PULMONER VERİLİŞE YÖNELİK TETANOZ TOKSOİD-MANNİTOL KURU TOZ İNHALASYON FORMÜLASYONU VE İN VİTRO - İN VİVO DEĞERLENDİRİLMESİ

Öz

Amaç: Konvansiyonel aşılara ağrı sınırlamaları, soğuk zincir depolama ve sterilite sorunları eşlik ettiğinden, pulmoner yolla uygulanan mukozal bir aşı üretimiştir. Tetanoz toksoidi (TT) ve mannitolün kuru toz inhalasyonu hazırlanmış ve konvansiyonel tetanoz toksoid aşısına kıyasla stabilite ve immünojenisite açısından değerlendirilmiştir. Gereç ve Yöntem: TT ve mannitol kuru toz inhalasyonu hazırlandı ve partikül boyutu analizi, FTIR, akış özellikleri, kapsülleme etkinliği, flokülasyon ve in vitro ilaç aşısı salım çalışmaları açısından değerlendirildi. Formülasyonun immünolojik çalışmaları, BALB/c fareleri üzerinde gerçekleştirildi. Sonuç ve Tartışma: TT ve mannitolün toz karışımı, işlem koşulları altında ve depolama sonrasında stabil kalmıştır. Sonuç, bir flokülasyon testi ile doğrulanmıştır. FTIR analizine göre hiçbir etkileşim tespit edilmemiştir. Homojenizasyon işlemi ile, 1312 ± 1310.9 nm'lik geometrik partikül boyutuna sahip, pulmoner uygulama için uygun bulunan bir toz elde edilmiştir. Zeta potansiyeli ve polidispersite indeksi (PDI) sırasıyla -22.6 ± 0.16 mV ve 0.499 ± 0.015 olarak bulunmuştur. Difüzyon çalışmaları, difüzyon mekanizması ve sıfır derece kinetiği takiben 2 saat içinde açığa çıkan etkin maddenin %82.4 ±% 6.7'si ile TT'nin derhal salındığını göstermiş ve mannitolün tetanoz toksoidinin salınımını geciktirmediği bulunmuştur. Ek olarak, kuru toz inhalasyonunun akış özelliklerinin iyi akış özelliklerine sahip olduğu rapor edilmiştir. Daha da önemlisi, immünolojik çalışmalar, geleneksel aşılara göre yüksek sistemik ve mukozal bağışıklığın indüklendiğini ortaya çıkarmıştır.

Anahtar Kelimeler

• Aşı
• kuru toz inhalasyonu
• mukozal bağışıklık
• pulmoner uygulama
• Tetanoz toksoidi

FORMULATION AND IN VITRO - IN VIVO EVALUATION OF TETANUS TOXOID-MANNITOL DRY POWDER INHALATION FOR PULMONARY DELIVERY

Öz

Objective: As the conventional vaccines were accompanied by the limitations of pain, cold chain storage and sterility issues, a mucosal vaccine which is administered through pulmonary route was fabricated. A dry powder inhalation of tetanus toxoid (TT) and mannitol was prepared and evaluated for stability and immunogenicity in comparison to the conventional TT vaccine. Material and Method: TT and mannitol dry powder inhalation was prepared and evaluated for particle size analysis, scanning electron microscopy, FTIR, flow properties, TT content estimation, flocculation, and in vitro drug vaccine release studies. Immunological studies of the formulation were performed on BALB/c mice. Result and Discussion: The powder blend of tetanus toxoid and mannitol remained stable under the process conditions and after storage. The result was confirmed through a flocculation test. The FTIR analysis indictated no interactions between the components. The homogenization process yielded a powder with a geometrical particle size diameter of 1312 ± 1310.9 nm which was found suitable for pulmonary administration. The zeta potential and polydispersity index {PDI} were found to be -22.6 ± 0.16 mV and 0.499 ± 0.015, respectively. The diffusion studies indicated immediate release of the TT with 82.4 ± 6.7% of drug released within 2 h following the diffusion mechanism and zero order kinetics and it was found that mannitol didn’t retard the release of tetanus toxoid. Additionally, the flow properties of the dry powder inhalation were reported to have good flow properties. More importantly, the immunological studies inferred the induction of high systemic and mucosal immunity over conventional vaccines.

Anahtar Kelimeler

• Dry powder inhalation
• mucosal immunity
• Tetanus toxoid
• pulmonary administration
• vaccine

Kaynakça

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