接注射到关节来了解该病毒是否具有预防骨关节炎（OA）的作用。将病毒载体注射到膝关节或兔OA模型中。然后，在HAS病毒注射后6周，评价关节液中透明质酸（HA）浓度、关节软骨的大体观察结果和组织学。结果表明，HA病毒注射组（HA病毒组）关节液中HA浓度与野生型病毒注射组（野生组）和载体载体注射组（载体组）无统计学差异。肉眼和组织学检查也显示三组之间无明显差异。然后，我们确定HAS 2病毒是否具有加速正常（非OA）兔膝关节HA合成的作用。HAS2病毒注射入 ...更多信息 家兔正常膝关节。结果表明，HAS 2病毒组血凝素浓度有略高于野生组，但低于携带组的趋势。这些结果表明，可能存在一种负反馈机制或病毒浓度太低而不能表现出HA合成的加速作用。2、我们将体内实验策略改为体外实验策略，以关节软骨细胞为研究对象。HA在软骨组织中也有重要作用，软骨细胞自身合成HA。将HAS 2病毒感染胎牛关节软骨分离的软骨细胞，在5个时间点收集条件培养基，直至7 d。HAS 2病毒感染软骨细胞的条件培养基中HA浓度在任何时间点都高于未感染软骨细胞，直到7天。证实了HAS病毒对体外培养的关节软骨细胞具有促进HA合成的作用。这种加速作用可用于关节软骨的组织工程技术或预防OA进展。少

1, Through the courtesy of the Medical Science Center of longevity, the adenovirus vector encording hyaluronic acid synthetase type-2 gene (HAS2 virus) was presented and used to know whether the virus had the prevention effects or osteoarthritis (OA) by injection directly to the joint. The virus vector was injected in the knee joints or rabbit OA model. Then, hyaluronic acid (HA) concentration in joint fluid, macroscopic findings and histology of articular cartilage were evaluated at 6 weeks after HAS virus injection. The result showed that HA concentration in joint fluid of HA virus injected group (HA virus group) had no statistical difference between wild type virus injected group (wild group) and carrier vehicle injected group (carrier group). Macroscopic and histological examination also showed no apparent difference between three groups. Then, we ascertained whether HAS2 virus had the acceleration effects of HA synthesis for normal (non-OA) rabbit knee. HAS2 virus was injected in … More the rabbit normal knee joints. The results showed that HA concentration of HAS2 virus group had the tendency towards a little bit higher than that of wild group, but lower than that of carrier group. Thus, these results indicated that a kind of negative feedback mechanisms might affect the results or the concentration of the virus was too low to express the acceleration effects of HA synthesis.2, We changed the strategy from in vivo study to in vitro study, in which articular chondrocytes were used. HA has some important roles also in cartilage tissue, and chondrocytes themselves synthesize HA.HAS2 virus was infected to the chondrocytes isolated from fetal cow articular cartilage, and condition medium was collected at 5 time points till 7 days. HA concentration in condition medium of HAS2 virus infected chondrocytes was higher than that of non-infeced chondrocytes at any time points till 7 days. Therefore it was verified that HAS virus had the acceleration effects of HA synthesis for articular chondrocytes in vitro. This acceleration effects can be used for tissue engineering technique of articular cartilage or prevention of OA progression. Less