AKTIVITAS MATRIX METALLOPROTEINASE-2 DAN TISSUE INHIBITORS OF METALLOPROTEINASE-2 DIHUBUNGKAN DENGAN DERAJAT INVASI JARINGAN ENDOMETRIOMA YANG DITRANSPLANTASIKAN PADA CHORIOALLANTOIC MEMBRANE (CAM)

TRI AGUNG WAHYU CAHYONO, DR. dr.Shofwal Widad, SpOG Subsp.F.E.R, Dr. dr. Cahyono Hadi, SpOG. Subsp. F.E.R

2023 | Tesis-Subspesialis | SUBSPESIALIS OBSTETRI DAN GINEKOLOGI

Latar belakang, Endometrioma adalah lesi kistik yang muncul dari proses endometriosis. Endometrioma umumnya ditemukan di ovarium. Proses perkembangan awal pembentukan endometrioma dimulai ketika jaringan endometrium melekat dan tumbuh pada epitel ovarium. Invaginasi progresif korteks ovarium pada jaringan yang melekat ini dapat menyebabkan pembentukan endometrioma. Salah satu patogenesis mendasar endometriosis untuk tumbuh dan berkembang adalah melakukan invasi seluler ke jaringan di sekitarnya di mana ia menempel. Proses invasi fokus endometriotik terkait dengan aktivitas matriks metalloptroteinases-2 (MMP-2) dan inhibitor jaringan metalloproteinases-2 (TIMP-2). Dalam penelitian ini, kami menggunakan jaringan endometrioma untuk ditanamkan pada membran chorioallantoic (CAM) sebagai media kultur untuk merekonstruksi proses invasi endometriosis, dan kemudian MMP-2, TIMP-2 dan pola invasi dapat diamati secara invitro.

Bahan dan Cara. Penelitian ini merupakan penelitian laboratorium observasional dengan tujuan untuk menilai hubungan antara aktivitas MMP-2, ekspresi TIMP-2 dengan tingkat invasi jaringan endometrioma yang ditransplantasikan pada CAM. Fragmen jaringan endometrioma diambil dari pasien endometrioma yang menjalani operasi laparoskopi. Fragmen jaringan dibagi menjadi 4-8 strip dengan dimensi masing-masing 2 x 2 mm2 . Dua strip dinilai untuk aktivitas MMP-2. Dua strip yang tersisa ditransplantasikan ke CAM. Setelah inkubasi selama lima hari, jaringan yang ditransplantasikan dipanen untuk penilaian MMP-2, TIMP-2 dan penilaian histologis. Penilaian histologis untuk tingkat invasi dilakukan dengan pewarnaan hematoksilin-eosin, penilaian MMP-2 dengan metode gelatin zymography dan TIMP-2 dengan western blot . Level invasi 1, non invasi, level 2,invasi parsial < 50> 50%, dan level 4 untuk total invasi.

Hasil. Dua puluh empat fragmen endometrioma didapatka dari 24 pasien endometrioma yang menjalani operasi laparoskopi. Usia rata-rata pasien adalah 31 ? 5 tahun, BMI adalah 22 ? 3 kg/m2 , lama infertilitas adalah 58 ? 3 bulan dan rASRM adalah 67 ? 37. Jaringan endometrioma yang ditransplantasikan pada CAM, aktivitas MMP-2 berkorelasi positif dengan TIMP-2 dengan hubungan sedang (r? = 0,56, p-value < 0 xss=removed>0,05).. Ekspresi TIMP-2 berkorelasi negatif dengan skor invasi dengan hubungan kuat (r? = -0,35, p-value <0 xss=removed>

Kesimpulan: Rasio MMP-2/TIMP-2 memiliki hubungan positif, MMP-2 memiliki hubungan positif dan TIMP-2 memiliki hubungan negative dengan level invasi jaringan endometrioma yang ditransplantasikan pada CAM

Kata kunci: Endometrioma, MMP-2, TIMP-2, invasi, CAM

Background. Endometrioma are cystic lesion which arises from endometriosis process. Endometrioma are commonly found in the ovary. The early developmental process of endometrioma formation is started when endometrial tissue attached and grew on ovarian epithelium. Progressive invagination of the ovarian cortex on to these attached tissue might lead to the formation of an endometrioma. Other study hypothesized that in endometrioma formation, the viable endometrial cells spread via hematogenous or lymphatic to develop metaplasia and differentiation into endometrial cells. One of the fundamental pathogenesis of endometriosis to grow and develop is to carry out cellular invasion of the surrounding tissue where it attaches. The invasion process of endometriotic focus is linked with the activity of matrix metalloptroteinases-2 (MMP-2) and tissue inhibitors of metalloproteinases2 (TIMP-2). In this study, we used endometrioma tissue to be implanted in chick chorioallantoic membrane (CAM) as a culture medium to reconstruct the invasion process of endometriosis, and then the MMP-2, TIMP-2 and invasion pattern can be invitro observed.

Materials and methods. This study was an observational laboratory study. The objective is to observe activity of MMP-2, TIMP-2 expression in different level invasion of endometrioma tissue transplanted onto CAM. Endometrioma tissue fragments taken from patients with ovarian endometrioma by laparoscopic surgery. The tissue fragments were divided into 4-8 strips with dimension each of 2 x 2 mm. Two strips were assessed for the activity of MMP-2. Two remaining strips were transplanted onto the CAM. After incubation for five days, the transplanted tissues were harvested for assessment of MMP-2, TIMP-2 and histological assessment. Histological assessment for the invasion level was carried out by hematoxylin-eosin staining, the assessment of MMP-2 by gelatin zymography and TIMP-2 by western blot. The invasion level number 1 is for non invasion, level 2 for partial invasion < 50> 50%, and level 4 for total invasion.

Result. Twenty four fragments of endometrioma have been collected from 24 endometrioma patients who underwent laparoscopy surgery. The average age was 31 ? 5 years, BMI was 22 ? 3 kg/m2 , duration of infertility was 58 ? 3 month and r-ASRM was 67 ? 36. In CAM of transplanted endometriomas tissue, MMP-2 activity was positively correlated with TIMP-2 with moderate relationship (r? = 0,56, p-value < 0 xss=removed>0,05). TIMP-2 expression was negatively correlated with invasion score 4 with strong relationship (r? = -0,35, p-value <0 xss=removed>

Conclusion. The ratio of MMP-2/TIMP-2 had a positive relationship, MMP-2 had a positive relationship and TIMP-2 had a negative relationship with the level of endometrioma invasion transplanted onto CAM

Keywords: Endometrioma, MMP-2, TIMP-2, invasion, CAM

Kata Kunci : Keywords: Endometrioma, MMP-2, TIMP-2, invasion, CAM