Hyaluronik asit - kollagen ile enkapsüle edilmiş mezenkimal kök hücrelerin, trombositten zengin plazmanın ve sinerjistik etkilerinin temporomandibular eklem osteoartritinde tedavi potensiyellerinin yurtiçi paten başvurusu desteği
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Tarih
2022
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info:eu-repo/semantics/closedAccess
Özet
Çalışmamızda hyaluronik asit (HA)-kollagen ile enkapsüle edilmiş mezenkimal kök hücre, trombositten zengin plazma (PRP) ve her ikisinin sinerjistik etkileri in vitro ve in vivo tavşan temporomandibuler eklem (TME) osteoartrit modelinde araştırılmıştır. Çalışmanın in vitro kısmında; karakterize edilen ve PRP, kök hücre ve her ikisini içeren HAkollagen enkapsülasyon malzemesinin kondrojenik kapasite üzerindeki etkileri hücre morfolojisi, glikozaminoglikan sentezi, kollagen tip I, kollagen tip II sentezi, apoptoz, nekroz, proliferasyon, sitotoksisite ve kondrosit canlılığı, PCR, Western Blot, Alamar Blue ve gerçek zamanlı hücre analiz sistemi yöntemleri ile değerlendirilmiştir. Çalışmanın in vivo kısmında 90 tavşan kullanılarak 6 grup (n=18) oluşturulmuştur: Grup 1; Enkapsülasyon malzemesi, Grup 2; Enkapsüle edilmiş kök hücre, Grup 3; Enkapsüle edilmiş PRP, Grup 4; Enkapsüle edilmiş kök hücre+PRP, Grup 5; Salin uygulanmış pozitif kontrol (sol TME sayısı), Grup 6; Sham kontrol. Grup 6 hariç diğer gruplarda tavşanların mandibular kondilinde subkondral defekt oluşturularak osteoartrit indüklendi ve 3 haftayı takiben tedavi protokolleri uygulandı. Her gruptan 6 tavşan tedaviyi takip eden 4., 12. ve 24. haftalarda sakrifiye edilerek TME' ler makroskopik, histolojik, histokimyasal, immunohistokimyasal olarak incelenmiştir. Verilerin değerlendirmelerinde Kruskal Wallis, Friedman ve Wilcoxon testleri uygulanmıştır Enkapsülasyon malzemesinin mezenkimal kök hücre ve primer kondrositler üzerinde sitotoksik olmadığı ve hücrelerin glikozaminoglikan sentezini sürdürdükleri bulunmuştur. PCR sonuçları; kollagen tip I ve II, Aggrecan, SOX9 ifadelerinin 3., 5. ve 8. günlerde artmış olduğunu, 15. günde ortalama 8'lik azalma olduğunu göstermiştir. Çalışmanın in vivo kısmında 4.haftada tedavi gruplarının salin grubundan daha iyi sonuçlar verdiği ancak aralarında istatistiksel olarak fark olmadığı, 12. haftada ise Grup 4?de eklem hasarının daha az olduğu bulunmuştur. 24. haftada Grup 4'ün sonuçları Grup 2'ninkine benzerlik gösterirken, diğer gruplardan olumlu yönde istatistiksel fark gösterdiği bulunmuştur. HA-kollagen ile enkapsüle edilmiş kök hücre ve PRP sinerjistik uzayan etki yaratarak TME osteoartritinde kartilaj rejenerasyonunu olumlu etkilediği ve potansiyel bir tedavi yöntemi olabileceğini göstermiştir.
In this study, synergistic effects of hyaluronic acid (HA)-collagen encapsulated mesenchymal stem cells, platelet-rich plasma (PRP) and both were investigated in vitro and in vivo rabbit temporomandibular joint (TME) osteoarthritis model. In the in vitro part of the study effect of characterized HA-collagen encapsulation material and encapsulated PRP, stem cells and both on chondrogenic capacity, cell morphology, glycosaminoglycan synthesis, collagen types I and II synthesis, apoptosis, necrosis, proliferation, cytotoxicity and chondrocyte viability were evaluated using PCR, Western Blot , Alamar Blue and real-time cell analysis system. For in vivo studies, 90 rabbits were allocated to 6 groups (n = 18); Group 1; Encapsulation material, Group 2; Encapsulated stem cell, Group 3; Encapsulated PRP, Group 4; Encapsulated stem cell+PRP, Group 5; Saline treated positive control (left TMJ number), Group 6; Sham control. Osteoarthritis was induced by creating subchondral defect in mandibular condyle of rabbits except group 6 and treatment protocols were applied after 3 weeks. Six rabbits from each group were sacrificed at the 4th, 12th and 24th weeks following treatment and TMJs were examined macroscopically, histologically, histochemically and immunohistochemically. Kruskal Wallis, Friedman and Wilcoxon tests were used to evaluate data (p≤0.05). It was found that the encapsulation material was not cytotoxic on mesenchymal stem cells and primary chondrocytes and cells maintained glycosaminoglycan synthesis. PCR results; collagen types I and II, Aggrecan, SOX9 expression was increased on the 3rd, 5th and 8th days and 0.5-2% decrease on the 15th day. In in vivo study, it was found that treatment groups gave better results than saline group at 4th week but there was no statistical difference between them and joint damage was less in Group 4 at 12th week (p<0.01). Results of Group 4 were similar to those of Group 2 at 24th week, but there was a statistically significant difference from the other groups (p<0.01). Intraarticular injection of HA-collagen encapsulated stem cells and PRP has shown that it can be a potential treatment modality by positively contributing to cartilage regeneration in TMJ osteoarthritis.
In this study, synergistic effects of hyaluronic acid (HA)-collagen encapsulated mesenchymal stem cells, platelet-rich plasma (PRP) and both were investigated in vitro and in vivo rabbit temporomandibular joint (TME) osteoarthritis model. In the in vitro part of the study effect of characterized HA-collagen encapsulation material and encapsulated PRP, stem cells and both on chondrogenic capacity, cell morphology, glycosaminoglycan synthesis, collagen types I and II synthesis, apoptosis, necrosis, proliferation, cytotoxicity and chondrocyte viability were evaluated using PCR, Western Blot , Alamar Blue and real-time cell analysis system. For in vivo studies, 90 rabbits were allocated to 6 groups (n = 18); Group 1; Encapsulation material, Group 2; Encapsulated stem cell, Group 3; Encapsulated PRP, Group 4; Encapsulated stem cell+PRP, Group 5; Saline treated positive control (left TMJ number), Group 6; Sham control. Osteoarthritis was induced by creating subchondral defect in mandibular condyle of rabbits except group 6 and treatment protocols were applied after 3 weeks. Six rabbits from each group were sacrificed at the 4th, 12th and 24th weeks following treatment and TMJs were examined macroscopically, histologically, histochemically and immunohistochemically. Kruskal Wallis, Friedman and Wilcoxon tests were used to evaluate data (p≤0.05). It was found that the encapsulation material was not cytotoxic on mesenchymal stem cells and primary chondrocytes and cells maintained glycosaminoglycan synthesis. PCR results; collagen types I and II, Aggrecan, SOX9 expression was increased on the 3rd, 5th and 8th days and 0.5-2% decrease on the 15th day. In in vivo study, it was found that treatment groups gave better results than saline group at 4th week but there was no statistical difference between them and joint damage was less in Group 4 at 12th week (p<0.01). Results of Group 4 were similar to those of Group 2 at 24th week, but there was a statistically significant difference from the other groups (p<0.01). Intraarticular injection of HA-collagen encapsulated stem cells and PRP has shown that it can be a potential treatment modality by positively contributing to cartilage regeneration in TMJ osteoarthritis.
