Disertasi

Peran Faktor Pertumbuhan pada Platelet Rich Fibrin Matrix dan Platelet Rich Plasma Autologus terhadap Percepatan Proses Penyembuhan Luka Tandur Kulit. = The Role of Growth Factors in Autologous Platelet Rich Fibrin Matrix and Platelet Rich Plasma in the Acceleration of Wound Healing Process in Skin Graft.

Latar Belakang: Produk biologi yang merupakan rekayasa jaringan dapat membantu memper- cepat proses penyembuhan luka. Salah satunya yang saat ini banyak digunakan dalam proses penyembuhan luka adalah Platelet Rich Plasma (PRP). Meskipun demikian PRP tidak memberikan hasil optimal, karena berbentuk cairan dan proses pembuatannya membutuhkan bovine thrombin yang bersifat xenologus. Selain itu sebagian besar faktor pertumbuhan telah dilepaskan saat awal aplikasi di luka. Produk biologi lainnya adalah Platelet Rich Fibrin Matrix (PRFM), yang merupakan generasi terbaru konsentrat trombosit yang menghasilkan fibrin alami. Pada operasi THT-KL khususnya plastik rekonstruksi, tandur kulit banyak digunakan untuk defek yang tidak dapat ditutup primer dengan jabir lokal. Dengan cara tersebut penyembuhan luka tandur kulit berlangsung lama, tandur kulit kontraktur dan hasilnya tidak optimal. Aplikasi PRFM pada implantasi tandur kulit diharapkan dapat meningkatkan mutu kesintasan tandur. Walaupun mekanisme kerja PRFM dalam mempercepat proses penyembuhan luka tandur kulit belum diketahui, secara in vitro mengarah pada adanya peran faktor pertumbuhan. Tujuan: Mendapatkan PRFM yang lebih baik dari PRP dalam mempercepat penyembuhan luka tandur kulit, tanpa menggunakan perangkat komersial, serta mengetahui peran faktor pertumbuhan dalam mempercepat penyembuhan luka tandur in vivo. Metode: Sebanyak 150 jaringan biopsi tandur kulit full thickness skin graft (FTSG) dan split thickness skin graft (STSG) yang diperoleh dari 5 ekor babi, dibagi menjadi tiga perlakuan yaitu tandur-kontrol, tandur-PRP, dan tandur-PRFM. Biopsi jaringan tandur dilakukan pada hari ke- 1, -3, -7, -14 dan ke-30 sesuai fase penyembuhan luka. Jaringan biopsi dievaluasi secara histologi dengan pewarnaan hematoksilin eosin, trichrome Masson dan Picrosirius red; juga kadar TGFβ1 dan PDGF. Pemeriksaan makroskopik terhadap luka tandur kulit dilakukan dengan mengevaluasi hasil dokumentasi fotografi menggunakan program ImageJ. Pada awal penelitian diukur kadar faktor pertumbuhan TGFβ1 dan PDGF di PRP dan PRFM, serta dinilai karakteristik struktur mikroskopik, ukuran serat fibrin dan diameter trombosit PRFM menggunakan SEM. PRFM dibuat dari PRP tanpa menggunakan perangkat komersial. Hasil: Kadar faktor pertumbuhan, kepadatan sel PMN, sel makrofag, fibroblas, kepadatan kolagen tipe 1 dan kesintasan tandur-PRFM lebih baik dibandingkan tandur-PRP dan tandur- kontrol. Diperoleh PRFM babi dan manusia yang memiliki struktur mikrokopik serat fibrin seperti jala dengan trombosit tersebar di serat fibrin. Karakter tersebut menyerupai PRFM yang diperoleh menggunakan perangkat komersial. Simpulan: Pemberian PRFM sebagai preparat trombosit autologus meningkatkan percepatan penyembuhan luka tandur kulit karena mengandung faktor pertumbuhan yang diperlukan pada penyembuhan luka. PRFM dapat dibuat tanpa perangkat komersial.
Kata Kunci: faktor pertumbuhan, PRFM, PRP, rekayasa jaringan, trombosit.


Background: Biological products that are tissue engineered can help accelerate the wound healing process. One of the mostly used biological products for wound healing process is Platelet Rich Plasma (PRP). However, it has not provided optimal results, because of its liquid form, the development process that use xenologous bovine thrombin, and most of the growth factors will be released prior to its application on a wound. Another biological product is Platelet Rich Fibrin Matrix (PRFM), a new generation of concentrated blood platelets that produce natural fibrin. For facial plastic and reconstructive surgery, skin graft is often used on defects that cannot be covered primarily by local flap. By this method the wound healing of skin graft is slow, skin graft contracture occured and the results were not optimal. Application of PRFM in the skin graft implants is expected to increase the survival of the graft. Furthermore the mechanism of PRFM in accelerating wound healing process of skin graft is still unknown; reported in vitro studies showed the important role of growth factor. Objective: To obtain PRFM that is better then PRP in accelerating the healing process of skin graft wound, without using comersial devices. Also to investigate the role of growth factors in accelerating the healing process of skin graft wound by in vivo study. Methods:. 150 tissue biopsies of full thickness (FTSG) and split thickness (STSG) skin grafts were obtained from 5 porcines with three different treatments, control-graft, PRP-graft and PRFM-graft. Biopsy of each tissue-graft was done on day-1, -3, -7, -14 and -30 according to the phases of wound healing. Each tissue biopsy was evaluated by histopathology using hematoxyilin eosin, trichrome Masson and Picrosirius red stainings; measurement of the TGFβ1 and the PDGF levels was done by ELISA. Macroscopic examination towards skin graft wound was done by evaluating photographic documentation results using ImageJ program. The amount of TGFβ1 and PDGF in PRP and PRFM was determined at the beginning of this research, as well as evaluation of the microscopic structure characteristic, fibrin fiber size and platelets diameter in PRFM by using SEM Results: TGFβ1 and PDGF levels, PMN cell, macrophage, fibroblast, and collagen type 1 density, as well as survival graft of PRFM addition were better compared to PRP and control. Porcine and human PRFM has microscopic fibrin fiber structure like nets with the platelets spread on fibrin fibers. This character of the PRFM is similar to the PRFM prepared by using a commercial device. Conclusion: Application of PRFM as an autologous thrombocyte preparation increase the acceleration of skin graft wound healing because it contains growth factors that are needed in wound healing.
Keywords: growth factor, platelets, PRFM, PRP, tissue engineering.