Hemoroidal yaralarda lazer sonrası enfeksiyonu önlemek için antimikrobiyal büyüme faktörü füzyon proteinlerinin bilgisayar simülasyonlu tasarımı

Öz

Amaç: Lazer sonrası yara enfeksiyonu, hemoroid cerrahisi sonrası tekrarlayan bir komplikasyondur, çünkü anorektal ortamda polimikrobiyal flora kolonize olur ve mukozal onarım bozulur. Bu çalışma, mevcut tedavi seçeneklerinin dezavantajlarını ortadan kaldırmayı ve anorektal yara onarımının mikroçevresinin özel koşullarını karşılamak üzere izlenen, antimikrobiyal aktiviteye sahip yeni nesil bir profilaktik biyomolekül sağlamayı amaçlamaktadır. Gereç ve Yöntemler: Enfeksiyon ve yavaş yara iyileşmesi gibi bu iki sorunu aşmak için, mevcut bir çalışmada, seçilen antimikrobiyal peptidleri (AMP'ler) fibroblast büyüme faktörü 10 (FGF10) alanıyla birleştiren yeni bir antimikrobiyal-büyüme faktörü füzyon proteini tasarlamak ve karakterize etmek için bir in-silico biyomühendislik müdahalesi geliştirilmiştir. Bulgular: Antimikrobiyal Peptid Veritabanı, AP00608, AP02570, AP03795 ve AP00952 dahil olmak üzere dört katyonik AMP ile taranmış ve AllerTOP v2.0 ve ToxiPred kullanılarak taranarak alerjenik ve toksik olmadıkları doğrulanmıştır. Elde edilen mühendislik füzyon yapısı, istenen fizikokimyasal özelliklere (32,84 kDa; pI 10,19; kararsızlık indeksi 38,52; GRAVY -0,33; çözünürlük 0,48) ve bu da yüksek stabilite ve hidrofiliklik anlamına gelmektedir. İkincil yapı tahmini %27,34 alfa sarmal ve %16,26 beta sarmal gösterirken, AlphaFold olasılığı ve ERRAT (%94,28 yapısal doğruluk kanıtladı). ClusPro 2.0, FGFR2b (1NUN) ve FGFR2 (1EV2) ile sırasıyla -1161,9 kcal mol -1 ve -1144,4 kcal mol -1 enerjilerle iyi bağlanma afiniteleri gösterdi. 500 ikonik Desmond moleküler dinamik simülasyonu, termodinamik stabiliteyi, kompakt katlanmayı ve kompleksler içindeki hidrojen bağlarının korunmasını belirledi. Sonuç: Tüm bu bulgular, önerilen füzyon yapısının, lazer sonrası hemoroidal yaralarda mikrobiyal kolonizasyonu önleyen ve epitel onarımını uyaran çift etkili bir tedavi yöntemi olarak kullanılma potansiyeline sahip olduğunu göstermektedir. Bu hesaplamalı araştırma, bu nedenle gelecekte çok işlevli yara iyileştirici biyolojik ürünlerin deneysel olarak doğrulanması ve translasyonel geliştirilmesinin temelini oluşturmaktadır.

Anahtar Kelimeler

• Antimikrobiyal peptitler
• fibroblast büyüme faktörü 10
• moleküler kenetlenme
• desmond simülasyonu
• in-silico füzyon proteini
• yara iyileşmesi
• lazer sonrası hemoroid enfeksiyonu

In-silico engineering of an antimicrobial growth factor fusion protein to prevent post-laser infection in hemorrhoidal wounds

Öz

Aim: Post laser wound infection is a recurrent complication after hemorrhoidal surgery mainly because polymicrobial flora is colonized in the anorectal environment and the mucosal repair is impaired. This study aims to overcome the drawbacks of current therapeutic options and to provide a novel generation of prophylactic biomolecule with antimicrobial activities, monitored to meet the specific conditions of the microenvironment of anorectal wound repair. Material and Methods: To overcome this two-fold problem of infection and slow wound healing, a current study developed an in-silico bioengineering intervention to design and characterize a new antimicrobial- growth factor fusion protein fusing selected antimicrobial peptide (AMPs) with the fibroblast growth factor 10 (FGF10) domain. Results: The Antimicrobial Peptide Database was searched with four cationic AMPs, which included AP00608, AP02570, AP03795, and AP00952, and screened using AllerTOP v2.0 and ToxiPred, which confirmed their non-allergenicity and non-toxicity. The obtained engineered fusion construct possessed desirable physicochemical properties (32.84 kDa; pI 10.19; instability index 38.52; GRAVY -0.33; solubility 0.48) which implies high stability and hydrophilicity. Secondary structure prediction showed 27.34% alpha-helices and 16.26% beta-strands, whereas AlphaFold probability and ERRAT (94.28% proved structural accuracy). ClusPro 2.0 showed good binding affinities to FGFR2b (1NUN) and FGFR2 (1EV2) with energies of -1161.9 kcal mol -1 and -1144.4 kcal mol -1 respectively. Following 500 Desmond molecular dynamics simulations determined the thermodynamic stability, compact folding, and the maintenance of hydrogen bonds within the complexes. Conclusion: All these findings suggest that the proposed fusion construct has a potential to serve as a dual-acting therapeutic - preventing microbial colonization and stimulating epithelial repair in post-laser hemorrhoidal wounds. This computational research therefore provides the basis of experimental validation and translational development of multifunctional wound-healing biologics in the future.

Anahtar Kelimeler

• Antimicrobial peptides
• fibroblast growth factor 10
• molecular docking
• desmond simulation
• in-silico fusion protein
• wound healing
• post-laser hemorrhoid infection

Kaynakça

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