EFEK PENAMBAHAN ASAM HIALURONAT PADA LISAT PLATELET RICH FIBRIN TERHADAP PERBAIKAN AKTIVITAS SELULAR FIBROBLAS TUA

Rika Azyenela, Dr. dr. Indah Julianto, Sp.KK(K).; Dr. dr. Yohanes Widodo Wirohadidjojo, Sp.KK(K).

2016 | Tesis-Spesialis | S2 Ilmu Kesehatan Kulit dan Kelamin

Latar Belakang: Kegagalan sembuh pada luka kronis dan keriput pada kulit menua dini disebabkan oleh adanya fibroblas tua. Fibroblas tua mengalami penurunan kemampuan berproliferasi dan mensintesis kolagen karena tidak berespon terhadap pacuan transforming growth factor1 (TGF-). Platelet derived growth factor (PDGF) yang berasal dari lisat platelet rich fibrin (PRF) dapat memulihkan ekspresi reseptor TGF-beta. Peningkatan kekuatan mekanik matriks ekstraselular dengan penambahan asam hialuronat (HA) dapat memperbaiki signaling TGF-beta Tujuan: Penelitian ini bertujuan untuk membuktikan apakah penambahan asam hialuronat ke dalam lisat PRF dapat memperbaiki aktivitas selular sel fibroblas tua dibandingkan lisat PRF dan asam hialuronat tanpa kombinasi. Metode: Dengan rancangan eksperimental kuasi, fibroblas kulit manusia pasase 3 yang berasal dari kulit preputium 6 orang anak, dibuat berpasangan dan dibagi menjadi dua kelompok: kelompok I yaitu fibroblas normal tanpa puasa dan kelompok II yaitu fibroblas yang dipuasakan selama 48 jam dan diberikan terapi lisat PRF 50%; HA 20,83 mM; HA 41,67 mM; HA 83,33 mM dan penambahan dosis HA tersebut diatas pada lisat PRF 50% ( lisat PRF 50%+ HA 20,83mM; lisat PRF 50%+ HA 41,67 mM; lisat PRF 50%+ HA83,33mM). Setelah perlakuan dilakukan pengukuran indeks proliferasi dan deposisi kolagen. Hasil: Biakan fibroblas tua dalam campuran lisat PRF 50% dengan HA 20,83 mM menunjukkan peningkatan proliferasi fibroblas secara bermakna (p<0,05) dibandingkan fibroblas normal, kontrol negatif, asam hialuronat/lisat PRF 50% tanpa kombinasi. Sedangkan, biakan fibroblas tua dalam campuran lisat PRF 50% dengan HA 41,67 mM menunjukkan peningkatan deposisi kolagen yang bermakna (p<0,05) dibandingkan fibroblas normal, kontrol negatif, asam hialuronat/lisat PRF 50% tanpa kombinasi. Morfologi fibroblas terbaik dan terpadat juga terlihat pada fibroblas yang dibiakan dalam medium campuran lisat PRF 50% dengan asam hialuronat. Kesimpulan: Penambahan asam hialuronat kedalam lisat PRF terbukti dapat meningkatkan proliferasi dan deposisi kolagen secara bermakna.

Background: Failure of chronic ulcer healing and wrinkle lines in photoaged skin are both resulting from the presence of senescent fibroblasts. Senescent fibroblasts have diminished ability in proliferation and collagen synthesis due to unresponsive of transforming growth factor-beta(TGF-beta) stimulation. Platelet derived growth factor (PDGF) obtained from platelet rich fibrin (PRF) lysate can restore gene expression of TGF-beta receptors. Enhancement of extracellular matrix mechanical force with hyaluronic acid can improve fibroblasts functions. Objective: This study was performed to determine whether hyaluronic addition to PRF lysate has a better activity than PRF-lysate or hyaluronic acid alone in restoration of senescent human dermal fibroblasts (HDFs) activities. Methods: A quasi experimental design was done in artifcially senescent human dermal fibroblasts. Human dermal fibroblasts passage 3 cultivated from 6 boys foreskins were paired and devided into two groups. Group I was normal fibroblasts without serum starvation, Group II was 48 hours serum starvation which then treated with 50% PRF lysate, 20,83 mM HA; 41,67 mM HA; 83,33 mm HA and addition of those level o HA into 50% of PRF lysate (50% PRF lysate+20,83mM HA; 50% PRF lysate+41,67 mMHA; 50% PRF lysate+83,33 mMHA). After treatments, the proliferation index and collagen deposition were analyzed. Results: Cultivation of senescent human dermal fibroblasts in combination of 50% PRF lysate with 20,83 mM HA showed significant (p<0,05) elevation of proliferation compared to normal fibroblasts, negative control, 50% PRF lysate/HA alone. Whereas, cultivation of senescent human dermal fibroblasts in combination of 50% PRF lysate with 41,67 mM HA showed significant (p<0,05) elevation of collagen deposition compared to normal fibroblasts, negative control, 50% PRF lysate/HA alone. The best and densest fibroblas morphology was also found in this medium combination. Conclusion: The addition of hyaluronic acid into PRF lysate improve proliferation and collagen deposition of senescen human dermal fibroblasta significantly.

Kata Kunci : collagen deposition, hyaluronic acid, plateletrich fibrin lysate, proliferation index, senescent human dermal fibroblasts