Anti-Inflammatory Effect of Rice Bran-Fortified Ready-to-Use Therapeutic Food Digestates in Caco-2/HT29-MTX/THP-1 Triculture Model

Abstract

Severe Acute Malnutrition is frequently associated with Environmental Enteric Dysfunction, necessitating therapeutic foods that provide both high nutritional density and immunomodulatory benefits. Ready-to-Use Therapeutic Food contain ingredients such as skim milk powder, wheat flour, and the addition of 5% Rice Bran that are hoping to give additional benefits. This study aimed to evaluate anti-inflammatory effects of RUTF + 5% Rice Bran formulation by comparing them to standard RUTF samples supplied by the SEAFAST Center IPB. The experiment focused on monitoring cellular immune responses in vitro, where the production of pro-inflammatory cytokine human Interleukin-8 (IL-8) and Tumor Necrosis Factor-alpha (TNFa) was challenged using Lipopolysaccharide (LPS) and Interferon-gamma (IFN-?) stimulation. The Neutral Red cell viability assay test revealed that In Vitro Digestion (IVD) product formulations were non-toxic and preserved lysosomal integrity at a 1/5 dilution. In the Phenol Red permeability test, the addition of RUTF + 5% Rice Bran digestate has shown to maintain tight junction integrity and low diffusion coefficients despite exposure to the inflammatory environment. Furthermore, ELISA test results revealed that pre-treatment with RUTF + 5% Rice Bran digestate has shown to have higher levels of pro-inflammatory cytokine biomarkers secretion of IL-8 but lower TNFa secretion than Inflammatory digestate, the difference in results begs the need for more comprehensive experiments.

Severe Acute Malnutrition sering berkaitan erat dengan Environmental Enteric Dysfunction, sehingga membutuhkan pangan terapi yang tidak hanya berkepadatan gizi tinggi tetapi juga memberikan manfaat immunomodulatory. Ready-to-Use Therapeutic Food / RUTF memanfaatkan bahan-bahan seperti susu bubuk skim, tepung terigu, serta penambahan 5% rice bran yang diharapkan mampu memberikan manfaat ekstra. Penelitian ini bertujuan untuk mengevaluasi efek anti-inflamasi dari formulasi RUTF + 5% rice bran dengan membandingkannya terhadap sampel RUTF standar buatan SEAFAST Center IPB. Pengujian berfokus pada pemantauan respon imun seluler secara in vitro, di mana produksi sitokin pro-inflamasi human Interleukin-8 (IL-8) dan Tumor Necrosis Factor-alpha (TNFa) dipicu menggunakan stimulasi Lipopolysaccharide (LPS) dan Interferon-gamma (IFN-?). Uji viabilitas sel Neutral Red menunjukkan bahwa hasil cerna In Vitro Digestion (IVD) dari formulasi ini bersifat tidak toksik serta mampu menjaga integritas lisosom pada pengenceran 1/5. Pada uji permeabilitas Phenol Red, penambahan hasil cerna RUTF + 5% rice bran terbukti dapat mempertahankan integritas tight junction dan menjaga koefisien difusi tetap rendah meskipun dipaparkan pada kondisi inflamasi. Lebih lanjut, hasil uji ELISA mengungkapkan bahwa pra-perlakuan dengan hasil cerna RUTF + 5% rice bran menghasilkan sekresi biomarker sitokin pro-inflamasi IL-8 yang lebih tinggi, namun sekresi TNFa yang lebih rendah dibandingkan dengan kontrol inflamasi. Perbedaan hasil ini mengindikasikan perlunya penelitian yang lebih komprehensif ke depannya.