Tesis

Efek Sekretom pada Model Tikus Sprague Dawley dengan Anastomosis Usus = Effects of Secretome on a Sprague Dawley Rat Model with Intestinal Anastomosis.

Latar belakang : Anastomosis usus pada bedah pediatri, khususnya pasca-reseksi sering mengalami komplikasi seperti kebocoran, striktur, dan ileus yang meningkatkan morbiditas dan mortalitas. Sekretom sel punca mesenkimal tali pusat mengandung faktor pertumbuhan dan sitokin yang berpotensi meningkatkan angiogenesis, modulasi inflamasi, dan regenerasi jaringan. Penelitian ini mengevaluasi efek sekretom sel punca mesenkimal yang diaplikasikan dengan Surgicel pada penyembuhan anastomosis jejunum tikus Sprague Dawley, melalui pengukuran fibrosis (Masson’s trichrome), skor histologi (HE), serta ekspresi VEGF dan PDGF pada hari ke-3 dan ke-7 pascaoperasi. Metode : Penelitian eksperimental post-test only control group menggunakan 18 tikus jantan (8-12 minggu, 250-350 g), dibagi menjadi tiga kelompok: kontrol (anastomosis saja), Surgicel, dan Surgicel-sekretom (150 μL). Prosedur meliputi laparotomi midline, reseksi jejunum 1 cm, anastomosis end-to-end 6-0 absorbable, dengan terminasi hari ke-3 (inflamasi) dan ke-7 (proliferasi). Evaluasi imunohistokimia dilakukan untuk menilai ekspresi VEGF dan PDGF (H‑score), pewarnaan Masson’s trichrome untuk persentase fibrosis, serta pewarnaan hematoksilin eosin untuk skor histologis (fibroblas, kolagen, neovaskularisasi, inflamasi, dan integritas mukosa). Analisis menggunakan ANOVA/Kruskal-Wallis (p < 0,05). Hasil : Pada hari ke-7, fibrosis signifikan lebih tinggi pada kelompok Surgicelsekretom (49,90±7,77%) dibandingkan kontrol (28,18±6,19%; p=0,027) dan Surgicel (38,86±1,18%), sementara VEGF menurun (2,13±1,81 vs. 36,44±2,87; p=0,070) dan PDGF meningkat (139,60±12,59 vs. 32,86±26,16 Surgicel; p=0,001). Skor histologi tidak berbeda signifikan antar kelompok (p > 0,05). Kesimpulan : Kombinasi surgicel-sekretom meningkatkan fibrosis dan PDGF, menunjukkan efek pro-regeneratif awal melalui modulasi faktor pertumbuhan, meskipun tanpa perbedaan histologi klinis terukur. Temuan ini mendukung potensi sekretom sebagai adjuvan bedah untuk mencegah kebocoran anastomosis, dengan rekomendasi optimasi dosis dan evaluasi fungsional lanjutan.
Kata Kunci : Sekretom, anastomosis usus, VEGF, PDGF, skor histologi, fibrosis


Background: Intestinal anastomosis in pediatric surgery, particularly after bowel resection, is prone to complications such as leakage, stricture, and ileus, which increase morbidity and mortality. The secretome of umbilical cord mesenchymal stem cells contains growth factors and cytokines that may enhance angiogenesis, modulate inflammation, and promote tissue regeneration. This study evaluated the effect of mesenchymal stem cell secretome applied with Surgicel on jejunal anastomosis healing in Sprague Dawley rats by assessing fibrosis (Masson’s trichrome), histological score (HE), and VEGF and PDGF expression on postoperative days 3 and 7. Methods: This post-test only control group experimental study used 18 male rats (8–12 weeks, 250–350 g) divided into three groups: control (anastomosis only), Surgicel, and Surgicel-secretome (150 μL). The procedures included a midline laparotomy, 1 -cm jejunal resection, and end-to-end anastomosis with 6-0 absorbable sutures, followed by termination on day 3 (inflammatory phase) and day 7 (proliferative phase). Immunohistochemistry was performed to evaluate VEGF and PDGF expression (H-score), Masson’s trichrome staining for fibrosis percentage, and hematoxylin-eosin staining for histological scoring (fibroblast proliferation, collagen deposition, neovascularization, inflammation, and mucosal integrity). Data were analyzed using ANOVA or Kruskal–Wallis tests (p < 0.05). Results: On day 7, fibrosis was significantly higher in the Surgicel-secretome group (49.90 ± 7.77%) compared with the control (28.18 ± 6.19%; p = 0.027) and Surgicel (38.86 ± 1.18%) groups, while VEGF expression decreased (2.13 ± 1.81 vs. 36.44 ± 2.87; p = 0.070), and PDGF expression increased (139.60 ± 12.59 vs. 32.86 ± 26.16 in Surgicel; p = 0.001). Histological scores did not differ significantly among groups (p > 0.05). Conclusion: The combination of Surgicel and secretome increased fibrosis and PDGF expression, indicating early pro-regenerative effects through growth factor modulation, although no significant histological differences were observed. These findings support the potential of secretome as a surgical adjuvant to prevent anastomotic leakage, with recommendations for further dose optimization and functional evaluation.
Keywords : Secretome, intestinal anastomosis, VEGF, PDGF, histology score, fibrosis