Perubahan Morfologis Kulit Tikus Tua Pasca Xenotranplantasi Human Wharton’s Jelly Mesenchymal Stem Cell

Abstract

Penuaan seringkali menyebabkan keadaan patologis pada tubuh. Hal itu terjadi karena adanya kerusakan yang terakumulasi pada tingkat seluler maupun makromolekuler. Kulit merupakan salah satu organ yang juga mengalami penurunan kualitas pada kondisi tua. Terapi sel menjadi upaya dalam memperbaiki kualitas organ tersebut. Mesenchymal Stem Cells yang berasal dari Wharton’s Jelly (hWJ-MSCs) diharapkan mampu mengembalikan kerja sel-sel yang berperan dalam regenerasi kulit. Penelitian ini bertujuan menganalisis efek terapi tersebut yang meliputi perubahan fisiologis morfologi kulit tikus tua, perkembangan kulit tikus tua pascapemberian hWj-MSCs intravena, perkembangan sel fibroblas, serta populasi sel mast pada kulit. Penelitian ini menggunakan 12 tikus jantan yang masing-masing terdiri dari 3 tikus muda tanpa perlakuan, 3 tikus kontrol tua fisiologis (diinjeksi NaCl 0.9% volume 0.4 ml), 3 tikus tua perlakuan 1 (diinjeksi hWJ-MSCs dosis 1x106 sel/kg BB volume 0.4 ml), dan 3 tikus tua perlakuan 2 (diinjeksi hWJ-MSCs dosis 10x106 sel/kg BB volume 0.4 ml). Penyuntikan dilakukan 4 kali dengan interval 3 bulan dalam satu tahun. Pada akhir penelitian, tikus dianestesi, dikorbankan, kemudian diproses secara histologis. Histomorfologi jaringan kulit dianalisis menggunakan pewarnaan Hematoksilin Eosin sedangkan kepadatan serabut kolagen dengan pewarnaan Cason’s Trichrome. Pewarnaan imunohistokimia digunakan untuk menganalisis perkembangan sel fibroblas dan populasi sel mast dianalisis menggunakan pewarnaan Toluidine Blue. Hasil penelitian menunjukkan perbaikan pada morfologi kulit tikus tua yang diinjeksikan hWJ-MSCs. Kepadatan serabut kolagen pada tikus tua dengan perlakuan dosis 10x106 sel/kg BB mengalami peningkatan secara signifikan (P<0.05). Hasil pewarnaan imunohistokimia tidak menunjukkan adanya proliferasi sel fibroblas. Sementara itu, populasi sel mast tikus tua dengan perlakuan cenderung mengalami peningkatan meskipun tidak signifikan (P<0.05). Pemberian hWJ-MSCs secara intravena dapat memperbaiki densitas serabut kolagen pada kulit tikus tua namun tidak terjadi melalui perbanyakan sel fibroblas. MSCs xenogenik yang diinjeksikan secara intravena dapat menuju jaringan kulit tikus tua tanpa direjeksi oleh sistem pertahanan tubuh resipien.

Aging often causes pathological conditions in the body. This matter occurs due to damage accumulated at the cellular to macromolecular level. In the aged condition, the quality of body organs will decrease, including the skins. Cell therapy is an effort to improve quality of the skin. Human Mesenchymal Stem Cells derived from Wharton's Jelly (hWJ-MSCs) are expected to be able to refunction cells in skin regeneration. This study was aimed to analyze the therapeutic effects which includes physiological changes in the skin morphology of aged rats, their development after intravenous hWj-MSCs administration, fibroblast cell development, and mast cell population on the skin. This study used 12 male rats which consists of 3 untreated young rats, 3 physiological aged rats as a control (injected with 0.9% volume of 0.4 ml NaCl), 3 aged rats for first treatment (injected with hWJ-MSCs at a dose of 1x106 cells/kg BW volume 0.4 ml), and 3 aged rats for second treatment (injected with hWJMSCs dose of 10x106 cells/kg BW 0.4 ml volume). Injections are carried out 4 times at intervals of 3 months in a year. Histomorphology of skin tissue was analyzed using Hematoxylin Eosin staining, while the density of collagen fibers was stained with Cason's Trichrome. Immunohistochemical staining was used to analyze the development of fibroblasts and mast cell populations were analyzed using Toluidine Blue stain. The results showed the skin morphology of old rats was improved after hWJ-MSCs injection. Collagen fiber density in aged rats treated with a dose of 10x106 cells/kg BW increased significantly (p<0.05). On the other hand, the immunohistochemical staining did not show the presence of fibroblast cell proliferation. Meanwhile, the mast cell population of treated rats tended to increase, although it was not significant (p>0.05). Intravenous administration of hWJ-MSCs can improve the density of collagen fibers in the skin of aged rats, but did not occur through fibroblast cell multiplication. Xenogenic MSCs also reach the skin tissue of aged rat without being rejected by the recipient's immune system.