以植入 S.D.rat*s 大腿肌肉模式評估豬皮膠原蛋白膜誘發之組織病理變化 與免疫反應

1. 以植入S.D.rat*s大腿肌肉模式評估豬皮膠原蛋白膜誘發之組織病理變化 與免疫反應 本研究之目的為針對已改良之豬皮膠原蛋白膜（porcine dermal collagen membrane--PDCM），經四種不同濃度(0%,0.01%, 0.05%, 3%)之 戊二醛（GA）交鏈一小時後之成品，以Sprague-Dawley rats(S.D. rats) 實驗動物模式植入其肌肉層內，經 1,3,5,7,10,14,21,28,42日，共九個 階段後，利用傳統 Hematoxylin & Eosin stain(H-E stain), Alcian blue－PAS(AB－PAS) stain，及免疫染色等技術，評估豬皮膠原蛋白膜之 活體吸收速率及所引發組織病理變化與免疫生成性(immunogenecity)。結 果發現：PDCM之活體吸收速率：0％ GA-PDCM<三週內被吸收0.01％ GA- PDCM<六週內被吸收0.05％及3％ GA-PDCM>六週內仍未被完全吸收。 而PDCM維持細胞隔離效果之時間則為： 3% GA-PDCM可維持21日細胞隔離 ；而0. 05%、0.01%、0% GA-PDCM 僅可維持七?十日 就被細胞及新生微 血管增生、穿透。 在組織病理變化方面：豬皮膠原蛋白膜會誘發S.D. rats出 現慢性異物反應，並逐漸發展為DTH反應；其激烈程度隨 著膜片 存在時間延長而加劇，亦隨著膜片被吸收而減緩。 亦即為3％GA-PDCM之 異物反應最持久且能發展較為較 成熟的DTH反應。 而經由免疫染色證實 ：豬皮膠原蛋白膜確實會誘發以 macrophage 主導之DTH反應。且此反應 與CD4 T lymphocytes 相關，而與CD 8 T lymphocytes 無關；且植 入 區域內之肌肉及修復膜片之結締組織並未出現抗體 (IgG,IgM)大量沉積於 此處，表示PDCM 誘發type II hypersensitivity 之可能性極低。 因 此推測PDCM所引發之免疫反應屬於Thl cell主導之反應。若應用PDCM於牙 周治療，則除了可能具有引導組織再生之效果外；且可能以Th1 cell 分 泌之IL-2、IFN-r等cytokines，使發炎牙周組織朝向Thl主導之組織修復 的方向進行?

2. Assays of Histopathologic Change and Immune Response Induced by Porcine Dermal Collagen Membrane, Evaluated by S.D. Rats Animal Model • 。The histopathologic change and immune response, induced by porcine dermal collagen membrane（PDCM）, was evaluated by Sprague-Dawley rats animal model. The PDCM, which was cross- linked by 3%, 0.05%, 0.01%, or 0% glutaraldehyde (GA) for 1 hour, was implanted into muscle tissue of S.D. rats. Following 1, 3, 5, 7, 10, 14, 21, 28, 42 days, the rats were sacrificed； PDCM with the surroundingmuscle tissue in implanted sites were prepared for (1) Hematoxylin & Eosin stain, (2) Alcian blue-PAS stain, (3) immunohistochemistry stain（avidin-biotin complex method）. According to microscropic observation, the resorption rates of PDCM in vivo were: 0% GA-PDCM less than 3 weeks, 0.01% GA-PDCM less than 6 weeks, 0.05% and 3% GA-PDCM longer than 6 weeks.However, the sustenance to maintain cellular seperating effect was: 3% GA-PDCM maintaining cellular seperating effect for 21 day, and 0.05%、0.01%、0% GA- PDCM maintaining for 7 to 10 days. After this period, PDCM were invaded by fibroblasts and capillaries. PDCM could induced foreign body reaction in implanted muscle tissue, which would progressed to delayed type hypersensitivity (DTH). The intensity of induced DTH was corelated to the persistence of PDCM; the longer PDCM persised , the more intensively the DTH would be. The DTH induced by 3% GA-PDCM was most intensively and progressed to most mature reaction, but those of 0%GA-PDCM was most mild. According to the result of immunohistochemistric stain, the activated macrophages were the primary effector cells of DTH. This reaction was corelated to CD4 T lymphocytes, but not to CD8 T lymphocytes. In implanted sites, there was no significant amount of IgG、IgM antibody deposition. In other words, the possibility of PDCM inducing type II hypersensitivity was very low. According to above finding, we suggest that the immune response induced by PDCM belongesto reaction managed by T helper 1(Th 1) cells. In the furture, if PDCM is utilized in periodontal therapy, there would be not only guded tissue regenerative effect, but also changing immune response from destructive pathway to repair one,which was managed by Th 1 cells.